Tau-binding compounds

Abstract

The present disclosure provides anti-tau antibodies and methods of using anti-tau antibodies for the treatment and diagnosis of neurological indications.

Description

[Technical Field]

[0001] Related applications This international patent application claims priority to U.S. Provisional Patent Application No. 63 / 126,024, filed on 16 December 2020, the entire contents of which are incorporated herein by reference.

[0002] Sequence List This application includes an electronically submitted sequence listing in ASCII format, which is incorporated herein by reference in its entirety. The above ASCII copy, created on December 14, 2021, is named 2057_1314USPRO_SL.txt and has a size of 135,153 bytes.

[0003] This disclosure presents tau-binding compounds. In particular, this disclosure provides antibodies that bind to tau (e.g., human tau), such as antibodies that bind to phosphorylated epitopes on human tau. [Background technology]

[0004] Tauopathy is a group of neurodegenerative diseases characterized by dysfunction and / or aggregation of the microtubule-associated protein tau. Tau is a normally highly soluble protein known to associate with microtubules based on its phosphorylation level. Given its distinctive and elongated structure, tau is considered a crucial component of intracellular transport processes, particularly in nerve cells. Hyperphosphorylation of tau inhibits its binding to microtubules and its microtubule assembly activity. Furthermore, hyperphosphorylation of tau makes it more prone to misfolding and aggregation. In tauopathy, tau is hyperphosphorylated, misfolds, and aggregates as neurofibrillary tangles (NFTs) of paired helical fibrils (PHFs), twisted ribbons, or straight filaments. These NFTs are widely considered to indicate that nerve cell death is imminent and are thought to contribute to widespread neuronal loss, leading to various behavioral and cognitive impairments.

[0005] The first genetically defined tauopathy was reported when it was shown that mutations in the tau gene led to an autosomal dominant tauopathy known as frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17). This was the first evidence that changes in tau could lead to neurodegenerative changes in the brain. These molecules are thought to be more amyloid-forming, meaning they are more easily hyperphosphorylated and more likely to aggregate into NFTs (Hutton, M. et al., 1998, Nature 393(6686):702-5).

[0006] Several approaches have been proposed to therapeutically interfere with the progression of tau lesions and prevent subsequent molecular and cellular consequences. Given that NFTs are composed of hyperphosphorylated, misfolded, and aggregated tau, a range of eagerly pursued targets have emerged through interference at each of these stages. The introduction of agents that restrict phosphorylation, block misfolding, or prevent aggregation has all yielded promising results. Passive and active immunization with late-stage anti-phosphorylated tau antibodies in mouse models resulted in a dramatic reduction in tau aggregation and improvement in cognitive parameters. It has also been suggested that the introduction of anti-tau antibodies can prevent transneuronal diffusion of tau lesions.

[0007] Anti-tau antibodies remain needed for the treatment, diagnosis, and other applications of tauopathy. This disclosure addresses this need through the relevant compounds and methods described herein. [Overview of the project]

[0008] This specification provides isolated, for example, recombinant antibodies that bind to tau (e.g., human tau). In particular, this specification provides anti-tau antibodies that are not only structurally unique but also exhibit different binding patterns to various phosphorylated epitopes in tau, despite binding to similar / overlapping regions within tau, compared to phosphorylated epitopes recognized by anti-tau antibodies (e.g., PT3 and AT8) that are widely used to study the pathology of tau. In addition to therapeutic applications, the anti-tau antibodies provided herein can also function as diagnostic tools for detecting unique phosphorylation states of tau.

[0009] In some embodiments, the disclosure provides antibodies comprising heavy chain variable domains (VH) having complementarity-determining regions (CDR) H1, CDRH2, and CDRH3 containing any of the amino acid sequences listed in Table 4, or fragments thereof, and light chain variable domains (VL) having CDRL1, CDRL2, and CDRL3 containing any of the amino acid sequences listed in Table 1, or fragments thereof. The antibody may also comprise a set of variable domain CDR amino acid sequences, in which case the set of variable domain CDR amino acid sequences is selected from any of the amino acid sequences listed in Table 1. The antibody may also comprise a pair of variable domain CDR amino acid sequence sets, in which case the pair of variable domain CDR amino acid sequence sets is selected from any of the amino acid sequences listed in Table 1. The VH may comprise framework regions (FR) H1, FRH2, FRH3, and FRH4 selected from any of the amino acid sequences or fragments listed in Table 1. The VL may comprise FRL1, FRL2, FRL3, and FRL4 containing amino acid sequences selected from any of the amino acid sequences or fragments listed in Table 1. VH may contain an amino acid sequence selected from any of the amino acid sequences listed in Table 1, and / or may be encoded by a nucleic acid sequence selected from any of the nucleic acid sequences listed in Table 1. VL may contain an amino acid sequence selected from any of the amino acid sequences listed in Table 1, and / or may be encoded by a nucleic acid sequence selected from any of the nucleic acid sequences listed in Table 1. The antibody may contain a pair of variable domains selected from any of the variable domains listed in Table 1. The antibody may include monoclonal antibodies, multispecific antibodies, chimeric antibodies, antibody mimetic forms, single-chain Fv(scFv) forms, and forms selected from antibody fragments. The antibody may include an antibody class selected from IgA, IgD, IgE, IgG, and IgM. The antibody may contain one or more non-human constant domains. The antibody may contain one or more human constant domains. One or more human constant domains may be selected from any of the domains listed in Table 5. The antibody may contain human IgG, and the human IgG may include an isotype selected from IgG1, IgG2, IgG3, or IgG4.The antibody may be a human antibody. The antibody may bind to a tau protein epitope. The tau protein epitope may contain, or be contained within, an amino acid sequence selected from any of the amino acid sequences listed in Table 4. The tau protein epitope may contain a region formed by a complex of at least two tau proteins. When measured by direct enzyme-linked immunosorbent assay (ELISA), the antibody may bind to concentrated pair-helical fibrillary tau protein (ePHF) at a maximum half-volume effective concentration (EC50) of approximately 0.01 nM to approximately 100 nM. The antibody may not bind to non-pathological tau. The antibody may bind to pathological tau tangles. The antibody may inhibit tau aggregation. The antibody may contain a conjugate. The conjugate may contain a therapeutic agent. The conjugate may contain a detectable label.

[0010] In some embodiments, this disclosure provides constructs encoding antibodies according to any of the above or as described herein.

[0011] In some embodiments, the Disclosure provides methods for treating therapeutic indications in a subject, the methods comprising administering an antibody to the subject by any of the foregoing or as herein. Therapeutic indications may include neurological indications. Neurological indications may include one or more of the following: neurodegenerative diseases, Alzheimer's disease (AD), frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17), frontotemporal lobar degeneration (FTLD), frontotemporal dementia (FTD), chronic traumatic encephalopathy (CTE), progressive supranuclear palsy (PSP), Down syndrome, Pick's disease, corticobasal degeneration (CBD), corticobasal syndrome, amyotrophic lateral sclerosis (ALS), prion diseases, Creutzfeldt-Jakob disease (CJD), multiple system atrophy, tangle-only dementia, stroke, or progressive subcortical gliosis.

[0012] In some embodiments, the Disclosure provides a method for diagnosing therapeutic indications in a subject, the method comprising any of the above or the use of antibodies as herein. The therapeutic indications may include neurological indications. Neurological indications may include one or more of the following: neurodegenerative diseases, AD, FTDP-17, FTLD, FTD, CTE, PSP, Down syndrome, Pick's disease, CBD, corticobasal syndrome, ALS, prion diseases, CJD, multiple system atrophy, neurofibrillary senile dementia, stroke, and progressive subcortical gliosis. Antibodies may be used to detect pathological tau in the tissue of interest. The tissue of interest may include CNS tissue. The tissue of interest may include tissue strips. The tissue strips may include cryopreserved tissue pieces.

[0013] Those skilled in the art will be able to recognize or confirm many equivalents to the specific embodiments of the invention described herein simply by using standard experiments. Such equivalents are intended to be encompassed in the embodiments listed below.

[0014] List of embodiments E1. An isolated, for example, recombinant antibody that binds to human tau (e.g., SEQ ID NO: 274), wherein the antibody comprises a heavy chain variable region (VH) containing one, two, or three of the heavy chain complementarity-determining regions 1 (HCDR1), 2 (HCDR2), and 3 (HCDR3), and / or a light chain variable region (VL) containing one, two, or three of the light chain complementarity-determining regions 1 (LCDR1), 2 (LCDR2), and 3 (LCDR3), for example, a CDR sequence according to the Chothia numbering scheme. (i) The HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each contain the amino acid sequences of SEQ ID NOs. 82, 97, 115, 127, 141, and 159, respectively. (ii) The HCDR1, the HCDR2, the HCDR3, the LCDR1, the LCDR2, and the LCDR3 each contain the amino acid sequence of SEQ ID NO: 79, 94, 111, 127, 141, and 156, or (iii) The HCDR1, the HCDR2, the HCDR3, the LCDR1, the LCDR2, and the LCDR3 each contain the amino acid sequence of SEQ ID NO: 80, 95, 112, 129, 143, and 157, or (iv) The HCDR1, the HCDR2, the HCDR3, the LCDR1, the LCDR2, and the LCDR3 each contain the amino acid sequence of SEQ ID NO: 81, 94, 114, 127, 141, and 156, or (v) The HCDR1, the HCDR2, the HCDR3, the LCDR1, the LCDR2, and the LCDR3 each contain the amino acid sequence of SEQ ID NO: 82, 101, 119, 132, 149, and 164, or (vi) The HCDR1, the HCDR2, the HCDR3, the LCDR1, the LCDR2, and the LCDR3 each contain the amino acid sequence of SEQ ID NO: 77, 92, 109, 127, 141, and 154, or (vii) The HCDR1, the HCDR2, the HCDR3, the LCDR1, the LCDR2, and the LCDR3 each contain the amino acid sequence of SEQ ID NO: 78, 93, 110, 128, 142, and 155, or (viii) The HCDR1, the HCDR2, the HCDR3, the LCDR1, the LCDR2, and the LCDR3 each contain the amino acid sequence of SEQ ID NO: 78, 96, 113, 130, 144, and 158, or (xi) The HCDR1, the HCDR2, the HCDR3, the LCDR1, the LCDR2, and the LCDR3 each contain the amino acid sequence of SEQ ID NO: 83, 98, 116, 131, 145, and 160, or (x) The HCDR1, the HCDR2, the HCDR3, the LCDR1, the LCDR2, and the LCDR3 each contain the amino acid sequence of SEQ ID NO: 84, 99, 117, 132, 146, and 161, or (xi) The heavy chain complementarity-determining region 1 (HCDR1), the heavy chain complementarity-determining region 2 (HCDR2), the heavy chain complementarity-determining region 3 (HCDR3), the light chain complementarity-determining region 1 (LCDR1), the light chain complementarity-determining region 2 (LCDR2), and the light chain complementarity-determining region 3 (LCDR3) each contain the amino acid sequences of SEQ ID NO: 85, 100, 118, 133, 147, and 162, or (xii) The heavy chain complementarity-determining region 1 (HCDR1), the heavy chain complementarity-determining region 2 (HCDR2), the heavy chain complementarity-determining region 3 (HCDR3), the light chain complementarity-determining region 1 (LCDR1), the light chain complementarity-determining region 2 (LCDR2), and the light chain complementarity-determining region 3 (LCDR3) each contain the amino acid sequences of SEQ ID NO: 85, 100, 118, 134, 148, and 163, or (xiii) The heavy chain complementarity-determining region 1 (HCDR1), the heavy chain complementarity-determining region 2 (HCDR2), the heavy chain complementarity-determining region 3 (HCDR3), the light chain complementarity-determining region 1 (LCDR1), the light chain complementarity-determining region 2 (LCDR2), and the light chain complementarity-determining region 3 (LCDR3) each contain the amino acid sequences of SEQ ID NO: 86, 102, 120, 127, 141, and 156, or (xiv) The heavy chain complementarity-determining region 1 (HCDR1), the heavy chain complementarity-determining region 2 (HCDR2), the heavy chain complementarity-determining region 3 (HCDR3), the light chain complementarity-determining region 1 (LCDR1), the light chain complementarity-determining region 2 (LCDR2), and the light chain complementarity-determining region 3 (LCDR3) each contain the amino acid sequences of SEQ ID NO: 87, 103, 121, 132, 149, and 165, or (xv) The heavy chain complementarity-determining region 1 (HCDR1), the heavy chain complementarity-determining region 2 (HCDR2), the heavy chain complementarity-determining region 3 (HCDR3), the light chain complementarity-determining region 1 (LCDR1), the light chain complementarity-determining region 2 (LCDR2), and the light chain complementarity-determining region 3 (LCDR3) each contain the amino acid sequences of SEQ ID NO: 78, 104, 122, 135, 143, and 166, or (xvi) The heavy chain complementarity-determining region 1 (HCDR1), the heavy chain complementarity-determining region 2 (HCDR2), the heavy chain complementarity-determining region 3 (HCDR3), the light chain complementarity-determining region 1 (LCDR1), the light chain complementarity-determining region 2 (LCDR2), and the light chain complementarity-determining region 3 (LCDR3) each contain the amino acid sequences of SEQ ID NO: 78, 104, 122, 136, 150, and 167, or (xvii) The heavy chain complementarity-determining region 1 (HCDR1), the heavy chain complementarity-determining region 2 (HCDR2), the heavy chain complementarity-determining region 3 (HCDR3), the light chain complementarity-determining region 1 (LCDR1), the light chain complementarity-determining region 2 (LCDR2), and the light chain complementarity-determining region 3 (LCDR3) each contain the amino acid sequences of SEQ ID NO: 88, 105, 123, 137, 151, and 168, or (xviii) The heavy chain complementarity-determining region 1 (HCDR1), the heavy chain complementarity-determining region 2 (HCDR2), the heavy chain complementarity-determining region 3 (HCDR3), the light chain complementarity-determining region 1 (LCDR1), the light chain complementarity-determining region 2 (LCDR2), and the light chain complementarity-determining region 3 (LCDR3) each contain the amino acid sequences of SEQ ID NO: 89, 106, 124, 138, 152, and 169, or (xix) The HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each contain the amino acid sequences of SEQ ID NOs. 90, 107, 125, 139, 151, and 170, respectively. (xx) The HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each contain the amino acid sequences of SEQ ID NOs. 91, 108, 126, 140, 153, and 171, respectively. (xxi) The HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 contain any of the amino acid sequences of the HCDR and LCDR sequences shown in Table 1, or The antibody comprising a variant of any one antibody from (xxii)(i) to (xxi), e.g., a functional variant, wherein one, two, three, four, five, or all of HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and / or LCDR3 contain one, two, or at most three substitutions (e.g., conservative substitutions), or one, two, three, four, five, or all of HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and / or LCDR3 contain one, two, or at most three amino acids different from any of the sequences of (i)-(xxi).

[0015] E2. The antibody according to Embodiment E1, comprising any one of the HCDR1, HCDR2, and HCDR3 sequences (i) to (xxii).

[0016] E3. The antibody according to Embodiment E1 or E2, comprising any one of the LCDR1, LCDR2, and LCDR3 sequences of (i) to (xxii).

[0017] E4. The antibody according to any one of Embodiments E1 to E3, comprising one of the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 sequences from (i) to (xxii).

[0018] E4. Isolated, for example, recombinant antibodies that bind to human tau (e.g., SEQ ID NO: 274), wherein the antibodies are: (i) SEQ ID NOs: 7 and 25, respectively; (ii) SEQ ID NOs: 3 and 21, respectively; (iii) SEQ ID NOs: 4 and 22, respectively; (iv) SEQ ID NOs: 6 and 24, respectively; (v) SEQ ID NOs: 11 and 30, respectively; (vi) SEQ ID NOs: 1 and 19, respectively; (vii) SEQ ID NOs: 2 and 20, respectively; (viii) SEQ ID NOs: 5 and 23, respectively; (ix) (x) Sequence IDs 8 and 26 respectively, (x) Sequence IDs 9 and 27 respectively, (xi) Sequence IDs 10 and 28 respectively, (xii) Sequence IDs 10 and 29 respectively, (xiii) Sequence IDs 12 and 31 respectively, (xiv) Sequence IDs 13 and 32 respectively, (xv) Sequence IDs 14 and 33 respectively, (xvi) Sequence IDs 14 and 34 respectively, (xvii) Sequence IDs 15 and 35 respectively, (xviii) Sequence IDs 16 and 36 respectively, (xix) Sequence IDs 17 and 37 respectively, or (xx) Sequence IDs 18 and 38 respectively The antibody comprises heavy chain CDR1, CDR2, and CDR3, and / or light chain CDR1, CDR2, and CDR3, which include a heavy chain variable region (VH) and a light chain variable region (VL).

[0019] E5. The antibody according to Embodiment E4, wherein the CDR sequence is based on the Kabat numbering system, the Chothia numbering system, or the IMGT numbering system.

[0020] E6. (i) Any VH amino acid sequence shown in Table 1, or an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto, (ii) An amino acid sequence that has at least 1, 2, or 3 modifications compared to any VH shown in Table 1, but with no more than 30, 20, or 10 modifications, (iii) An amino acid sequence that, compared to any one of the amino acid sequences of any VH sequence shown in Table 1, contains at least 1, 2, or 3 different amino acids, but no more than 30, 20, or 10 different amino acids, or (iv) Any VH nucleotide sequence shown in Table 1X, or an amino acid sequence encoded by a nucleotide sequence having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto. The antibody according to any one of the prior embodiments, comprising VH containing the above.

[0021] E7. (i) any amino acid sequence of sequence numbers 7, 3, 4, 6, and 11, or an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto, (ii) an amino acid sequence of any of sequence numbers 7, 3, 4, 6, and 11 with at least one, two, or three modifications, provided that the amino acid sequence includes 30, 20, or 10 or fewer modifications. (iii) an amino acid sequence that, compared to any one of the amino acid sequences of SEQ ID NOs. 7, 3, 4, 6, and 11, contains at least 1, 2, or 3 different amino acids, but no more than 30, 20, or 10 different amino acids, or (iv) an amino acid sequence encoded by any of the base sequences of SEQ ID NOs. 51, 55, 54, 52, 47, 39, 56, 41, 50, 49, 48, 46, 45, 44, 43, 42, 53, or 40, or by a nucleotide sequence having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% sequence identity to it. The antibody according to any one of the prior embodiments, comprising VH containing the above.

[0022] E8. (i) Any VL amino acid sequence shown in Table 1, or an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto, (ii) An amino acid sequence that has at least 1, 2, or 3 modifications compared to any VL shown in Table 1, but with no more than 30, 20, or 10 modifications, (iii) An amino acid sequence that, compared to any one of the amino acid sequences of any VL sequence shown in Table 1, contains at least 1, 2, or 3 different amino acids, but no more than 30, 20, or 10 different amino acids, or (iv) Any VL nucleotide sequence shown in Table 1, or an amino acid sequence encoded by a nucleotide sequence having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto. The antibody according to any one of the prior embodiments, comprising a VL containing the above.

[0023] E9. (i) any amino acid sequence of sequence numbers 25, 21, 22, 24, and 30, or an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto, (ii) At least one, two, or three modifications to any of the amino acid sequences of SEQ ID NOs. 25, 21, 22, 24, and 30, provided that the amino acid sequence includes 30, 20, or 10 or fewer modifications. (iii) an amino acid sequence that, compared to any one of the amino acid sequences of sequence numbers 25, 21, 22, 24, and 30, contains at least 1, 2, or 3 different amino acids, but not more than 30, 20, or 10 different amino acids, or (iv) an amino acid sequence encoded by any of the base sequences of SEQ ID NOs. 67, 75, 74, 72, 66, 57, 76, 59, 70, 69, 68, 65, 64, 62, 63, 61, 60, 73, or 58, or a nucleotide sequence having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto. The antibody according to any one of the prior embodiments, comprising a VL containing the above.

[0024] E10. (i) (a) Any VH amino acid sequence shown in Table 1, or an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto. (b) An amino acid sequence having at least 1, 2, or 3 modifications, but no more than 30, 20, or 10 modifications, compared to any VH amino acid sequence shown in Table 1. (c) An amino acid sequence that contains at least 1, 2, or 3, 30, 20, or 10 different amino acids compared to any one of the amino acid sequences of any VH sequence shown in Table 1, or (d) Any VH nucleotide sequence shown in Table 1, or an amino acid sequence encoded by a nucleotide sequence having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto. VH including, and (ii) (a) Any VL amino acid sequence shown in Table 1, or an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto. (b) Amino acid sequences that, compared to any VL amino acid sequence shown in Table 1, have at least 1, 2, or 3 modifications, but no more than 30, 20, or 10 modifications, (c) Compared to any one of the amino acid sequences of any VL sequence shown in Table 1, an amino acid sequence containing at least 1, 2, or 3 different amino acids, but not more than 30, 20, or 10 different amino acids, or (d) Any VL nucleotide sequence shown in Table 1, or an amino acid sequence encoded by a nucleotide sequence having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto. VL including The antibody described in any one of the prior embodiments, including the antibody described in any one of the prior embodiments.

[0025] E11. The antibody according to any one of the prior embodiments, comprising the amino acid sequence of any VH of the antibody shown in Table 1, and the amino acid sequence of VL of the antibody shown in Table 1.

[0026] E12. (i) an amino acid sequence of SEQ ID NO: 7, an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto, an amino acid sequence containing at least 1, 2, or 3 different amino acids, but not more than 30, 20, or 10, compared to SEQ ID NO: 7, or a VH containing an amino acid sequence containing at least 1, 2, or 3 modifications, e.g., substitutions (e.g., conservative substitutions), compared to SEQ ID NO: 7, and (ii) an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto, an amino acid sequence having at least 1, 2, or 3 different amino acids compared to SEQ ID NO: 25, but no more than 30, 20, or 10 different amino acids, or an amino acid sequence having at least 1, 2, or 3 modifications, e.g., substitutions (e.g., conservative substitutions) compared to SEQ ID NO: 25 The antibody described in any one of the prior embodiments, including the antibody described in any one of the prior embodiments.

[0027] E13. (i) an amino acid sequence of SEQ ID NO: 3, an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto, an amino acid sequence containing at least 1, 2, or 3 different amino acids compared to SEQ ID NO: 3, but no more than 30, 20, or 10 different amino acids, or a VH containing at least 1, 2, or 3 modifications, e.g., substitutions (e.g., conservative substitutions) compared to SEQ ID NO: 3, and (ii) The amino acid sequence of SEQ ID NO: 21, or an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto, an amino acid sequence containing at least 1, 2, or 3 different amino acids compared to SEQ ID NO: 21, but no more than 30, 20, or 10 different amino acids, or an amino acid sequence containing at least 1, 2, or 3 modifications, e.g., substitutions (e.g., conservative substitutions) compared to SEQ ID NO: 21, VL The antibody according to any one of embodiments E1 to E11, including the antibody described in any one of embodiments E1 to E11.

[0028] E14. (i) an amino acid sequence of SEQ ID NO: 4, an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto, an amino acid sequence containing at least 1, 2, or 3 different amino acids, but not more than 30, 20, or 10, compared to SEQ ID NO: 4, or an amino acid sequence containing at least 1, 2, or 3 modifications, e.g., substitutions (e.g., conservative substitutions), compared to SEQ ID NO: 4, and VH, and (ii) an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% sequence identity with respect to the amino acid sequence of SEQ ID NO: 22, an amino acid sequence containing at least 1, 2, or 3 different amino acids, but not more than 30, 20, or 10, compared to SEQ ID NO: 22, or an amino acid sequence containing at least 1, 2, or 3 modifications, e.g., substitutions (e.g., conservative substitutions), compared to SEQ ID NO: 22, but not more than 30, 20, or 10 modifications, e.g., substitutions (e.g., conservative substitutions) in a VL The antibody according to any one of embodiments E1 to E11, including the antibody described in any one of embodiments E1 to E11.

[0029] E15. (i) an amino acid sequence of SEQ ID NO: 6, an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto, an amino acid sequence having at least 1, 2, or 3 different amino acids compared to SEQ ID NO: 6, but no more than 30, 20, or 10 different amino acids, or a VH having at least 1, 2, or 3 modifications, e.g., substitutions (e.g., conservative substitutions) compared to SEQ ID NO: 6, and (ii) The amino acid sequence of SEQ ID NO: 22, or an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto, an amino acid sequence containing at least 1, 2, or 3 different amino acids compared to SEQ ID NO: 24, but not more than 30, 20, or 10 different amino acids, or an amino acid sequence containing at least 1, 2, or 3, 30, 20, or 10 modifications, e.g., substitutions (e.g., conservative substitutions) compared to SEQ ID NO: 24, VL The antibody according to any one of embodiments E1 to E11, including the antibody described in any one of embodiments E1 to E11.

[0030] E16. (i) an amino acid sequence of SEQ ID NO: 11, an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto, an amino acid sequence containing at least 1, 2, or 3 different amino acids compared to SEQ ID NO: 11, but no more than 30, 20, or 10 different amino acids, or a VH containing at least 1, 2, or 3 modifications, e.g., substitutions (e.g., conservative substitutions) compared to SEQ ID NO: 11, and (ii) The amino acid sequence of SEQ ID NO: 30, or an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto, an amino acid sequence containing at least 1, 2, or 3 different amino acids compared to SEQ ID NO: 30, but no more than 30, 20, or 10 different amino acids, or an amino acid sequence containing at least 1, 2, or 3 modifications, e.g., substitutions (e.g., conservative substitutions) compared to SEQ ID NO: 30, including VL The antibody according to any one of embodiments E1 to E11, including the antibody described in any one of embodiments E1 to E11.

[0031] E17. An isolated, for example, recombinant antibody that binds to human tau (e.g., SEQ ID NO: 274), wherein the antibody comprises a heavy chain variable region (VH) and / or a light chain variable region (VL), which are (i) SEQ ID NO: 7 and / or 25, respectively, (ii) SEQ ID NO: 3 and / or 21, respectively, (iii) SEQ ID NO: 4 and / or 22, respectively, (iv) SEQ ID NO: 6 and / or 24, respectively, (v) SEQ ID NO: 11 and / or 30, respectively, (vi) SEQ ID NO: 1 and / or 19, (vi i) Sequence IDs 2 and / or 20 respectively, (viii) Sequence IDs 5 and / or 23 respectively, (ix) Sequence IDs 8 and / or 26 respectively, (x) Sequence IDs 9 and / or 27 respectively, (xi) Sequence IDs 10 and / or 28 respectively, (xii) Sequence IDs 10 and / or 29 respectively, (xiii) Sequence IDs 12 and / or 31 respectively, (xiv) Sequence IDs 13 and / or 32 respectively, (xv) Sequence IDs 14 and / or 33 respectively, (xvi) Sequence ID 14 and / or 34, (xvii) SEQ ID NOs. 15 and / or 35 respectively, (xviii) SEQ ID NOs. 16 and / or 36 respectively, (xix) SEQ ID NOs. 17 and / or 37 respectively, (xx) SEQ ID NOs. 18 and / or 38 respectively, (xxi) The VH and / or VL have at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% sequence identity with respect to any one of (i) to (xx), any of (i) to (xx) An antibody comprising a variant of one antibody, e.g., a functional variant, or (xxii) the VH and / or VL having at least 1, 2, or 3 modifications, but no more than 30, 20, or 10 modifications (e.g., amino acid substitutions, e.g., conservative substitutions), or the VH and / or VL having at least 1, 2, or 3 different amino acids, but no more than 30, 20, or 10, comprising a variant of any one of the antibodies (i) to (xx), e.g., a functional variant.

[0032] E18. The antibody according to any one of Embodiments E1 to E17, wherein the antibody comprises one of the VH sequence and VL sequence of Embodiment E17(i) to (xxii).

[0033] E19. For example, an antibody according to any one of Embodiments E1 to E18 that binds to tau protein at a maximum half-volume effective concentration (EC50) of approximately 0.001 nM to approximately 10 nM, or approximately 0.01 nM to approximately 2 nM, when evaluated by direct enzyme-linked immunosorbent assay (ELISA).

[0034] E20. For example, the antibody according to any one of Embodiments E1 to E19, which binds to concentrated pair-helical fibrillary tau protein (ePHF) at a maximum half-volume effective concentration (EC50) of approximately 0.001 nM to approximately 100 nM, or approximately 0.01 nM to approximately 20 nM, when evaluated by direct enzyme-linked immunosorbent assay (ELISA).

[0035] E21. For example, when evaluated by biolayer interferometry, the dissociation constant (K) is approximately 0.1 to 10 nM, or approximately 0.2 to 5 nM. D An antibody according to any one of Embodiments E1 to E20, which binds to iPHF.

[0036] E22. An antibody according to any one of the prior embodiments, which binds to a tau protein epitope comprising a region formed by a complex of at least two tau proteins.

[0037] E23. The antibody according to any one of the prior embodiments, wherein the antibody is conjugated to all or part of tau amino acid residues selected from (a) 183-212, (b) 187-218, (c) 33-82, 159-182, 197-226, and 229-246, (d) 217-242, (e) 35-76 and 187-218, (f) 5-34, (g) 187-218, (h) 33-82, 159-188, and 191-230, (i) 35-62, 107-124, and 203-220, (j) 35-82, 159-188, and 197-224, and (k) 53-78, 329-348, or 381-408, and the human tau is numbered by Sequence ID No. 274.

[0038] E24. Isolated, for example, recombinant antibodies that bind to human tau (e.g., SEQ ID NO: 274), wherein the antibodies are (a) 183-212, (b) 187-218, (c) 33-82, 159-182, 197-226, and 229-246, (d) 217-242, (e) 35-76 and 187-218, (f) 5-34, (g) 187-218, (h) 33- The antibody is characterized by binding to all or part of tau amino acid residues selected from (i) 82, 159-188, and 191-230, (j) 35-82, 159-188, and 197-224, or (k) 53-78, 329-348, and 381-408, wherein the human tau is numbered by Sequence ID No. 274.

[0039] E25. The antibody according to Embodiment E23 or E24, wherein one or more of serine, threonine, and / or tyrosine in a stretch of amino acids selected from (a) to (k) are phosphorylated.

[0040] E26. The antibody according to any one of Embodiments E23 to E25, wherein all of the serine, threonine, and / or tyrosine in the stretch of amino acids selected from (a) to (k) are phosphorylated.

[0041] E27. An antibody according to any one of Embodiments E24 to E26, comprising one of the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 sequences from any one of Embodiments E3(i) to (xxii).

[0042] E28. An antibody according to any one of Embodiments E24 to E27, comprising one VH sequence and a VL sequence from any one of Embodiments E17(i) to (xxii).

[0043] E29. For example, when evaluated by biolayer interferometry, the dissociation constant (K) is approximately 1 pM to 50 pM, or approximately 1 to 25 pM. D An antibody according to any one of Embodiments E1 to E28, wherein the antibody binds to all or part of amino acids 195 to 215 of tau.

[0044] E30. For example, when evaluated by biolayer interferometry, the dissociation constant (K) is approximately 0.1 nM to 10 nM, or approximately 0.5 to 5 nM. D An antibody according to any one of Embodiments E1 to E28, wherein the antibody binds to all or part of amino acids 191 to 214 of tau phosphorylated at S199.

[0045] E31. For example, when evaluated by biolayer interferometry, the dissociation constant (K) is approximately 0.1 nM to 10 nM, or approximately 0.1 to 5 nM. D The antibody according to any one of Embodiments E1 to E28, wherein the antibody binds to all or part of amino acids 217-234 of tau phosphorylated at T217, T220, and T231.

[0046] E32. For example, when evaluated by biolayer interferometry, the dissociation constant (K) is approximately 0.1 nM to 25 nM, or approximately 0.1 to 15 nM. D An antibody according to any one of Embodiments E1 to E28, wherein the antibody binds to all or part of amino acids 225-240 of tau phosphorylated at T231.

[0047] E33. Isolated, for example, recombinant antibodies that bind to human tau phosphorylated at amino acid residue S404, or to peptides containing or consisting of the amino acid sequence DHGAEIVYKSPVVSGDT(pS)PRHLSNVSSTG (SEQ ID NO: 281) (wherein p(S) corresponds to a phosphorylated serine residue).

[0048] E34. The antibody according to Embodiment E33, wherein the antibody comprises a heavy chain variable region (VH) containing CDR1, CDR2, and CDR3 sequences, respectively, including SEQ ID NOs. 89, 106, and 124, and a light chain variable region (VL) containing CDR1, CDR2, and CDR3 sequences, respectively, including SEQ ID NOs. 138, 152, and 169.

[0049] E35. The antibody according to Embodiment E33 or E34, wherein the antibody comprises a VH sequence and a VL sequence, respectively, comprising SEQ ID NOs. 16 and 36.

[0050] E36. Isolated, for example, recombinant antibodies, (a) Human tau, which is phosphorylated at amino acid residue S199 but not at amino acid residues S202 and T205, (b) Human tau that is phosphorylated at amino acid residue S202 but not at amino acid residues S199 and T205, (c) Human tau, which is phosphorylated at amino acid residue T205 but not at amino acid residues S199 and S202, (d) Binding to human tau that is phosphorylated at amino acid residues S199 and T205 but not at amino acid residue S202 (in which case, for example, binding to tau phosphorylated at S199 and T205 is at least 3 times stronger (e.g., at least 4 times stronger) than background (e.g., nonspecific) binding, e.g., binding by hIgG1 isotype control), (e) Human tau that is phosphorylated at amino acid residues S202 and T205 but not at amino acid residue S199 is phosphorylated; however, human tau that is phosphorylated at residues S199 and S202 but not at T205 is not phosphorylated. (f) Human tau phosphorylated with amino acid residues (i) S202 and T205 (excluding S119), and (ii) S199 and T205 (excluding S202), binds at least twice as strongly as background (e.g., nonspecific) binding, e.g., at least three times, at least four times, at least five times, 2-6 times, 2-5 times, 2-4 times, 2-3 times, 3-5 times, or 4-5 times stronger than the binding level by the hIgG1 isotype control. (g) amino acid residues (i) S199 and S202 (excluding T205), (ii) S202 and T205 (excluding S199), (iii) S199 and T205 (excluding S202), and (iv) human tau phosphorylated with combinations of S199, S202, and T205, in which case the binding to phosphorylated tau is at least 1.6 times stronger (e.g., at least 1.7 times, at least 1.8 times, at least 1.9 times, at least 2 times, at least 3 times, 1.6 to 3 times, 1.6 to 2 times stronger) than background (e.g., nonspecific) binding, e.g., binding by hIgG1 isotype control), (h) A peptide containing or consisting of the amino acid sequence SGDRSGYS(pS)PGSPGTPGSRSRTPS (SEQ ID NO: 284) is bound to the peptide. (i) A peptide containing or consisting of the amino acid sequence SGDRSGYSSPG(pS)PGTPGSRSRTPS (SEQ ID NO: 285) is bound to it. (j) A peptide containing or consisting of the amino acid sequence SGDRSGYSSPGSPG(pT)PGSRSRTPS (SEQ ID NO: 286), or (k) Binding to a peptide containing or consisting of the amino acid sequence SGDRSGYS(pS)PGSPG(pT)PGSRSRTPS (SEQ ID NO: 290) (for example, in which case the binding to the peptide is at least 3 times stronger (for example, at least 4 times stronger) than the level of background (e.g., nonspecific) binding, e.g., binding by an hIgG1 isotype control), (l) It binds to peptides containing or consisting of the amino acid sequence SGDRSGYSSPG(pS)PG(pT)PGSRSRTPS (SEQ ID NO: 289), but does not bind to peptides containing or consisting of the amino acid sequence SGDRSGYS(pS)PG(pS)PGTPGSRSRTPS (SEQ ID NO: 288), (m) Binds to peptides containing or consisting of the amino acid sequences SGDRSGYSSPG(pS)PG(pT)PGSRSRTPS (SEQ ID NO: 289) and SGDRSGYS(pS)PGSPG(pT)PGSRSRTPS (SEQ ID NO: 290) (in which case the binding to the latter peptide is at least twice as strong (e.g., at least three times, at least four times, at least five times, 2-6 times, 2-5 times, 2-4 times, 2-3 times, 3-5 times, or 4-5 times) than background (e.g., nonspecific) binding, e.g., binding by an hIgG1 isotype control), or (n) Binding to peptides containing or consisting of the amino acid sequences SGDRSGYS(pS)PG(pS)PGTPGSRSRTPS (SEQ ID NO: 288), SGDRSGYSSPG(pS)PG(pT)PGSRSRTPS (SEQ ID NO: 289), SGDRSGYS(pS)PGSPG(pT)PGSRSRTPS (SEQ ID NO: 290), and SGDRSGYS(pS)PG(pS)PG(pT)PGSRSRTPS (SEQ ID NO: 287) (for example, in this case, the binding to the peptide is at least 1.6 times stronger than the level of background (e.g., nonspecific) binding, e.g., binding by an hIgG1 isotype control (e.g., at least 1.7 times, at least 1.8 times, at least 1.9 times, at least 2 times, at least 3 times, 1.6 to 4 times, 1.6 to 3 times stronger)), p(S) and p(T) correspond to phosphorylated serine and phosphorylated threonine, respectively. Optionally, the binding may be evaluated using, for example, the one-point ELISA described in Example 7, and optionally, the human tau antibody may have the sequence described in SEQ ID NO: 274.

[0051] E37. The aforementioned antibody (a) Heavy chain variable region (VH) containing CDR1, CDR2, and CDR3 sequences including sequence numbers 82, 97, and 115, respectively, and light chain variable region (VL) containing CDR1, CDR2, and CDR3 sequences including sequence numbers 127, 141, and 159, respectively, (b) Heavy chain variable region (VH) containing CDR1, CDR2, and CDR3 sequences including sequence numbers 79, 94, and 111, respectively, and light chain variable region (VL) containing CDR1, CDR2, and CDR3 sequences including sequence numbers 127, 141, and 156, respectively, (c) Heavy chain variable region (VH) containing CDR1, CDR2, and CDR3 sequences containing sequence numbers 81, 94, and 114 respectively, and light chain variable region (VL) containing CDR1, CDR2, and CDR3 sequences containing sequence numbers 127, 141, and 156 respectively, (d) Heavy chain variable regions (VH) containing CDR1, CDR2, and CDR3 sequences containing sequence numbers 77, 92, and 109 respectively, and light chain variable regions (VL) containing CDR1, CDR2, and CDR3 sequences containing sequence numbers 127, 141, and 154 respectively, or (e) Heavy chain variable regions (VH) containing CDR1, CDR2, and CDR3 sequences containing sequence numbers 86, 102, and 120, respectively, and light chain variable regions (VL) containing CDR1, CDR2, and CDR3 sequences containing sequence numbers 127, 141, and 156, respectively. The antibody according to embodiment E36, including the antibody described above.

[0052] E38. The antibody comprises VH and VL, and the VH and VL are (a) Sequence IDs 7 and 25, respectively (b) Sequence IDs 8 and 21, respectively (c) Sequence IDs 6 and 24 respectively, (d) Sequence IDs 1 and 19, respectively, or (e) Sequence IDs 12 and 31 respectively The antibody according to embodiment E36 or E37, comprising the amino acid sequence.

[0053] E39. Isolated, for example, recombinant antibodies, (a) Conjugates to human tau that is phosphorylated at amino acid residue S199 but not at amino acid residues S202 and T205, wherein the antibody comprises a heavy chain variable region (VH) containing CDR1, CDR2, and CDR3 sequences including SEQ ID NOs. 82, 97, and 115, respectively, and a light chain variable region (VL) containing CDR1, CDR2, and CDR3 sequences including SEQ ID NOs. 127, 141, and 159, respectively. (b) Human tau that is phosphorylated at amino acid residue S202 but not at amino acid residues S199 and T205, and the antibody contains (i) a heavy chain variable region (VH) containing CDR1, CDR2, and CDR3 sequences containing SEQ ID NOs. 79, 94, and 111, respectively, and a light chain variable region (VL) containing CDR1, CDR2, and CDR3 sequences containing SEQ ID NOs. 127, 141, and 156, respectively, or (ii) a CDR containing SEQ ID NOs. 81, 94, and 114, respectively. 1. A heavy chain variable region (VH) containing CDR2 and CDR3 sequences, and a light chain variable region (VL) containing CDR1, CDR2, and CDR3 sequences containing SEQ ID NOs. 127, 141, and 156, respectively, or (iii) a heavy chain variable region (VH) containing CDR1, CDR2, and CDR3 sequences containing SEQ ID NOs. 86, 102, and 120, respectively, and a light chain variable region (VL) containing CDR1, CDR2, and CDR3 sequences containing SEQ ID NOs. 127, 141, and 156, respectively. (c) Human tau that is phosphorylated at amino acid residue T205 but not at amino acid residues S199 and S202, and the antibody contains (i) a heavy chain variable region (VH) containing CDR1, CDR2, and CDR3 sequences containing SEQ ID NOs. 79, 94, and 111, respectively, and a light chain variable region (VL) containing CDR1, CDR2, and CDR3 sequences containing SEQ ID NOs. 127, 141, and 156, respectively, or (ii) a CDR containing SEQ ID NOs. 81, 94, and 114, respectively. 1. A heavy chain variable region (VH) containing CDR2 and CDR3 sequences, and a light chain variable region (VL) containing CDR1, CDR2, and CDR3 sequences containing SEQ ID NOs. 127, 141, and 156, respectively, or (iii) a heavy chain variable region (VH) containing CDR1, CDR2, and CDR3 sequences containing SEQ ID NOs. 86, 102, and 120, respectively, and a light chain variable region (VL) containing CDR1, CDR2, and CDR3 sequences containing SEQ ID NOs. 127, 141, and 156, respectively. (d) Binds to human tau that is phosphorylated at amino acid residues S199 and T205 but not at amino acid residue S202 (for example, in which case binding to tau phosphorylated at S199 and T205 is at least 3 times stronger (for example, at least 4 times stronger) than background (e.g., nonspecific) binding, e.g., binding by hIgG1 isotype control), wherein the antibody comprises (i) a heavy chain variable region (VH) comprising CDR1, CDR2, and CDR3 sequences comprising SEQ ID NOs. 79, 94, and 111, respectively, and a light chain variable region (VL) comprising CDR1, CDR2, and CDR3 sequences comprising SEQ ID NOs. 127, 141, and 156, respectively, or (ii) CDR1 (iii) a heavy chain variable region (VH) containing CDR2 and CDR3 sequences, and a light chain variable region (VL) containing CDR1, CDR2 and CDR3 sequences containing sequence numbers 127, 141 and 156, respectively; or (iv) a heavy chain variable region (VH) containing CDR1, CDR2 and CDR3 sequences containing sequence numbers 82, 97 and 115, respectively, and a light chain variable region (VL) containing CDR1, CDR2 and CDR3 sequences containing sequence numbers 127, 141 and 159, respectively; (e) Human tau that is phosphorylated at amino acid residues S202 and T205 but not at amino acid residue S199, and is not human tau that is phosphorylated at amino acid residues S199 and S202 but not at T205, and the antibody binds to such human tau, and the antibody comprises a heavy chain variable region (VH) containing CDR1, CDR2, and CDR3 sequences including SEQ ID NOs. 77, 92, and 109, respectively, and a light chain variable region (VL) containing CDR1, CDR2, and CDR3 sequences including SEQ ID NOs. 127, 141, and 154, respectively. (f) Human tau phosphorylated with amino acid residues (i) S202 and T205 (excluding S119), and (ii) S199 and T205 (excluding S202), binds at least twice as strongly to background (nonspecific) binding, for example, at least three times, at least four times, at least five times, 2-6 times, 2-5 times, 2-4 times, 2-3 times, 3-5 times, or 4-5 times more strongly than the binding level by the hIgG1 isotype control, wherein the antibody comprises (i) a heavy chain variable region (VH) comprising CDR1, CDR2, and CDR3 sequences including SEQ ID NOs. 79, 94, and 111, respectively, and a light chain variable region (VL) comprising CDR1, CDR2, and CDR3 sequences including SEQ ID NOs. 127, 141, and 156, respectively, or (ii) SEQ ID NOs. 81, 94, and 11 (iii) a heavy chain variable region (VH) containing CDR1, CDR2, and CDR3 sequences containing 4, and a light chain variable region (VL) containing CDR1, CDR2, and CDR3 sequences containing SEQ ID NOs. 127, 141, and 156, respectively; or (iv) a heavy chain variable region (VH) containing CDR1, CDR2, and CDR3 sequences containing SEQ ID NOs. 82, 97, and 115, respectively, and a light chain variable region (VL) containing CDR1, CDR2, and CDR3 sequences containing SEQ ID NOs. 127, 141, and 159, respectively; (g) Binding to phosphorylated human tau with amino acid residues (i) S199 and S202 (excluding T205), (ii) S202 and T205 (excluding S199), (iii) S199 and T205 (excluding S202), and (iv) combinations of S199, S202, and T205 (for example, in that case, the binding to phosphorylated tau is background (non-specific) binding, e.g., the level of binding by hIgG1 isotype control) The antibody is at least 1.6 times stronger (for example, at least 1.7 times, at least 1.8 times, at least 1.9 times, at least 2 times, at least 3 times, 1.6 to 3 times, 1.6 to 2 times stronger), and the antibody contains (i) heavy chain variable regions (VH) containing CDR1, CDR2, and CDR3 sequences including SEQ ID NOs. 79, 94, and 111, respectively, and CDR1, CDR2, and CDR3 sequences including SEQ ID NOs. 127, 141, and 156, respectively. (ii) a light chain variable region (VL) containing a heavy chain variable region (VH) containing CDR1, CDR2, and CDR3 sequences containing sequence numbers 81, 94, and 114 respectively, and a light chain variable region (VL) containing CDR1, CDR2, and CDR3 sequences containing sequence numbers 127, 141, and 156 respectively, or (iii) a heavy chain variable region containing CDR1, CDR2, and CDR3 sequences containing sequence numbers 82, 97, and 115 respectively. (iv) comprising a heavy chain variable region (VH) and a light chain variable region (VL) comprising CDR1, CDR2, and CDR3 sequences including sequence numbers 127, 141, and 159, respectively, and a heavy chain variable region (VH) comprising CDR1, CDR2, and CDR3 sequences including sequence numbers 86, 102, and 120, respectively, and a light chain variable region (VL) comprising CDR1, CDR2, and CDR3 sequences including sequence numbers 127, 141, and 156, respectively, (h) Conjugated to a peptide containing or comprising the amino acid sequence SGDRSGYS(pS)PGSPGTPGSRSRTPS (SEQ ID NO: 284), the antibody comprises a heavy chain variable region (VH) containing CDR1, CDR2, and CDR3 sequences, respectively, including SEQ ID NOs: 82, 97, and 115, and a light chain variable region (VL) containing CDR1, CDR2, and CDR3 sequences, respectively, including SEQ ID NOs: 127, 141, and 159. (i) Conjugated to a peptide containing or comprising the amino acid sequence SGDRSGYSSPG(pS)PGTPGSRSRTPS (SEQ ID NO: 285), wherein the antibody contains (i) a heavy chain variable region (VH) containing CDR1, CDR2, and CDR3 sequences, respectively, including SEQ ID NOs: 79, 94, and 111, and a light chain variable region (VL) containing CDR1, CDR2, and CDR3 sequences, respectively, including SEQ ID NOs: 127, 141, and 156, respectively, or (ii) containing SEQ ID NOs: 81, 94, and 114, respectively (iii) A heavy chain variable region (VH) containing CDR1, CDR2, and CDR3 sequences, and a light chain variable region (VL) containing CDR1, CDR2, and CDR3 sequences containing SEQ ID NOs. 127, 141, and 156, respectively, or (iii) a heavy chain variable region (VH) containing CDR1, CDR2, and CDR3 sequences containing SEQ ID NOs. 86, 102, and 120, respectively, and a light chain variable region (VL) containing CDR1, CDR2, and CDR3 sequences containing SEQ ID NOs. 127, 141, and 156, respectively. (j) Conjugated to a peptide containing or comprising the amino acid sequence SGDRSGYSSPGSPG(pT)PGSRSRTPS (SEQ ID NO: 286), wherein the antibody (i) comprises a heavy chain variable region (VH) comprising CDR1, CDR2, and CDR3 sequences comprising SEQ ID NOs: 79, 94, and 111, respectively, and a light chain variable region (VL) comprising CDR1, CDR2, and CDR3 sequences comprising SEQ ID NOs: 127, 141, and 156, respectively, or (ii) comprising SEQ ID NOs: 81, 94, and 114, respectively (iii) A heavy chain variable region (VH) containing CDR1, CDR2, and CDR3 sequences, and a light chain variable region (VL) containing CDR1, CDR2, and CDR3 sequences containing SEQ ID NOs. 127, 141, and 156, respectively, or (iii) a heavy chain variable region (VH) containing CDR1, CDR2, and CDR3 sequences containing SEQ ID NOs. 86, 102, and 120, respectively, and a light chain variable region (VL) containing CDR1, CDR2, and CDR3 sequences containing SEQ ID NOs. 127, 141, and 156, respectively. (k) Binding to a peptide containing or consisting of the amino acid sequence SGDRSGYS(pS)PGSPG(pT)PGSRSRTPS (SEQ ID NO: 290) (for example, in which case the binding to the peptide is at least 3 times stronger (e.g., at least 4 times stronger) than the level of background (nonspecific) binding, e.g., binding by an hIgG1 isotype control), wherein the antibody contains (i) a heavy chain variable region (VH) containing CDR1, CDR2, and CDR3 sequences, respectively, including SEQ ID NOs: 79, 94, and 111, and a light chain variable region (VL) containing CDR1, CDR2, and CDR3 sequences, respectively, including SEQ ID NOs: 127, 141, and 156, respectively, or (ii) CDR1, CDR2, and CDR3 sequences, respectively, including SEQ ID NOs: 81, 94, and 114 (iii) a heavy chain variable region (VH) containing a DR3 sequence, and a light chain variable region (VL) containing CDR1, CDR2, and CDR3 sequences containing sequence numbers 127, 141, and 156, respectively; or (iv) a heavy chain variable region (VH) containing CDR1, CDR2, and CDR3 sequences containing sequence numbers 82, 97, and 115, respectively, and a light chain variable region (VL) containing CDR1, CDR2, and CDR3 sequences containing sequence numbers 127, 141, and 159, respectively; (l) It binds to peptides containing or consisting of the amino acid sequence SGDRSGYSSPG(pS)PG(pT)PGSRSRTPS (SEQ ID NO: 289), but does not bind to peptides containing or consisting of the amino acid sequence SGDRSGYS(pS)PG(pS)PGTPGSRSRTPS (SEQ ID NO: 288), wherein the antibody comprises a heavy chain variable region (VH) containing CDR1, CDR2, and CDR3 sequences, respectively, including SEQ ID NOs: 77, 92, and 109, and a light chain variable region (VL) containing CDR1, CDR2, and CDR3 sequences, respectively, including SEQ ID NOs: 127, 141, and 154. (m) Binds to a peptide containing or consisting of the amino acid sequences SGDRSGYSSPG(pS)PG(pT)PGSRSRTPS (SEQ ID NO: 289) and SGDRSGYS(pS)PGSPG(pT)PGSRSRTPS (SEQ ID NO: 290), in which case the binding to the latter peptide is at least twice as strong (e.g., at least three times, at least four times, at least five times, 2-6 times, 2-5 times, 2-4 times, 2-3 times, 3-5 times, or 4-5 times) than background (non-specific) binding, for example, binding by an hIgG1 isotype control, and the antibody contains (i) a heavy chain variable region (VH) containing CDR1, CDR2, and CDR3 sequences, respectively, including SEQ ID NOs: 79, 94, and 111, and a light chain variable region (VL) containing CDR1, CDR2, and CDR3 sequences, respectively, including SEQ ID NOs: 127, 141, and 156. (ii) comprising a heavy chain variable region (VH) comprising CDR1, CDR2, and CDR3 sequences including sequence numbers 81, 94, and 114 respectively, and a light chain variable region (VL) comprising CDR1, CDR2, and CDR3 sequences including sequence numbers 127, 141, and 156 respectively, (iii) comprising a heavy chain variable region (VH) comprising CDR1, CDR2, and CDR3 sequences including sequence numbers 82, 97, and 115 respectively, and a light chain variable region (VL) comprising CDR1, CDR2, and CDR3 sequences including sequence numbers 127, 141, and 159 respectively, or (iv) comprising a heavy chain variable region (VH) comprising CDR1, CDR2, and CDR3 sequences including sequence numbers 86, 102, and 120 respectively, and a light chain variable region (VL) comprising CDR1, CDR2, and CDR3 sequences including sequence numbers 127, 141, and 156 respectively, or (n) A peptide containing or composed of the amino acid sequences SGDRSGYS(pS)PG(pS)PGTPGSRSRTPS (SEQ ID NO: 288), SGDRSGYSSPG(pS)PG(pT)PGSRSRTPS (SEQ ID NO: 289), SGDRSGYS(pS)PGSPG(pT)PGSRSRTPS (SEQ ID NO: 290), and SGDRSGYS(pS)PG(pS)PG(pT)PGSRSRTPS (SEQ ID NO: 287) (for example, in that case, the peptide The binding to the plutide is at least 1.6 times stronger (at least 1.7 times, at least 1.8 times, at least 1.9 times, at least 2 times, at least 3 times, 1.6 to 4 times, 1.6 to 3 times stronger) than the background (non-specific) binding, for example, the binding level by the hIgG1 isotype control, and the antibody (i) heavy chain variable regions (VH) containing the CDR1, CDR2, and CDR3 sequences, respectively, including SEQ ID NOs. 79, 94, and 111, and SEQ ID NOs. 127 (ii) a light chain variable region (VL) containing CDR1, CDR2, and CDR3 sequences including 141, and 156 respectively, (ii) a heavy chain variable region (VH) containing CDR1, CDR2, and CDR3 sequences including SEQ ID NOs. 81, 94, and 114 respectively, and a light chain variable region (VL) containing CDR1, CDR2, and CDR3 sequences including SEQ ID NOs. 127, 141, and 156 respectively, (iii) CDR1, CDR2, and (iv) A heavy chain variable region (VH) containing the CDR3 sequence, and a light chain variable region (VL) containing the CDR1, CDR2, and CDR3 sequences containing SEQ ID NOs. 127, 141, and 159, respectively, or (iv) a heavy chain variable region (VH) containing the CDR1, CDR2, and CDR3 sequences containing SEQ ID NOs. 86, 102, and 120, respectively, and a light chain variable region (VL) containing the CDR1, CDR2, and CDR3 sequences containing SEQ ID NOs. 127, 141, and 156, respectively. p(S) and p(T) correspond to phosphorylated serine and phosphorylated threonine, respectively. Optionally, the binding is evaluated using, for example, the one-point ELISA described in Example 7, and optionally, the antibody has the sequence described in SEQ ID NO: 274 for human tau.

[0054] E40. Isolated, for example, recombinant antibodies, (a) It binds to tau that is phosphorylated at T217 but not at T212 or T214, or (b) Binds to peptides containing or consisting of the sequences GTPGSRSRTPSLP(pT)PPTRE (SEQ ID NO: 293) and GTPGSSRSRTP(pS)LP(pT)PPTRE (SEQ ID NO: 296), but does not bind to peptides containing or consisting of the sequences GTPGSSRSR(pT)PSLPTPPTRE (SEQ ID NO: 291), GTPGSSRSRTP(pS)LPTPPTRE (SEQ ID NO: 292), and GTPGSSRSR(pT)P(pS)LPTPPTRE (SEQ ID NO: 294). p(S) and p(T) correspond to phosphorylated serine and phosphorylated threonine, respectively. Optionally, the binding of the antibody to tau or the peptide is at least 1.5 times stronger than the background (non-specific) binding level, e.g., the binding level by the hIgG1 isotype control (e.g., at least 1.6 times, at least 1.7 times, at least 1.8 times, at least 1.9 times, at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 6 times, 1.5 to 4 times, 1.5 to 3 times, 4 to 6 times stronger), Optionally, the binding of the antibody to tau or the peptide is evaluated using, for example, the one-point ELISA described in Example 8, and optionally, the human tau antibody has the sequence described in SEQ ID NO: 274.

[0055] E41. The aforementioned antibody (a) Heavy chain variable region (VH) containing CDR1, CDR2, and CDR3 sequences including sequence numbers 80, 95, and 112, respectively, and light chain variable region (VL) containing CDR1, CDR2, and CDR3 sequences including sequence numbers 129, 143, and 157, respectively, (b) Heavy chain variable regions (VH) containing CDR1, CDR2, and CDR3 sequences containing SEQ ID NOs. 78, 104, and 122 respectively, and light chain variable regions (VL) containing CDR1, CDR2, and CDR3 sequences containing SEQ ID NOs. 136, 150, and 167 respectively, or (c) Heavy chain variable region (VH) containing CDR1, CDR2, and CDR3 sequences containing SEQ ID NOs. 90, 107, and 125, respectively, and light chain variable region (VL) containing CDR1, CDR2, and CDR3 sequences containing SEQ ID NOs. 139, 151, and 170, respectively. The antibody according to Embodiment E40, including the antibody described above.

[0056] E42. The antibody comprises VH and VL, and the VH and VL are (a) Sequence IDs 4 and 22, respectively (b) Sequence IDs 14 and 34, respectively, or (c) Sequence IDs 17 and 37 respectively The antibody according to embodiment E40 or E41, comprising the amino acid sequence.

[0057] E43. The antibody according to any one of the prior embodiments, wherein the antibody is an IgA, IgD, IgE, IgG, or IgM antibody.

[0058] E44. The antibody according to Embodiment E43, wherein the antibody is an IgG antibody.

[0059] E45. The antibody according to any one of the prior embodiments, wherein the IgG is an isotype selected from IgG1, IgG2, IgG3, and IgG4.

[0060] E46. The antibody according to Embodiment E45, wherein the antibody is an IgG1 antibody.

[0061] E47. An antibody according to any of the prior embodiments, comprising a heavy chain constant region selected from human IgG1, human IgG2, human IgG3, human IgG4, mouse IgG1, mouse IgG2a, mouse IgG2b, mouse IgG2c, and mouse IgG3, and / or a light chain constant region selected from κ or λ light chain constant regions.

[0062] E48. The antibody according to Embodiment E47, wherein the antibody comprises the heavy chain constant region of human IgG1.

[0063] E49. The aforementioned antibody (i) Heavy chain constant region (CH), for example, any amino acid sequence of any of the heavy chain constant regions in Table 5, or a sequence having at least 80% (e.g., 85, 90, 95, 96, 97, 98, or 99%) sequence identity with the heavy chain constant region sequence in Table 5, an amino acid sequence having at least 1, 2, or 3 modifications, but no more than 30, 20, or 10 modifications, compared to the amino acid sequence of the heavy chain constant region sequence in Table 5, or a CH having at least 1, 2, or 3 different amino acids, but no more than 30, 20, or 10, compared to the amino acid sequence of the heavy chain constant region sequence in Table 5, and / or (ii) Light chain constant region (CL), for example, any amino acid sequence of any of the CL sequences in Table 5, or a sequence having at least 80% (e.g., 85, 90, 95, 96, 97, 98, or 99%) sequence identity with any of the CL sequences in Table 5, an amino acid sequence having at least 1, 2, or 3 modifications, but no more than 30, 20, or 10 modifications, compared to the amino acid sequence of the light chain constant region sequence in Table 5, or an amino acid sequence having at least 1, 2, or 3, but no more than 30, 20, or 10 different amino acids, compared to the amino acid sequence of the light chain constant region sequence in Table 5 The antibody described in any one of the prior embodiments, including the antibody described in any one of the prior embodiments.

[0064] E50. The antibody according to any one of the prior embodiments, wherein the antibody molecule comprises an Fc region or a variant thereof, such as a functional variant.

[0065] E51. The antibody according to any one of the prior embodiments, wherein the antibody molecule comprises an Fc region that has been modified, for example, increased or decreased (e.g., removed), in terms of affinity to the Fc receptor compared to a reference, and the reference is a wild-type Fc receptor.

[0066] E52. The antibody according to any one of Embodiments E1 to E51, wherein the antibody molecule comprises an Fc region having mutations at one, two, or all of the following positions, numbered according to the EU index, such as in Kabat: I253 (e.g., I235A), H310 (e.g., H310A), and / or H435 (e.g., H435A).

[0067] E53. The antibody according to any one of the prior embodiments, wherein the antibody is a full-length antibody, a bispecific antibody, an internal antibody, Fab, F(ab')2, Fv, a single-chain Fv fragment (scFv), a single-domain antibody, or an antibody from a camelid.

[0068] E54. In the antibody, (i) The VH and VL are connected directly, for example, without a linker, or (ii) The antibody according to any one of the prior embodiments, wherein the VH and VL are connected via a linker.

[0069] E55. The antibody according to any one of the prior embodiments, wherein the antibody is a human antibody, a humanized antibody, or a chimeric antibody.

[0070] E56. The antibody is the antibody according to any one of the prior embodiments, wherein the antibody comprises a signal sequence.

[0071] E57. In the antibody, (i) The signal sequence is located at 5' relative to the VH and / or heavy chain, and / or (ii) The signal sequence is located at 5' relative to the VL and / or light chain, The antibody described in Embodiment E56.

[0072] E58. The antibody according to any one of the prior embodiments, wherein the antibody molecule includes a second antigen-binding region having a binding specificity different from that of the antigen-binding region that binds to tau.

[0073] E59. The antibody according to any one of the prior embodiments, wherein the antibody molecule is a multispecific antibody molecule, for example, a bispecific antibody molecule, comprising at least a first antigen-binding domain and a second antigen-binding domain.

[0074] E60. The antibody according to any one of the prior embodiments, wherein the antibody does not bind to non-pathological tau.

[0075] E61. The antibody according to any one of the prior embodiments, wherein the antibody binds to pathological tau entanglement.

[0076] E62. The antibody described above is the antibody according to any one of the prior embodiments, wherein the antibody inhibits tau aggregation.

[0077] E63. An isolated, for example, recombinant antibody that competes with the antibody described in any one of the prior embodiments in terms of binding to human tau.

[0078] E64. An isolated, for example, recombinant antibody that binds to the same epitope as, substantially the same, overlapping, or substantially overlapping epitopes of the antibody described in any one of the prior embodiments.

[0079] E65. The antibody according to any one of the prior embodiments, wherein the antibody comprises a conjugate, such as a therapeutic agent or a detectable label.

[0080] E66. A composition (e.g., a pharmaceutical composition) comprising the antibody and carrier (e.g., a pharmaceutically acceptable carrier) described in any one of the prior embodiments.

[0081] E67. An isolated, for example, recombinant nucleic acid, or combination of nucleic acids encoding the antibody described in any one of Embodiments E1 to E65.

[0082] E68. (a) Any VH nucleotide sequence shown in Table 1, or nucleic acid sequences having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto, and / or (b) Any VL nucleotide sequence shown in Table 1, or a nucleotide sequence having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto. A nucleic acid or combination of nucleic acids as described in Embodiment E67, including the following.

[0083] E69. (a) Any one nucleotide sequence of sequence numbers 51, 55, 54, 52, 47, 39, 56, 41, 50, 49, 48, 46, 45, 44, 43, 42, 53, 40, or nucleic acid sequences having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto, and / or (b) Any one of the nucleotide sequences of SEQ ID NOs: 67, 75, 74, 72, 66, 57, 76, 59, 70, 69, 68, 65, 64, 62, 63, 61, 60, 73, 58, or any nucleotide sequence having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto. A nucleic acid or combination of nucleic acids as described in either embodiment E67 or E68, including the following:

[0084] E70. The isolated nucleic acid sequence according to any one of embodiments E67 to E69, wherein the nucleic acid sequence encoding the heavy chain variable region and / or the light chain variable region is codon-optimized.

[0085] E71. An isolated, for example, recombinant antibody encoded by a nucleic acid as described in any one of Embodiments E67 to E70.

[0086] E72. A vector (e.g., an expression vector) or a combination of vectors (e.g., a combination of expression vectors) comprising a nucleic acid or a combination of nucleic acids as described in any one of Embodiments E67 to E70.

[0087] E73. A host cell containing a nucleic acid or combination of nucleic acids as described in any one of Embodiments E67 to E70, or a vector or combination of vectors as described in Embodiment E62.

[0088] E74. The host cell according to embodiment E73, wherein the host cell is a bacterial cell or a mammalian cell.

[0089] E75. A method for producing an antibody that binds to human tau, wherein the method comprises culturing a host cell described in Embodiment E73 or E74 under conditions suitable for gene expression.

[0090] E76. A method for delivering an exogenous antibody bound to human tau to a target, wherein the method comprises administering to the target an effective amount of the antibody described in any one of Embodiments E1 to E65, or the composition described in Embodiment E66 (for example, a pharmaceutical composition).

[0091] E77. The method according to Embodiment E76, wherein the subject has, has been diagnosed with, or is at risk of developing, a disease related to the expression of tau.

[0092] E78. The method according to Embodiment E76 or E77, wherein the subject has, has been diagnosed with, or is at risk of having, a neurological disorder, such as a neurodegenerative disorder.

[0093] E79. The method according to any one of embodiments E76 to E78, wherein the subject has, has been diagnosed with, or is at risk of developing tauopathy.

[0094] E80. A method for treating a subject who has or has been diagnosed with a disease related to the expression of tau, the method comprising administering to the subject an effective amount of an antibody according to any one of embodiments E1 to E65, or a composition according to embodiment E66 (e.g., a pharmaceutical composition).

[0095] E81. A method for treating a subject suffering from or diagnosed with a neurological disorder, such as a neurodegenerative disorder, the method comprising administering to the subject an effective amount of an antibody described in any one of embodiments E1 to E65, or a composition described in embodiment E66 (e.g., a pharmaceutical composition).

[0096] E82. A method for treating a subject suffering from or diagnosed with tauopathy, the method comprising administering to the subject an effective amount of an antibody described in any one of embodiments E1 to E65, or a composition described in embodiment E66 (e.g., a pharmaceutical composition).

[0097] E83. The method according to any one of Embodiments E77 to E82, wherein the disease associated with the expression of tau, the neurological disorder, or the tauopathy includes Alzheimer's disease (AD), frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17), frontotemporal lobar degeneration (FTLD), frontotemporal dementia (FTD), chronic traumatic encephalopathy (CTE), progressive supranuclear palsy (PSP), Down syndrome, Pick's disease, corticobasal degeneration (CBD), corticobasal syndrome, amyotrophic lateral sclerosis (ALS), prion disease, Creutzfeldt-Jakob disease (CJD), multiple system atrophy, neurofibrillary senile dementia, or progressive subcortical gliosis.

[0098] E84. The method according to any one of embodiments E80 to E83, wherein the treatment includes preventing the progression of the disease or disorder in the subject.

[0099] E85. The method according to any one of embodiments E76 to E84, wherein the subject is a human.

[0100] E86. The method according to any one of embodiments E76 to E85, wherein the antibody is administered intravenously.

[0101] E87. The method according to any one of embodiments E76 to E86, wherein administration of the antibody reduces the presence, level, and / or activity of tau protein.

[0102] E88. The method according to any one of Embodiments E76 to E87, further comprising the administration of an additional therapeutic agent and / or therapy suitable for treating or preventing disorders, neurological disorders, such as neurodegenerative disorders, associated with tau expression.

[0103] E89. The method according to Embodiment E88, wherein the additional therapeutic agent and / or therapy includes cholinesterase inhibitors (e.g., donepezil, rivastigmine, and / or galantamine), N-methyl D-aspartate (NMDA) antagonists (e.g., memantine), antipsychotics, anxiolytics, anticonvulsants, dopamine agonists (e.g., pramipexole, ropinirole, rotigotine, and / or apomorphine), MAO B inhibitors (e.g., selegiline, rasagiline, and / or safinamide), catechol O-methyltransferase (COMT) inhibitors (entacapone, opicapone, and / or tolcapone), anticholinergics (e.g., benztropine and / or trihexyphenidyl), amantadine, carbidopa-levodopa, deep brain stimulation (DBS), or a combination thereof.

[0104] E90. A method for diagnosing neurological disorders, neurodegenerative disorders, diseases related to the expression or activity of tau, or tau-related diseases (e.g., tauopathies) in a subject, wherein the method includes the use of an antibody described in any one of Embodiments E1 to E65.

[0105] E91. A method for detecting tau, the method comprising contacting a sample (for example, a biological sample such as human tissue, for example, human CNS tissue) with an antibody described in any one of embodiments E1 to E65, and detecting the formation of a complex between the antibody and tau.

[0106] E92. The method according to Embodiment E91, wherein the tissue is a thin tissue section or a cryopreserved tissue section.

[0107] E93. An antibody according to any one of Embodiments E1 to E65 for use in a method of treating neurological disorders, neurodegenerative disorders, diseases related to the expression or activity of tau, or diseases related to tau (e.g., tauopathy).

[0108] E94. An antibody according to any one of Embodiments E1 to E65, or a composition according to Embodiment E56, for use in the manufacture of a pharmaceutical product.

[0109] E95. An antibody according to any one of Embodiments E1 to E65, or a composition according to Embodiment E56, for use in the manufacture of a pharmaceutical product for treating neurological disorders, neurodegenerative disorders, diseases related to the expression or activity of tau, or diseases related to tau (e.g., tauopathy).

[0110] E96. Use of an antibody described in any one of Embodiments E1 to E65, or a composition described in Embodiment E66, in the manufacture of a pharmaceutical product.

[0111] E97. Use of an antibody according to any one of Embodiments E1 to E65, or a composition according to Embodiment E66, in the manufacture of a pharmaceutical product for treating neurological disorders, neurodegenerative disorders, diseases related to the expression or activity of tau, or diseases related to tau (e.g., tauopathy).

[0112] Further embodiments 1. (i) A heavy chain variable domain (VH) wherein the VH comprises a complementarity-determining region (CDR) H1 containing an amino acid sequence selected from the group consisting of any of the amino acid sequences listed in Table 1, or a fragment thereof; a CDRH2 containing an amino acid sequence selected from the group consisting of any of the amino acid sequences listed in Table 1, or a fragment thereof; and a CDRH3 containing an amino acid sequence selected from the group consisting of any of the amino acid sequences listed in Table 1, or a fragment thereof; and (ii) A light chain variable domain (VL) wherein the VL comprises a CDRL1 containing an amino acid sequence selected from the group consisting of any of the amino acid sequences listed in Table 1, or a fragment thereof; a CDRL2 containing an amino acid sequence selected from the group consisting of any of the amino acid sequences listed in Table 1, or a fragment thereof; and a CDRL3 containing an amino acid sequence selected from the group consisting of any of the amino acid sequences listed in Table 1, or a fragment thereof.

[0113] 2. The antibody according to Embodiment 1, wherein the antibody comprises a variable domain CDR amino acid sequence set, and the variable domain CDR amino acid sequence set is selected from the group consisting of any of the amino acid sequences listed in Table 1.

[0114] 3. The antibody according to Embodiment 1 or 2, wherein the antibody comprises a pair of variable domain CDR amino acid sequence sets, and the pair of variable domain CDR amino acid sequence sets is selected from the group consisting of any of those listed in Table 1.

[0115] 4. (i) The antibody according to any one of Embodiments 1 to 3, wherein the VH comprises a framework region (FR) H1 comprising an amino acid sequence selected from the group consisting of any of the amino acid sequences listed in Table 1, or a fragment thereof; FRH2 comprising an amino acid sequence selected from the group consisting of any of the amino acid sequences listed in Table 1, or a fragment thereof; FRH3 comprising an amino acid sequence selected from the group consisting of any of the amino acid sequences listed in Table 1, or a fragment thereof; and (ii) The antibody according to any one of Embodiments 1 to 3, wherein the VL comprises FRL1 comprising an amino acid sequence selected from the group consisting of any of the amino acid sequences listed in Table 1, or a fragment thereof; FRL2 comprising an amino acid sequence selected from the group consisting of any of the amino acid sequences listed in Table 1, or a fragment thereof; FRL3 comprising an amino acid sequence selected from the group consisting of any of the amino acid sequences listed in Table 1, or a fragment thereof; and FRL4 comprising an amino acid sequence selected from the group consisting of any of the amino acid sequences listed in Table 1, or a fragment thereof.

[0116] 5. The antibody according to any one of Embodiments 1 to 4, wherein the VH comprises an amino acid sequence selected from the group consisting of any of the amino acid sequences listed in Table 1, and / or an amino acid sequence encoded by a nucleic acid sequence selected from the group consisting of any of the nucleic acid sequences listed in Table 1.

[0117] 6. The antibody according to any one of Embodiments 1 to 5, wherein the VL comprises an amino acid sequence encoded by an amino acid sequence selected from the group consisting of any of the amino acid sequences listed in Table 1, and / or a nucleic acid sequence selected from the group consisting of any of the nucleic acid sequences listed in Table 1.

[0118] 7. The antibody according to any one of Embodiments 1 to 6, wherein the antibody comprises a pair of variable domains, and the pair of variable domains is selected from the group consisting of any of the variable domains listed in Table 1.

[0119] 8. The antibody according to any one of Embodiments 1 to 7, wherein the antibody includes a form selected from the group consisting of a monoclonal antibody, a multispecific antibody, a chimeric antibody, an antibody mimetic, a single-chain Fv(scFv) form, and an antibody fragment.

[0120] 9. The antibody according to any one of Embodiments 1 to 7, wherein the antibody comprises an antibody class selected from the group consisting of IgA, IgD, IgE, IgG, and IgM.

[0121] 10. The antibody according to any one of Embodiments 1 to 7, wherein the antibody comprises one or more non-human constant domains.

[0122] 11. The antibody according to any one of Embodiments 1 to 7, wherein the antibody comprises one or more human constant domains.

[0123] 12. The antibody according to Embodiment 11, wherein one or more human constant domains are selected from the group consisting of any of the constant domains listed in Table 5.

[0124] 13. The antibody according to Embodiment 12, wherein the antibody comprises human IgG, and the human IgG comprises an isotype selected from the group consisting of IgG1, IgG2, IgG3, and IgG4.

[0125] 14. The antibody is the antibody according to any one of Embodiments 1 to 7, wherein the antibody includes a human antibody.

[0126] 15. The antibody according to any one of Embodiments 1 to 14, wherein the antibody binds to a tau protein epitope.

[0127] 16. The antibody according to Embodiment 15, wherein the tau protein epitope comprises, or is contained within, an amino acid sequence selected from the group consisting of any of the amino acid sequences listed in Table 4.

[0128] 17. The antibody according to embodiment 15 or 16, wherein the tau protein epitope includes a region formed by a complex of at least two tau proteins.

[0129] 18. The antibody according to any one of Embodiments 15 to 17, wherein the antibody binds to concentrated pair helical fibrillary tau protein (ePHF) at a maximum half-volume effective concentration (EC50) of approximately 0.01 nM to approximately 100 nM when measured by direct enzyme immunosorbent assay (ELISA).

[0130] 19. The antibody according to any one of embodiments 15 to 18, wherein the antibody does not bind to non-pathological tau.

[0131] 20. The antibody according to any one of embodiments 15 to 19, wherein the antibody binds to pathological tau entanglement.

[0132] twenty one. The antibody according to any one of embodiments 15 to 20, wherein the antibody inhibits tau aggregation.

[0133] twenty two. The antibody according to any one of Embodiments 1 to 21, wherein the antibody includes a conjugate.

[0134] twenty three. The antibody according to Embodiment 22, wherein the conjugate comprises a therapeutic agent.

[0135] twenty four. The antibody according to Embodiment 22, wherein the conjugate includes a detectable label.

[0136] twenty five. A construct encoding the antibody described in any one of Embodiments 1 to 21.

[0137] 26. A method for treating a therapeutic indication in a subject, comprising administering an antibody described in any one of Embodiments 1 to 24 to the subject.

[0138] 27. The method according to Embodiment 26, wherein the therapeutic indications include neurological indications.

[0139] 28. The method according to Embodiment 27, wherein the neurological indication includes one or more of the following: neurodegenerative diseases, Alzheimer's disease (AD), frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17), frontotemporal lobar degeneration (FTLD), frontotemporal dementia (FTD), chronic traumatic encephalopathy (CTE), progressive supranuclear palsy (PSP), Down syndrome, Pick's disease, corticobasal degeneration (CBD), corticobasal syndrome, amyotrophic lateral sclerosis (ALS), prion diseases, Creutzfeldt-Jakob disease (CJD), multiple system atrophy, neurofibrillary senile dementia, stroke, or progressive subcortical gliosis.

[0140] 29. A method for diagnosing a therapeutic indication in a subject, wherein the method includes the use of an antibody described in any one of Embodiments 1 to 24.

[0141] 30. The method according to Embodiment 29, wherein the therapeutic indications include neurological indications.

[0142] 31. The method according to Embodiment 30, wherein the neurological indications include one or more of the following: neurodegenerative diseases, AD, FTDP-17, FTLD, FTD, CTE, PSP, Down syndrome, Pick's disease, CBD, corticobasal syndrome, ALS, prion diseases, CJD, multiple system atrophy, neurofibrillary type senile dementia, stroke, and progressive subcortical gliosis.

[0143] 32. The method according to any one of embodiments 29 to 31, wherein the antibody is used to detect pathological tau in a target tissue.

[0144] 33. The method according to Embodiment 32, wherein the target organization includes a CNS organization.

[0145] 34. The method according to embodiment 32 or 33, wherein the target tissue includes thin tissue sections.

[0146] 35. The method according to Embodiment 34, wherein the thin tissue sections include cryopreserved tissue sections. [Brief explanation of the drawing]

[0147] [Figure 1A] The results of a competitive ELISA assay regarding the binding of the indicated antibody (± competing substance) to peptide 12 (corresponding to SEQ ID NO: 277) are shown. [Figure 1B] The results of a competitive ELISA assay regarding the binding of the indicated antibodies (± competing substances) to the TauS404 peptide are shown. [Figure 1C] The results of a competitive ELISA assay regarding the binding of the indicated antibodies (± competing substances) to the AC04 peptide are shown. [Figure 2] The results of competitive ELISA assays for the binding of AT8 to PepScan fragment 97 (corresponding to SEQ ID NO: 283) in the presence or absence of VY003, VY007, VY006, VY001, or isotype antibody controls are shown. [Figure 3]The results of a one-point ELISA assay for the binding of the indicated antibodies to tau peptides containing the following phosphorylated residues are shown: pT231 (left bar), pS235 (right bar), or pT231 / pS235 (center bar). [Modes for carrying out the invention]

[0148] I. Composition In some embodiments, this disclosure provides compositions that interact with the human microtubule-associated protein tau. Such compositions may be antibodies that bind to the tau protein epitope, referred herein as “anti-tau antibodies.” Dysfunction and / or aggregation of tau is observed in a class of neurodegenerative diseases called tauopathies. Hyperphosphorylation of tau leads to aggregation and inhibition of tau-dependent microtubule assembly. In tauopathies, tau aggregates form paired helical fibrils (PHFs) seen in neurofibrillary tangles (NFTs). These aggregates lead to neuronal loss and cognitive decline. The anti-tau antibodies of this disclosure may be useful for the treatment and / or diagnosis of tauopathies, as well as for other uses described herein.

[0149] antibody In some embodiments, the compounds (e.g., anti-tau antibodies) and compositions of this disclosure comprise antibodies or fragments thereof. As used herein, the term “antibody” is used in its broadest sense and specifically encompasses a variety of embodiments, including but not limited to monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies formed from at least two intact antibodies), single-chain Fv(scFv) formats, and antibody fragments (e.g., Fab, F(ab'), F(ab')2, or Fv), provided that they exhibit functional or biological activity. Antibodies are primarily amino acid-based molecules, but may include one or more modifications (including, but not limited to, the addition of sugar moieties, fluorescent moieties, and chemical tags).

[0150] The antibodies of this disclosure may include, but are not limited to, polyclonal, monoclonal, multispecific, bispecific, trispecific, human, humanized, chimeric, single-chain, diabody, linear, Fab fragment, F(ab') fragment, F(ab')2 fragment, Fv fragment, single-chain Fv fragment (scFv), fragments produced by Fab expression libraries, variable domains, anti-idiotype (anti-Id) antibodies (e.g., anti-Id antibodies against the antibodies of this invention), intracellularly produced antibodies (i.e., intrabodies), codon-optimized antibodies, tandem scFv antibodies, bispecific T cell engagers, mAb2 antibodies, chimeric antigen receptors (CARs), tetravalent bispecific antibodies, biosynthetic antibodies, native antibodies, miniaturized antibodies, unibody, maxibody, intrabody, camelid antibodies, and any of the epitope-binding fragments described above.

[0151] As used herein, the term “antibody fragment” refers to a portion of an intact antibody or its fusion protein, which may include at least one antigen-binding region. Examples of antibody fragments include Fab, Fab', F(ab')2, Fv fragments, single-chain variable fragments (scFv), diabodies, tri(a)bodies, linear antibodies, single-chain antibody molecules, and multispecific antibodies formed from antibody fragments. Papain digestion of an antibody produces two identical antigen-binding fragments called “Fab” fragments, each having a single antigen-binding site. A residual “Fc” fragment is also produced, its name reflecting its ability to readily crystallize. Pepsin treatment yields an F(ab')2 fragment having two antigen-binding sites and still being able to crosslink to an antigen. The antibodies of this disclosure may include one or more of these fragments and may be produced, for example, by enzymatic digestion of the whole antibody or by recombinant expression.

[0152] "Natural antibodies" are typically heterotetrameric glycoproteins of approximately 150,000 daltons, composed of two identical light (L) chains and two identical heavy (H) chains. The genes encoding the heavy and light chains of antibodies are publicly known, and the segments constituting each are well-characterized and described (Matsuda, F. et al., 1998. The Journal of Experimental Medicine. 188(11); 2151-62 and Li, A. et al., 2004. Blood. 103 (12: 4602-9; the contents of each reference are incorporated herein by reference in their entirety). Each light chain is linked to the heavy chain by one disulfide covalent bond, but the number of disulfide bonds differs between heavy chains of different immunoglobulin isotypes. In addition, each heavy and light chain has intrachain disulfide bridges at regular intervals. Each heavy chain has a variable domain (V) at one end. H It has a variable domain (V) at one end. L It has a constant domain at the other end. The constant domain of the light chain is aligned with the first constant domain of the heavy chain, and the variable domain of the light chain is aligned with the variable domain of the heavy chain.

[0153] As used herein, the term “variable domain” refers to a specific antibody domain found in both the heavy and light chains of an antibody, which exhibits a significantly different sequence within the antibody and is used for the binding and specificity of each particular antibody to a particular antigen. Variable domains include hypervariable regions. As used herein, the term “hypervariable region” refers to a region within a variable domain that contains amino acid residues involved in antigen binding. The amino acids present within the hypervariable region determine the structure of the complementarity-determining region (CDR), which forms part of the antibody’s antigen-binding site. As used herein, the term “CDR” refers to a region of the antibody that contains a structure complementary to its target antigen or epitope. Other parts of the variable domain that do not interact with the antigen are each called “framework regions” (FRs). The antigen-binding site (also known as the antigen-combining site or paratope) contains the amino acid residues necessary to interact with a particular antigen. The exact residues constituting the antigen-binding site can be determined by CDR analysis. As used herein, the term “CDR analysis” refers to any process used to determine which antibody variable domain residues constitute the CDR. CDR analysis can be performed by cocrystal structure analysis with the bound antigen. In some embodiments, CDR analysis may include computer evaluation based on comparison with other antibodies (Strohl, WR Therapeutic Antibody Engineering. Woodhead Publishing, Philadelphia PA. 2012. Ch.3, pp. 47-54; the contents of this document are incorporated herein by reference in their entirety).CDR analysis was performed by Kabat [Wu, TT et al., 1970, JEM, 132(2):211-50 and Johnson, G. et al., 2000, Nucleic Acids Res. 28(1):214-8; the contents of each reference are incorporated herein by reference in their entirety], Chothia [Chothia and Lesk, J. Mol. Biol. 196, 901 (1987), Chothia et al., Nature 342, 877 (1989), and Al-Lazikani, B. et al., 1997, J. Mol. Biol. 273(4):927-48; the contents of each reference are incorporated herein by reference in their entirety], Lefranc (Lefranc, M. et al., 2005, Immunome Res. 1:3), and Honegger (Honegger, A. and This may include, but is not limited to, the use of numbering schemes taught by Pluckthun, A. 2001. J. Mol. Biol. 309(3): 657-70 (the entire contents of which are incorporated herein by reference).

[0154] The precise amino acid sequence boundaries of a given CDR can be determined using one of several well-known schemes, including those described in Kabat et al. (1991), “Sequences of Proteins of Immunological Interest,” 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD (Kabat numbering scheme) and Al-Lazikani et al., (1997) JMB 273, 927-948 (Chothia numbering scheme). In some embodiments, a CDR defined according to the Chothia numbering scheme is sometimes referred to as a hypervariable loop.

[0155] For example, in Kabat, the CDR amino acid residues of the heavy chain variable domain (VH) are numbered 31-35 (HCDR1), 50-65 (HCDR2), and 95-102 (HCDR3), and the CDR amino acid residues of the light chain variable domain (VL) are numbered 24-34 (LCDR1), 50-56 (LCDR2), and 89-97 (LCDR3). In Chothia, the CDR amino acids of VH are numbered 26-32 (HCDR1), 52-56 (HCDR2), and 95-102 (HCDR3), and the amino acid residues of VL are numbered 26-32 (LCDR1), 50-52 (LCDR2), and 91-96 (LCDR3). Combining the CDR definitions of both Kabat and Chothia, the amino acid residues of the CDR are numbered as follows: in human VH, amino acid residues 26-35 (HCDR1), 50-65 (HCDR2), and 95-102 (HCDR3); and in human VL, amino acid residues are numbered as follows: 24-34 (LCDR1), 50-56 (LCDR2), and 89-97 (LCDR3).

[0156] The VH and VL domains each have three CDRs. The VL CDRs are referred to herein as CDRL1, CDRL2, and CDRL3, in order of appearance when moving from the N-terminus to the C-terminus along the variable domain polypeptide. The VH CDRs are referred herein as CDRH1, CDRH2, and CDRH3, in order of appearance when moving from the N-terminus to the C-terminus along the variable domain polypeptide. Each CDR has a preferred canonical structure, with the exception of CDRH3, which contains an amino acid sequence that is highly variable in sequence and length among antibodies and can result in various three-dimensional structures of the antigen-binding domain (Nikoloudis, D. et al., 2014. PeerJ.2:e456). In some cases, CDRH3 may be analyzed in a group of related antibodies to assess antibody diversity. Various methods for determining CDR sequences are known in the art and can be applied to known antibody sequences (Strohl, WR Therapeutic Antibody Engineering. Woodhead Publishing, Philadelphia PA. 2012. Ch.3, pp. 47-54; the contents of this document are incorporated herein by reference in their entirety).

[0157] The VH and VL domains each have four framework regions (FRs) located before, after, and between the CDR regions. The VH framework regions are referred to herein as FRH1, FRH2, FRH3, and FRH4, and the VL framework regions are referred to herein as FRL1, FRL2, FRL3, and FRL4. In the VH domain, the FRs and CDRs are typically in the order FRH1-CDRH1-FRH2-CDRH2-FRH3-CDRH3-FRH4 from the N-terminus to the C-terminus. In the VL domain, the FRs and CDRs are typically in the order FRL1-CDRL1-FRL2-CDRL2-FRL3-CDRL3-FRL4 from the N-terminus to the C-terminus.

[0158] As used herein, the term "Fv" refers to an antibody fragment containing the smallest fragment of antibody necessary to form a complete antigen-binding site. These regions consist of a dimer of one heavy chain variable domain and one light chain variable domain in a closely coupled, non-covalently associated state. Fv fragments can be produced by proteolytic cleavage, but are mostly unstable. Recombination methods for producing stable Fv fragments are known in the art, and these typically involve inserting a mobile linker between the light chain variable domain and the heavy chain variable domain [thus forming a single-chain Fv (scFv)] or introducing a disulfide crosslink between the heavy chain variable domain and the light chain variable domain (Strohl, WR Therapeutic Antibody Engineering. Woodhead Publishing, Philadelphia PA. 2012. Ch.3, pp. 46-47; the contents of this document are incorporated herein by reference in their entirety).

[0159] Antibodies derived from any vertebrate species can be assigned one of two distinct types, kappa or lambda, based on the amino acid sequence of their constant domain. Antibodies can be assigned different classes depending on the amino acid sequence of their heavy chain's constant domain.

[0160] As used herein, the terms “single-chain Fv” or “scFv” refer to a fusion protein of a VH antibody domain and a VL antibody domain, where these domains are linked together by a mobile peptide linker to form a single polypeptide chain. In some embodiments, the Fv polypeptide linker enables the scFv to form a structure desirable for antigen binding. In some embodiments, the scFv is used in conjunction with phage display, yeast display, or other display methods, where the scFv is expressed in association with a surface member (e.g., a phage coat protein) and can be used to identify a high-affinity peptide for a given antigen. In some embodiments, the antibodies of this disclosure are prepared as scFvFc antibodies. The term “scFvFc” refers to an antibody format comprising a fusion of one or more scFvs with an antibody Fc domain.

[0161] The term "chimeric antibody" refers to an antibody that has a portion derived from two or more sources. Chimeric antibodies may contain portions derived from different species. For example, a chimeric antibody may contain an antibody having a mouse variable domain and a human constant domain. Further examples of chimeric antibodies and methods for producing them include all of the following: Morrison, SL, Transfectomas provide novel chimeric antibodies. Science. 1985 Sep 20;229(4719):1202-7; Gillies, S et al., High-level expression of chimeric antibodies using adapted cDNA variable region cassettes. J Immunol Methods. 1989 Dec 20;125(1-2):191-202; and all of the following U.S. Patents No. 5,807,715, No. 4,816,567, and No. 4,816,397, the contents of which are incorporated herein by reference in their entirety.

[0162] The term "diabody" refers to a small antibody fragment having two antigen-binding sites, comprising a light-chain variable domain V L connected to a heavy-chain variable domain V H in the same polypeptide chain. By using a linker that is too short to allow pairing between the two domains on the same chain, these domains are paired with the complementary domains on another chain to create two antigen-binding sites. Diabodies are described in more detail, for example, in EP404,097, WO93 / 11161, and Hollinger et al., Proc. Natl. Acad. Sci. USA, 90:6444-6448 (1993), the contents of each of which are hereby incorporated by reference in their entirety.

[0163] The term "intrabody" refers to a form of antibody that targets one or more intracellular protein(s) instead of being secreted from the cell in which it is produced. Intrabodies can be used to affect a number of cellular processes, including but not limited to intracellular trafficking, transcription, translation, metabolic processes, proliferative signaling, and cell division. In some embodiments, the methods of the present invention can include intrabody-based therapies. In some such embodiments, the variable domain sequences and / or CDR sequences disclosed herein can be incorporated into one or more constructs for intrabody-based therapies. In some cases, the intrabodies of the present invention may target one or more glycosylated intracellular proteins or may modulate the interaction between one or more glycosylated intracellular proteins and alternative proteins.

[0164] As used herein, the terms “chimeric antigen receptor” or “CAR” refer to an artificial receptor expressed on the surface of an immune effector cell, which has been engineered to specifically target cells expressing entities that bind to the artificial receptor with high affinity. A CAR may be designed to include an antibody, an antibody variable domain, and / or one or more segments of an antibody CDR, such that when the CAR is expressed on an immune effector cell, the immune effector cell binds to and eliminates cells recognized by the antibody portion of the CAR. In some cases, a CAR may be designed to specifically bind to cancer cells to result in immunomodulatory clearance of cancer cells.

[0165] As used herein, the term “monoclonal antibody” refers to an antibody obtained from a population of substantially homogeneous cells (or clones), i.e., the individual antibodies constituting that population are identical and / or bind to the same epitope, except for any variants that may occur during the production of the monoclonal antibody, which are generally present in trace amounts. Unlike polyclonal antibody preparations, which typically contain different antibodies against different determinants (epitopes), each monoclonal antibody is against a single determinant on an antigen.

[0166] The modifier “monoclonal” indicates a characteristic of the antibody that it is obtained from a substantially homogeneous population of antibodies, and should not be interpreted as requiring the production of the antibody by any particular method. Monoclonal antibodies as used herein include “chimeric” antibodies (immunoglobulins) in which a portion of the heavy chain and / or light chain is identical or identical to a corresponding sequence in an antibody derived from a particular species or belonging to a particular antibody class or subclass, but the remainder of the chain is identical or identical to a corresponding sequence in an antibody derived from a different species or belonging to a different antibody class or subclass, as well as fragments of such antibodies.

[0167] The antibodies of this disclosure may originate from any animal species, including mammals, birds, reptiles, and insects. Mammalian antibodies may originate from, for example, humans, murids (e.g., mice or rats), donkeys, sheep, rabbits, goats, guinea pigs, camels, cattle, or horses.

[0168] In some embodiments, the antibodies of this disclosure may be antibody mimics. The term “antibody mimic” refers to any molecule that mimics the function or effect of an antibody and binds specifically and with high affinity to its molecular target. In some embodiments, an antibody mimic may be a monobody designed to incorporate a fibronectin type III domain (Fn3) as a protein scaffold (US6,673,901, US6,348,584). In some embodiments, an antibody mimic may include, but not be limited to, afibody molecules, affilins, afitins, antikalins, avimers, DARPin, Fynomers, and Kunitz, as well as domain peptides known in the art. In other embodiments, an antibody mimic may comprise one or more non-peptide regions.

[0169] As used herein, the term “antibody variant” refers to a biomolecule that is similar in structure, sequence, and / or function to a given antibody, but has some differences in its amino acid sequence, composition, or structure compared to another antibody or a native antibody.

[0170] Intrabody In some embodiments, the antibodies described herein are intrabodies. In some embodiments, an intrabody is a form of antibody that is not secreted from the cell that produces it, but instead targets one or more intracellular proteins. Intrabodies can be expressed and function within cells and used to influence a number of cellular processes, including but not limited to intracellular transport, transcription, translation, metabolic processes, proliferation signaling, and cell division. In some embodiments, the methods described herein include treatments based on intrabodies.

[0171] In some embodiments, intrabodies are single-chain variable fragments (scFv) expressed from recombinant nucleic acid molecules and engineered to be retained within cells (e.g., in the cytoplasm, endoplasmic reticulum, or periplasm). Intrabodies can be used, for example, to eliminate the function of the protein to which they bind. Exemplary intrabodies include (Marasco et al.,1993 Proc. Natl. Acad. Sci. USA,90:7889-7893;Chen et al.,1994,Hum.Gene Ther.5:595-601;Chen et al. al.,1994,Proc.Natl.Acad.Sci.USA,91:5932-5936;Maciejewski et al.,1995,Nature Med.,1:667-673;Marasco,1995,Immunotech,1:1-19;Mhashilkar,et al.,1995,EMBO J.14:1542-51;Chen et al.,1996,Hum.Gene Therap.,7:1515-1525;Marasco,Gene Ther.4:11-15,1997;Rondon and Marasco,1997,Annu.Rev.Microbiol.51:257-283;Cohen, et al.,1998,Oncogene 17:2445-56;Proba et al.,1998,J.Mol.Biol.275:245-253;Cohen et al. al.,1998,Oncogene 17:2445-2456;Hassanzadeh,et al.,1998,FEBS Lett.437:81-6;Richardson et al.,1998,Gene Ther.5:635-44;Ohage and Steipe,1999,J.Mol.Biol.291:1119-1128;Ohage et al. al.,1999,J.Mol.Biol.291:1129-1134;Wirtz and Steipe,1999,Protein Sci.8:2245-2250;Zhu et al.,1999,J.Immunol.Methods 231:207-222;Arafat et al.,2000,Cancer Gene Ther.(7:1250-6; der Maur et al., 2002, J. Biol. Chem. 277:45075-85; Mahashilkar et al., 2002, Gene Ther. 9:307-19; and Wheeler et al., 2003, FASEB J. 17:1733-5 and the references cited therein are described and outlined). In particular, the CCR5 intrabody is produced by Steinberger et al., 2000, Proc. Natl. Acad. Sci. USA 97:805-810. Generally, see Marasco, WA, 1998, “Intrabodies: Basic Research and Clinical Gene Therapy Applications” Springer: New York. For an overview of scFv, see Pluckthun in “The Pharmacology of Monoclonal Antibodies,” 1994, vol. 113, Rosenburg and Moore eds. Springer-Verlag, New York, pp. 269–315 (all of these are incorporated herein by reference).

[0172] Sequences derived from donor antibodies can be used to develop intrabodies. In many cases, intrabodies are recombinantly expressed as single-domain fragments, such as isolated VH and VL domains, or as single-chain variable fragment (scFv) antibodies in cells. For example, intrabodies are often expressed as a single polypeptide, forming a single-chain antibody containing heavy and light chain variable domains linked by a flexible linker polypeptide. Intrabodies typically lack disulfide bonds and can regulate the expression or activity of target genes through specific binding activity. Single-chain antibodies can also be expressed as single-chain variable region fragments bound to the light chain constant region.

[0173] In some embodiments, intrabodies can be incorporated into recombinant polynucleotide vectors to encode intracellular transport signals at their N-terminus or C-terminus, allowing for high-concentration expression in the intracellular compartment where the target protein is located. For example, intrabodies targeting the endoplasmic reticulum (ER) are engineered to incorporate a leader peptide and, optionally, a C-terminal ER retention signal such as a KDEL amino acid motif. Intrabodies intended to exert activity in the nucleus are engineered to include a nuclear localization signal. Lipid moieties are attached to the intrabodies to anchor them to the cytosolic side of the cell membrane. Intrabodies can also be targeted to exert function in the cytosol. For example, intracytosolic intrabodies are used to sequester factors within the cytosol, thereby preventing the factors from being transported to their original cellular destination.

[0174] Intrabodies possess a virtually limitless ability to specifically recognize different three-dimensional structures of proteins, including pathological isoforms, and can target potential aggregation sites (both intracellular and extracellular). Therefore, intrabodies may be promising therapeutic agents for the treatment of misfolding diseases, including tauopathies, prion diseases, Alzheimer's disease, Parkinson's disease, and Huntington's disease. These molecules can function as neutralizers for amyloid-forming proteins by preventing their aggregation, and / or as molecular shunters for intracellular transport by diverting proteins away from their potential aggregation sites (Cardinale, and Biocca, Curr. Mol. Med. 2008, 8:2-11).

[0175] Antibody development The antibodies described herein can be developed using methods standard in the art. Primary antibody preparation techniques include two approaches: immunization and antibody display. In either case, the desired antibody is identified from a larger candidate pool based on its affinity for a specific target or epitope. An immune response is characterized by the reaction of an organism's cells, tissues, and / or organs to the presence of a foreign substance. Such an immune response typically leads to the organism producing one or more antibodies against a foreign substance, such as an antigen or a part of an antigen.

[0176] antigen Antibodies can be developed (e.g., via immunization) or selected (e.g., from a pool of candidates) using, for example, any naturally occurring or synthetic antigen. As used herein, “antigen” is an entity that induces or triggers an immune response in an organism and may also refer to an antibody-binding partner. An immune response is characterized by the reaction of an organism’s cells, tissues, and / or organs to the presence of a foreign substance. Such an immune response typically leads to the organism producing one or more antibodies against the foreign substance. In some embodiments, the antigen includes a tau protein. As used herein, the term “tau protein” refers to a protein or protein complex containing microtubule-associated protein tau or a peptide fragment thereof. Tau proteins may include enriched paired helical fibrillary tau protein (ePHF), also called “sarcosyl-insoluble tau,” or fragments thereof. Tau proteins may contain one or more phosphorylated residues. Such phosphorylated residues may correspond to disease-associated tau proteins (also referred herein as “pathological tau”).

[0177] immunization In some embodiments, antibodies can be prepared by immunizing a host with the antigen of interest. Immunizing a host animal (e.g., mouse, rabbit, goat, or llama) with an antigenic protein can induce lymphocytes that specifically bind to the antigen. Lymphocytes may be collected and fused with immortalized cell lines to produce hybridomas, which may then be cultured in a suitable culture medium for promoting growth (see, for example, Kohler, G. et al., Continuous cultures of fused cells secreting antibody of predefined specificity. Nature. 1975 Aug 7;256(5517):495-7, the contents of which are incorporated herein by reference in their entirety). Alternatively, lymphocytes may be immunized in vitro.

[0178] In some embodiments, the antibodies of this disclosure may be prepared through immunization using mouse host animals. Such host animals may include transgenic mice. Transgenic mice may include mice that have been engineered to express human antibody sequences by optionally replacing mouse antibody sequences. Transgenic mice may express human variable domain sequences and / or constant domain sequences. In some embodiments, the mouse host animals used for immunization may include any of the transgenic mice described in U.S. Patents 7,435,871, 7,547,817, 9,346,873, 9,580,491, or 10,555,506, the contents of which are incorporated herein by reference in whole.

[0179] Lymphocytes may be fused with an immortalized cell line using a suitable flux (e.g., polyethylene glycol) to form hybridoma cells (see, for example, Goding, JW, Monoclonal Antibodies: Principles and Practice. Academic Press. 1986; 59-1031, the contents of which are incorporated herein by reference in their entirety). The immortalized cell line may be transformed mammalian cells, particularly myeloma cells of rodent, rabbit, bovine, or human origin. In some embodiments, rat or mouse myeloma cell lines are used. Hybridoma cells may be cultured in a suitable culture medium containing one or more substances that typically inhibit the growth or survival of unfused cells. For example, parental cells lacking the enzyme hypoxanthine guanine phosphoribosyltransferase (HGPRT or HPRT) may be used, and the culture medium for the resulting hybridoma cells may be supplemented with hypoxanthine, aminopterin, and thymidine ("HAT medium") to prevent the growth of HGPRT-deficient (unfused) cells.

[0180] Desirable properties of immortalized cell lines may include, but are not limited to, efficient fusion, support for high levels of antibody expression by selected antibody-producing cells, and sensitivity to unfused cell inhibitory media (e.g., HAT medium). In some embodiments, the immortalized cell line is a mouse myeloma cell line. Such cell lines can be obtained, for example, from the Salk Institute Cell Distribution Center (San Diego, CA) or the American Type Culture Collection (Manassas, VA). Human myeloma and mouse-human heterozygous myeloma cell lines can also be used for the production of human monoclonal antibodies (see, for example, Kozbor, D. et al., A human hybrid myeloma for production of human monoclonal antibodies. J Immunol. 1984 Dec;133(6):3001-5 and Brodeur, B. et al., Monoclonal Antibody Production Techniques and Applications. Marcel Dekker, Inc., New York. 1987;33:51-63. The contents of each reference are incorporated herein by reference in their entirety).

[0181] Hybridoma cell culture media can be assayed for the presence of monoclonal antibodies with desired binding specificity. Assays may include, but are not limited to, immunoprecipitation assays, in vitro binding assays, radioimmunoassays (RIAs), surface plasmon resonance (SPR) assays, and / or enzyme-linked immunosorbent assays (ELISAs). In some embodiments, the binding specificity of monoclonal antibodies may be determined by Scatchard analysis (Munson, P.Jet al., Ligand: a versatile computerized approach for characterization of ligand-binding systems. Anal Biochem. 1980 Sep 1;107(1):220-39; the contents of this document are incorporated herein by reference in their entirety).

[0182] Antibodies produced by cultured hybridomas may be analyzed to determine their binding specificity to the target antigen. Once antibodies with desirable characteristics are identified, the corresponding hybridomas can be subcloned using limiting dilution procedures and grown by standard methods. Antibodies produced by hybridomas can be isolated and purified using standard immunoglobulin purification procedures such as protein A-Sepharose, hydroxyapatite chromatography, gel electrophoresis, dialysis, or affinity chromatography. Alternatively, hybridoma cells may be grown in vivo as ascites in mammals. In some embodiments, antibodies may be isolated directly from the serum of an immunized host.

[0183] In some embodiments, recombinant versions of antibodies produced by immunization may be prepared. Such antibodies may be prepared from selected hybridomas using genomic antibody sequences. The genomic antibody sequence of a hybridoma can be obtained by extracting RNA molecules from antibody-producing hybridoma cells and producing cDNA by reverse transcriptase polymerase chain reaction (PCR). PCR may be used to amplify the cDNA using primers specific to the heavy and light chains of the antibody. The PCR product may then be subcloned into a plasmid for sequence analysis. The antibody may be produced by inserting the obtained antibody sequence into an expression vector. Some recombinant antibodies may be prepared using synthetic nucleic acid constructs that encode amino acid sequences corresponding to amino acid sequences obtained from isolated hybridoma antibodies.

[0184] entire antibody In some embodiments, antibodies may be developed using antibody display technology. “Display technology” refers to a system and method for expressing a nucleic acid encoding an amino acid-based candidate compound, with the compound linked to it, in a format accessible to a target or ligand. Candidate compounds are expressed on a host capsid or cell surface in most systems, although some hostless systems exist (e.g., ribosome displays). Display technology allows for the generation of a display “library” containing a set of candidate compound library members. A display library containing antibodies (or their variants or fragments) as library members is referred to herein as an “antibody display library.” Antibodies can be designed, selected, or optimized by screening target antigens using an antibody display library. An antibody display library may contain millions to billions of members, each expressing a unique antibody domain. The presented antibody fragments are V H and V L The antibody domain may be an scFv antibody fragment, which is a fusion protein in which the antibody domain is linked by a mobile linker. The display library may contain antibody fragments with different levels of diversity between the variable domain framework region and the CDR. The CDR of the display library antibody fragment may contain unique variable loop lengths and / or sequences. The antibody variable domains or CDRs obtained from the display library selection may be directly incorporated into the antibody sequence for recombinant antibody production, or they may be mutated and used for further optimization via in vitro affinity maturation.

[0185] Antibody display libraries may include antibody-phage display libraries. Antibody-phage display libraries utilize phage virus particles as hosts, along with millions to billions of members, each expressing a unique antibody domain. Such libraries can provide a diverse source that can be used to select, in some cases, hundreds of antibody fragments with varying levels of affinity to one or more target antigens (McCafferty, et al., 1990. Nature. 348:552-4; Edwards, B. et al., 2003. JMB. 334:103-18; Schofield, D. et al., 2007. Genome Biol. 8, R254; and Pershad, K. et al., 2010. Protein Engineering Design and Selection. 23:279-88; the contents of each reference are incorporated herein by reference in their entirety). The antibody fragments presented may be scFv antibody fragments. Phage display library members can be expressed as fusion proteins linked to viral coat proteins (e.g., the N-terminus of the viral pIII coat protein). L By expressing the chains separately, V is expressed within the periplasm. H After assembling with the chain, the complex may be incorporated into the viral coat. The precipitated library members may be sequenced from the bound phage to obtain cDNA encoding the desired antibody domain.

[0186] In some embodiments, antibody display libraries may be generated using yeast surface display techniques. Antibody yeast display libraries consist of yeast cells having surface-presented antibodies or antibody fragments. Antibody yeast display libraries may include antibody-variable domains expressed on the surface of Saccharomyces cerevisiae cells. Yeast display libraries may be developed by presenting the desired antibody fragment as a fusion protein with a yeast surface protein (e.g., Aga2p protein). Yeast cells presenting antibodies or antibody fragments with affinity for a specific target can be isolated according to standard methods. Such methods may include, but are not limited to, magnetic separation and flow cytometry.

[0187] Recombinant synthesis The antibodies of this disclosure may be prepared using recombinant DNA technology and related processes. A construct encoding the antibody (e.g., a DNA expression plasmid) may be prepared and used to synthesize a complete antibody or a portion thereof. In some embodiments, a DNA sequence encoding the antibody variable domain of this disclosure may be inserted into an expression vector encoding another antibody domain (e.g., a mammalian expression vector), and this may be used to prepare an antibody having the inserted variable domain. The DNA sequence encoding the antibody variable domain may be inserted downstream of an upstream expression vector region having a promoter / enhancer element and / or encoding an immunoglobulin signaling sequence. The DNA sequence encoding the antibody variable domain may be inserted upstream of a downstream expression vector region encoding a constant immunoglobulin domain. The encoded constant domain may originate from any class (e.g., IgG, IgA, IgD, IgE, and IgM) or species (e.g., human, mouse, rabbit, rat, and non-human primates). In some embodiments, the encoded constant domain encodes a constant domain of human IgG (e.g., IgG1, IgG2a, IgG2b, IgG2c, IgG3, or IgG4). In some embodiments, the encoded constant domain encodes a constant domain of mouse IgG (e.g., IgG1, IgG2a, IgG2b, or IgG3).

[0188] The antibody-encoding expression vectors of this disclosure may be used to transfect cells for antibody production. Such cells may be mammalian cells. The antibody expression vector may be used to prepare cell lines that have been stably transfected and to establish stable cell lines. The antibody-producing cell lines may be expanded to express the antibody, which may then be isolated or purified from the cell culture medium.

[0189] Anti-tau antibody sequence This specification describes antibodies, such as recombinant antibodies, characterized by specific functional and structural features or properties. For example, the antibody specifically binds to human tau (e.g., human tau having the sequence shown in SEQ ID NO: 274). The specific antibodies described herein are antibody VY011, VY007, VY004, VY006, VY018, VY003, VY016, VY017, VY012, VY009, VY010, VY022, VY001, VY019, VY020, VY005, VY002, VY014, VY008, and VY013, and antibodies having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto, and antibodies having variable regions with at least 1, 2, or 3 modifications, provided 30, 20, or 10 or fewer modifications (e.g., 1 - 30, 1 - 20, 1 - 10, 1 - 5, 1 - 4, 1 - 3, 1 - 2, or 1 amino acid modifications, such as amino acid substitutions (e.g., conservative substitutions)) compared to the VH and / or VL sequences of the listed VY antibodies. The ability of variant antibodies to bind to tau (e.g., wild - type tau (e.g., SEQ ID NO: 274), ePHF, iPHF) can be measured using binding assays recognized in the art, such as the binding assays described in the examples. Table 1 is an overview of the amino acid sequences and nucleotide sequences encoding the VY antibodies.

Table 1 - 1

Table 1 - 2

Table 1 - 3

Table 1 - 4

Table 1 - 5

[0190] The anti-tau antibodies of this disclosure may comprise a variable domain amino acid sequence, which may be one of the variable domain amino acid sequences listed in Table 1. In some embodiments, the anti-tau antibody variable domain comprises a fragment or variant of the listed variable domain amino acid sequences. For example, in some embodiments, the anti-tau antibody may include an amino acid sequence selected from SEQ ID NOs. 7, 3, 4, 6, 11, 1, 2, 5, 8, 9, 10, 12, 13, 14, 15, 16, 17, or 18, or an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto, or a VH having at least 1, 2, or 3 modifications, but no more than 30, 20, or 10 modifications (e.g., 1-30, 1-20, 1-10, 1-5, 1-4, 1-3, 1-2, or 1 amino acid modification, e.g., amino acid substitution (e.g., conservative substitution)) compared to the VH sequence. In some embodiments, the anti-tau antibody comprises an amino acid sequence selected from SEQ ID NOs. 25, 21, 22, 24, 30, 19, 20, 23, 26, 27, 28, 29, 31, 32, 33, 34, 35, 36, 37, or 38, or an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto, or a VL comprising an amino acid sequence having at least 1, 2, or 3 modifications, but no more than 30, 20, or 10 modifications (e.g., 1-30, 1-20, 1-10, 1-5, 1-4, 1-3, 1-2, or 1 amino acid modification, e.g., amino acid substitution (e.g., conservative substitution)) compared to the VL sequence.In some embodiments, the anti-tau antibody may comprise VH and VL, (a) VH comprising an amino acid sequence selected from SEQ ID NOs. 7, 3, 4, 6, 11, 1, 2, 5, 8, 9, 10, 12, 13, 14, 15, 16, 17, or 18, or an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto, or an amino acid sequence having at least 1, 2, or 3 modifications, but no more than 30, 20, or 10 modifications (e.g., amino acid substitutions such as conservative substitutions), and / or (b) VL comprising SEQ ID NOs. 25, An amino acid sequence selected from 21, 22, 24, 30, 19, 20, 23, 26, 27, 28, 29, 31, 32, 33, 34, 35, 36, 37, or 38, or an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto, or an amino acid sequence having at least 1, 2, or 3 modifications, but no more than 30, 20, or 10 modifications (e.g., 1-30, 1-20, 1-10, 1-5, 1-4, 1-3, 1-2, or 1 amino acid modification, e.g., amino acid substitution (e.g., conservative substitution)) compared to the VL sequence.

[0191] In this specification, (i) SEQ ID NOs: 7 and / or 25, respectively; (ii) SEQ ID NOs: 3 and / or 21, respectively; (iii) SEQ ID NOs: 4 and / or 22, respectively; (iv) SEQ ID NOs: 6 and / or 24, respectively; (v) SEQ ID NOs: 11 and / or 30, respectively; (vi) SEQ ID NOs: 1 and / or 19, respectively; (vii) SEQ ID NOs: 2 and / or 20, respectively; (viii) SEQ ID NOs: 5 and / or 23, respectively; (ix) SEQ ID NOs: 8 and / or 26, respectively; (x) SEQ ID NOs: 9 and / or 27, respectively; (xi) SEQ ID NOs: 10 and / or 28, respectively; (xii) SEQ ID NOs: 10 and / or 29 The present invention provides isolated, for example, recombinant antibodies that conjugate to human tau (e.g., human tau having the sequence described in SEQ ID NO: 274), comprising VH and / or VL sequences selected from (xiii) SEQ ID NO: 12 and / or 31, (xiv) SEQ ID NO: 13 and / or 32, (xv) SEQ ID NO: 14 and / or 33, (xvi) SEQ ID NO: 14 and / or 34, (xvii) SEQ ID NO: 15 and / or 35, (xviii) SEQ ID NO: 16 and / or 36, (xix) SEQ ID NO: 17 and / or 37, or (xx) SEQ ID NO: 18 and / or 38, respectively. In some embodiments, the antibody comprises VH and / or VL sequences having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% sequence identity to any one of the sequences (i) to (xx). In some embodiments, the antibody comprises a VH and / or VL sequence having at least 1, 2, or 3 modifications, or 30, 20, or 10 or fewer modifications (e.g., 1-30, 1-20, 1-10, 1-5, 1-4, 1-3, 1-2, or 1 amino acid modification, e.g., amino acid substitution (e.g., conservative substitution)) compared to any one of the VH and / or VL sequences of (i) to (xx).

[0192] In some embodiments, the antibody that binds to human tau comprises VH and VL sequences selected from (i) SEQ ID NOs. 7 and 25, respectively, (ii) SEQ ID NOs. 3 and 21, respectively, (iii) SEQ ID NOs. 4 and 22, respectively, (iv) SEQ ID NOs. 6 and 24, respectively, (v) SEQ ID NOs. 11 and 30, respectively, (vi) SEQ ID NOs. 1 and 19, respectively, (vii) SEQ ID NOs. 2 and 20, respectively, (viii) SEQ ID NOs. 5 and 23, respectively, (ix) SEQ ID NOs. 8 and 26, respectively, (x) SEQ ID NOs. 9 and 27, respectively, (xi) SEQ ID NOs. 10 and 28, respectively, (xii) SEQ ID NOs. 10 and 29, respectively, (xiii) SEQ ID NOs. 12 and 31, respectively, (xiv) SEQ ID NOs. 13 and 32, respectively, (xv) SEQ ID NOs. 14 and 33, respectively, (xvi) SEQ ID NOs. 14 and 34, respectively, (xvii) SEQ ID NOs. 15 and 35, respectively, (xviii) SEQ ID NOs. 16 and 36, respectively, (xix) SEQ ID NOs. 17 and 37, respectively, or (xx) SEQ ID NOs. 18 and 38, respectively. In some embodiments, the antibody comprises a VH sequence and a VL sequence having at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% sequence identity to any one of the sequences (i) to (xx). In some embodiments, the antibody comprises a VH and / or VL sequence having at least 1, 2, or 3 modifications, but no more than 30, 20, or 10 modifications (e.g., 1 to 30, 1 to 20, 1 to 10, 1 to 5, 1 to 4, 1 to 3, 1 to 2, or 1 amino acid modification, amino acid substitution (e.g., conservative substitution)) compared to any one of the VH and VL sequences (i) to (xx).

[0193] The anti-tau antibody variable domains of this disclosure may be encoded by nucleic acid sequences listed in Table 1. In some embodiments, the nucleic acid sequences encoding the anti-tau antibody variable domains of this disclosure may include fragments or variants of the listed nucleic acid sequences. For example, in some embodiments, the nucleic acid sequence encoding VH may include a nucleotide sequence selected from SEQ ID NOs: 51, 55, 54, 52, 47, 39, 56, 41, 50, 49, 48, 46, 45, 44, 43, 42, 53, or 40, or a nucleic acid sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto. In some embodiments, the nucleic acid sequence encoding VL may include a nucleotide sequence selected from SEQ ID NOs. 67, 75, 74, 72, 66, 57, 76, 59, 70, 69, 68, 65, 64, 62, 63, 61, 60, 73, or 58, or a nucleic acid sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto. In some embodiments, the nucleic acids described herein may encode an anti-tau antibody. In some embodiments, the anti-tau antibody is (a) a nucleotide sequence encoding VH selected from SEQ ID NOs: 51, 55, 54, 52, 47, 39, 56, 41, 50, 49, 48, 46, 45, 44, 43, 42, 53, or 40, or a nucleic acid sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto. The sequence may include a nucleotide sequence encoding a VL selected from (b) SEQ ID NOs: 67, 75, 74, 72, 66, 57, 76, 59, 70, 69, 68, 65, 64, 62, 63, 61, 60, 73, or 58, or a nucleic acid sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto.In some embodiments, the anti-tau antibody is (i) SEQ ID NOs. 51 and 71, respectively, (ii) SEQ ID NOs. 55 and 75, respectively, (iii) SEQ ID NOs. 54 and 74, respectively, (iv) SEQ ID NOs. 52 and 72, respectively, (v) SEQ ID NOs. 47 and 66, respectively, (vi) SEQ ID NOs. 39 and 57, respectively, (vii) SEQ ID NOs. 56 and 76, respectively, (viii) SEQ ID NOs. 41 and 59, respectively, (ix) SEQ ID NOs. 50 and 70, respectively, (x) SEQ ID NOs. 49 and 69, respectively, (xi) SEQ ID NOs. 48 and 67, (xii) SEQ ID NOs. The domains may be encoded by nucleic acids, or combinations of nucleic acids, comprising nucleotide sequences encoding VH and nucleotide sequences encoding VL, selected from (xiii) SEQ ID NOs. 48 and 68, (xiii) SEQ ID NOs. 46 and 65 respectively, (xiv) SEQ ID NOs. 45 and 64 respectively, (xv) SEQ ID NOs. 44 and 62 respectively, (xvi) SEQ ID NOs. 44 and 63 respectively, (xvii) SEQ ID NOs. 43 and 61 respectively, (xviii) SEQ ID NOs. 42 and 60 respectively, (ix) SEQ ID NOs. 53 and 73 respectively, or (xx) SEQ ID NOs. 40 and 58 respectively. In some embodiments, the nucleic acid sequences encoding the anti-tau antibody variable domains of the present disclosure include codon-optimized variants of the enumerated nucleic acid sequences.

[0194] In some embodiments, the anti-tau antibody of this disclosure comprises one or more CDRs (e.g., CDRs 1, 2, 3, 4, 5, or all 6) having amino acid sequences derived from one or more variable domain amino acid sequences shown in Table 1, e.g., CDRs based on the Chothia numbering scheme. In some embodiments, the anti-tau antibody of this disclosure comprises one or more CDRs (e.g., CDRs 1, 2, 3, 4, 5, or all 6). The anti-tau antibody CDR is encoded by a nucleic acid sequence derived from one or more variable domain nucleic acid sequences shown in Table 1. The anti-tau antibody CDR may contain one or more amino acid residues involved in antigen binding (e.g., determined by cocrystallography with the bound antigen). The anti-tau antibodies of this disclosure may include CDRs identified by cocrystallography with the bound antigen, computational evaluation based on comparison with other antibodies (see, for example, Strohl, WR Therapeutic Antibody Engineering. Woodhead Publishing, Philadelphia PA. 2012. Ch.3, pp. 47-54), or by CDR analysis of the variable domain sequences presented herein using the Kabat, Chothia, Al-Lazikani, Lefranc, IMGT, or Honegger numbering schemes recognized in the art as described above. In some embodiments, the CDR amino acid sequence of the anti-tau antibody may include any of the amino acid sequences shown in Table 1, or fragments thereof. In some embodiments, the anti-tau antibodies of this disclosure include CDRs containing the listed amino acid sequence variants. The amino acid fragments or variants contained in the anti-tau antibody CDR may have approximately 50% to approximately 99.9% sequence identity with one or more CDR sequences (e.g., approximately 50% to approximately 60%, approximately 55% to approximately 65%, approximately 60% to approximately 70%, approximately 65% ​​to approximately 75%, approximately 70% to approximately 80%, approximately 75% to approximately 85%, approximately 80% to approximately 90%, approximately 85% to approximately 95%, approximately 90% to approximately 99.9%, approximately 95% to approximately 99.9%, approximately 97%, approximately 97.5%, approximately 98%, approximately 98.5%, approximately 99%, approximately 99.5%, approximately 99.6%, approximately 99.7%, or approximately 99.8%).

[0195] The anti-tau antibodies described herein include VH containing one, two, or three of the heavy chain complementarity-determining region 1 (HCDR1), heavy chain complementarity-determining region 2 (HCDR2), and heavy chain complementarity-determining region 3 (HCDR3), and / or one, two of the light chain complementarity-determining region 1 (LCDR1), light chain complementarity-determining region 2 (LCDR2), and light chain complementarity-determining region 3 (LCDR3), or VLs including three, for example, may include one of the following CDRs based on the Chothia numbering scheme: VY011, VY007, VY004, VY006, VY018, VY003, VY016, VY017, VY012, VY009, VY010, VY022, VY001, VY019, VY020, VY005, VY002, VY014, VY008, and VY013.

[0196] In some embodiments, recombinant antibodies that bind to human tau are provided herein, (i) SEQ ID NOs. 7 and 25, (ii) SEQ ID NOs. 3 and 21, (iii) SEQ ID NOs. 4 and 22, (iv) SEQ ID NOs. 6 and 24, (v) The antibody comprises heavy chain CDR1, CDR2, and CDR3 and / or light chain CDR1, CDR2, and CDR3, each containing a heavy chain variable region (VH) and a light chain variable region (VL) including (vi) SEQ ID NOs: 11 and 30, (vi) SEQ ID NOs: 1 and 19, (vii) SEQ ID NOs: 2 and 20, (viii) SEQ ID NOs: 5 and 23, (ix) SEQ ID NOs: 8 and 26, (x) SEQ ID NOs: 9 and 27, (xi) SEQ ID NOs: 10 and 28, (xii) SEQ ID NOs: 10 and 29, (xiii) SEQ ID NOs: 12 and 31, (xiv) SEQ ID NOs: 13 and 32, (xv) SEQ ID NOs: 14 and 33, (xvi) SEQ ID NOs: 14 and 34, (xvii) SEQ ID NOs: 15 and 35, (xviii) SEQ ID NOs: 16 and 36, (xix) SEQ ID NOs: 17 and 37, or (xx) SEQ ID NOs: 18 and 38, respectively. In some embodiments, the CDR sequence is based on the Kabat numbering scheme, the Chothia numbering scheme, or the IMGT numbering scheme.

[0197] In some embodiments, as used herein, (a) one, two, or all three HCDRs selected from HCDR1 containing an amino acid sequence selected from SEQ ID NOs: 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, or 91; HCDR2 containing an amino acid sequence selected from SEQ ID NOs: 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, or 108; and HCDR3 containing an amino acid sequence selected from SEQ ID NOs: 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, or 126; and / or (b) The present invention provides isolated, for example, recombinant antibodies that bind to human tau, comprising one, two, or all three LCDRs selected from LCDR1 containing an amino acid sequence selected from sequence numbers 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, or 140; LCDR1 containing an amino acid sequence selected from sequence numbers 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, or 153; and LCDR1 containing an amino acid sequence selected from sequence numbers 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, or 171. In some embodiments, one or more of HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 (1, 2, 3, 4, 5, or all 6 CDRs) have 1, 2, or up to 3 amino acid substitutions (e.g., conservative substitutions).

[0198] In some embodiments, the anti-tau antibodies described herein include VH and VL, where VH includes the combination of HCDR1, HCDR2, and HCDR3 listed in Table 2. In some embodiments, the anti-tau antibodies described herein include VH and VL, where VL includes the combination of LCDR1, LCDR2, and LCDR3 listed in Table 2. In some embodiments, one or more (one, two, or all three) of HCDR1, HCDR2, and HCDR3, and / or one or more (one, two, or all three) of LCDR1, LCDR2, and LCDR3 have one, two, or at most three amino acid substitutions (e.g., conservative substitutions). [Table 2]

[0199] In some embodiments, the anti-tau antibody of this disclosure comprises a pair of variable domain CDR amino acid sequence sets presented herein. In some embodiments, the anti-tau antibody of this disclosure comprises a pair of variable domain CDR amino acid sequence sets shown in Table 3. [Table 3] In this specification, isolated, for example, recombinant antibodies conjugated to human tau are provided, each antibody comprising VH containing one, two, or all three of the heavy chain complementarity-determining regions 1 (HCDR1), 2 (HCDR2), and 3 (HCDR3), and / or VL containing one, two, or all three of the light chain complementarity-determining regions 1 (LCDR1), 2 (LCDR2), and 3 (LCDR3), in which case (i) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LC (ii) DR3 contains the amino acid sequences of SEQ ID NOs. 82, 97, 115, 127, 141, and 159, respectively; (ii) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 contain the amino acid sequences of SEQ ID NOs. 79, 94, 111, 127, 141, and 156, respectively; (iii) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 contain the amino acid sequences of SEQ ID NOs. 80, 95, 112, 129, 143, and 157, respectively; (iv) HCDR1, HCDR2, HCDR3, LCDR1, L (v) CDR2 and LCDR3 contain the amino acid sequences of SEQ ID NOs. 81, 94, 114, 127, 141, and 156, respectively; (v) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 contain the amino acid sequences of SEQ ID NOs. 82, 101, 119, 132, 149, and 164, respectively; (vi) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 contain the amino acid sequences of SEQ ID NOs. 77, 92, 109, 127, 141, and 154, respectively; (vii) HCDR1, HCDR2, HCD (viii) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 contain the amino acid sequences of SEQ ID NOs. 78, 93, 110, 128, 142, and 155, respectively; (xi) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 contain the amino acid sequences of SEQ ID NOs. 78, 96, 113, 130, 144, and 158, respectively; (xi) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 contain the amino acid sequences of SEQ ID NOs. 83, 98, 116, 131, 145, and 160, respectively; (x) HCDR1,(xi) HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 contain the amino acid sequences of SEQ ID NOs. 84, 99, 117, 132, 146, and 161, respectively; (xi) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 contain the amino acid sequences of SEQ ID NOs. 85, 100, 118, 133, 147, and 162, respectively; (xii) HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 contain the amino acid sequences of SEQ ID NOs. 85, 100, 118, 134, 148, and 163, respectively. (xiii)HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each contain the amino acid sequences of SEQ ID NOs. 86, 102, 120, 127, 141, and 156, respectively; (xiv)HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each contain the amino acid sequences of SEQ ID NOs. 87, 103, 121, 132, 149, and 165, respectively; (xv)HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each contain SEQ ID NOs. 7 (xvi)HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 contain amino acid sequences 78, 104, 122, 136, 150, and 167, respectively; (xvii)HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 contain amino acid sequences 88, 105, 123, 137, 151, and 168, respectively; (xviii)HCDR1, HCDR2, HCDR3, LCD R1, LCDR2, and LCDR3 contain the amino acid sequences of SEQ ID NOs. 89, 106, 124, 138, 152, and 169, respectively; (xix)HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 contain the amino acid sequences of SEQ ID NOs. 90, 107, 125, 139, 151, and 170, respectively; or (xx)HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 contain the amino acid sequences of SEQ ID NOs. 91, 108, 126, 140, 153, and 171, respectively.

[0200] In some embodiments, the antibody comprises one of the HCDR1, HCDR2, and HCDR3 sequences from (i) to (xx). In some embodiments, the antibody comprises one of the LCDR1, LCDR2, and LCDR3 sequences from (i) to (xx). In some embodiments, the antibody comprises one of the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 sequences from (i) to (xx).

[0201] In some embodiments, one or more of the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 antibodies of (i) to (xx) (1, 2, 3, 4, 5, or all 6 CDRs) have 1, 2, or up to 3 amino acid substitutions (e.g., conservative substitutions).

[0202] In some embodiments, the anti-tau antibody of this disclosure comprises one or more framework regions (FRs). The FR may comprise an amino acid sequence derived from the amino acid sequences of the variable domains shown in Table 1. The FR may be encoded by a nucleic acid sequence derived from the nucleic acid sequences of one or more variable domains shown in Table 1. In some embodiments, the anti-tau antibody FR may comprise an amino acid sequence or a fragment thereof, which is one of the amino acid sequences shown in Table 1. In some embodiments, the anti-tau antibody of this disclosure comprises an FR comprising the listed amino acid sequence variants. The amino acid fragments or variants contained in the anti-tau antibody FR include one or more listed amino acid sequences with approximately 50% to approximately 99.9% sequence identity (e.g., approximately 50% to approximately 60%, approximately 55% to approximately 65%, approximately 60% to approximately 70%, approximately 65% ​​to approximately 75%, approximately 70% to approximately 80%, approximately 75% to approximately 85%, approximately 80% to approximately 90%, approximately 85% to approximately 95%, approximately 90% to approximately 99.9%, approximately 95% to approximately 99.9%, approximately 97%, approximately 97.5%, approximately 98%, approximately 98.5%, approximately 99%, approximately 99.5%, approximately 99.6%, approximately 99.7%, or approximately 99.8%). In some embodiments, the FR has at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% sequence identity with respect to the FR sequences listed in Table 1, or includes at least 1, 2, or 3 modifications, but no more than 30, 20, or 10 modifications (e.g., 1-30, 1-20, 1-10, 1-5, 1-4, 1-3, 1-2, or 1 amino acid modification, e.g., amino acid substitution (e.g., conservative substitution)) compared to the FRs listed in Table 1.

[0203] This specification also provides anti-tau antibodies that bind to tau protein antigens. The tau protein antigen may include human microtubule-associated protein tau, isoform 2 (SEQ ID NO: 274) or a fragment thereof. The tau protein antigen may include ePHF or a fragment thereof. The tau protein antigen may include one or more phosphorylated residues. Such phosphorylated residues may correspond to residues found in pathological tau. In some embodiments, the tau protein antigen includes one of the antigens listed in Table 4. In the table, the phosphorylated residues associated with each antigen are indicated as (pS) for phosphorylated serine and (pT) for phosphorylated threonine. In some embodiments, the tau protein may include variants (e.g., phosphorylated variants or non-phosphorylated variants) or fragments of the listed sequences. [Table 4-1] [Table 4-2]

[0204] In some embodiments, the anti-tau antibody of this disclosure binds to a tau protein epitope on a tau protein antigen described herein. Such a tau protein epitope includes, or may include, the tau protein antigen amino acid sequences listed in Table 4. In some embodiments, the anti-tau antibody of this disclosure binds to a tau protein epitope that includes a region formed by a complex of at least two tau proteins.

[0205] In some embodiments, the anti-tau antibodies of this disclosure exhibit binding to regions of tau recognized by antibodies known in the art, such as AT8 and PT3, but show different binding patterns to phosphorylated tau compared to antibodies known in the art.

[0206] Therefore, in one embodiment, the Specified provides isolated, for example, recombinant antibodies that bind to human tau, the antibodies being (a) 183-212, (b) 187-218, (c) 33-82, 159-182, 197-226, and 229-246, (d) 217-242, (e) 35-76 and 187-218, (f) 5-34, (g) 187-218, ( (a)-(k) is phosphorylated to all or part of the amino acid residues of tau selected from (a)-(k), (i) 35-62, 107-124, and 203-220, (j) 35-82, 159-188, and 197-224, or (k) 53-78, 329-348, and 381-408, and human tau is numbered by SEQ ID NO: 274. In some embodiments, one or more serine, threonine, and / or tyrosine in the stretch of amino acids selected from (a)-(k) are phosphorylated. In some embodiments, all of the serine, threonine, and / or tyrosine in the stretch of amino acids selected from (a)-(k) are phosphorylated. In some embodiments, the antibody comprises the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 sequences, or the VH and VL sequences, of the antibodies listed in Table 1.

[0207] In some embodiments, the anti-tau antibody has a dissociation constant (K) of approximately 1 pM to approximately 50 pM, or approximately 1 to 25 pM, when evaluated by, for example, biolayer interferometry. D ) binds to all or part of amino acids 195-215 of human tau (for example, all serine, threonine, and / or tyrosine present in the stretch of these amino acids are phosphorylated).

[0208] In some embodiments, the anti-tau antibody has a dissociation constant (K) of approximately 0.1 nM to approximately 10 nM, or approximately 0.5 to 5 nM, when evaluated by, for example, biolayer interferometry. D ) binds to all or part of amino acids 191-214 of human tau that are phosphorylated at S199 (for example, phosphorylated only at S199 in this amino acid stretch or the entire tau protein).

[0209] In some embodiments, the anti-tau antibody has a dissociation constant (K) of approximately 0.1 nM to approximately 10 nM, or approximately 0.1 to 5 nM, when evaluated by, for example, biolayer interferometry. D ) binds to all or some of the amino acids 217-234 of human tau that are phosphorylated at T217, T220, and T231 (for example, in this stretch only phosphorylated at T217, T220, and T231).

[0210] In some embodiments, the anti-tau antibody has a dissociation constant (K) of approximately 0.1 nM to approximately 25 nM, or approximately 0.1 to 15 nM, when evaluated by, for example, biolayer interferometry. D ) binds to all or part of amino acids 225-240 of tau that are phosphorylated at T231 (for example, phosphorylated only at T231 in this amino acid stretch or the entire tau protein).

[0211] In another embodiment, the Specified Materials provide an isolated, for example, recombinant antibody bound to human tau phosphorylated at amino acid residue S404, or a peptide comprising or consisting of the amino acid sequence DHGAEIVYKSPVVSGDT(pS)PRHLSNVSSTG (SEQ ID NO: 281), where p(S) corresponds to a phosphorylated serine residue. In some embodiments, the antibody comprises a heavy chain variable region (VH) comprising CDR1, CDR2, and CDR3 sequences, respectively, comprising SEQ ID NOs: 89, 106, and 124, and a light chain variable region (VL) comprising CDR1, CDR2, and CDR3 sequences, respectively, comprising SEQ ID NOs: 138, 152, and 169. In some embodiments, the antibody comprises VH and VL sequences, respectively, comprising SEQ ID NOs: 16 and 36.

[0212] In another embodiment, this specification provides isolated, for example, recombinant antibodies that bind to: (a) human tau phosphorylated at amino acid residue S199 but not at amino acid residues S202 and T205; (b) human tau phosphorylated at amino acid residue S202 but not at amino acid residues S199 and T205; (c) human tau phosphorylated at amino acid residue T205 but not at amino acid residues S199 and S202; (d) amino acid residues S199 and T205 (e) Human tau that is phosphorylated in combination but not at amino acid residue S202 (for example, the binding of tau phosphorylated in combination with S199 and T205 is at least 3 times stronger (e.g., at least 4 times stronger) than background (e.g., nonspecific) binding, e.g., binding by hIgG1 isotype control), (e) Human tau that is phosphorylated in combination with amino acid residues S202 and T205 but not at amino acid residue S199 (the binding of tau phosphorylated in combination with residues S199 and S202) (g) Human tau phosphorylated with amino acid residues (i) S202 and T205 (excluding S119), and (ii) S199 and T205 (excluding S202) in combination (at least twice as strong as background (e.g., nonspecific) binding, e.g., at least three times, at least four times, at least five times, 2-6 times, 2-5 times, 2-4 times, 2-3 times, 3-5 times, or 4-5 times stronger than the level of binding by hIgG1 isotype control) (i) S19 (ii) S202 and S202 (excluding T205), (iii) S199 and T205 (excluding S199), and (iv) human tau phosphorylated with combinations of S199, S202, and T205 (e.g., at least 1.6 times stronger than background (e.g., nonspecific) binding, e.g., binding by hIgG1 isotype controls (e.g., at least 1.7 times, at least 1.8 times, at least 1.9 times, at least 2 times, at least 3 times, 1.6 to 3 times, 1.6 to 2 times stronger)),(h) A peptide containing or consisting of the amino acid sequence SGDRSGYS(pS)PGSPGTPGSRSRTPS (SEQ ID NO: 284), (i) A peptide containing or consisting of the amino acid sequence SGDRSGYSSPG(pS)PGTPGSRSRTPS (SEQ ID NO: 285), (j) A peptide containing or consisting of the amino acid sequence SGDRSGYSSPGSPG(pT)PGSRSRTPS (SEQ ID NO: 286), (k) A peptide containing the amino acid sequence SGDRSGYS(pS)PGSPG(pT)PGSRSRTPS (SEQ ID NO: 290) (i) peptides containing or consisting thereof (for example, the binding to the peptide is at least 3 times (e.g., at least 4 times) stronger than the level of background (e.g., nonspecific) binding, e.g., binding by an hIgG1 isotype control), (l) peptides containing or consisting of the amino acid sequence SGDRSGYSSPG(pS)PG(pT)PGSRSRTPS (SEQ ID NO: 289) (except peptides containing or consisting of the amino acid sequence SGDRSGYS(pS)PG(pS)PGTPGSRSRTPS (SEQ ID NO: 288)). , (m) peptides containing or consisting of the amino acid sequences SGDRSGYSSPG(pS)PG(pT)PGSRSRTPS (SEQ ID NO: 289) and SGDRSGYS(pS)PGSPG(pT)PGSRSRTPS (SEQ ID NO: 290) (the binding to the latter peptide is at least twice (e.g., at least three times, at least four times, at least five times, 2-6 times, 2-5 times, 2-4 times, 2-3 times, 3-5 times, or 4-5 times) background (e.g., nonspecific) binding, e.g., binding by an hIgG1 isotype control). (strong), or peptides containing or consisting of the (n) amino acid sequences SGDRSGYS(pS)PG(pS)PGTPGSRSRTPS (SEQ ID NO: 288), SGDRSGYSSPG(pS)PG(pT)PGSRSRTPS (SEQ ID NO: 289), SGDRSGYS(pS)PGSPG(pT)PGSRSRTPS (SEQ ID NO: 290), and SGDRSGYS(pS)PG(pS)PG(pT)PGSRSRTPS (SEQ ID NO: 287) (for example, the binding to the peptide is background (e.g., nonspecific) binding, e.g.,The binding level is at least 1.6 times stronger than that of the hIgG1 isotype control (e.g., at least 1.7 times, at least 1.8 times, at least 1.9 times, at least 2 times, at least 3 times, 1.6 to 4 times, 1.6 to 3 times stronger), where p(S) and p(T) correspond to phosphorylated serine and phosphorylated threonine, respectively, and binding is optionally evaluated using, for example, the one-point ELISA described in Example 7. In some embodiments, the antibody comprises: (a) a heavy chain variable region (VH) containing CDR1, CDR2, and CDR3 sequences including SEQ ID NOs. 82, 97, and 115, respectively, and a light chain variable region (VL) containing CDR1, CDR2, and CDR3 sequences including SEQ ID NOs. 127, 141, and 159, respectively; (b) a heavy chain variable region (VH) containing CDR1, CDR2, and CDR3 sequences including SEQ ID NOs. 79, 94, and 111, respectively, and a light chain variable region (VL) containing CDR1, CDR2, and CDR3 sequences including SEQ ID NOs. 127, 141, and 156, respectively; (c) a heavy chain variable region (VH) containing CDR1, CDR2, and CDR3 sequences including SEQ ID NOs. 81, 94, and 114, respectively, and a light chain variable region (VL) containing CDR1, CDR2, and CDR3 sequences including SEQ ID NOs. 127, 141, and 156, respectively; and (d) (e) A heavy chain variable region (VH) containing CDR1, CDR2, and CDR3 sequences including 77, 92, and 109, and a light chain variable region (VL) containing CDR1, CDR2, and CDR3 sequences including sequence numbers 127, 141, and 154, respectively, (e) a heavy chain variable region (VH) containing CDR1, CDR2, and CDR3 sequences including sequence numbers 86, 102, and 120, respectively, and including sequence numbers 127, 141, and 156, respectively (f) a light chain variable region (VL) containing CDR1, CDR2, and CDR3 sequences, (g) a VH sequence and a VL sequence containing sequence numbers 7 and 25, respectively, (h) a VH sequence and a VL sequence containing sequence numbers 6 and 24, respectively, (i) a VH sequence and a VL sequence containing sequence numbers 1 and 19, respectively, or (j) a VH sequence and a VL sequence containing sequence numbers 12 and 31, respectively.

[0213] In another embodiment, as used herein, (a) binds to tau that is phosphorylated at T217 but not at T212 or T214, or (b) binds to peptides containing or comprising the sequences GTPGSRSRTPSLP(pT)PPTRE (SEQ ID NO: 293) and GTPGSSRSRTP(pS)LP(pT)PPTRE (SEQ ID NO: 296), but does not bind to peptides containing or comprising the sequences GTPGSSRSR(pT)PSLPTPPTRE (SEQ ID NO: 291), GTPGSSRSRTP(pS)LPTPPTRE (SEQ ID NO: 292), and GTPGSSRSR(pT)P(pS)LPTPPTRE (SEQ ID NO: 294), isolated, for example. For example, recombinant antibodies are provided, where p(S) and p(T) correspond to phosphorylated serine and phosphorylated threonine, respectively, and optionally, the binding of the antibody to tau or peptide is at least 1.5 times stronger (e.g., at least 1.6 times, at least 1.7 times, at least 1.8 times, at least 1.9 times, at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 6 times, 1.5 to 4 times, 1.5 to 3 times, 4 to 6 times stronger) than background (e.g., nonspecific) binding, e.g., binding by an hIgG1 isotype control, and optionally, the binding of the antibody to tau or peptide is evaluated using, for example, the one-point ELISA described in Example 8.In some embodiments, the antibody comprises (a) a heavy chain variable region (VH) containing CDR1, CDR2, and CDR3 sequences including SEQ ID NOs. 80, 95, and 112, respectively, and a light chain variable region (VL) containing CDR1, CDR2, and CDR3 sequences including SEQ ID NOs. 129, 143, and 157, respectively; (b) a heavy chain variable region (VH) containing CDR1, CDR2, and CDR3 sequences including SEQ ID NOs. 78, 104, and 122, respectively, and CDR1, CDR2, and CD (c) a light chain variable region (VL) containing the R3 sequence, (d) a VH sequence and a VL sequence containing the CDR1, CDR2, and CDR3 sequences containing sequence numbers 90, 107, and 125 respectively, and a light chain variable region (VL) containing the CDR1, CDR2, and CDR3 sequences containing sequence numbers 139, 151, and 170 respectively, (e) a VH sequence and a VL sequence containing sequence numbers 4 and 22 respectively, (f) a VH sequence and a VL sequence containing sequence numbers 14 and 24 respectively, or (f) a VH sequence and a VL sequence containing sequence numbers 17 and 37 respectively.

[0214] As described in the examples, the specific binding to the phosphorylated residues or peptides can be measured by comparing the background binding level in an assay (e.g., one-point ELISA) or the binding level with a negative control such as an isotype control antibody (e.g., human IgG1 isotype control antibody).

[0215] In some embodiments, this specification provides anti-tau antibodies that compete with any of the anti-tau antibodies described herein in terms of binding to human tau or human tau peptides.

[0216] Furthermore, this specification also provides anti-tau antibodies that exhibit similar epitope-binding properties to the anti-tau antibodies described herein, for example, the ability to bind to phosphorylated epitopes on human tau. Thus, in some embodiments, the antibodies provided herein provide antibodies that bind to the same epitopes (e.g., phosphorylated epitopes), substantially the same epitopes, overlapping epitopes, or substantially overlapping epitopes recognized by the anti-tau antibodies described herein. In some embodiments, the antibodies described herein bind to discontinuous epitopes, such as structural epitopes.

[0217] Exemplary methods for determining whether an antibody binds to the same epitope (or substantially the same epitope, overlapping epitope, or substantially overlapping epitope) on human tau as the antibody described herein include, for example, X-ray spectroscopy epitope mapping of the antigen:antibody complex crystal, which provides atomic resolution of the epitope. Other methods involve monitoring the binding of the antibody to an antigen fragment or a variant of the antigen. In this case, loss of binding due to modification of amino acid residues in the antigen sequence is often considered an indicator of the epitope component. Computational combinatorial methods can also be used for epitope mapping. The method may also depend on the ability of the antibody of interest to affinity isolate specific short peptides (either in their native three-dimensional form or a modified form) from a combinatorial phage display peptide library. Epitope mapping can also be performed using MS-based protein footprints (e.g., HDX-MS, FPOP). Overlap phosphorylated peptide scanning can also be used for epitope mapping, as described in the examples.

[0218] Anti-tau antibodies according to this disclosure may be prepared using any of the antibody sequences presented herein (e.g., variable domain amino acid sequences, variable domain amino acid sequence pairs, CDR amino acid sequences, variable domain CDR amino acid sequence sets, pairs of variable domain CDR amino acid sequence sets, and / or framework region amino acid sequences), any of which may be prepared, for example, as monoclonal antibodies, multispecific antibodies, chimeric antibodies, antibody mimes, scFvs, or antibody fragments. In some embodiments, anti-tau antibodies using any of the antibody sequences presented herein may be prepared as IgA, IgD, IgE, IgG, or IgM antibodies. When prepared as mouse IgG antibodies, anti-tau antibodies may be prepared as IgG1, IgG2a, IgG2b, or IgG3 isotypes. When prepared as human IgG antibodies, anti-tau antibodies may be prepared as IgG1, IgG2, IgG3, or IgG4 isotypes. Anti-tau antibodies prepared as human or humanized antibodies may contain one or more human constant domains. The human constant domains included in the anti-tau antibody of this disclosure include, but are not limited to, any of the domains listed in Table 5.

[0219] In some embodiments, the anti-tau antibody has (i) a heavy chain constant region (CH), such as the amino acid sequence of a human CH, such as the CH of human IgG1, IgG2, IgG3, or IgG4 (e.g., the CHs listed in Table 5), or a mouse CH, or a sequence having at least 80% (e.g., 85, 90, 95, 96, 97, 98, or 99%) sequence identity thereto, or at least 1, 2, or 3 modifications compared to the constant region sequence (e.g., the CHs listed in Table 5), but no more than 30, 20, or 10 modifications (e.g., 1-30, 1-20, 1-10, 1-5, 1-4, 1-3, 1-2, or 1 amino acid modification, e.g., substitution (e.g., conservative substitution)). CH containing an amino acid sequence, and / or (ii) a light chain constant region (CL), for example, an amino acid sequence of a human CL or mouse CL such as a human λ light chain or κ light chain CL (e.g., the CLs listed in Table 5), or a sequence having at least 80% (e.g., 85, 90, 95, 96, 97, 98, or 99%) sequence identity thereto, or a CL containing an amino acid sequence having at least 1, 2, or 3 modifications, but 30, 20, or 10 or fewer modifications (e.g., 1-30, 1-20, 1-10, 1-5, 1-4, 1-3, 1-2, or 1 amino acid modification, e.g., substitution (e.g., conservative substitution)) compared to a constant region sequence (e.g., the CHs listed in Table 5).

[0220] In some embodiments, the heavy chain constant region sequence (e.g., the human heavy chain constant region sequence) has a C-terminal lysine (K), a C-terminal glycine (G), or a C-terminal glycine and lysine (GK). In some embodiments, the heavy chain constant region sequence (e.g., the human heavy chain constant region sequence) lacks a C-terminal lysine (K), a C-terminal glycine (G), or a C-terminal glycine and lysine (GK).

[0221] In some embodiments, the anti-tau antibodies of this disclosure include a constant domain comprising amino acid sequence variants and / or fragments of the amino acid sequences listed in Table 5. The amino acid fragments or variants contained in the constant domain of the anti-tau antibody may have about 50% to about 99.9% sequence identity with one or more of the amino acid sequences listed in Table 5 (e.g., about 50% to about 60%, about 55% to about 65%, about 60% to about 70%, about 65% to about 75%, about 70% to about 80%, about 75% to about 85%, about 80% to about 90%, about 85% to about 95%, about 90% to about 99.9%, about 95% to about 99.9%, about 97%, about 97.5%, about 98%, about 98.5%, about 99%, about 99.5%, about 99.6%, about 99.7%, or about 99.8%). In some embodiments, the constant domain has at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% sequence identity with respect to the amino acid sequences listed in Table 5, or at least 1, 2, or 3 modifications, provided that there are no more than 30, 20, or 10 modifications (e.g., amino acid substitutions such as conservative substitutions).

[0222] The VH domains described herein, or one or more CDRs thereof, can be linked to a constant domain to form a heavy chain, such as a full-length heavy chain. Similarly, the VL domains described herein, or one or more CDRs thereof, can be linked to a constant domain to form a light chain, such as a full-length light chain. A full-length heavy chain (whose C-terminus is not lysine (K), whose C-terminus is not glycine (G), or whose C-terminus is not glycine and lysine (GK), and which may or may not be present) and a full-length light chain can bind to form a full-length antibody. [Table 5-1] [Table 5-2] [Table 5-3] [Table 5-4]

[0223] In some embodiments, the anti-tau antibody may include an Fc region or a variant thereof, such as a functional variant.

[0224] In some embodiments, the antibody comprises an Fc region that has been modified, for example, increased or decreased (e.g., removed), to have affinity for the Fc receptor compared to a reference, where the reference is a wild-type Fc receptor.

[0225] In some embodiments, the antibody includes an Fc region containing mutations at one, two, or all of the following positions, numbered according to the EU index, such as in Kabat: I253 (e.g., I235A), H310 (e.g., H310A), and / or H435 (e.g., H435A).

[0226] Antibody Characterization In some embodiments, antibodies of this disclosure may be identified, selected, or excluded based on different characteristics. Such characteristics may include, but are not limited to, physical and functional characteristics. Physical characteristics may include properties of the antibody structure [e.g., amino acid sequence or residues; secondary, tertiary, or quaternary protein structure; post-translational modifications (e.g., glycosylation); chemical bonding, and stability]. Functional characteristics may include, but are not limited to, antibody affinity (i.e., against a particular epitope and / or antigen) and antibody activity (e.g., the ability of an antibody to activate or inhibit a target, process, or pathway).

[0227] Antibody binding and affinity In some embodiments, the antibodies of this disclosure may be identified, selected, or excluded based on their level of binding and / or affinity to a particular epitope and / or antigen. The level of antibody binding and / or affinity may be evaluated using different antigen formats. In some embodiments, antibody affinity to different antigen formats may be tested in vitro (e.g., by ELISA). In vitro testing of anti-tau antibodies may be performed using brain samples or fragments. Such samples or fragments may be obtained from subjects with AD (e.g., human AD patients). In some embodiments, brain samples or fragments may be obtained from non-human subjects. Such non-human subjects may include non-human animals (e.g., mice, rats, and primates) used in AD disease model studies. In some embodiments, the brain samples or fragments used in antibody affinity testing may be derived from the TG4510 / P301S mouse strain. Antibody affinity may be compared against a control sample lacking the specific antigen for which affinity is to be analyzed. In some embodiments, the control sample used in anti-tau antibody testing may include brain samples or fragments derived from unaffected human subjects. In some embodiments, brain samples or fragments derived from wild-type and / or tau knockout mouse strains may be used as control samples.

[0228] In vitro affinity testing may be performed using recombinant or isolated protein antigens (e.g., by ELISA). For example, recombinant or isolated ePHF may be used for anti-tau antibody affinity testing. In some embodiments, the anti-tau antibodies of this disclosure may exhibit a maximum half-volume effective concentration (EC50) of about 0.01 nM to about 100 nM for binding to ePHF when evaluated by ELISA. In some embodiments, the exhibited EC50 may be less than about 50 nM, less than about 20 nM, less than about 10 nM, or less than about 1 nM. In some embodiments, the anti-tau antibodies of this disclosure may exhibit an EC50 of about 0.01 nM to about 100 nM for binding to any of the antigens listed in Table 4, or to epitopes (including but not limited to structural epitopes) containing or included in any of the antigens, when evaluated by ELISA. In some embodiments, the exhibited EC50 may be less than about 50 nM, less than about 20 nM, less than about 10 nM, or less than about 1 nM.

[0229] In some embodiments, the anti-tau antibodies of this disclosure bind to pathological tau but not to non-pathological tau. Such antibodies may be referred to herein as "selective" to pathological forms of tau. In some embodiments, the anti-tau antibodies of this disclosure bind to tau concentrates.

[0230] In some embodiments, antibody affinity analysis may be used to identify, select, or exclude multispecific antibodies. As used herein, the term “multispecific antibody” refers to an antibody that has affinity for multiple epitopes or antigens. In some embodiments, multispecific antibodies may be identified, selected, or excluded based on their relative affinity for each recognized epitope or antigen. For example, a multispecific antibody may be selected for use or further development based on the fact that its affinity for a certain epitope or antigen is higher than that for a second epitope or antigen.

[0231] In some embodiments, anti-tau antibodies may be tested for competition with other anti-tau antibodies. Such testing can be performed to provide information about specific epitopes recognized by the antibody and may yield information related to the level of epitope affinity compared to competing antibodies. In some embodiments, the anti-tau antibody used for antibody binding and / or affinity analysis may include anti-tau antibody PT3 as described in U.S. Patent No. 9,371,376, anti-tau antibody C10.2 as described in U.S. Patent No. 10,196,439 (referred to as antibody "C10-2" in the same document), anti-tau antibody IPN002 as described in U.S. Patent No. 10,040,847, anti-tau antibody AT8 (ThermoFisher, Waltham, MA), anti-tau antibody AT100 (ThermoFisher, Waltham, MA), anti-tau antibody AT120 as described in U.S. Patent No. 5,843,779, or anti-tau antibody PT76 as described in Vandermeeren, M. et al., J Alzheimers Dis. 2018; 65(1): 265-281.

[0232] antibody activity In some embodiments, antibodies of the present disclosure may be identified, selected, or excluded based on their ability to promote or reduce certain activities. Antibody activity can be evaluated using analytical assays. Such assays may be selected or designed to detect, screen, measure, and / or rank antibodies based on such antibody activity.

[0233] Anti-tau antibodies can be characterized by their ability to inhibit tau aggregation. Inhibition may be based on the physical disruption of tau aggregation or on the anti-tau antibody-dependent depletion (immunodepletion) of tau protein. Characterization based on tau aggregation inhibition can be evaluated using one or more tau aggregation assays. In some embodiments, anti-tau antibodies can be characterized by a tau seeding assay. A tau seeding assay typically involves the initiation of tau aggregation in vitro and the evaluation of aggregation inhibition by the candidate compound under test. A tau seeding assay may be performed using tau-aggregating biosensor cells. Tau-aggregating biosensor cells produce a detectable signal (e.g., a fluorescent signal) in response to tau aggregation. Tau-aggregating biosensor cells may be cultured with recombinant or isolated tau, or with high-tau brain tissue or fluid-derived samples (to promote tau aggregation), and treated with or without the candidate compound to evaluate tau aggregation inhibition. In some embodiments, an anti-tau antibody may be used to deplete tau from the culture medium before incubation with biosensor cells. The inhibitory function of the anti-tau antibody can be evaluated by comparing the level of aggregation in the depleted medium with the level of aggregation in the non-depleted medium. Tau-aggregating biosensor cells may include, but are not limited to, tau RD biosensor cells. In some embodiments, neurons expressing human tau may be used.

[0234] In some embodiments, the anti-tau antibodies of this disclosure can inhibit tau aggregation at a maximum half-mass inhibitory concentration (IC50) of about 1 nM to about 30 nM, as determined by an immunodepletion assay (e.g., using tau RD biosensor cells).

[0235] Antibody structure and variations The antibodies of this disclosure may exist as whole polypeptides, multiple polypeptides, or polypeptide fragments, which may independently be encoded by one or more nucleic acids, multiple nucleic acids, nucleic acid fragments, or variants of any of the foregoing. As used herein, “polypeptide” means a polymer of amino acid residues (natural or unnatural) linked together most often by peptide bonds. As used herein, this term refers to proteins, polypeptides, and peptides of any size, structure, or function. Polypeptides smaller than about 50 amino acids may be referred to using the term “peptide.” Peptides may be at least about 2, 3, 4, or at least 5 amino acid residues in length. The polypeptides of this disclosure may include gene products, naturally occurring polypeptides, synthetic polypeptides, homologs, orthologues, paralogs, fragments, or other equivalents, variants, and analogues of the foregoing. Polypeptides may be single molecules or multimolecular complexes such as dimers, trimers, or tetramers. Polypeptides may include single-chain or multi-chain polypeptides, which may be associated or linked. Polypeptides may contain amino acid polymers in which one or more amino acid residues are artificial chemical analogs of the corresponding naturally occurring amino acids.

[0236] The term "polypeptide variant" refers to a molecule whose amino acid sequence differs from that of the native sequence or a reference sequence. Amino acid sequence variants may have substitutions, deletions, and / or insertions at specific positions within the amino acid sequence when compared to the native sequence or a reference sequence. Typically, variants have at least about 50% identity (homology) to the native sequence or a reference sequence, preferably at least about 80%, and more preferably at least about 90% identity (homology) to the native sequence or a reference sequence.

[0237] In some embodiments, “variant mimetic” is provided. As used herein, the term “variant mimetic” comprises one or more amino acids that mimic an activated sequence. For example, glutamic acid may function as a mimetic for phosphorylated threonine and / or phosphorylated serine. Alternatively, a variant mimetic may result in an inactivated product that is inactivated or contains the mimetic; for example, phenylalanine may act as an inactivating substitution for tyrosine, or alanine may act as an inactivating substitution for serine.

[0238] The term "amino acid sequence variant" refers to a molecule that has several differences in its amino acid sequence compared to the natural or starting sequence. Amino acid sequence variants may have substitutions, deletions, and / or insertions at specific positions within the amino acid sequence. "Natural" or "starting" sequences should not be confused with wild-type sequences. As used herein, "natural" or "starting" sequences are relative terms referring to the original molecule from which comparisons may be made. A "natural" or "starting" sequence or molecule may represent the wild type (the sequence found in nature), but does not necessarily have to be a wild-type sequence.

[0239] Typically, variants have at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.8%, or at least 99.9% sequence identity compared to the natural sequence.

[0240] In the context of amino acid sequences, "homogram" refers to a corresponding sequence of another species that has substantial identity with respect to a second sequence of the second species.

[0241] The term "analog" is intended to include polypeptide variants that differ by one or more amino acid changes, such as substitution, addition, or deletion of amino acid residues, but still retain the properties of the parent polypeptide.

[0242] This disclosure envisions variants and derivatives of antibodies presented herein. These include substitutions, insertions, deletions, and covalent variants and derivatives. For example, sequence tags or one or more amino acids, such as lysine, may be added to an antibody peptide sequence (e.g., at the N-terminus or C-terminus). Sequence tags may be used for the purification or localization of the peptide. Lysine may be used to increase the solubility of the peptide or to enable biotinylation. Alternatively, amino acid residues located in the carboxyl-terminal and amino-terminal regions of the amino acid sequence of a peptide or polypeptide may be optionally deleted to result in a truncated sequence. Alternatively, certain amino acids (e.g., C-terminal or N-terminal residues) may be deleted depending on the use of the sequence, for example, as part of a larger, soluble sequence or in the expression of a sequence linked to a solid support.

[0243] When referring to polypeptides, a "substituted variant" is one in which at least one amino acid residue is removed from the natural or starting sequence and replaced by a different amino acid inserted at the same position. The substitution can be a single substitution, where only one amino acid in the molecule is replaced, or a multiple substitution, where two or more amino acids are replaced in the same molecule.

[0244] As used herein, the term “conservative amino acid substitution” refers to the substitution of an amino acid normally present in a sequence with a different amino acid that is similar in size, charge, or polarity. Examples of conservative substitutions include the substitution of a nonpolar (hydrophobic) residue, such as isoleucine, valine, and leucine, with another nonpolar residue. Similarly, examples of conservative substitutions include the substitution of one polar (hydrophilic) residue with another, such as arginine and lysine, glutamine and asparagine, and glycine and serine. Further examples of conservative substitutions include the substitution of a basic residue, such as lysine, arginine, or histidine, with another, or the substitution of one acidic residue, such as aspartic acid or glutamic acid, with another acidic residue. Examples of non-conservative substitutions include the substitution of a nonpolar (hydrophobic) amino acid residue, such as isoleucine, valine, leucine, alanine, or methionine, with a polar (hydrophilic) residue, such as cysteine, glutamine, glutamic acid, or lysine, and / or the substitution of a polar residue with a nonpolar residue.

[0245] The term "functional variant" refers to a polypeptide variant or polynucleotide variant that has at least one activity of the reference sequence.

[0246] When referring to polypeptides, an "insertion variant" is one that has one or more amino acids inserted immediately next to an amino acid at a specific position in the natural or starting sequence. "Immediately next to" an amino acid means that it is attached to either the alpha-carboxyl or alpha-amino functional group of the amino acid.

[0247] When referring to polypeptides, a "deletion variant" is one in which one or more amino acids have been removed from the native or starting amino acid sequence. Typically, a deletion variant is the absence of one or more amino acids within a specific region of the molecule.

[0248] As used herein, the term “derivative” is used synonymously with the term “variant” and refers to a molecule that has been modified or altered in some way relative to a reference molecule or starting molecule. In some embodiments, derivatives include native or starting polypeptides modified by proteinaceous or nonproteinaceous organic derivatizers and post-translational modifications. Covalent modifications are conventionally introduced by reacting a target amino acid residue in a polypeptide with an organic derivatizer that can react with selected side-chain or terminal residues, or by utilizing a post-translational modification mechanism that functions in selected recombinant host cells. The resulting covalent derivatives are useful in programs aimed at identifying residues important for biological activity, immunoassays, or the preparation of antibodies for immunoaffinity purification.

[0249] Certain post-translational modifications are the result of the recombinant host cell's action on the expressed polypeptide. Glutaminyl and asparaginyl residues are frequently deamidated post-translation to their corresponding glutamyl and aspartyl residues. Alternatively, these residues are deamidated under weakly acidic conditions. Any of these residue forms may be present in the polypeptide used in accordance with this disclosure.

[0250] Other post-translational modifications include hydroxylation of proline and lysine, phosphorylation of hydroxyl groups of ceryl or threonyl residues, and methylation of alpha-amino groups of lysine, arginine, and histidine side chains (TECreighton, Proteins: Structure and Molecular Properties, WH Freeman & Co., San Francisco, pp. 79-86 (1983)).

[0251] Covalent derivatives specifically include fusion molecules in which a polypeptide is covalently bonded to a non-proteinaceous polymer. Non-proteinaceous polymers may include hydrophilic synthetic polymers, i.e., polymers otherwise not found in nature. However, naturally occurring polymers produced by recombinant or in vitro methods are just as useful as polymers isolated from nature. Hydrophilic polyvinyl polymers may include polyvinyl alcohol and / or polyvinylpyrrolidone. Particularly useful are polyvinyl alkylene ethers such as polyethylene glycol and polypropylene glycol. Polypeptides may be bonded to various non-proteinaceous polymers such as polyethylene glycol, polypropylene glycol, or polyoxyalkylene in the manner described in U.S. Patents 4,640,835, 4,496,689, 4,301,144, 4,670,417, 4,791,192, or 4,179,337, the contents of which are incorporated herein by reference in their entirety.

[0252] When referring to polypeptides, "characteristics" are defined as constituent elements based on the different amino acid sequences of the molecule. Polypeptide characteristics may include surface expression, local three-dimensional shape, folds, loops, semi-loops, domains, semi-domains, sites, terminals, or any combination thereof.

[0253] In this specification, when referring to polypeptides, the term "surface-expressed" refers to the amino acid-based components of the polypeptide that appear on the outermost surface.

[0254] In this specification, when referring to polypeptides, the term “local three-dimensional shape” means the amino acid-based structural expression of the polypeptide located within the definable space of the polypeptide.

[0255] As used herein when referring to polypeptides, the term "fold" means the three-dimensional structure of the amino acid sequence obtained when energy is minimized. Folds may occur at the secondary or tertiary levels of the folding process. Examples of secondary-level folds include β-sheets and α-helices. Examples of tertiary folds include domains and regions formed by the aggregation or separation of energy forces. Regions thus formed include hydrophobic pockets and hydrophilic pockets, among others.

[0256] As used herein, the term “turn” in relation to polypeptide stereochemistry means a curve that changes the orientation of the main chain of a peptide or polypeptide, and may include one, two, three or more amino acid residues.

[0257] As used herein when referring to polypeptides, the term “loop” refers to a structural feature of a peptide or polypeptide that reverses the orientation of the peptide or polypeptide backbone and contains four or more amino acid residues. Oliva et al. have identified at least five classes of polypeptide loops (J.Mol Biol 266(4):814-830;1997, the contents of which are incorporated herein by reference in their entirety).

[0258] When used herein in reference to polypeptides, the term “half-loop” refers to a portion of an identified loop that contains at least half of the amino acid residues present in the original loop. It is understood that a loop does not necessarily contain an even number of amino acid residues. Therefore, if a loop contains, or is identified as containing, an odd number of amino acids, the half-loops of an odd-numbered loop will contain the integer part of the loop or the next integer part (the number of amino acids in loop / 2 ± 0.5 amino acids). For example, a loop identified as a 7-amino acid loop may produce 3-amino acid or 4-amino acid half-loops (7 / 2 = 3.5 ± 0.5 is 3 or 4).

[0259] As used herein in reference to polypeptides, the term “domain” refers to a polypeptide motif having one or more identifiable structural and / or functional features or properties (e.g., binding ability) that function, for example, as a site for protein-protein interactions.

[0260] When referring to polypeptides, as used herein, the term “half-domain” means a portion of an identified domain having at least half the number of amino acid residues as the original domain. It is understood that a domain does not necessarily contain an even number of amino acid residues. Therefore, if a domain contains, or is identified as containing, an odd number of amino acids, the half-domains of an odd-numbered domain contain an integer part of the domain or the next integer part (the number of amino acids in domain / 2 ± 0.5 amino acids). For example, a domain identified as a 7-amino acid domain may produce 3-amino acid or 4-amino acid half-domains (7 / 2 = 3.5 ± 0.5 is 3 or 4). It is also understood that subdomains may be identified within a domain or half-domain, and these subdomains do not necessarily possess all of the structural or functional properties identified in the original domain or half-domain. It is also understood that any amino acids in any of the domain types herein do not need to be contiguous along the backbone of the polypeptide (i.e., non-adjacent amino acids can structurally fold to produce a domain, half-domain, or subdomain).

[0261] As used herein in reference to polypeptides, the term “site” is synonymous with “amino acid residue” and “amino acid side chain.” A site represents a location on a polypeptide that can be modified, manipulated, altered, derivatized, or changed within that polypeptide.

[0262] As used herein, the term “terminus” refers to the end of a peptide or polypeptide when referring to a polypeptide. Such an end is not limited to the first or last site of a peptide or polypeptide and may include further amino acids within the terminal region. Polypeptide-based molecules of this disclosure may be characterized as having both an N-terminus (terminating with an amino acid having a free amino group) and a C-terminus (terminating with an amino acid having a free carboxyl group). Proteins of this disclosure may optionally consist of multiple polypeptide chains held together by disulfide bonds or non-covalent bonds (multimers, oligomers). These types of proteins may have multiple N-terminuses and C-terminuses. Alternatively, the terminus of a polypeptide may optionally be modified to begin or end with a non-polypeptide-based portion, such as an organic conjugate.

[0263] Antibody modification Antibodies can be modified to obtain variants having one or more altered properties. Such properties may include, or relate to, the structure, function, affinity, specificity, protein folding, stability, manufacturing, expression, and / or immunogenicity (i.e., the immune response in a subject treated with such antibody). In some embodiments, antibody fragments or variants may be used to modify another antibody or incorporated into a synthetic antibody.

[0264] Antibody modifications may include amino acid sequence modifications. Such modifications may include, but are not limited to, amino acid deletions, additions, and / or substitutions. Information about modifications may be obtained by amino acid sequence analysis. Such analysis may include amino acid sequence alignment between different antibodies or antibody variants. Two or more antibodies can be compared to identify residues or regions suitable for modification. The antibodies being compared may include those that bind to the same epitope. The antibodies being compared may bind to different (separate or overlapping) epitopes of the same protein or target (for example, this may identify residues or regions that give specificity to a particular epitope). Comparisons may include analysis of light chain and / or heavy chain sequence variations, CDR sequence variations, germline sequence analysis, and / or framework sequence analysis. Information obtained from such analyses can be used to identify amino acid residues, amino acid segments, amino acid side chains, CDR lengths, and / or other properties or characteristics that are conserved or variable between antibodies that bind to the same or different epitopes.

[0265] Functional modification In some embodiments, the antibodies of this disclosure may be modified to optimize one or more functional properties (e.g., antibody affinity or activity). Non-limiting examples of functional properties of an antibody include epitope or antigen affinity, ability to recruit or immobilize a target, and ability to activate or inhibit a target, process, or pathway. In some embodiments, functional properties include, or relate to, the ability to modulate protein-protein interactions, protein aggregation, enzyme activity, receptor-ligand interactions, cell signaling pathways, proteolytic cascades, and / or biological or physiological responses.

[0266] Antibody modification can optimize antibodies by adjusting epitope affinity. Such modifications can be performed by affinity maturation. Affinity maturation techniques are used to identify sequences encoding CDRs with the highest affinity to the target antigen. In some embodiments, antibody display techniques (e.g., phages or yeast) may be used. Such methods may involve mutating the nucleotide sequence encoding the parent antibody to be optimized. The nucleotide sequence can be mutated randomly as a whole, or by altering expression at specific amino acid residues, to produce millions to billions of variants. Sites or residues may be selected for mutation based on the frequency of sequences or amino acids observed in the native human antibody repertoire. Variants may be subjected to multiple iterations of affinity screening for target antigen binding [e.g., using display library screening techniques, surface plasmon resonance techniques, fluorescence-associated cell sorting (FACS) analysis, enzyme-linked immunosorbent assay (ELISA), etc.]. Multiple iterations of selection, mutation, and expression may be performed to identify antibody fragment sequences with the highest affinity to the target antigen. Such sequences may be directly incorporated into the antibody sequence for production. In some cases, the goal of affinity maturation is to increase the antibody affinity by at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 100, 1000, 500, 1000, or more than 1000 times compared to the affinity of the original antibody or starting antibody. If the affinity is lower than desired, this process may be repeated.

[0267] In some embodiments, antibody affinity may be evaluated using different antigen formats. In some embodiments, antibody affinity to different antigen formats may be tested in vitro (e.g., by ELISA). In vitro testing may be performed using brain samples or fragments. Such samples or fragments may be obtained from subjects with AD (e.g., human AD patients). In some embodiments, brain samples or fragments may be obtained from non-human subjects. Such non-human subjects may include non-human animals (e.g., mice, rats, and primates) used in AD disease model studies. In some embodiments, brain samples or fragments used in antibody affinity testing may be derived from the TG4510 / P301S mouse strain. Antibody affinity may be compared against a control sample lacking the specific antigen for which affinity is to be analyzed. In some embodiments, the control sample may include brain samples or fragments derived from non-affected human subjects. In some embodiments, brain samples or fragments derived from wild-type and / or tau knockout mouse strains may be used as control samples. In vitro affinity testing may be performed using recombinant or isolated protein antigens (e.g., by ELISA). In some embodiments, recombinant or isolated ePHF is used for antibody affinity testing. In some embodiments, the antigens listed in Table 8 may be used.

[0268] In some embodiments, antibody affinity analysis may be used to adjust the multispecificity of an antibody (for example, to reduce or enhance its multispecificity). Such adjustment may involve adjusting the relative affinity to two or more epitopes or antigens. For example, an antibody may be optimized so that its affinity to one epitope or antigen is higher than that to a second epitope or antigen.

[0269] Antibodies can be modified to optimize their functional properties. Such functional properties can be evaluated or engineered based on analytical assay results related to one or more functional properties of the antibody. Assays may be used to screen multiple antibodies and identify or rank them based on functional criteria. Anti-tau antibodies can be modified to optimize tau agglutination inhibition. Such inhibition may be based on the physical disruption of tau agglutination or on the ability of the anti-tau antibody to deplete tau protein from the assay sample. Optimization based on tau agglutination inhibition can be evaluated using one or more tau agglutination assays (e.g., by a tau seeding assay).

[0270] Modification of production In some embodiments, modifications may be made to optimize antibody production. Such modifications may include, or relate to, one or more of the following: protein folding, stability, expression, and / or immunogenicity. Modifications may be made to address one or more antibody properties that adversely affect production. Such properties may include, but are not limited to, unpaired cysteine ​​or irregular disulfide; glycosylation sites (e.g., N-linked NXS / T sites); acid cleavage sites, amino acid oxidation sites, compatibility with mouse germline sequences; asparagine deamidation sites; aspartate isomerization sites; N-terminal pyroglutamate formation sites; and easily aggregated amino acid sequence regions (e.g., within CDR sequences).

[0271] Production method In some embodiments, the antibodies of this disclosure may be prepared using recombinant DNA technology (see, for example, U.S. Patent No. 4,816,567, which is incorporated herein by reference in whole). The DNA encoding the antibody can be isolated and sequenced using conventional procedures (for example, by using oligonucleotide probes that can specifically bind to the genes encoding the heavy and light chains of the mouse antibody). In some embodiments, hybridoma cells may be used as a preferred DNA source. Once isolated, the DNA can be placed into an expression vector, which is then transfected into host cells. The host cells may include, but are not limited to, HEK293 cells, HEK293T cells, monkey COS cells, Chinese hamster ovary (CHO) cells, and myeloma cells, which otherwise do not produce immunoglobulin proteins, for the synthesis of monoclonal antibodies in recombinant host cells. DNA can also be modified, for example, by substituting the coding sequences of human heavy and light chain constant domains for homologous mouse sequences (U.S. Patent No. 4,816,567), or by covalently bonding all or part of the coding sequence of a non-immunoglobulin polypeptide to an immunoglobulin coding sequence.

[0272] The method for producing anti-tau antibodies described herein includes, for example, expressing the heavy and light chains of an antibody in a cell line containing a nucleic acid sequence or vector (or more) expressing the heavy and light chains (e.g., an expression vector (or more)). Host cells containing these nucleic acid sequences (e.g., the nucleic acid sequences described herein, such as in Table 1) are included herein.

[0273] After obtaining DNA fragments encoding the VH and VL segments, these DNA fragments can be further manipulated using standard recombinant DNA techniques to convert, for example, a variable region gene into a full-length antibody chain gene, a Fab fragment gene, or an scFv gene. These operations involve ligating the VL or VH-encoding DNA fragment into an operational structure to another DNA fragment encoding another protein, such as an antibody constant region (e.g., the constant regions listed in Table 5) or a flexible linker. In this context, the term "ligated into an operational structure" is intended to mean joining two DNA fragments in such a way that the amino acid sequences encoded by the two DNA fragments are maintained in frame.

[0274] The isolated DNA encoding the VH region can be converted into a full-length heavy chain gene by ligating this VH-encoding DNA into another DNA molecule encoding the heavy chain constant region (hinge, CH1, CH2, and / or CH3), for example, the heavy chain constant region listed in Table 5. The sequences of human heavy chain constant region genes are known in the art (see, for example, Kabat, EA, el al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, USD Department of Health and Human Services, NIH Publication No. 91-3242), and DNA fragments containing these regions can be obtained by standard PCR amplification. The heavy chain constant region may be the IgG1, IgG2, IgG3, IgG4, IgA, IgE, IgM, or IgD constant region, for example, the IgG1 region as described above. In the case of Fab fragment heavy chain genes, the DNA encoding VH can be ligated into another DNA molecule encoding only the heavy chain CH1 constant region.

[0275] Isolated DNA encoding the VL region can be converted into a full-length light chain gene (and Fab light chain gene) by ligating this VL-encoding DNA to another DNA molecule encoding the light chain constant region CL, for example, the light chain constant regions listed in Table 5. The sequences of human light chain constant region genes are publicly known in the art (see, for example, Kabat, EA, et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, USD Department of Health and Human Services, NIH Publication No. 91-3242), and DNA fragments containing these regions can be obtained by standard PCR amplification. The light chain constant region may be a κ or λ constant region, as described above.

[0276] To express the antibodies described herein, nucleic acids encoding partial or full-length light and heavy chains, or combinations thereof, can be obtained by standard molecular biology techniques (e.g., PCR amplification or cDNA cloning using a hybridoma expressing the antibody of interest), and the DNA can be inserted into an expression vector so that the gene is functionally linked to transcriptional and translational regulatory sequences. In this regard, the term “functionally linked” is intended to mean that the antibody gene is linked to the vector so that the transcriptional and translational regulatory sequences in the vector perform their intended functions of regulating the transcription and translation of the antibody gene. The expression vector and expression regulatory sequences are selected to be compatible with the expression host cells used. The antibody light chain gene and antibody heavy chain gene can be inserted into separate vectors, or both genes can be inserted into the same expression vector. The antibody gene is inserted into the expression vector(s) by standard methods (e.g., ligation of the antibody gene fragment and complementary restriction enzyme sites on the vector, or blunt-end ligation if restriction enzyme sites are absent). Using the light chain and heavy chain variable regions of the antibodies described herein, HThe segment is C in the vector H The segment is operably connected, V L The segment is C in the vector L By inserting segments into an expression vector that already encodes the heavy chain constant region and light chain constant region of the desired isotype, so that they are linked to the activating mechanism, a full-length antibody gene of any antibody isotype can be constructed.

[0277] Furthermore, recombinant expression vectors can encode signal peptides that promote the secretion of antibody chains from host cells. Antibody chain genes can be cloned into the vector so that the signal peptide is in-frame ligated to the amino terminus of the antibody chain gene. The signal peptide may be an immunoglobulin signal peptide or a heterologous signal peptide (i.e., a non-immunoglobulin protein-derived signal peptide).

[0278] For light and heavy chain expression, an expression vector (e.g., where the nucleic acids encoding the light and heavy chains are in a single vector) or a combination of expression vectors (e.g., where the nucleic acid encoding the light chain is in a single vector and the nucleic acid encoding the heavy chain is in a separate vector) is transfected into host cells using standard techniques. The various forms of the term "transfection" are intended to encompass a variety of techniques commonly used to introduce exogenous DNA into prokaryotic (bacterial host cells) or eukaryotic host cells, such as electroporation, calcium phosphate precipitation, and DEAE dextran transfection.

[0279] Exemplary mammalian cells for expressing the recombinant antibodies described herein include Chinese hamster ovary (CHO cells) (e.g., including dhfr-CHO cells described in Urlaub and Chasin, (1980) Proc. Natl. Acad. Sci. USA 77:4216-4220, used with a DHFR selection marker such as those described in RJ Kaufman and PA Sharp (1982) Mol. Biol. 159:601-621), NSO myeloma cells, COS cells, and SP2 cells. When a recombinant expression vector encoding an antibody gene is introduced into mammalian host cells, antibody expression occurs within the host cells, or more preferably, the host cells are cultured in a medium for a period sufficient to allow antibody secretion. The antibody can be recovered from the medium using standard protein purification methods.

[0280] Humanization of antibodies In some embodiments, the anti-tau antibodies of this disclosure may be prepared as humanized antibodies. A “humanized” antibody is a chimeric antibody containing a minimal sequence (e.g., a variable domain or CDR) derived from a non-human immunoglobulin (e.g., mouse immunoglobulin). Humanized antibodies may be prepared from human (recipient) immunoglobulin in which residues in the hypervariable region are substituted with hypervariable region residues derived from one or more non-human “donor” antibodies (e.g., mouse, rat, rabbit, or non-human primate). The donor antibody may be selected based on desired specificity, affinity, and / or ability. Humanized antibodies may contain one or more reverse mutations, including the return of one or more amino acids to amino acids found in the donor antibody. Conversely, residues derived from the donor antibody contained in the humanized antibody may be mutated to match residues present in the human recipient antibody. Reverse mutations may be introduced to reduce the human immune response to the humanized antibody. In some embodiments, reverse mutations are introduced to avoid problems related to antibody production (e.g., protein aggregation or post-translational modification).

[0281] To construct an expression plasmid encoding a fully humanized antibody with a human constant region, the DNA sequence encoding the antibody variable region may be inserted between the upstream promoter / enhancer and immunoglobulin signaling sequences and the downstream immunoglobulin constant region gene in an expression vector (e.g., a mammalian expression vector). The DNA sample may then be transfected into mammalian cells for antibody production. A constant domain derived from any class of human antibody can be used. Intact human antibodies have five major classes: IgA, IgD, IgE, IgG, and IgM, some of which can be further classified into subclasses (isotypes), e.g., IgG1, IgG2a, IgG2b, IgG2c, IgG3, IgG4, IgA, and IgA2.

[0282] Cell lines in which DNA encoding humanized antibodies has been stably transfected may be prepared and used to establish stable cell lines. Humanized antibodies may also be expressed by expanding cell lines that produce humanized antibodies, and these may be collected from the cell culture medium and purified.

[0283] In some embodiments, the humanized antibodies of this disclosure may exhibit cross-reactivity with non-human species. Species cross-reactivity may allow the antibody to be used in different animals for various purposes. For example, cross-reactive antibodies can be used in preclinical animal experiments to provide information on the efficacy and / or toxicity of the antibody. Non-human species may include, but are not limited to, mice, rats, rabbits, dogs, pigs, goats, sheep, and non-human primates (e.g., cynomolgus monkeys).

[0284] Antibody conjugate In some embodiments, the antibodies of this disclosure may be antibody conjugates or may be prepared as antibody conjugates. As used herein, the term “conjugate” refers to any drug, cargo, or chemical moiety attached to a receptor entity, or the process of attaching such drug, cargo, or chemical moiety. As used herein, the term “antibody conjugate” refers to any antibody having an attached drug, cargo, or chemical moiety. The conjugate used to prepare an antibody conjugate may include a therapeutic agent. Such a therapeutic agent may include a drug. An antibody conjugate containing a conjugate drug is referred herein to as an “antibody-drug conjugate.” An antibody-drug conjugate may be used to direct the conjugate drug to a target based on the affinity of the associated antibody to a protein or epitope associated with a particular target. Such an antibody-drug conjugate may be used to localize the biological activity associated with such a conjugate drug to a target cell, tissue, organ, or other target entity. In some embodiments, the conjugate used to prepare an antibody conjugate includes a detectable label. Antibodies may be conjugated with detectable labels for detection purposes. Such detectable labels may include, but are not limited to, radioisotopes, fluorophores, chromophores, chemiluminescent compounds, enzymes, enzyme cofactors, dyes, metal ions, ligands, biotin, avidin, streptavidin, haptens, quantum dots, or any other detectable labels known in the art or described herein.

[0285] The conjugate may be attached to the antibody directly or via a linker. Direct attachment may be by covalent bonding or by non-covalent association (e.g., ionic bonding, hydrostatic bonding, hydrophobic bonding, hydrogen bonding, hybridization, etc.). The linker used for conjugate attachment may contain any chemical structure that can connect the antibody to the conjugate. In some embodiments, the linker contains a polymer (e.g., nucleic acids, polypeptides, polyethylene glycol, carbohydrates, lipids, or combinations thereof). The antibody conjugate linker may be cleavable (e.g., via contact with enzymes, changes in pH, or changes in temperature).

[0286] II. Formulation and Delivery Pharmaceutical composition The compounds disclosed herein may be prepared as pharmaceutical compositions. As used herein, the term “pharmaceutical composition” means a composition comprising at least one active ingredient and, in most cases, pharmaceutically acceptable excipients.

[0287] The relative amounts of the active ingredient (e.g., antibody), pharmaceutically acceptable excipients, and / or any further ingredients in the pharmaceutical compositions according to this disclosure may vary depending on the characteristics, size, and / or condition of the object being treated, and further depending on the route through which the composition is administered. For example, a composition may contain 0.1% to 99% (w / w) of the active ingredient. For example, a composition may contain 0.1% to 100%, e.g., 0.5 to 50%, 1 to 30%, 5 to 80%, or at least 80% (w / w) of the active ingredient.

[0288] The descriptions of pharmaceutical compositions provided herein primarily concern pharmaceutical compositions suitable for administration to humans, but those skilled in the art will understand that such compositions are generally suitable for administration to any other animal, such as non-human animals, such as non-human mammals. It is well understood that pharmaceutical compositions suitable for administration to humans can be modified to be suitable for administration to various animals, and such modifications can be designed and / or carried out by a veterinary pharmacologist of ordinary skill, if any, by ordinary experimentation. The subjects to which the pharmaceutical compositions are intended to be administered include, but are not limited to, humans and / or other primates, mammals including commercially valuable mammals such as cattle, pigs, horses, sheep, cats, dogs, mice, and rats, and birds including commercially valuable birds such as poultry, chickens, ducks, geese, and / or turkeys.

[0289] In some embodiments, the composition is administered to a human, a human patient, or a subject.

[0290] formulation The compounds of the present disclosure may be formulated with one or more excipients for (1) increasing stability, (2) increasing cell permeability, (3) enabling sustained or delayed release (e.g., from sustained-release formulations), and / or (4) altering in vivo distribution (e.g., targeting antibodies to specific tissues or cell types). In addition to conventional excipients such as any solvent, dispersion medium, diluent, or other liquid vehicle, dispersion or suspension aids, surfactants, isotonic agents, thickeners or emulsifiers, and preservatives, formulations of the present disclosure may include, but are not limited to, liposomes, lipid nanoparticles, polymers, lipoplexes, core-shell nanoparticles, peptides, proteins, transfected cells (e.g., for transplantation into a subject), and combinations thereof.

[0291] The pharmaceutical compositions described herein may be prepared by methods known or to be developed in the field of pharmacology. Such preparation methods may include the step of associating the active ingredient with excipients and / or one or more other auxiliary components.

[0292] The pharmaceutical compositions described herein may be prepared, packaged, and / or sold in bulk as single unit doses and / or as multiple single unit doses. As used herein, “unit dose” refers to an individual amount of a pharmaceutical composition containing a predetermined amount of the active ingredient. The amount of the active ingredient is generally equal to the dose of the active ingredient administered to a subject, and / or a convenient portion of such a dose, for example, half or one-third of such a dose.

[0293] The relative amounts of the active ingredient (e.g., antibody), pharmaceutically acceptable excipients, and / or any further ingredients in the pharmaceutical compositions according to this disclosure may vary depending on the characteristics, size, and / or condition of the object being treated, and further depending on the route of administration. For example, a composition may contain 0.1% to 99% (w / w) of the active ingredient. For example, a composition may contain 0.1% to 100%, for example, 0.5 to 50%, 1 to 30%, 5 to 80%, or at least 80% (w / w) of the active ingredient.

[0294] According to this disclosure, compounds may be formulated for CNS delivery. Drugs that can cross the blood-brain barrier may be used. For example, several cell-permeable peptides capable of targeting the blood-brain barrier endothelium may be used in the formulation (e.g., Mathupala, Expert Opin Ther Pat., 2009, 19, 137-140; the contents of that document are incorporated herein by reference in their entirety).

[0295] Excipients and diluents In some embodiments, pharmaceutically acceptable excipients may be at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% pure. In some embodiments, the excipients are approved for human and veterinary use. In some embodiments, the excipients may be approved by the U.S. Food and Drug Administration. In some embodiments, the excipients may be pharmaceutical grade. In some embodiments, the excipients may meet the standards of the United States Pharmacopeia (USP), European Pharmacopeia (EP), British Pharmacopeia, and / or International Pharmacopoeia.

[0296] Excipients, as used herein, include, but are not limited to, any solvent, dispersion medium, diluent, or other liquid vehicle, dispersing or suspending aid, surfactant, isotonic agent, thickener or emulsifier, preservative, etc., suitable for the specific dosage form of the desired invention. A variety of excipients and preparation techniques for formulating pharmaceutical compositions are known in the art (see Remington: The Science and Practice of Pharmacy, 21st Edition, ARGennaro, Lippincott, Williams & Wilkins, Baltimore, MD, 2006, which is incorporated herein by reference in its entirety). Unless any conventional excipient medium may be incompatible with a particular substance or its derivatives, for example, by causing some undesirable biological effect or by interacting in a harmful manner with any other component(s) of the pharmaceutical composition of the Disclosure, the use of conventional excipient mediums may be intended to be within the scope of the Disclosure.

[0297] Exemplary diluents include, but are not limited to, calcium carbonate, sodium carbonate, calcium phosphate, dicalcium phosphate, calcium sulfate, calcium hydrogen phosphate, sodium lactose, sucrose, cellulose, microcrystalline cellulose, kaolin, mannitol, sorbitol, inositol, sodium chloride, dry starch, corn starch, powdered sugar, and / or combinations thereof.

[0298] inactive ingredients In some embodiments, the formulations of the Disclosure may include at least one inactive component. As used herein, the term “inactive component” refers to a drug that does not contribute to the activity of the pharmaceutical composition. In some embodiments, all or some of the inactive components that may be used in the formulations of the Disclosure may or may not be approved by the U.S. Food and Drug Administration (FDA).

[0299] The formulations disclosed herein may contain cations or anions. The formulations may contain Zn 2+ Ca 2+ Cu 2+ Mn 2+ Mg + , or combinations thereof. Non-limiting examples include polymers and complexes containing metal cations (see, for example, U.S. Patent Nos. 6,265389 and 6,555525; each of these documents is incorporated herein by reference in its entirety).

[0300] III. Administration and Medication Administration The compounds and compositions of this disclosure may be administered by any delivery route that yields a therapeutically effective outcome. Delivery routes include: intra-intestinal (into the intestines), gastrointestinal, epidural (into the dura mater), oral (through the mouth), percutaneous, intracerebral (into the cerebrum), intraventricular (into the ventricles), intracutaneous (application to the skin), intradermal (into the skin itself), subcutaneous (under the skin), nasal administration (through the nose), intravenous (into a vein), intravenous bolus, intravenous infusion, intra-arterial (into an artery), intramuscular (into the muscle), intracardiac (into the heart), intraosseous (into the bone marrow), intrasacral (into the spinal canal), intraparenchymal (into tissue, e.g., in the parenchyma of brain tissue), intraperitoneal (injection or injection into the peritoneum), intravesical infusion, and nitrate. Intrauterine (through the eye), intracavitary injection (into the pathological cavity), intracavitary (into the base of the penis), vaginal administration, intrauterine, extraamniotic administration, transdermal (diffusion through intact skin for systemic distribution), transmucosal (diffusion through mucous membrane), transvaginal, suction, sublingual, sublabial, enema, eye drops (on the conjunctiva), nasal drops, transaural (in or through the ear), buccal (directed towards the cheek), transconjunctival, cutaneous, transdental (into one or more teeth), electroosmosis, intracervical, intrasinusial, intratracheal, extracorporeal, hemodialysis, infiltration, intrastitium, intraabdominal, intraamniotic, intraarticular, intrabile , within the bronchi, within the capsule, within the cartilage (inside the cartilage), within the caudal (inside the cauda equina), within the cisterna magna (inside the cisterna magna of the cerebellum and medulla oblongata), within the cornea (inside the cornea), within the tooth crown, within the coronary arteries (inside the coronary arteries), within the corpus cavernosum (inside the expansive space of the corpus cavernosum of the penis), within the intervertebral disc (inside the intervertebral disc), within the glandular tubules (inside the glandular tubules), within the duodenum (inside the duodenum), within or beneath the dura mater, within the epidermis (into the epidermis), within the esophagus (into the esophagus), within the stomach (inside the stomach), within the gingiva (inside the gingiva), within the ileum (inside the distal part of the small intestine), within the lesion (inside the local lesion or directly introduced into the local lesion), within the lumen (inside the lumen), within the lymphatic vessels (inside the lymph), medulla Internal (in the medullary cavity of bone), intrameningeal (in the meninges), intracardiac (in the myocardium), intraocular (in the eye), intraovarian (in the ovary), intrapericardial (in the pericardial), intrapleural (in the pleura), intraprostate (in the prostate), intralungal (in the lung or its bronchi), intrasinal (in the nasal sinuses or perioboribular sinuses), intraspinal (in the spinal column), synovial (in the synovial fluid space of a joint), intratendinous (in the tendon), intratesticular (in the testis), intrasacral (in the cerebrospinal fluid at any level of the cerebrospinal axis), intrathoracic (in the thoracic cavity), intratubular (in the tubules of organs), intratumoral (in the tumor), intratympanic cavity (in the middle ear),Intravascular (in one or more blood vessels), intraventricular (inside the ventricle), ionophoresis (by electric current to move ions of soluble salts into body tissues), irrigation (to immerse or wash out open wounds or body cavities), larynx (directly into the larynx), transnasogastrostomy (through the nose into the stomach), occlusive dressing techniques (covering and sealing the site with a bandage after local administration), ocular (in the external eye), oropharyngeal (directly into the mouth and throat), parenteral, percutaneous, periarticular, supradural, perineurial, periodontal, rectal, respiratory (locally or For systemic effects, it can be administered orally or nasally by inhalation into the airways, retroocular (behind the pons or behind the eyeball), soft tissue, subarachnoid, subconjunctival, submucosa, topical, transplacental (through or across the placenta), transtracheal (through the tracheal wall), transtympanic (cross or through the tympanic cavity), ureter (into the ureter), urethra (into the urethra), vagina, sacral block, diagnostic, nerve block, biliary perfusion, cardiac perfusion, photopheresis, and spinal cord, including but not limited to these.

[0301] In some embodiments, the compositions may be administered in a manner that allows them to cross the blood-brain barrier, the vascular barrier, or other epithelial barriers. The compounds and compositions of the Disclosure may be administered in any preferred form, including but not limited to, as solutions, suspensions, or solid forms suitable for solutions or suspensions in solutions.

[0302] In some embodiments, delivery to the target may be by a single route of administration. In some embodiments, delivery to the target may be by multiple administration routes. Administration may include bolus infusion. Administration may include sustained delivery over periods of minutes, hours, or days. Administration by infusion may include infusion rates that may vary depending on the target, distribution, formulation, or other delivery parameters. Administration may be by multiple administration routes. In non-limiting examples, concomitant administration may include intra-sheath and intraventricular administration, or intravenous and intraparenchymal administration.

[0303] Intravenous administration The compounds and compositions of this disclosure may be administered to subjects by systemic administration. Systemic administration may include intravenous administration. Systemic administration may include intra-arterial administration.

[0304] The compounds and compositions of this disclosure may be administered to subjects by intravenous administration. Intravenous administration may be achieved by subcutaneous delivery. Intravenous administration may be achieved by tail vein injection (e.g., in a mouse model). Intravenous administration may be achieved by retroorbital injection.

[0305] Administration to the CNS The compounds and compositions of this disclosure may be administered to a subject by direct injection into the brain. In non-limiting examples, delivery to the brain may be by intrahippocampal administration. Administration may also be by intraparenchymal administration. In one embodiment, intraparenchymal administration is to the tissues of the central nervous system. Administration may also be by intracranial delivery (see, for example, U.S. Patent No. 8,119611; the contents of that document are incorporated herein by reference in their entirety). Administration may also be by injection into the CSF pathway. Non-limiting examples of delivery into the CSF pathway include intrasacral and intraventricular administration. Administration to the brain may also be by systemic delivery. In non-limiting examples, systemic delivery may be by intravascular administration. In non-limiting examples, systemic or intravascular delivery may be intravenous. Administration may also be by an intraocular delivery route. Non-limiting examples of intraocular delivery include intravitreous injection.

[0306] Dosage and regimen This disclosure provides methods for administering the compounds and compositions described herein to subjects in need. Administration may be carried out by any route of administration in any amount effective for the prevention, treatment, management, or diagnosis of a disease, disorder, and / or condition. The exact amount required may vary from subject to subject, depending on the species, age, general condition, severity of the disease, specific composition, mode of administration, mode of activity, etc. Subjects may be, but are not limited to, humans, mammals, or animals. Compositions may be formulated in unit dosage forms for ease of administration and uniformity of dosage. However, it should be understood that the total daily dose of the compositions described herein may be determined by the attending physician within reasonable medical judgment. The specific therapeutically effective dose level, prophylactically effective dose level, or appropriate diagnostic dose level for any particular individual may vary depending on a variety of factors, including the disorder being treated and its severity, the activity of the particular payload used, the particular composition used, the patient's age, weight, overall health, sex, and diet, the timing of administration, route of administration, and excretion rate of the compound and composition used, the duration of treatment, drugs used in combination with or concurrently with the compound and composition used, and similar factors well known in the medical field.

[0307] In certain embodiments, the compounds and compositions according to the present disclosure may be administered once or more times daily at dose levels sufficient to deliver approximately 0.0001 mg / kg to approximately 100 mg / kg, approximately 0.001 mg / kg to approximately 0.05 mg / kg, approximately 0.005 mg / kg to approximately 0.05 mg / kg, approximately 0.001 mg / kg to approximately 0.005 mg / kg, approximately 0.05 mg / kg to approximately 0.5 mg / kg, approximately 0.01 mg / kg to approximately 50 mg / kg, approximately 0.1 mg / kg to approximately 40 mg / kg, approximately 0.5 mg / kg to approximately 30 mg / kg, approximately 0.01 mg / kg to approximately 10 mg / kg, approximately 0.1 mg / kg to approximately 10 mg / kg, or approximately 1 mg / kg to approximately 25 mg / kg per day, based on the body weight of the subject, to obtain the desired therapeutic, diagnostic, or prophylactic effect.

[0308] In certain embodiments, the desired dose may be delivered using multiple doses (e.g., two, three, four, or more than four doses). When multiple doses are used, a divided dosing regimen, such as those described herein, may be used. As used herein, “divided dose” means dividing a “single unit dose” or total daily dose into two or more doses, for example, two or more “single unit doses.” As used herein, “single unit dose” means a dose of any therapeutic agent administered in one dose / in one instance / through a single route / at a single point of contact, i.e., in a single dosing event.

[0309] The compounds and compositions of this disclosure may be administered as “pulse doses” or as “continuous flow.” As used herein, “pulse dose” is a series of single doses of any therapeutic agent administered at a constant frequency over a period of time. As used herein, “continuous flow” is a dose of a therapeutic agent administered continuously over a period of time via a single route / single point of contact, i.e., in a continuous administration event. The total daily dose, which is the amount administered or prescribed over a 24-hour period, may be administered by any of these methods, a combination of these methods, or by any other method suitable for pharmacopoeias.

[0310] combination The compounds and compositions of this disclosure may be used in combination with one or more other therapeutic, prophylactic, research, or diagnostic agents. The phrase "in combination with" is not intended to imply that the agents must be administered simultaneously and / or formulated together for delivery, but these delivery methods are within the scope of this disclosure. The compositions may be administered simultaneously with, before, or after one or more other desired therapeutic or medical procedures. Generally, each agent is administered in a dose and / or time schedule determined for that agent. In some embodiments, this disclosure encompasses the delivery of pharmaceutical compositions, prophylactic compositions, research compositions, or diagnostic compositions in combination with agents capable of improving their bioavailability, reducing and / or modifying their metabolism, inhibiting their excretion, and / or modifying their distribution in the body.

[0311] IV. Method and Use of Composition In some embodiments, the present disclosure provides methods relating to the use and evaluation of compounds and compositions for therapeutic and diagnostic applications.

[0312] Therapeutic applications In some embodiments, the methods of the Disclosure include methods for treating therapeutic indications using compounds and / or compositions disclosed herein. As used herein, the term “therapeutic indication” means any symptom, condition, disorder, or disease that can be alleviated, stabilized, improved, cured, or otherwise addressed by any form of treatment or other therapeutic intervention. In some embodiments, the methods of the Disclosure include treating therapeutic indications by administering antibodies disclosed herein. In some embodiments, therapeutic indications are neurological disorders, e.g., neurodegenerative disorders, diseases associated with tau expression or activity, and / or diseases related to tau (e.g., tauopathies).

[0313] As used herein, terms such as “to treat” and “treatment” refer to the alleviation or reduction of a pathological process. In the context of this disclosure, as far as terms such as “to treat” and “treatment” are concerned with any of the other conditions listed herein below, they mean alleviating or reducing at least one symptom associated with such condition, or slowing or reversing the progression or expected progression of such condition.

[0314] In the context of disease markers or symptoms, “decrease” or “reduction” means a significant, often statistically significant, decrease at such a level. This decrease may be, for example, at least 10%, at least 20%, at least 30%, or at least 40% or more, and preferably a decrease to a level that is acceptable as within the normal range for individuals without such disorder.

[0315] In the context of disease markers or symptoms, “increase” or “rise” means a significant, often statistically significant, increase at such a level. This increase may be, for example, at least 10%, at least 20%, at least 30%, or at least 40% or more, and preferably to a level that is acceptable as within the normal range for individuals without such disorder.

[0316] The effectiveness of a treatment or improvement of a disease can be evaluated, for example, by measuring disease progression, disease remission, symptom severity, pain reduction, quality of life, the dose of drug required to maintain the effect of the treatment, the level of disease markers, or any other measurable parameter appropriate to a given disease being treated or targeted for prevention. Monitoring the effectiveness of a treatment or prevention by measuring any one of such parameters or any combination of parameters is well within the capabilities of those skilled in the art. In relation to the administration of the compounds or compositions described herein, "effective against" a disease or disorder means that administration in a clinically appropriate manner results in beneficial effects in at least some patients, such as improvement of symptoms, cure, reduction of disease burden, reduction of protein aggregation, reduction of neurofibrillary tangles, reduction of neurodegeneration, extension of lifespan, improvement of quality of life, or other effects generally recognized as positive by physicians familiar with the treatment of a particular type of disease or disorder.

[0317] The effectiveness of a treatment or preventative measure is evident when there is a significant, often statistically significant, improvement in one or more parameters of the disease, or when the symptoms do not worsen or develop as would otherwise be expected. For example, a favorable change of at least 10%, preferably at least 20%, 30%, 40%, or 50% or more, in a measurable parameter of the disease may indicate an effective treatment. The effectiveness of a given compound or composition may be determined using an experimental animal model for a given disease known in the art. When using an experimental animal model, the effectiveness of the treatment is demonstrated when a statistically significant adjustment in a marker or symptom is observed.

[0318] The compounds of this disclosure and further therapeutic agents and / or therapies can be administered in combination. Such combinations may be in the same composition, or the further therapeutic agent may be administered as part of a separate composition, or by other methods described herein. In some embodiments, the additional therapeutic agents and / or therapies are therapeutic agents and / or therapies suitable for the treatment or prevention of neurological disorders, such as neurodegenerative disorders, diseases associated with the expression or activity of tau, and / or diseases related to tau (e.g., tauopathies). In some embodiments, additional therapeutic agents and / or therapies are cholinesterase inhibitors (e.g., donepezil, rivastigmine, and / or galantamine), N-methyl-D-aspartate (NMDA) antagonists (e.g., memantine), antipsychotics, anxiolytics, anticonvulsants, dopamine agonists (e.g., pramipexole, ropinirole, rotigotine, and / or apomorphine), MAO B inhibitors (e.g., selegiline, rasagiline, and / or safinamide), catechol O-methyltransferase (COMT) inhibitors (entacapone, opicapone, and / or tolcapone), anticholinergics (e.g., benztropine and / or trihexyphenidyl), amantadine, carbidopa-levodopa, deep brain stimulation (DBS), or combinations thereof.

[0319] In some embodiments, therapeutic indications that can be addressed by the methods of the present disclosure include neurological indications. As used herein, “neurological indication” refers to any therapeutic indication relating to the central nervous system (CNS). Methods of addressing neurological indications according to the present disclosure may include administering compounds (e.g., antibodies) and / or compositions described herein. Neurological indications may include neurological disorders and / or neurological conditions involving irregular expression or aggregation of tau. Such indications may include, but are not limited to, neurodegenerative diseases, Alzheimer's disease (AD), frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17), frontotemporal lobar degeneration (FTLD), frontotemporal dementia (FTD), chronic traumatic encephalopathy (CTE), progressive supranuclear palsy (PSP), Down syndrome, Pick's disease, corticobasal degeneration (CBD), corticobasal syndromes, amyotrophic lateral sclerosis (ALS), prion diseases, Creutzfeldt-Jakob disease (CJD), multiple system atrophy, neurofibrillary senile dementia, stroke, and progressive subcortical gliosis.

[0320] In some embodiments, a method for treating neurological diseases and / or neurological disorders in a subject requiring such treatment may include one or more of the following steps: (1) deriving, generating, and / or selecting an anti-tau antibody or a fragment or composition thereof; and (2) administering the anti-tau antibody or a fragment or composition thereof to the subject. Administration to the subject may slow, halt, or reverse disease progression. In a non-limiting example, disease progression may be measured by cognitive tests such as, but not limited to, the Mini-Mental State Examination (MMSE) or other similar diagnostic tools known to those skilled in the art. In another non-limiting example, disease progression may be measured by changes such as, but not limited to, a decrease in the level of tau (either soluble or insoluble) in the pathological characteristics of the subject's brain, CSF, or other tissues. In some embodiments, the level of insoluble hyperphosphorylated tau decreases. In some embodiments, the level of soluble tau decreases. In some embodiments, both soluble and insoluble tau decrease. In some embodiments, the level of insoluble hyperphosphorylated tau increases. In some embodiments, the level of soluble tau increases. In some embodiments, levels of both insoluble and soluble tau increase. In some embodiments, neurofibrillary tangles decrease in size, number, density, or a combination thereof. In other embodiments, neurofibrillary tangles increase in size, number, density, or a combination thereof.

[0321] Neurodegeneration Neurodegenerative diseases refer to a group of conditions characterized by the progressive loss of neuronal structure and function, ultimately leading to neuronal death. Neurons are components of the nervous system and are generally unable to regenerate and / or replace; therefore, neuronal damage and / or death are particularly devastating. Other nondegenerative diseases, such as stroke, which lead to neuronal loss, also result in similarly debilitating outcomes. Targeting molecules that contribute to the deterioration of cellular structure or function may generally prove beneficial in the treatment of neurological indications, including neurodegenerative diseases and stroke.

[0322] Certain molecules are thought to have an inhibitory effect on neurite outgrowth, which contributes to the limited ability of the central nervous system to repair damage. Such molecules include, but are not limited to, myelin-related proteins, such as, but are not limited to, RGM (repulsion guidance molecules), NOGO (neurite outgrowth inhibitors), NOGO receptors, MAG (myelin-related glycoproteins), and MAI (myelin-related inhibitors). In some embodiments, the anti-tau antibodies of this disclosure can be used to target the aforementioned antigens (e.g., neurite outgrowth inhibitors).

[0323] Many neurodegenerative diseases are associated with the aggregation of misfolded proteins, including but not limited to alpha-synuclein, tau (in cases such as tauopathy), amyloid-beta, prion proteins, TDP-43, and huntingtin (see, for example, De Genst et al., 2014, Biochim Biophys Acta; 1844(11): 1907-1919, and Yu et al., 2013, Neurotherapeutics.; 10(3): 459-472, references in the same literature; all of these are incorporated herein by reference in their entirety). Aggregation arises from a disease-specific conversion of soluble proteins into insoluble, highly ordered fibrous deposits. This conversion is thought to prevent the proper disposal or degradation of misfolded proteins, thereby leading to further aggregation. Conditions associated with alpha-synuclein misfolding and aggregation are called "synuclein diseases." In some embodiments, the anti-tau antibodies of this disclosure can be used to target misfolded or aggregated proteins.

[0324] Alzheimer's disease Alzheimer's disease (AD) is a debilitating neurodegenerative disease that currently affects more than 35 million people worldwide, and that number is expected to double in the coming decades. Symptomatic treatments have been used for many years, but these treatments do not address the underlying pathophysiology. Most recent clinical trials using these and other treatments have failed, and no known cure has been identified to date.

[0325] The brain of Alzheimer's disease (AD) is characterized by the presence of two forms of pathological aggregates: extracellular plaques composed of beta-amyloid (Aβ) and intracellular neurofibrillary tangles (NFTs) composed of hyperphosphorylated microtubule-associated protein tau. Based on early genetic findings, it was thought that β-amyloid alterations initiated the disease, while tau alterations were considered downstream. Therefore, most clinical trials focused on Aβ. Although mutations in the tau gene have not been associated with AD, alterations in the tau gene have been shown to result in a group of dementias known as tauopathy, indicating that tau alterations may contribute to the neurodegenerative process. Tau is a normally highly soluble protein known to associate with microtubules based on its phosphorylation level. Hyperphosphorylation of tau inhibits its binding to microtubules and microtubule assembly activity. In tauopathy, tau is hyperphosphorylated, misfolds, and aggregates as NFTs in the form of paired helical fibrils (PHFs), twisted ribbons, or straight filaments. In Alzheimer's disease (AD), neuronal fatigue tissue (NFT) pathology correlates more closely with neuropathological markers such as neuronal loss, synaptic deficiencies, disease severity, and cognitive decline than plaque pathology. NFT pathology progresses typologically within the brain, and animal studies suggest a transcellular propagation mechanism along neuronal connections.

[0326] Several approaches have been proposed to therapeutically interfere with the progression of tau lesions and prevent subsequent molecular and cellular consequences. Given that NFTs are composed of hyperphosphorylated, misfolded, and aggregated forms of tau, interference at each of these stages presents a set of most eagerly pursued targets. The introduction of agents that restrict phosphorylation, block misfolding, or prevent aggregation has all yielded promising results. Passive and active immunization with late-stage anti-phosphotau antibodies in mouse models has led to a dramatic reduction in tau aggregation and improvement in cognitive parameters. It has also been suggested that the introduction of anti-tau antibodies can prevent transneuronal diffusion of tau lesions.

[0327] In some embodiments, the anti-tau antibodies of the Disclosure may be used in accordance with the methods presented herein to treat subjects suffering from AD and other tauopathies. In some cases, the methods of the Disclosure may be used to treat subjects suspected of developing AD or other tauopathies.

[0328] Frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17) While Alzheimer's disease is partly characterized by the presence of tau lesions, no known mutations in the tau gene have been linked to the disease as a cause. Mutations in the tau gene have been shown to lead to an autosomal dominant tauopathy known as frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17), indicating that changes in tau can lead to neurodegenerative changes in the brain. Mutations in the tau gene linked to FTDP-17 are thought to affect the splicing pattern, thereby leading to an increased proportion of tau with four microtubule-binding domains (instead of three). These molecules are thought to be more amyloidogenic, meaning they are more easily hyperphosphorylated and more likely to aggregate into NFTs (Hutton, M. et al., 1998, Nature 393(6686):702-5, the entire content of which is incorporated herein by reference). Although FTDP-17 patients may appear physically and behaviorally very similar to Alzheimer's disease patients, autopsies reveal that FTDP-17 brains do not exhibit the prominent Aβ plaque pathology seen in AD brains (Gotz, J. et al., 2012, British Journal of Pharmacology 165(5):1246-59; the entire content of this article is incorporated herein by reference). Therapeutic targeting of tau protein aggregates may improve and prevent degenerative changes in the brain, potentially leading to improved cognitive abilities.

[0329] Currently, there are no treatments to prevent, slow the progression of, or cure FTDP-17. Medications may be prescribed to reduce aggressive, agitated, or dangerous behaviors. Therapies that address the underlying pathophysiology, such as antibody therapies targeting the tau protein, remain necessary.

[0330] In some embodiments, the anti-tau antibody of the present disclosure may be used to treat subjects suffering from FTDP-17. In some cases, the method of the present disclosure may be used to treat subjects suspected of developing FTDP-17.

[0331] Chronic traumatic encephalopathy Unlike genetically linked tauopathies, chronic traumatic encephalopathy (CTE) is a degenerative tauopathy associated with repeated head injuries. The disease was first reported in a boxer exhibiting "punch-drunk" behavior and has since been primarily observed in athletes participating in American football, ice hockey, wrestling, and other contact sports. The brains of those with CTE are characterized by a distinctive pattern of brain atrophy accompanied by the accumulation of hyperphosphorylated aggregate tau species in the neurofibromatosis (NFT). In CTE, pathological changes in tau are associated with several other pathobiological processes, such as inflammation (Daneshvar, DH et al., 2015 Mol Cell Neurosci 66(Pt B):81-90; the content of this paper is incorporated herein by reference in its entirety). Targeting tau aggregates may temporarily mitigate disease progression and potentially lead to improved cognitive function.

[0332] Currently, there is no medical treatment to treat or cure CTE. This condition is only diagnosed post-mortem because there are no in vivo techniques to identify CTE-specific biomarkers. Therapies that address the underlying pathophysiology, such as antibody therapies targeting tau protein, remain necessary.

[0333] In some embodiments, the anti-tau antibodies of the present disclosure may be used to treat subjects suffering from CTE. In some cases, the methods of the present disclosure may be used to treat subjects suspected of developing CTE.

[0334] Prion disease Prion diseases, also known as transmissible spongiform encephalopathy (TSE), are a group of rare, progressive conditions affecting the nervous system. The associated conditions are rare and are typically caused by mutations in the PRNP gene, which enables the production of prion proteins. These gene mutations result in prion proteins with abnormal structures. Alternatively, abnormal prions may be acquired through exposure to external sources, such as the consumption of beef products containing abnormal prion proteins. Abnormal prions misfold and rapidly degenerate brain tissue. Prion diseases include, but are not limited to, Creutzfeldt-Jakob disease (CJD), Gerstmann-Streussler-Scheinker syndrome (GSS), fatal insomnia (FFI), variable protease-sensitive prionopathy (VPSPr), and kuru. Prion diseases are rare; approximately 350 cases are diagnosed annually in the United States.

[0335] CJD is a degenerative brain disorder characterized by problems with muscle coordination, personality changes including mental dysfunction, visual impairment, involuntary muscle spasms, weakness, and ultimately, coma. The most common categories of CJD are sporadic, hereditary (due to genetic mutation), and acquired. Sporadic CJD is the most common form affecting people who do not have known risk factors for the disease. Acquired CJD is transmitted when brain and nervous system tissues are exposed to prions. For example, variant CJD (vCDJ) is associated with bovine spongiform encephalopathy (BSE), also known as "mad cow disease." CJD is fatal, and patients usually die within one year of diagnosis.

[0336] Prion diseases are associated with infectious agents consisting of PrPSc, an alternative structural isoform of the prion protein. PrPSc replication is thought to occur through the induction of infectious prions in the normal prion protein (PrPC). Replication occurs without nucleic acid.

[0337] Currently, there are no therapies to manage or cure CJD or other prion diseases. Treatment typically aims to alleviate symptoms and improve patient comfort, for example, by using analgesics. Therapies that address the underlying pathophysiology are still needed.

[0338] In some embodiments, the anti-tau antibodies of the present disclosure may be used to treat subjects suffering from prion diseases. In some cases, the methods of the present disclosure may be used to treat subjects suspected of developing prion diseases.

[0339] Diagnostic application In some embodiments, the compounds (e.g., antibodies) and compositions of this disclosure may be used as diagnostic agents. Anti-tau antibodies may be used to identify, label, or stain cells, tissues, organs, etc., that express tau protein. Anti-tau antibodies may also be used to identify tau protein present in tissue sections (e.g., histological tissue sections) containing tissue known or suspected to have tau protein aggregates. Such antibodies may, in some cases, be used to identify subjects with neurological diseases and / or neuropathy. Tissue sections may be derived from CNS tissue.

[0340] In some embodiments, the diagnostic methods of this disclosure may include the analysis of one or more cells or tissues using immunohistochemical techniques. Such methods may include the use of one or more anti-tau antibodies described herein. The immunohistochemical method may include staining tissue sections to determine the presence and / or levels of one or more tau proteins or other markers. Tissue sections may be derived from the CNS tissue of interest (e.g., the CNS of a patient, the CNS of an animal, and the CNS of an animal model of the disease). Tissue sections may be derived from formalin-fixed tissue or unfixed fresh-frozen tissue. In some cases, tissue sections may be derived from formalin-fixed paraffin-embedded (FFPE) tissue. The anti-tau antibodies described herein may be used as primary antibodies. The primary antibody is used to contact the tissue section directly and to bind to a target epitope. The primary antibody may be directly conjugated to a detectable label or may be detected by the use of a detection agent such as a secondary antibody. In some embodiments, the primary antibody or detection agent includes an enzyme that can be used to react with a substrate to produce a visible product (e.g., a precipitate). Such enzymes may include, but are not limited to, horseradish peroxidase, alkaline phosphatase, beta-galactosidase, and catalase.

[0341] The anti-tau antibodies described herein may be used in accordance with the immunohistochemical methods of this disclosure to detect tau protein in tissues or cells. In some cases, these antibodies are used to detect and / or determine the level of tau protein in tissues. The level of anti-tau antibody used in immunohistochemical staining techniques may be varied to increase visible staining or to decrease the background level of staining. In some embodiments, antibody concentrations of about 0.01 μg / ml to about 50 μg / ml are used. For example, antibody concentrations of about 0.01 μg / ml to about 1 μg / ml, about 0.05 μg / ml to about 5 μg / ml, about 0.1 μg / ml to about 3 μg / ml, about 1 μg / ml to about 10 μg / ml, about 2 μg / ml to about 20 μg / ml, about 3 μg / ml to about 25 μg / ml, about 4 μg / ml to about 30 μg / ml, or about 5 μg / ml to about 50 μg / ml may be used.

[0342] The level and / or identity of tau protein can be determined according to any method known in the Art for protein identification and / or quantification of protein levels. In some embodiments, such methods may include, but are not limited to, mass spectrometry, array analysis (e.g., antibody arrays or protein arrays), Western blotting, flow cytometry, immunoprecipitation, surface plasmon resonance analysis, and ELISA. Tau protein may optionally be immunoprecipitated from the sample before analysis. Such immunoprecipitation may be performed using anti-tau antibodies disclosed herein. In some embodiments, tau protein is immunoprecipitated from a biological sample using an anti-tau antibody and then identified and / or quantified using mass spectrometry.

[0343] In some embodiments, a method for detecting tau (e.g., human tau) using the antibodies described herein includes (a) contacting a sample (e.g., a biological sample such as a tissue section) with the anti-tau antibody described herein for a time sufficient to allow the anti-tau antibody to specifically bind to tau in the sample, and (b) contacting the sample with a detection reagent, e.g., an anti-tau antibody, e.g., an antibody that specifically binds to the Fc region of the anti-tau antibody, thereby detecting the tau to which the anti-tau antibody binds.

[0344] The Specified Anti-Tau Antibody also provides a method for detecting the presence of tau (e.g., human tau) in a sample, or a method for measuring the amount of tau, which includes contacting the sample (e.g., a biological sample such as a tissue sample) with the anti-tau antibody described herein under conditions that allow for the formation of a complex between the antibody and tau, and detecting the formation of the complex. In some embodiments, the method may also include contacting the sample with a control antibody (e.g., an isotype control antibody) in parallel, and the difference in complex formation between the anti-tau antibody and the sample, and between the control antibody and the sample, indicates the presence of tau in the sample. In some embodiments, the anti-tau antibody described herein can be used to purify tau (e.g., various phosphorylated tau species recognized by the anti-tau antibody described herein) via immunoaffinity purification.

[0345] In some embodiments, treatment information is obtained from diagnostic information generated using an anti-tau antibody. Accordingly, this disclosure provides a method for treating neurological diseases and / or neuropathy, comprising obtaining a sample from a subject, diagnosing one or more neurological diseases and / or neuropathy using an anti-tau antibody, and administering a treatment selected based on the diagnosis. Such treatment may include treatment with an anti-tau antibody. The anti-tau antibody administered according to such a method may include any of those described herein.

[0346] In some embodiments, this disclosure provides methods for detecting and / or quantifying tau protein in a sample through the use of capture antibodies and detection antibodies. As used herein, “capture antibody” is an antibody that binds to an analyte so that the analyte can be isolated or detected. The capture antibody may be associated with a surface or other carrier (e.g., beads). The detection antibody is an antibody that facilitates the observation of the presence or absence of an analyte. According to some methods for detecting and / or quantifying tau protein, both the capture antibody and the detection antibody bind to the tau protein. The capture antibody and the detection antibody may bind to different epitopes or regions of the tau protein to avoid binding competition. In some embodiments, the detection antibody may be conjugated with a detectable label for direct detection. In some embodiments, the binding of the detection antibody may be evaluated using a secondary antibody that binds to the constant domain of the detection antibody or to a detectable label of the detection antibody. The capture antibody, detection antibody, and / or secondary antibody may originate from different species. This can prevent the secondary antibody from binding to both the capture antibody and the detection antibody.

[0347] V. Kits and Devices kit In some embodiments, the compounds and compositions of this disclosure may be included in a kit. Such compounds and compositions may include anti-tau antibodies disclosed herein. In non-limiting examples, a kit may include reagents for generating anti-tau antibodies, comprising a tau protein antigen. A kit may include further reagents and / or instructions for use, for example, for producing or synthesizing anti-tau antibodies. A kit may include one or more buffers. A kit may include further components, such as solid supports or substrates for attaching antibodies or antigens.

[0348] In some embodiments, the disclosure includes a kit for screening, monitoring, and / or diagnosing a target, comprising one or more anti-tau antibodies. Such a kit may be used alone or in combination with one or more other screening, monitoring, and / or diagnostic methods. The kit may comprise one or more of the following: buffers, biological standards, secondary antibodies, detection reagents, and compositions for sample preparation (e.g., for antigen recovery, blocking, etc.).

[0349] The kit components may be packaged. In some embodiments, the kit components are packaged in an aqueous medium or in a lyophilized form. The packaging may include one or more vials, test tubes, flasks, bottles, syringes, or other containers that can hold and / or suitably dispense the components. If there are multiple kit components (labeled reagents and labels may be packaged together), the kit may include a second, third, or other additional container that can hold the additional components separately.

[0350] If the kit components are provided in one and / or more solutions, the solutions may be aqueous solutions. The solutions may be prepared sterile. The kit components may be provided as dry powders. The dry powder components may be prepared so that they can be reconstituted by the kit user, for example, by adding a suitable solvent. The solvent may be provided in the kit in one or more separate containers. In some embodiments, the labeling dye is provided in dry powder format.

[0351] The kit may include instructions for using the kit components as well as other reagents not included in the kit. The instructions may include feasible variations.

[0352] device Any of the compounds and compositions described herein can be combined with a device, coated onto a device, embedded in a device, or delivered by a device. The device may include, but is not limited to, implants, stents, bone replacements, artificial joints, valves, pacemakers, or other implantable therapeutic devices.

[0353] VI.Definitions Throughout this specification, substituents of the compounds of the Disclosure are disclosed in multiple groups or ranges. This disclosure is specifically intended to include all individual partial combinations of members of such groups and ranges.

[0354] Approximately: As used herein, the term "approximately" means ±10% of the stated value.

[0355] Activity: As used herein, the term “activity” refers to a state in which something is happening or being done. A composition may have activity, and this activity may involve one or more biological events.

[0356] Combined administration: As used herein, the terms “combined administration” or “combined administration” mean that two or more drugs are administered to a target simultaneously or at intervals in which there may be overlap in the effects of each drug on the patient. In some embodiments, these are administered within approximately 60, 30, 15, 10, 5, or 1 minute of each other. In some embodiments, the administration of drugs is carried out at intervals close enough to achieve a combinatorial effect (e.g., a synergistic effect).

[0357] Improvement: As used herein, the terms “improvement” or “to improve” refer to a reduction in the severity of at least one indicator of a condition or disease. For example, in the context of neurodegenerative disorders, improvement includes a reduction in neuronal loss.

[0358] Animals: As used herein, the term “animal” refers to any member of the animal kingdom. In some embodiments, “animal” refers to a human at any developmental stage. In some embodiments, “animal” refers to a non-human animal at any developmental stage. In certain embodiments, a non-human animal is a mammal (e.g., rodents, mice, rats, rabbits, monkeys, dogs, cats, sheep, cattle, primates, or pigs). In some embodiments, an animal includes, but is not limited to, mammals, birds, reptiles, amphibians, fish, and insects. In some embodiments, an animal is a transgenic animal, a genetically engineered animal, or a clone.

[0359] Approximately: As used herein, the terms “approximately” or “about” applied to one or more values ​​for purposes refer to a value similar to the given reference value. In certain embodiments, the terms “approximately” or “about” refer to a range of values ​​that fall within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less in any direction (above or below) the given reference value, except where otherwise stated or where a different meaning is evident from the context (unless such a number exceeds 100% of the possible value).

[0360] Association: As used herein, the terms “association,” “conjugate,” “linking,” “attached,” and “moored” mean that multiple entities physically associate or connect with one another, either directly or via a linker, to form a structure that is sufficiently stable to maintain a physically associated state, for example, under working conditions, or for example, under physiological conditions. “Association” does not have to be by covalent chemical bonding and may include other forms of association or bonding that are sufficiently stable to maintain a physically associated state of the “associated” entities, such as ionic bonding, hydrostatic bonding, hydrophobic bonding, hydrogen bonding, or bonding based on hybridization.

[0361] Difunctionality: As used herein, the term “difunctionality” refers to any substance, molecule, or part that is capable of or maintains at least two functions. These functions may result in the same or different outcomes. The structures that produce the functions may be the same or different.

[0362] Biocompatibility: As used herein, the term “biocompatible” means that a product is compatible with living cells, tissues, organs, or systems and presents little to no risk of immune system injury, toxicity, or rejection.

[0363] Biodegradability: As used herein, the term "biodegradable" means that a material can be broken down into harmless products by biological action.

[0364] Biologically active: As used herein, the term “biologically active” refers to any substance that is active in a biological system and / or in an organism. For example, a substance that, when administered to an organism, has a biological effect on that organism is considered biologically active.

[0365] Chimeric Antigen Receptor (CAR): As used herein, the terms “chimeric antigen receptor” or “CAR” refer to an artificial chimeric protein comprising at least one antigen-specific targeting region (ASTR), a transmembrane domain, and an intracellular signaling domain, wherein the antigen-specific targeting region comprises a full-length antibody or a fragment thereof. As a non-limiting example, the ASTR of a CAR may be any of the antibodies or fragments thereof presented herein. Any molecule capable of binding to a target antigen with high affinity can be used as the ASTR of a CAR. CARs may optionally have an extracellular spacer domain and / or a costimulatory domain. CARs can also be used to generate cytotoxic cells that carry the CAR.

[0366] Compounds: The compounds of this disclosure include all isotopes of atoms that result in intermediates or final compounds. "Isotopes" refer to atoms that have the same atomic number but different mass numbers due to a difference in the number of neutrons in their nuclei. For example, tritium and deuterium are isotopes of hydrogen.

[0367] The compounds and salts of this disclosure can be prepared by conventional methods in combination with solvents or water molecules to form solvates and hydrates.

[0368] Comprehensive Positional Evolution (CPE®): As used herein, the term “comprehensive positional evolution” refers to an antibody evolution technique that enables mapping the effects of amino acid changes at any position along the sequence of an antibody variable domain. This comprehensive mutagenesis technique can be used to enhance one or more properties or characteristics of an antibody.

[0369] Comprehensive Protein Synthesis (CPS®): As used herein, the term “comprehensive protein synthesis” refers to combinatorial protein synthesis techniques that can be used to optimize the properties or characteristics of an antibody by combining the best properties to create a new, high-performance antibody.

[0370] Conditionally active: As used herein, the term “conditionally active” refers to a variant or offshoot of a wild-type polypeptide that is more or less active than the parent polypeptide under physiological conditions. Furthermore, conditionally active polypeptides may have increased or decreased activity under abnormal conditions compared to the parent polypeptide. Conditionally active polypeptides may be reversibly or irreversibly inactivated under normal or abnormal physiological conditions.

[0371] Conserved: As used herein, “conserved” means a nucleotide or amino acid residue in a polynucleotide or polypeptide sequence that occurs unchanged at the same position in two or more sequences being compared. Relatively conserved nucleotides or amino acids are those that are conserved between sequences that are more relevant than they appear elsewhere in the sequence.

[0372] In some embodiments, two or more sequences are said to be "fully conserved" if they are 100% identical to each other. In some embodiments, two or more sequences are said to be "highly conserved" if they are at least 70% identical, at least 80% identical, at least 90% identical, or at least 95% identical to each other. In some embodiments, two or more sequences are said to be "highly conserved" if they are approximately 70% identical, at about 80% identical, at about 90% identical, at about 95% identical, at about 98% identical, or at about 99% identical to each other. In some embodiments, two or more sequences are said to be "conserved" if they are at least 30% identical, at least 40% identical, at least 50% identical, at least 60% identical, at least 70% identical, at least 80% identical, at least 90% identical, or at least 95% identical to each other. In some embodiments, two or more sequences are said to be “conserved” if they are approximately 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, or 99% identical to each other. Sequence conservation may apply to the entire length of a polynucleotide or polypeptide, or to a portion, region, or characteristic thereof.

[0373] Cytotoxicity: As used herein, “cytotoxicity” means killing or causing a damaging, toxic, or lethal effect on cells (e.g., mammalian cells (e.g., human cells)), bacteria, viruses, fungi, protozoa, parasites, prions, or combinations thereof.

[0374] Delivery: As used herein, “delivery” means the act or manner of delivering a compound, substance, entity, part, cargo, or payload to a target or destination.

[0375] Detectable Labels: As used herein, “detectable labels” means one or more markers, signals, or parts that attach to, incorporate into, or associate with another entity and are readily detectable by methods known in the art, including radiography, fluorescence, chemiluminescence, enzyme activity, absorbance, etc. Detectable labels include, but are not limited to, radioisotopes, fluorophores, chemiluminescent compounds, chromophores, enzymes, enzyme cofactors, dyes, metal ions, ligands, biotin, avidin, streptavidin, haptens, quantum dots, etc. Detectable labels may be located at any position in or on the entity to which they are conjugated, or otherwise attached, incorporated into, or associated. For example, when conjugated to, or otherwise attached to, incorporated into, or associated with a peptide or protein, detectable labels may be on, in, or between amino acids, or attached to or associated with the N-terminus or C-terminus.

[0376] Digestion: As used herein, the term “digestion” means breaking down into smaller fragments or components. When referring to polypeptides or proteins, digestion results in the production of peptides.

[0377] Distal: As used herein, the term “distal” means a location that is away from the center or away from the point or area of ​​interest.

[0378] Medication regimen: As used herein, “medication regimen” refers to a schedule of administration or a regimen of treatment, prevention, or palliative care determined by a physician.

[0379] Engineering: As used herein, an embodiment is "engineered" if its properties or characteristics are designed to differ, structurally or chemically, from a starting compound, starting material, or starting molecule (e.g., wild-type or natural molecule).

[0380] Effective dose: As used herein, the term “effective dose” of a drug means an amount sufficient to produce a beneficial or desired outcome, such as a clinical outcome, and therefore “effective dose” depends on the context in which it is applied. For example, in the context of administering a drug to treat cancer, the effective dose of the drug is, for example, an amount sufficient to achieve treatment of the therapeutic indication compared to the response obtained without administration of the drug.

[0381] Epitope: As used herein, “epitope” refers to a surface or region in one or more entities that can interact with an antibody or other bound biomolecule. For example, a protein epitope may comprise one or more amino acids and / or post-translational modifications (e.g., phosphorylated residues) that interact with an antibody. In some embodiments, an epitope may also be a “structural epitope,” which refers to an epitope having a specific three-dimensional arrangement of entities that have or form an epitope. For example, a structural epitope of a protein may comprise a combination of amino acids and / or post-translational modifications from a folded nonlinear segment of an amino acid chain.

[0382] EvoMap®: As used herein, EvoMap® refers to a polypeptide map that presents detailed informatics regarding the effects of single amino acid mutations throughout the entire length of a polypeptide and their impact on the properties and characteristics of the polypeptide.

[0383] Expression: As used herein, “expression” of a gene, nucleic acid, or protein means one or more of the following events: (1) production of an RNA template from a DNA sequence (e.g., by transcription); (2) processing of an RNA transcript (e.g., by splicing, editing, 5' cap formation, and / or 3' end processing); (3) translation of RNA into a polypeptide or protein; and (4) post-translational modification of a polypeptide or protein.

[0384] Characteristics: As used herein, “characteristics” refers to features, properties, or distinctive elements.

[0385] Formulation: As used herein, “formulation” means a substance or mixture prepared according to a manufacturing process. A formulation may include a compound (e.g., an antibody) or substance in combination with a carrier or excipient.

[0386] Fragment: As used herein, “fragment” refers to a portion of a protein. For example, a protein fragment may include a polypeptide obtained by digesting a full-length protein isolated from cultured cells.

[0387] Functional: As used herein, a “functional” biological molecule is a biological molecule in a form that exhibits characteristic properties and / or activity. For example, a “functional” antibody may include an antibody that binds to a specific target or an antibody that activates or inhibits a specific biological process.

[0388] Maximum Half-Volume Effective Concentration: As used herein, the term “maximum half-volume effective concentration” or “EC50” refers to the concentration of a substance required to increase a given reaction, activity, or process by half. For example, when measuring the binding of an antibody to a target in a sample using a binding assay (e.g., an ELISA assay), EC50 is the concentration of antibody in the sample required to produce 50% of the maximum binding that can be observed in that assay. Similarly, the term “maximum half-volume inhibitory concentration” or “IC50” refers to the concentration required to reduce a given reaction or process by half. For example, for an antibody that can inhibit a biological process, IC50 is the concentration of antibody in the sample required to reduce the biological process by 50%. The values ​​of EC50 and IC50 may vary depending on specific time constraints and / or conditions.

[0389] Homology: As used herein, the term “homology” refers to the overall relationship between macromolecules, such as polynucleotide molecules (e.g., DNA molecules and / or RNA molecules) and / or polypeptide molecules. In some embodiments, macromolecules are considered “homology” to one another if their sequences are identical or similar by at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99%. The term “homology” necessarily refers to a comparison between at least two sequences (polynucleotide sequences or polypeptide sequences). According to this disclosure, two polynucleotide sequences are considered homologous if the polypeptides they encode are at least about 50%, 60%, 70%, 80%, 90%, 95%, or even 99% of at least one interval consisting of at least about 20 amino acids. In some embodiments, homologous polynucleotide sequences are characterized by their ability to encode at least four to five uniquely designated amino acid intervals. For polynucleotide sequences less than 60 nucleotides in length, homology is determined by the ability to encode at least four to five uniquely designated amino acid intervals. According to this disclosure, two protein sequences are considered homologous if these proteins are at least about 50%, 60%, 70%, 80%, or 90% identical in at least one interval consisting of at least about 20 amino acids.

[0390] Heterogeneous domain: As used herein, the term “heterogeneous domain” refers to a domain that is not considered homologous.

[0391] Homologous Region: As used herein, the term “homologous region” refers to a region that is similar in location, structure, evolutionary origin, characteristics, morphology, or function.

[0392] Identity: As used herein, the term “identity” refers to the overall relationship between macromolecules, e.g., between polynucleotide molecules (e.g., DNA molecules and / or RNA molecules) and / or between polypeptide molecules. For example, the percentage of identity of two polynucleotide sequences can be calculated by aligning the two sequences for the purpose of optimal comparison (for example, gaps may be introduced in one or both of the first and second nucleic acid sequences for optimal alignment, and non-identical sequences may be ignored for comparison purposes). In certain embodiments, the length of the sequences aligned for comparison purposes is at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or 100% of the length of the reference sequence. Then, the nucleotides at the corresponding nucleotide positions are compared. If a position in the first sequence is occupied by the same nucleotide as the corresponding position in the second sequence, then these molecules are identical at that position. The percentage of identity between two arrays is a function of the number of identical positions common to the arrays, taking into account the number of gaps that need to be introduced for optimal alignment of the two arrays and the length of each gap. The comparison of arrays and the determination of the percentage of identity between two arrays can be achieved using mathematical algorithms.For example, the percentage of identity between two nucleotide sequences can be determined using methods such as those described in Computational Molecular Biology, Lesk, AM, ed., Oxford University Press, New York, 1988; Biocomputing: Informatics and Genome Projects, Smith, DW, ed., Academic Press, New York, 1993; Sequence Analysis in Molecular Biology, von Heinje, G., Academic Press, 1987; Computer Analysis of Sequence Data, Part I, Griffin, AM, and Griffin, HG, eds., Humana Press, New Jersey, 1994; and Sequence Analysis Primer, Gribskov, M. and Devereux, J., eds., M Stockton Press, New York, 1991, each of which is incorporated herein by reference. For example, the percentage of identity between two nucleotide sequences can be determined using the Meyers and Miller (CABIOS, 1989, 4:11-17) algorithm incorporated into the ALIGN program (version 2.0), using the PAM120 weight residue table, gap length penalty 12, and gap penalty 4. Alternatively, the percentage of identity between two nucleotide sequences may be determined using the GAP program in the GCG software package, using the NWSgapdna.CMP matrix. Commonly used methods for determining the percentage of identity between sequences include, but are not limited to, those disclosed in Carillo, H. and Lipman, D., SIAM J Applied Math., 48:1073 (1988), which are incorporated herein by reference. Techniques for determining identity are systematized in publicly available computer programs.Exemplary computer software for determining homology between two sequences includes, but is not limited to, the GCG program package (Devereux, J., et al., Nucleic Acids Research, 12(1), 387 (1984)), BLASTP, BLASTN, and FASTA (Altschul, SF et al., J. Molec. Biol., 215, 403 (1990)).

[0393] in vitro: As used herein, the term “in vitro” refers to events occurring in an artificial environment, such as a test tube or reaction vessel, cell culture, or petri dish, rather than in living organisms (e.g., animals, plants, or microorganisms).

[0394] in vivo: As used herein, the term “in vivo” refers to an event occurring in a living organism (for example, an animal, plant, or microorganism, or their cells or tissues).

[0395] Isolation: As used herein, the term “isolated” means a substance or entity separated from at least a portion of the associated components (whether in a natural or experimental environment). Isolated substances may have varying levels of purity compared to the substances from which they are associated. Isolated substances and / or entities may be separated from at least about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, or more of the other components that were originally associated with them. In some embodiments, isolated agents have a purity of about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or more than about 99%. As used herein, a substance is “pure” if it is substantially free of other components.

[0396] Substantially isolated: “Substantially isolated” means that a substance is substantially separated from the environment in which it was formed or detected. Substantially isolated can include compositions containing at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 97%, or at least about 99% by weight of the compound or salt thereof of the present disclosure. Methods for isolating compounds and salts thereof are commonplace in the art.

[0397] Linker: As used herein, “linker” refers to a molecule or group of molecules that connects two molecules. In some embodiments, linkers may be cleavable (e.g., by contact with an enzyme, a change in pH, or a change in temperature).

[0398] Modification: As used herein, “modification” refers to a change in the state or structure of a molecule. Molecules can be modified in many ways, including chemically, structurally, and functionally.

[0399] Natural origin: As used herein, “natural origin” or “wild type” means that it is present in nature without artificial assistance or human intervention.

[0400] Non-human vertebrates: As used herein, “non-human vertebrates” includes all vertebrates except Homo sapiens, including wild and domesticated species. Examples of non-human vertebrates include, but are not limited to, mammals such as alpacas, bantengs, bison, camels, cattle, cattle, deer, dogs, donkeys, gayars, goats, guinea pigs, horses, llamas, mules, pigs, primates, rabbits, reindeer, sheep, buffalo, and yaks.

[0401] Off-target: As used herein, “off-target” refers to unintended activity or binding to entities other than the expected target.

[0402] Operable linkage: As used herein, the term “operable linkage” refers to a functional connection between two or more molecules, constructs, transcripts, entities, parts, etc.

[0403] Patient: As used herein, “patient” means a person who may seek or require treatment for a particular disease or condition, a person who requires treatment, a person who is receiving treatment, a person who will receive treatment, or a person who is receiving care from a trained professional.

[0404] Peptide: As used herein, “peptide” is shorter than or equal to 50 amino acids in length, for example, about 5, 10, 15, 20, 25, 30, 35, 40, 45, or 50 amino acids in length.

[0405] Pharmacologically acceptable: The term "pharmaceutically acceptable" is used herein to mean a compound, substance, composition, and / or dosage form that, within reasonable medical judgment, is suitable for use in contact with human and animal tissues without causing excessive toxicity, irritation, allergic reactions, or other problems or complications, and that provides a reasonable benefit-to-risk ratio.

[0406] Pharmacovigilant excipients or pharmaceutically acceptable carriers: As used herein, the terms “pharmaceutically acceptable excipient” or “pharmaceutically acceptable carrier” refer to any component other than the compounds described herein (e.g., a vehicle capable of suspending or dissolving the active compound) that is substantially non-toxic and non-inflammatory in the patient. Examples of excipients include antifouling agents, antioxidants, binders, coatings, compression aids, disintegrants, dyes (colorants), emollients, emulsifiers, fillers (diluents), film-forming agents or coatings, flavoring agents, fragrances, flow enhancers, lubricants, preservatives, printing inks, adsorbents, suspending agents or dispersants, sweeteners, and hydration water. Exemplary excipients include, but are not limited to, butylated hydroxytoluene (BHT), calcium carbonate, dibasic calcium phosphate, calcium stearate, croscarmellose, cross-linked polyvinylpyrrolidone, citric acid, crospovidone, cysteine, ethylcellulose, gelatin, hydroxypropylcellulose, hydroxypropylmethylcellulose, lactose, magnesium stearate, maltitol, mannitol, methionine, methylcellulose, methylparaben, microcrystalline cellulose, polyethylene glycol, polyvinylpyrrolidone, povidone, pregelatinized starch, propylparaben, retinyl palmitate, shellac, silicon dioxide, sodium carboxymethylcellulose, sodium citrate, sodium starch glycolate, sorbitol, starch (corn), stearic acid, sucrose, talc, titanium dioxide, vitamin A, vitamin E, vitamin C, and xylitol.

[0407] pharmaceutically acceptable salts: This disclosure also includes pharmaceutically acceptable salts of the compounds described herein. As used herein, “pharmaceutically acceptable salt” means a derivative of a disclosed compound in which the parent compound has been modified by converting an existing acidic or base moiety to its salt form (for example, by reacting a free base with a suitable organic acid). Examples of pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of basic residues such as amines, and alkali or organic salts of acidic residues such as carboxylic acids. Typical acid addition salts include acetate, acetic acid, adipine, alginate, ascorbate, aspartate, benzenesulfonate, benzenesulfonic acid, benzoate, bisulfate, borate, butyrate, camphor salt, camphor sulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, fumarate, glucoheptonate, glycerophosphate, hemisulfate, heptoneate, hexanoate, hydrobromide, hydrochloride, hydroiodide, and 2-hydroxy-ethanol. Examples include sulfonates, lactobionates, lactates, laurates, lauryl sulfates, malates, maleates, malons, methanesulfons, 2-naphthalenesulfons, nicotinates, nitrates, oleates, oxalates, palmitates, pamoates, pectates, persulfates, 3-phenylpropionates, phosphates, picrates, pivalates, propions, stearates, succinates, sulfates, tartrates, thiocyans, toluenesulfons, undecanoates, and valersates. Typical alkali salts or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and non-toxic ammonium, quaternary ammonium, and amine cations, including but not limited to ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, and ethylamine. The pharmaceutically acceptable salts of this disclosure include, for example, conventional non-toxic salts of parent compounds formed from non-toxic inorganic or organic acids.The pharmaceutically acceptable salts of the present disclosure can be synthesized from parent compounds containing a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acidic or basic form of these compounds with a stoichiometric amount of a suitable base or acid in water, an organic solvent, or a mixture of the two, generally in non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile. A list of suitable salts is found in Remington's Pharmaceutical Sciences, 17. th This information is found in *Mack Publishing Company, Easton, Pa., 1985, p. 1418*, *Pharmaceutical Salts: Properties, Selection, and Use*, PHStahl and CGWermuth (eds.), Wiley-VCH, 2008, and *Berge et al., Journal of Pharmaceutical Science, 66, 1-19 (1977)*, each of which is incorporated herein by reference in its entirety.

[0408] Pharmaceutically acceptable solvates: As used herein, the term “chemically acceptable solvate” means a compound in which molecules of a suitable solvent are incorporated into the crystal lattice. The suitable solvent is physiologically tolerable at the administered dose. For example, solvates can be prepared by crystallization, recrystallization, or precipitation from solutions containing organic solvents, water, or mixtures thereof. Examples of suitable solvents include ethanol, water (e.g., monohydrate, dihydrate, and trihydrate), N-methylpyrrolidinone (NMP), dimethyl sulfoxide (DMSO), N,N'-dimethylformamide (DMF), N,N'-dimethylacetamide (DMAC), 1,3-dimethyl-2-imidazolidinone (DMEU), 1,3-dimethyl-3,4,5,6-tetrahydro-2-(1H)-pyrimidinone (DMPU), acetonitrile (ACN), propylene glycol, ethyl acetate, benzyl alcohol, 2-pyrrolidone, and benzyl benzoate. When water is the solvent, the solvate is called a "hydrate."

[0409] Pharmacokinetics: As used herein, “pharmacokinetics” refers to one or more properties of a molecule or compound relating to the determination of the fate of a substance administered to a living organism. Pharmacokinetics is divided into several areas, including the degree and rate of absorption, distribution, metabolism, and excretion. This is commonly referred to as ADME, and refers to (A) absorption, which is the process by which a substance enters the bloodstream; (D) distribution, which is the dispersion or diffusion of a substance throughout the body’s fluids and tissues; (M) metabolism (or biotransformation), which is the irreversible transformation of a parent compound into daughter metabolites; and (E) excretion (or elimination), which is the removal of a substance from the body. Rarely, some drugs accumulate irreversibly in body tissues.

[0410] Prevention: As used herein, the term “prevention” means partially or completely delaying the onset of an infection, disease, disorder, and / or condition; partially or completely delaying the onset of one or more symptoms, characteristics, or clinical signs of a particular infection, disease, disorder, and / or condition; partially or completely delaying the onset of one or more symptoms, characteristics, or signs of a particular infection, disease, disorder, and / or condition; partially or completely delaying the progression from an infection, a particular disease, disorder, and / or condition; and / or reducing the risk of developing a condition associated with an infection, disease, disorder, and / or condition.

[0411] Proliferation: As used herein, the term “proliferation” means growing, expanding, or increasing, or causing to grow, expand, or rapidly increase. “Proliferative” means having the ability to proliferate. “Antiproliferative” means having properties opposite to or unsuitable for the proliferative nature.

[0412] Preventive: As used herein, “preventive” refers to a therapeutic agent or process of action used to prevent the spread of disease.

[0413] Prevention: As used herein, “prevention” refers to measures taken to maintain health and prevent the spread of disease.

[0414] Target Protein: As used herein, the terms “target protein” or “desired protein” include those provided herein, as well as their fragments, variants, variants, and altered forms.

[0415] Purified: As used herein, “purify,” “refined,” and “refined” mean to make substantially pure, or to remove unwanted components, contaminants, impurities, or contaminants. “Purified” refers to the state of being pure. “Refined” refers to the process of making pure.

[0416] Region: As used herein, the term “region” refers to a zone or general area. In some embodiments, where referring to a polypeptide or protein, a region may include a linear sequence of amino acids along the polypeptide or protein, or a three-dimensional area, an epitope, or a cluster of epitopes. Where referring to a polynucleotide, a region may include a linear sequence of nucleic acids along the polynucleotide, or a three-dimensional area, a secondary structure, or a tertiary structure. A region may include a terminal region. As used herein, the term “terminal region” refers to a region located at the end or “terminus” of a given entity. Where referring to a polypeptide, a terminal region may include the N-terminus and / or C-terminus. The N-terminus refers to the end of a polypeptide having a free amino acid amino group. The C-terminus refers to the end of a polypeptide having a free amino acid carboxyl group. The N-terminal region and / or C-terminal region may refer to a single terminal functional group, a single amino acid, or multiple amino acids located at either end. Where referring to a polynucleotide, a terminal region may include the 5' and 3' ends. The 5' end refers to the end of a polynucleotide having a free nucleic acid phosphate group. The 3' end refers to the end of a polynucleotide that contains a free nucleic acid hydroxyl group. The polynucleotide terminal region may refer to a single terminal functional group, a single nucleotide, or multiple nucleotides located at the end.

[0417] RNA and DNA: As used herein, the terms “RNA,” “RNA molecule,” or “ribonucleic acid molecule” refer to polymers of ribonucleotides, and the terms “DNA,” “DNA molecule,” or “deoxyribonucleic acid molecule” refer to polymers of deoxyribonucleotides. DNA and RNA can be synthesized naturally, for example, by DNA replication and DNA transcription, or they can be synthesized chemically. RNA and DNA may be single-stranded (i.e., ssRNA or ssDNA, respectively) or multi-stranded (e.g., double-stranded, i.e., dsRNA and dsDNA, respectively). As used herein, the terms “messenger RNA” or “mRNA” refer to single-stranded RNA encoding an amino acid sequence of one or more polypeptide chains.

[0418] Sample: As used herein, the term “sample” refers to a part or subset of a larger entity. A sample from a biological organism or substance is referred herein to as a “biological sample” and may include, but is not limited to, tissues, cells, and bodily fluids (e.g., blood, mucus, lymph, synovial fluid, cerebrospinal fluid, saliva, amniotic fluid, amniotic blood, urine, vaginal fluid, and semen). A sample may further include a whole organism, or a subset of its tissues, cells, or components, or fractions or parts thereof, including but not limited to plasma, serum, cerebrospinal fluid, lymph, external skin sections, trachea, intestines, and urogenital organs, tears, saliva, milk, blood cells, tumors, and organs, or homogenates, lysates, or extracts prepared from such fractions or parts. A sample may further include a culture medium, such as a nutrient broth or gel, which may contain cellular components such as proteins or nucleic acid molecules.

[0419] Signal sequence: As used herein, the term “signal sequence” refers to a sequence that can direct the transport or localization of a protein.

[0420] Single-dose unit: As used herein, “single-dose unit" refers to a dose of any therapeutic agent administered in one dose / in one instance / through a single route / at a single point of contact, i.e., in a single administration event. In some embodiments, the single-dose unit is provided as a separate dosage form (e.g., tablet, capsule, patch, pre-filled syringe, vial, etc.).

[0421] Divided Dosage: As used herein, “divided dose” means dividing a single dose or total daily dose into two or more doses.

[0422] Stable: As used herein, “stable” refers to a state of an entity that is robust enough to withstand a certain degree of disturbance. For example, a stable compound or protein may remain intact while being isolated from a reaction mixture to a useful purity.

[0423] Stabilization: As used herein, the terms “stabilize” or “stabilized” mean to make stable or to become stable.

[0424] Subject: As used herein, the term “Subject” means any living organism to which a compound, composition, method, kit, or device relating to this disclosure may be administered or applied for experimental, diagnostic, prophylactic, and / or therapeutic purposes. Subjects may include animals (e.g., mammals such as mice, rats, rabbits, non-human primates, and humans) and plants. A subject receiving, needing, being eligible for, or seeking medical treatment is referred to herein as a “Patient.”

[0425] Substantially: As used herein, the term “substantially” refers to a qualitative condition that exhibits the desired characteristic or property to a complete or near-complete degree or extent. Those skilled in the art of biology will understand that it is rare, if any, for biological and chemical phenomena to reach completion and / or progress to completion, or to achieve or avoid absolute results. Thus, the term “substantially” is used herein to capture the potential lack of completeness inherent in many biological and chemical phenomena.

[0426] To suffer from: An individual who "suffers" from a disease, disorder, and / or condition is diagnosed with a disease, disorder, and / or condition, or exhibits one or more symptoms thereof.

[0427] Susceptibility: Individuals who are “suspicious” for a disease, disorder, and / or condition may not have been diagnosed with the disease, disorder, and / or condition and / or may not exhibit its symptoms, but have a tendency to develop the disease or its symptoms. In some embodiments, individuals susceptible to a disease, disorder, and / or condition (e.g., neurodegenerative disease) may be characterized by one or more of the following: (1) gene mutations associated with the development of the disease, disorder, and / or condition; (2) gene polymorphisms associated with the development of the disease, disorder, and / or condition; (3) increased and / or decreased or impaired expression and / or activity of proteins and / or nucleic acids associated with the disease, disorder, and / or condition; (4) habits and / or lifestyles associated with the development of the disease, disorder, and / or condition; (5) family history of the disease, disorder, and / or condition; and (6) exposure to and / or infection of microorganisms associated with the development of the disease, disorder, and / or condition. In some embodiments, individuals susceptible to a disease, disorder, and / or condition develop the disease, disorder, and / or condition. In some embodiments, individuals susceptible to disease, disorder, and / or condition do not develop the disease, disorder, and / or condition.

[0428] Sustained release: As used herein, the term “sustained release” refers to the release of a compound or drug over a specific period of time, typically at a relatively controlled or consistent rate.

[0429] Synthetic: The term "synthetic" means produced, prepared, and / or manufactured by human hands. The synthetic polynucleotides, polypeptides, or other molecules of this disclosure may be prepared using chemical or enzymatic processes.

[0430] Target: As used herein, the term “target” means an entity of interest or purpose, which may include an object, organ, tissue, cell, protein, nucleic acid, biomolecule, or any group, complex, or part of any of the foregoing. In some embodiments, the target may be a protein or its epitope to which an antibody has affinity, or which is desired, designed, or developed to have affinity. As used herein, the term “target” may also be used to refer to the activity of a drug directed towards a particular object. For example, an antibody that has affinity for a particular protein “X” may be said to target protein X, or to be called an antibody that targets protein X, or to be called a protein X-targeted antibody. Similarly, the object that is the target of a drug's activity may be called the “targeted” object. For example, if an antibody has affinity for a particular protein “X”, protein X may be called the target of the antibody.

[0431] Therapeutic agents: The term “therapeutic agent” refers to any agent that, when administered to a subject, has a therapeutic, diagnostic, and / or prophylactic effect, and / or induces a desired biological and / or pharmacological effect. A therapeutic agent capable of producing a biological effect in a living organism is referred to herein as a “drug.”

[0432] Therapeutic Dose: As used herein, the term “therapeutic dose” means an amount of a drug (e.g., an antibody or other therapeutic agent) delivered to a subject suffering from or susceptible to an infection, disease, disorder, and / or condition, such amount, when delivered or administered, is sufficient to treat, improve, diagnose, prevent, and / or delay the onset of the infection, disease, disorder, and / or condition. In some embodiments, the therapeutic dose is provided in a single dose. In some embodiments, the therapeutic dose is administered in a dosage regimen comprising multiple doses. Those skilled in the art will understand that in some embodiments, a unit dosage form may be considered to contain a therapeutic dose of a particular drug or entity if it contains an amount that is effective when administered as part of such a dosage regimen.

[0433] Therapeutic outcome: As used herein, the term “therapeutic outcome” means an outcome in which, in a subject suffering from or susceptible to an infection, disease, disorder, and / or condition, the infection, disease, disorder, and / or condition is adequate to treat, improve its symptoms, diagnose, prevent, and / or delay its onset.

[0434] Total daily dose: As used herein, “total daily dose” is the amount administered or prescribed over a 24-hour period. This may be administered as a single unit dose.

[0435] To treat: As used herein, the term “to treat” means to partially or completely reduce, improve, enhance, alleviate, delay the onset, inhibit the progression, reduce the severity, and / or reduce the incidence of one or more symptoms or characteristics of a particular infection, disease, disorder, and / or condition. For example, “to treat” a neurodegenerative disease of interest may mean inhibiting neurodegeneration of the interest, promoting nerve cell health, reversing, preventing, or reducing the formation of plaques or condensates in the brain, and / or reversing, preventing, or reducing memory loss or other loss of neurological function or activity. Treatment may be administered to subjects who do not show signs of the disease, disorder, and / or condition, and / or who show only early signs of the disease, disorder, and / or condition, for the purpose of reducing the risk of developing a condition associated with the disease, disorder, and / or condition.

[0436] Unmodified: As used herein, “unmodified” refers to any substance, compound, or molecule before it is altered in any way. Unmodified may also refer to the wild-type or native form of a biomolecule. A molecule may undergo a series of modifications, and each such modified molecule may serve as an “unmodified” starting molecule for subsequent modifications.

[0437] Vector: As used herein, “vector” is any molecule or part that transports, transduces, or otherwise acts as a carrier for a different molecule. The vectors of this disclosure may be produced by recombinant.

[0438] VII. Equivalents and Scope Those skilled in the art will be able to recognize or confirm many equivalents to specific embodiments of the invention described herein si...

Claims

1. An antibody that binds to tau, the antibody comprising a heavy chain variable region (VH) comprising heavy chain complementarity determining region 1 (HC CDR1), HC CDR2, and HC CDR3, and a light chain variable region (VL) comprising light chain complementarity determining region 1 (LC CDR1), LC CDR2, and LC CDR3; (i) the HC CDR1, the HC CDR2, the HC CDR3, the LC CDR1, the LC CDR2, and the LC CDR3 comprise the amino acid sequences of SEQ ID NOs: 82, 101, 119, 132, 149, and 164, respectively; (ii) the HC CDR1, the HC CDR2, the HC CDR3, the LC CDR1, the LC CDR2, and the LC CDR3 comprise the amino acid sequences of SEQ ID NOs: 86, 102, 120, 127, 141, and 156, respectively; (iii) the HC CDR1, the HC CDR2, the HC CDR3, the LC CDR1, the LC CDR2, and the LC CDR3 comprise the amino acid sequences of SEQ ID NOs: 82, 97, 115, 127, 141, and 159, respectively; (iv) the HC CDR1, the HC CDR2, the HC CDR3, the LC CDR1, the LC CDR2, and the LC CDR3 comprise the amino acid sequences of SEQ ID NOs: 79, 94, 111, 127, 141, and 156, respectively; (v) the HC CDR1, the HC CDR2, the HC CDR3, the LC CDR1, the LC CDR2, and the LC CDR3 comprise the amino acid sequences of SEQ ID NOs: 80, 95, 112, 129, 143, and 157, respectively; (vi) the HC CDR1, the HC CDR2, the HC CDR3, the LC CDR1, the LC CDR2, and the LC CDR3 comprise the amino acid sequences of SEQ ID NOs: 81, 94, 114, 127, 141, and 156, respectively; (vii) the HC CDR1, the HC CDR2, the HC CDR3, the LC CDR1, the LC CDR2, and the LC CDR3 comprise the amino acid sequences of SEQ ID NOs: 88, 105, 123, 137, 151, and 168, respectively; (viii) the HC CDR1, the HC CDR2, the HC CDR3, the LC CDR1, the LC CDR2, and the LC CDR3 comprise the amino acid sequences of SEQ ID NOs: 77, 92, 109, 127, 141, and 154, respectively; (ix) the HC CDR1, the HC CDR2, the HC CDR3, the LC CDR1, the LC CDR2, and the LC CDR3 comprise the amino acid sequences of SEQ ID NOs: 78, 93, 110, 128, 142, and 155, respectively; (x) the HC CDR1, the HC CDR2, the HC CDR3, the LC CDR1, the LC CDR2, and the LC CDR3 comprise the amino acid sequences of SEQ ID NOs: 78, 96, 113, 130, 144, and 158, respectively; (xi) the HC CDR1, the HC CDR2, the HC CDR3, the LC CDR1, the LC CDR2, and the LC CDR3 comprise the amino acid sequences of SEQ ID NOs: 83, 98, 116, 131, 145, and 160, respectively; (xii) the HC CDR1, the HC CDR2, the HC CDR3, the LC CDR1, the LC CDR2, and the LC CDR3 comprise the amino acid sequences of SEQ ID NOs: 84, 99, 117, 132, 146, and 161, respectively; (xiii) the HC CDR1, the HC CDR2, the HC CDR3, the LC CDR1, the LC CDR2, and the LC CDR3 comprise the amino acid sequences of SEQ ID NOs: 85, 100, 118, 133, 147, and 162, respectively; (xiv) the HC CDR1, the HC CDR2, the HC CDR3, the LC CDR1, the LC CDR2, and the LC CDR3 comprise the amino acid sequences of SEQ ID NOs: 85, 100, 118, 134, 148, and 163, respectively; (xv) the HC CDR1, the HC CDR2, the HC CDR3, the LC CDR1, the LC CDR2, and the LC CDR3 comprise the amino acid sequences of SEQ ID NOs: 87, 103, 121, 132, 149, and 165, respectively; (xvi) the HC CDR1, the HC CDR2, the HC CDR3, the LC CDR1, the LC CDR2, and the LC CDR3 comprise the amino acid sequences of SEQ ID NOs: 78, 104, 122, 135, 143, and 166, respectively; (xvii) the HC CDR1, the HC CDR2, the HC CDR3, the LC CDR1, the LC CDR2, and the LC CDR3 comprise the amino acid sequences of SEQ ID NOs: 78, 104, 122, 136, 150, and 167, respectively; (xviii) the HC CDR1, the HC CDR2, the HC CDR3, the LC CDR1, the LC CDR2, and the LC CDR3 comprise the amino acid sequences of SEQ ID NOs: 89, 106, 124, 138, 152, and 169, respectively; (xix) the HC CDR1, the HC CDR2, the HC CDR3, the LC CDR1, the LC CDR2, and the LC CDR3 comprise the amino acid sequences of SEQ ID NOs: 90, 107, 125, 139, 151, and 170, respectively; (xx) the HC CDR1, the HC CDR2, the HC CDR3, the LC CDR1, the LC CDR2, and the LC CDR3 comprise the amino acid sequences of SEQ ID NOs: 91, 108, 126, 140, 153, and 171, respectively; or (xxi) The antibody, wherein the HC CDR1, the HC CDR2, the HC CDR3, the LC CDR1, the LC CDR2, and the LC CDR3 comprise any of the amino acid sequences of the HC CDR and LC CDR sequences shown in Table 1.

2. The VH An amino acid sequence of any one of SEQ ID NOs: 1-18, or an amino acid sequence at least 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical thereto, or An amino acid sequence that contains at least 1, 2, or 3 modifications, but not more than 30, 20, or 10 modifications, compared to the amino acid sequence of any one of SEQ ID NOs: 1 to 18. The antibody of claim 1 , comprising:

3. The VL is An amino acid sequence of any one of SEQ ID NOs: 19-38, or an amino acid sequence at least 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical thereto; or The antibody of claim 1 or 2, comprising an amino acid sequence containing at least 1, 2, or 3 modifications, but not more than 30, 20, or 10 modifications, compared to the amino acid sequence of any one of SEQ ID NOs: 19 to 38. Claim 4: The VH comprises an amino acid sequence of SEQ ID NO:11, or an amino acid sequence at least 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical thereto; The VL comprises an amino acid sequence of SEQ ID NO: 30, or an amino acid sequence at least 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical thereto. The antibody according to any one of claims 1 to 3.

5. The VH comprises an amino acid sequence of SEQ ID NO:12, or an amino acid sequence at least 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical thereto; The VL comprises the amino acid sequence of SEQ ID NO: 31, or an amino acid sequence at least 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical thereto. The antibody according to any one of claims 1 to 3.

6. The VH comprises an amino acid sequence of SEQ ID NO:7, or an amino acid sequence at least 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical thereto; The VL comprises an amino acid sequence of SEQ ID NO:25, or an amino acid sequence at least 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical thereto. The antibody according to any one of claims 1 to 3.

7. The VH comprises an amino acid sequence of SEQ ID NO:3, or an amino acid sequence at least 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical thereto; The VL comprises the amino acid sequence of SEQ ID NO:21, or an amino acid sequence at least 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical thereto. The antibody according to any one of claims 1 to 3.

8. The VH comprises an amino acid sequence of SEQ ID NO:4, or an amino acid sequence at least 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical thereto; The VL comprises the amino acid sequence of SEQ ID NO:22, or an amino acid sequence at least 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical thereto. The antibody according to any one of claims 1 to 3.

9. (i) the VH comprises an amino acid sequence of SEQ ID NO: 6, or an amino acid sequence at least 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical thereto, and the VL comprises an amino acid sequence of SEQ ID NO: 22, or an amino acid sequence at least 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical thereto; or (ii) the VH comprises an amino acid sequence of SEQ ID NO: 15, or an amino acid sequence at least 90% identical thereto, and the VL comprises an amino acid sequence of SEQ ID NO: 35, or an amino acid sequence at least 90% identical thereto; The antibody according to any one of claims 1 to 3.

10. The antibody, (a) binds to tau protein at a half maximal effective concentration (EC50) of about 0.001 nM to about 10 nM, or about 0.01 nM to about 2 nM; (b) binds to concentrated paired helical fibrillar tau protein (ePHF) at a half maximal effective concentration (EC50), e.g., from about 0.001 nM to about 100 nM, or from about 0.01 nM to about 20 nM; (c) a dissociation constant (K) of about 0.1 to about 10 nM, or about 0.2 to 5 nM Ｄ ) and binds to iPHF, (d) binding to a tau protein epitope that includes a region formed by a complex of at least two tau proteins; (e) (a) 183-212, (b) 187-218, (c) 33-82, 159-182, 197-226, and 229-246, (d) 217-242, (e) 35-76, and 187-218, (f) 5-34, (g) 187-218, (h) 33-82, 159-188, and 191-230, (i) 35-62, 107-124, and 203-220, (j) 35-82, 159-188, and 197-224, or (k) 53-7 8, 329-348, and 381-408, wherein human tau is numbered in SEQ ID NO: 274, and optionally one or more of the serine, threonine, and / or tyrosine are phosphorylated within said stretch of amino acids selected from (a) to (k), and optionally all of the serine, threonine, and / or tyrosine are phosphorylated within said stretch of amino acids selected from (a) to (k); (f) binds to all or a portion of amino acids 195-215 of tau with a dissociation constant (K D ) of about 1 pM to about 50 pM, or about 1-25 pM; (g) binds to all or a portion of amino acids 191-214 of tau phosphorylated at S199 with a dissociation constant (K D ) of about 0.1 nM to about 10 nM, or about 0.5-5 nM; (h) binds to all or a portion of amino acids 217-234 of tau phosphorylated at T217, T220, and T231 with a dissociation constant (K D ) of about 0.1 nM to about 10 nM, or about 0.1-5 nM; (i) binds to all or a portion of amino acids 225-240 of tau phosphorylated at T231 with a dissociation constant (K D ) of about 0.1 nM to about 25 nM, or about 0.1-15 nM; (j) does not bind to non-pathological tau; (k) binds to pathological tau tangles; (l) binds to a tau protein that contains one or more phosphorylated residues; and / or (m) Inhibiting tau aggregation The antibody according to any one of claims 1 to 9.

11. The antibody according to any one of claims 1 to 10, which is a full-length antibody, an intrabody, a bispecific antibody, a Fab, an F(ab') 2 , an Fv, or a single-chain Fv fragment (scFv).

12. The antibody of any one of claims 1 to 11, wherein the antibody comprises an IgG1, IgG2, IgG3, or IgG4 heavy chain constant region, and / or a kappa or lambda light chain constant region.

13. the antibody comprises a second antigen-binding region that has a different binding specificity than the antigen-binding region that binds to tau; Optionally, the antibody is a multispecific antibody molecule, such as a bispecific antibody molecule, comprising at least a first antigen-binding domain and a second antigen-binding domain.

14. A nucleic acid encoding the antibody according to any one of claims 1 to 13.

15. The nucleic acid of claim 14, wherein the nucleotide sequence encoding the VH comprises any one of the nucleotide sequences of SEQ ID NOs: 39-56, or a nucleic acid sequence at least 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical thereto.

16. The nucleic acid of claim 14 or 15, wherein the nucleotide sequence encoding the VL comprises any one of the nucleotide sequences of SEQ ID NOs:57-76, or a nucleotide sequence at least 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical thereto.

17. (i) the nucleotide sequence encoding the VH comprises SEQ ID NO:47, or a nucleotide sequence at least 90% identical thereto, and the nucleotide sequence encoding the VL comprises SEQ ID NO:66, or a nucleotide sequence at least 90% identical thereto; (ii) the nucleotide sequence encoding the VH comprises SEQ ID NO: 46, or a nucleotide sequence at least 90% identical thereto, and the nucleotide sequence encoding the VL comprises SEQ ID NO: 65, or a nucleotide sequence at least 90% identical thereto; (iii) the nucleotide sequence encoding the VH comprises SEQ ID NO:51, or a nucleotide sequence at least 90% identical thereto, and the nucleotide sequence encoding the VL comprises SEQ ID NO:71, or a nucleotide sequence at least 90% identical thereto; (iv) the nucleotide sequence encoding the VH comprises SEQ ID NO:55, or a nucleotide sequence at least 90% identical thereto, and the nucleotide sequence encoding the VL comprises SEQ ID NO:75, or a nucleotide sequence at least 90% identical thereto; (v) the nucleotide sequence encoding the VH comprises SEQ ID NO:54, or a nucleotide sequence at least 90% identical thereto, and the nucleotide sequence encoding the VL comprises SEQ ID NO:74, or a nucleotide sequence at least 90% identical thereto; (vi) the nucleotide sequence encoding the VH comprises SEQ ID NO:52, or a nucleotide sequence at least 90% identical thereto, and the nucleotide sequence encoding the VL comprises SEQ ID NO:72, or a nucleotide sequence at least 90% identical thereto; or (vii) the nucleotide sequence encoding the VH comprises SEQ ID NO: 43, or a nucleotide sequence at least 90% identical thereto, and the nucleotide sequence encoding the VL comprises SEQ ID NO: 61, or a nucleotide sequence at least 90% identical thereto. The nucleic acid according to any one of claims 14 to 16.

18. A vector encoding an antibody described in any one of claims 1 to 13, or comprising a nucleic acid described in any one of claims 14 to 17.

19. A host cell comprising a nucleic acid according to any one of claims 14 to 17, or a vector according to claim 18.

20. The host cell of claim 19, which is a bacterial cell or a mammalian cell.

21. A method for producing an antibody that binds to tau, the method comprising culturing a host cell described in claim 19 or 20 under conditions suitable for gene expression.

22. A pharmaceutical composition comprising an antibody according to any one of claims 1 to 13 and a pharma- ceutically acceptable excipient.

23. A composition comprising an antibody described in any one of claims 1 to 13, or a pharmaceutical composition described in claim 22, for use in a method for delivering an antibody that binds to tau to a subject.

24. The object is (i) a disease associated with tau expression; (ii) a neurological or neurodegenerative disorder, and / or (iii) tauopathy The composition of claim 23, wherein the patient is suffering from, diagnosed as suffering from, or at risk of suffering from.

25. A composition comprising an antibody according to any one of claims 1 to 13, or a pharmaceutical composition according to claim 22, for use in treating a neurological or neurodegenerative disorder in a subject.

26. The composition of claim 24 or 25, wherein the neurological or neurodegenerative disorder is Alzheimer's disease (AD), frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17), frontotemporal lobar degeneration (FTLD), frontotemporal dementia (FTD), chronic traumatic encephalopathy (CTE), progressive supranuclear palsy (PSP), Down's syndrome, Pick's disease, corticobasal degeneration (CBD), corticobasal syndrome, amyotrophic lateral sclerosis (ALS), prion disease, Creutzfeldt-Jakob disease (CJD), multiple system atrophy, neurofibrillary tangle dementia, or progressive subcortical gliosis.

27. The composition according to any one of claims 23 to 26, which is administered intravenously.

28. Use of an antibody described in any one of claims 1 to 13 in the manufacture of a pharmaceutical agent for a method of delivering an antibody that binds to tau to a subject.

29. The object, (i) a disease associated with tau expression; (ii) a neurological or neurodegenerative disorder, and / or (iii) tauopathy 29. The use of claim 28, wherein the patient is suffering from, diagnosed with, or at risk of suffering from.

30. Use of an antibody described in any one of claims 1 to 13 in the manufacture of a medicament for the treatment of a neurological or neurodegenerative disorder in a subject.

31. The use of claim 29 or 30, wherein the neurological or neurodegenerative disorder is Alzheimer's disease (AD), frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17), frontotemporal lobar degeneration (FTLD), frontotemporal dementia (FTD), chronic traumatic encephalopathy (CTE), progressive supranuclear palsy (PSP), Down's syndrome, Pick's disease, corticobasal degeneration (CBD), corticobasal syndrome, amyotrophic lateral sclerosis (ALS), prion disease, Creutzfeldt-Jakob disease (CJD), multiple system atrophy, neurofibrillary tangle dementia, or progressive subcortical gliosis.

32. The use of any one of claims 28 to 30, wherein the antibody is administered intravenously.

33. The method of claim 32, further comprising the use of an additional therapeutic agent and / or therapy suitable for the treatment or prevention of a neurological or neurodegenerative disorder, 33. The use of any one of claims 28 to 32, wherein the additional therapeutic agent and / or therapy comprises a cholinesterase inhibitor, donepezil, rivastigmine, galantamine, N-methyl D-aspartate (NMDA) antagonist, memantine, an antipsychotic, anxiolytic, anticonvulsant, dopamine agonist, pramipexole, ropinirole, rotigotine, apomorphine, MAO B inhibitor, selegiline, rasagiline, safinamide, catechol O-methyltransferase (COMT) inhibitor, entacapone, opicapone, tolcapone, anticholinergic, benztropine, trihexyphenidyl, amantadine, carbidopa-levodopa, deep brain stimulation (DBS), or a combination thereof.

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