JP2022033757A - Compositions and methods for treating tauopathies - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide methods for treating tauopathies such as progressive supranuclear palsy and Alzheimer's disease.

SOLUTION: A method comprises intravenously administering a fixed dose of 2000 mg of an anti-human tau antibody once every four weeks, where the anti-human tau antibody comprises an immunoglobulin heavy chain variable region (VH) and an immunoglobulin light chain variable region (VL), where: (a) the VH comprises VH complementarity determining regions (VH-CDRs) such that VH-CDR1, VH-CDR2 and VH-CDR3 consist of specific SEQ ID NOs: 16, 17 and 18 consisting of amino acids, respectively; and (b) the VL comprises VL-CDRs such that VL-CDR1, VL-CDR2 and VL-CDR3 consist of specific SEQ ID NOs: 19, 20 and 21 consisting of amino acids, respectively.

SELECTED DRAWING: Figure 8

COPYRIGHT: (C)2022,JPO&INPIT

Description

The application generally relates to administration regimens and formulations for clinical use of anti-tau antibodies.

Protein accumulation, degeneration and aggregation are pathological aspects in many neurodegenerative diseases. Abnormally denatured and aggregated tau, including hyperphosphorylated tau conformers, is a uniform feature of tauopathy and correlates with disease severity.

Tau, a microtubule-associated protein, is abundant in the central nervous system and is produced primarily by neurons. Tau promotes the assembly of microtubules, maintains the structure of microtubules, and stabilizes microtubules. Tau protein is derived from alternative mRNA splicing variants resulting from a single gene to become mature proteins ranging in size from 352 to 441 amino acids. While the fetal brain contains a single tau isoform (tau 352), there are six tau isoforms in the adult human brain. The six tauisoforms differ from each other by having 3 or 4 31-32 amino acid microtubule binding repeats, respectively, and an amino-terminal insertion of 29 amino acids of 2, 1 or 0, respectively.

Tauopathy is a class of neurodegenerative diseases caused by abnormal aggregation of tau protein in so-called neurofibrillary tangles (NFTs) in the brain. Some examples of tauopathy include progressive supranuclear palsy, Alzheimer's disease, frontotemporal dementia (FTD), corticobasal degeneration, and frontotemporal lobar degeneration.

Methods for treating tauopathy are sought in the art. In order to treat patients with an increasing number of tauopathy, therapeutic antibodies against tau, as well as appropriate dosing regimens and formulations for clinical use of such anti-human tau antibodies are sought.

The present disclosure relates in part to administration regimens and formulations of anti-human tau antibodies or tau binding fragments thereof, as well as their use in the treatment of tauopathy.

In one aspect, the disclosure provides a method for performing it in a human subject in need of treatment for tauopathy. The method comprises administering an anti-human tau antibody to a human subject at a fixed dose of 2000 mg. The anti-human tau antibody comprises an immunoglobulin heavy chain variable region (VH) and an immunoglobulin light chain variable region (VL). VH comprises the VH complementarity determining regions (VH-CDR), VH-CDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 16, and VH-CDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 17. , VH-CDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 18. VL comprises or comprises VL-CDR, VL-CDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 19, VL-CDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 20, and VL-CDR3 comprises or comprises the amino acid sequence of SEQ ID NO: 20. : Contains or consists of an amino acid sequence of 21. In certain cases, the anti-human tau antibody is administered intravenously to a human subject. In certain cases, a fixed dose of 2000 mg of anti-human tau antibody is administered once every 4 weeks.

In another aspect, the present specification provides a method for performing it in a human subject in need of treatment for tauopathy. The method comprises administering an anti-human tau antibody to a human subject at a fixed dose of 150 mg. The anti-human tau antibody comprises an immunoglobulin heavy chain variable region (VH) and an immunoglobulin light chain variable region (VL). VH comprises the VH complementarity determining regions (VH-CDR), VH-CDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 16, and VH-CDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 17. , VH-CDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 18. VL comprises or comprises VL-CDR, VL-CDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 19, VL-CDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 20, and VL-CDR3 comprises or comprises the amino acid sequence of SEQ ID NO: 20. : Contains or consists of an amino acid sequence of 21. In certain cases, the anti-human tau antibody is administered intravenously to a human subject. In certain cases, a fixed dose of 150 mg of anti-human tau antibody is administered once every 4 weeks.

In another aspect, the present specification provides a method for performing it in a human subject in need of treatment for tauopathy. The method comprises administering an anti-human tau antibody to a human subject at a fixed dose of 210 mg. The anti-human tau antibody comprises an immunoglobulin heavy chain variable region (VH) and an immunoglobulin light chain variable region (VL). VH comprises the VH complementarity determining regions (VH-CDR), VH-CDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 16, and VH-CDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 17. , VH-CDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 18. VL comprises or comprises VL-CDR, VL-CDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 19, VL-CDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 20, and VL-CDR3 comprises or comprises the amino acid sequence of SEQ ID NO: 20. : Contains or consists of an amino acid sequence of 21. In certain cases, the anti-human tau antibody is administered intravenously to a human subject. In certain cases, a fixed dose of 210 mg of anti-human tau antibody is administered once every 4 weeks.

In another aspect, the present specification provides a method for performing it in a human subject in need of treatment for tauopathy. The method comprises administering an anti-human tau antibody to a human subject at a fixed dose of 2100 mg. The anti-human tau antibody comprises an immunoglobulin heavy chain variable region (VH) and an immunoglobulin light chain variable region (VL). VH comprises the VH complementarity determining regions (VH-CDR), VH-CDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 16, and VH-CDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 17. , VH-CDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 18. VL comprises or comprises VL-CDR, VL-CDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 19, VL-CDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 20, and VL-CDR3 comprises or comprises the amino acid sequence of SEQ ID NO: 20. : Contains or consists of an amino acid sequence of 21. In certain cases, the anti-human tau antibody is administered intravenously to a human subject. In certain cases, a fixed dose of 2100 mg of anti-human tau antibody is administered once every 4 weeks.

In one aspect, the present specification provides a method for performing it in a human subject in need of treatment for tauopathy. The method comprises administering an anti-human tau antibody to a human subject at a fixed dose of 4200 mg. The anti-human tau antibody comprises an immunoglobulin heavy chain variable region (VH) and an immunoglobulin light chain variable region (VL). VH comprises the VH complementarity determining regions (VH-CDR), VH-CDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 16, and VH-CDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 17. , VH-CDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 18. VL comprises or comprises VL-CDR, VL-CDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 19, VL-CDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 20, and VL-CDR3 comprises or comprises the amino acid sequence of SEQ ID NO: 20. : Contains or consists of an amino acid sequence of 21. In certain cases, the anti-human tau antibody is administered intravenously to a human subject. In certain cases, a fixed dose of 4200 mg of anti-human tau antibody is administered once every 4 weeks.

The following embodiments apply to all of the above embodiments. In some cases, tauopathy is a progressive nuclear palsy, Alzheimer's disease, muscle atrophic lateral sclerosis / Parkinson's dementia complex, silver granule dementia, British amyloid angiopathy, brain amyloid angiopathy, cortex. Basal nuclear degeneration, Kreuzfeld-Jakob disease, boxer dementia, diffuse neurofibrillary tangle with calcification, Down syndrome, frontotemporal dementia (FTD), frontotemporal dementia linked to chromosome 17 with parkinsonism Temporal dementia, frontotemporal degeneration, Gerstmannstroisler Shineker's disease, Hallerfoldenspatz's disease, inclusion myositis, polyline atrophy, muscle tonic dystrophy, C-type Niemannpic's disease, neurofibrillary tangle Non-Guam-type motor neuron disease with fibrotic changes, Pick's disease, post-encephalitis Parkinson's syndrome, prion protein cerebral amyloid angiopathy, progressive subcortical gliosis, globular gliatauopathy, subacute sclerosing panencephalitis, neurofibrillary senile Dementia, multiple infarct dementia, stroke, chronic traumatic encephalopathy, traumatic brain injury, agitation, epilepsy, epilepsy, or acute frontotemporal dementia. In one example, tauopathy is a progressive supranuclear palsy. In another example, tauopathy is Alzheimer's disease. In certain examples, the VH of the anti-human tau antibody comprises or consists of SEQ ID NO: 12, and the VL of the anti-human tau antibody comprises or consists of SEQ ID NO: 13. In certain examples, the anti-human tau antibody comprises or comprises heavy and light chains, the heavy chain comprising or consisting of SEQ ID NO: 14, and the light chain comprising or consisting of SEQ ID NO: 15.

In another aspect, the disclosure is characterized by a pharmaceutical composition comprising an anti-human tau antibody. The pharmaceutical composition comprises an anti-human tau antibody at a concentration of 50 mg / ml or 60 mg / ml, a histidine at a concentration of 20 mM, a sucrose at a concentration of 250 mM, and a polysorbate 80 at a concentration of 0.05% (weight / volume). In some examples, the pharmaceutical composition further comprises 50 μM diethylenetriamine pentaacetic acid (DTPA). This anti-human tau antibody comprises VH and VL. VH comprises or comprises VH-CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 16, VH-CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 17, and the amino acid sequence of SEQ ID NO: 18. VH-CDR3, which comprises it. VL comprises or comprises VL-CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 19, VL-CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 20, and the amino acid sequence of SEQ ID NO: 21. VL-CDR3, which comprises it. The pharmaceutical composition has a pH of 6.0.

In some embodiments, the VH of the anti-human tau antibody comprises or comprises SEQ ID NO: 12, and the VL of the anti-human tau antibody comprises or comprises SEQ ID NO: 13. In some embodiments, the anti-human tau antibody comprises or comprises heavy and light chains, the heavy chain comprising or consisting of SEQ ID NO: 14, and the light chain comprising or consisting of SEQ ID NO: 15. In some embodiments, the pharmaceutical composition is used to perform it in a human subject in need of treatment for tauopathy by intravenously administering any of the above pharmaceutical compositions to a human subject. .. In some embodiments, the anti-human tau antibody of the pharmaceutical composition is administered to a human subject at a fixed dose of 150 mg once every 4 weeks. In other embodiments, the anti-human tau antibody of the pharmaceutical composition is administered to a human subject at a fixed dose of 210 mg once every 4 weeks. In yet another embodiment, the anti-human tau antibody of the pharmaceutical composition is administered to a human subject at a fixed dose of 700 mg once every 4 weeks. In a further embodiment, the anti-human tau antibody of the pharmaceutical composition is administered to a human subject at a fixed dose of 2000 mg once every 4 weeks. In other embodiments, the anti-human tau antibody of the pharmaceutical composition is administered to a human subject at a fixed dose of 2100 mg once every 4 weeks. In another embodiment, the anti-human tau antibody of the pharmaceutical composition is administered to a human subject at a fixed dose of 4200 mg once every 4 weeks. In certain examples, the pharmaceutical composition is administered for at least 12 weeks (eg, 12 weeks, 16 weeks, 20 weeks, 24 weeks, 30 weeks, 32 weeks, 36 weeks, 40 weeks, 48 weeks, 52 weeks). .. In certain cases, tauopathy is known as progressive nuclear palsy, Alzheimer's disease, muscular atrophic lateral sclerosis / Parkinson's dementia complex, silver granule dementia, British amyloid angiopathy, cerebral amyloid angiopathy, cortical basal Nuclear degeneration, Kreuzfeld-Jakob disease, boxer dementia, diffuse neurofibrillary tangle with calcification, Down syndrome, frontotemporal dementia (FTD), frontotemporal dementia linked to chromosome 17 Head dementia, frontotemporal degeneration, Gerstmannstroisler Shineker's disease, Hallerfoldenspatz's disease, encapsulation myitis, polyline atrophy, muscle tonic dystrophy, C-type Niemannpic's disease, neurofibrillary tangles Non-Guam-type motor neuron disease with changes, Pick's disease, post-encephalitis Parkinson's syndrome, prion protein cerebral amyloid vasculopathy, progressive subcortical gliosis, globular gliatauopathy, subacute sclerosing panencephalitis, neurofibrillary tangles Disease, multiple infarct dementia, stroke, chronic traumatic encephalopathy, traumatic brain injury, agitation, epilepsy, epilepsy, or acute frontotemporal dementia. In one embodiment, tauopathy is a progressive supranuclear palsy. In another embodiment, tauopathy is Alzheimer's disease.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the present invention belongs. Methods and substances similar to or equivalent to those described herein can be used in the practice or testing of the present invention, but exemplary methods and substances are described below. All publications, patent applications, patents and other references referred to herein are incorporated by reference in their entirety. In the event of a contradiction, this application (including definitions) will prevail. Substances, methods and examples are examples only and are not intended to be limiting.

Other features and advantages of the present invention will be apparent from the embodiments and claims for carrying out the following inventions.

Different forms of extracellular tau (e-tau) sequences compared to the human tau 441 isoform (2N4R) sequence (SEQ ID NO: 6), e-tau 1 (SEQ ID NO: 7), e-tau 2 (SEQ ID NO: 8). , E-tau 3 (SEQ ID NO: 9), and e-tau 4 (SEQ ID NO: 10). FIG. 6 is a schematic diagram of a study design for a single escalating dose test according to Example 1. It is a graph which shows the exposure-response model (Bayes E _max ) of the concentration vs. e-tau suppression in CSF. FIG. 6 is a schematic diagram of a study design for a multiple escalating dose test according to Example 3. 6 is a table describing the basic demographic characteristics of patients in the multiple escalating dose study described in Example 3. It is a table which described the list of adverse events in the multiple escalation dose study described in Example 3. It is a table which described the list of the serum PK parameters (at the 57th day of a test) of BIIB092. It is a graph which shows the average change amount of e-tau concentration with time. There was a dose-dependent relationship between the dose of BIIB092 and the degree of e-tau suppression in CSF. It is a table describing free e-tau in CSF as a baseline percentage to which BIIB092 was administered.

The present disclosure relates to administration regimens and formulations of anti-human tau antibodies and tau binding fragments thereof, as well as tauopathy (eg, diseases associated with tau aggregation, such as progressive nuclear palsy, Alzheimer's disease, muscular atrophic lateral sclerosis). / Parkinson dementia complex, silver granule dementia, British amyloid angiopathy, brain amyloid angiopathy, cortical basal nucleus degeneration, Kreuzfeld-Jakob disease, boxer dementia, diffuse neurofibrillary tangle with calcification , Down Syndrome, Frontotemporal Dementia (FTD), Frontotemporal Dementia Linked to Chromium 17 with Parkinsonism, Frontotemporal Dementia, Gerstmannstroisler Shineker's Disease, Haller Fordensch Patz's disease, inclusion myopathy, multilineage atrophy, muscle tonic dystrophy, C-type Niemann-Pick's disease, non-Guam-type motor neuron disease with neurofibrillary tangles, Pick's disease, post-encephalitis Parkinson's syndrome, prion protein brain amyloid angiopathy , Progressive subcortical gliosis, subacute sclerosing panencephalitis, neurofibrillary tangle dementia, multiple infarct dementia, stroke, chronic traumatic encephalopathy, traumatic brain injury, tremor, epilepsy, epilepsy, and acute It is characterized by its use in the treatment of (such as lead encephalopathy).

Tau tau is a protein that plays an important role in the etiology of several diseases collectively called tauopathy. There are several different isoforms of microtubule-associated proteins, which are described below.
352aa of fetal type tau isoform MAEPRQEFEVMEDHAGTYGLGDRKDQGGYTMHQDQEGDTDAGLKAEEAGIGDTPSLEDEAAGHVTQARMVSKSKDGTGSDDKKAKGADGKTKIATPRGAAPPGQKGQANATRIPAKTPPAPKTPPSSGEPPKSGDRSGYSSPGSPGTPGSRSRTPSLPTPPTREPKKVAVVRTPPKSPSSAKSRLQTAPVPMPDLKNVKSKIGSTENLKHQPGGGKVQIVYKPVDLSKVTSKCGSLGNIHHKPGGGQVEVKSEKLDFKDRVQSKIGSLDNITHVPGGGNKKIETHKLTFRENAKAKTDHGAEIVYKSPVVSGDTSPRHLSNVSSTGSIDMVDSPQLATLADEVSASLAKQGL (SEQ ID NO: 1)
381aa of tau B isoforms MAEPRQEFEVMEDHAGTYGLGDRKDQGGYTMHQDQEGDTDAGLKESPLQTPTEDGSEEPGSETSDAKSTPTAEAEEAGIGDTPSLEDEAAGHVTQARMVSKSKDGTGSDDKKAKGADGKTKIATPRGAAPPGQKGQANATRIPAKTPPAPKTPPSSGEPPKSGDRSGYSSPGSPGTPGSRSRTPSLPTPPTREPKKVAVVRTPPKSPSSAKSRLQTAPVPMPDLKNVKSKIGSTENLKHQPGGGKVQIVYKPVDLSKVTSKCGSLGNIHHKPGGGQVEVKSEKLDFKDRVQSKIGSLDNITHVPGGGNKKIETHKLTFRENAKAKTDHGAEIVYKSPVVSGDTSPRHLSNVSSTGSIDMVDSPQLATLADEVSASLAKQGL (SEQ ID NO: 2)
410aa of tau C isoforms MAEPRQEFEVMEDHAGTYGLGDRKDQGGYTMHQDQEGDTDAGLKESPLQTPTEDGSEEPGSETSDAKSTPTAEDVTAPLVDEGAPGKQAAAQPHTEIPEGTTAEEAGIGDTPSLEDEAAGHVTQARMVSKSKDGTGSDDKKAKGADGKTKIATPRGAAPPGQKGQANATRIPAKTPPAPKTPPSSGEPPKSGDRSGYSSPGSPGTPGSRSRTPSLPTPPTREPKKVAVVRTPPKSPSSAKSRLQTAPVPMPDLKNVKSKIGSTENLKHQPGGGKVQIVYKPVDLSKVTSKCGSLGNIHHKPGGGQVEVKSEKLDFKDRVQSKIGSLDNITHVPGGGNKKIETHKLTFRENAKAKTDHGAEIVYKSPVVSGDTSPRHLSNVSSTGSIDMVDSPQLATLADEVSASLAKQGL (SEQ ID NO: 3)
383aa of tau D isoforms MAEPRQEFEVMEDHAGTYGLGDRKDQGGYTMHQDQEGDTDAGLKAEEAGIGDTPSLEDEAAGHVTQARMVSKSKDGTGSDDKKAKGADGKTKIATPRGAAPPGQKGQANATRIPAKTPPAPKTPPSSGEPPKSGDRSGYSSPGSPGTPGSRSRTPSLPTPPTREPKKVAVVRTPPKSPSSAKSRLQTAPVPMPDLKNVKSKIGSTENLKHQPGGGKVQIINKKLDLSNVQSKCGSKDNIKHVPGGGSVQIVYKPVDLSKVTSKCGSLGNIHHKPGGGQVEVKSEKLDFKDRVQSKIGSLDNITHVPGGGNKKIETHKLTFRENAKAKTDHGAEIVYKSPVVSGDTSPRHLSNVSSTGSIDMVDSPQLATLADEVSASLAKQGL (SEQ ID NO: 4)
412aa of tau E isoforms MAEPRQEFEVMEDHAGTYGLGDRKDQGGYTMHQDQEGDTDAGLKESPLQTPTEDGSEEPGSETSDAKSTPTAEAEEAGIGDTPSLEDEAAGHVTQARMVSKSKDGTGSDDKKAKGADGKTKIATPRGAAPPGQKGQANATRIPAKTPPAPKTPPSSGEPPKSGDRSGYSSPGSPGTPGSRSRTPSLPTPPTREPKKVAVVRTPPKSPSSAKSRLQTAPVPMPDLKNVKSKIGSTENLKHQPGGGKVQIINKKLDLSNVQSKCGSKDNIKHVPGGGSVQIVYKPVDLSKVTSKCGSLGNIHHKPGGGQVEVKSEKLDFKDRVQSKIGSLDNITHVPGGGNKKIETHKLTFRENAKAKTDHGAEIVYKSPVVSGDTSPRHLSNVSSTGSIDMVDSPQLATLADEVSASLAKQGL (SEQ ID NO: 5)
441aa Tau F Isoform (2N4R)
MAEPRQEFEVMEDHAGTYGLGDRKDQGGYTMHQDQEGDTDAGLKESPLQTPTEDGSEEPGSETSDAKSTPTAEDVTAPLVDEGAPGKQAAAQPHTEIPEGTTAEEAGIGDTPSLEDEAAGHVTQARMVSKSKDGTGSDDKKAKGADGKTKIATPRGAAPPGQKGQANATRIPAKTPPAPKTPPSSGEPPKSGDRSGYSSPGSPGTPGSRSRTPSLPTPPTREPKKVAVVRTPPKSPSSAKSRLQTAPVPMPDLKNVKSKIGSTENLKHQPGGGKVQIINKKLDLSNVQSKCGSKDNIKHVPGGGSVQIVYKPVDLSKVTSKCGSLGNIHHKPGGGQVEVKSEKLDFKDRVQSKIGSLDNITHVPGGGNKKIETHKLTFRENAKAKTDHGAEIVYKSPVVSGDTSPRHLSNVSSTGSIDMVDSPQLATLADEVSASLAKQGL (SEQ ID NO: 6)

Intracellular tau levels in patients with Alzheimer's disease are elevated compared to non-dementia controls (Barton, Am J. Pathol., 137: 497-502 (1990); Katoon, J. Neurochem., 59). : 750-753 (1992)). This increase in intracellular tau levels has been shown to protect the decrease in intracellular tau content in neurodegenerative mouse models (Raportort et al., Proc. Natl. Acad. Sci. USA, 99: 6364-). 6369 (2002); Robertson et al., Science, 316: 750-754 (2007)), which is considered to be toxic to neurons, and therefore reducing intracellular tau levels is therapeutically beneficial. Can be.

An N-terminally truncated secretory tau species called extracellular tau (e-tau) has been identified. As used herein, "e-tau" includes any tau polypeptide that can be detected in cerebrospinal fluid (CSF) or interstitial fluid (ISF). In some embodiments, the e-tau is a polypeptide having the sequence set forth in one of SEQ ID NOs: 7-10 in FIG. The e-tau species vary from 171 amino acids of e-tau 1 to 67 amino acids of e-tau 4. Although tau lacks a signal sequence, tau is released into the medium from neuroblastoma cells, nonneuronal cells expressing tau, human neurons derived from induced pluripotent stem cells, and primary mouse neurons. I know there is. Therefore, tau can be freely secreted or associated with exosomes or other membrane vesicles. e-tau has also been detected in the brain ISF of both wild-type and P301S tau-expressing mice in microdialysis tests. e-tau has also been identified in the brain ISF of patients after traumatic brain injury. It has been shown that e-tau regulates Aβ production and enhances neuronal overactivity (Bright et al., Neuroscience and Aging, 36: 693-709 (2015)). Treatment with e-tau neutralizing antibodies suppresses the overactivity of e-tau-mediated neurons. See, for example, WO2014 / 02877. Increased neuronal overactivity stimulated by e-tau leads not only to increased Aβ secretion but also to further increase in e-tau secretion, resulting in e-tau and Aβ creating a feed-forward disease mechanism that persists the disease. Is proposed. Therefore, by neutralizing e-tau, it suppresses the spread of tau and tau lesions in the brain, suppresses Aβ levels in the central nervous system and consequent neuronal overactivity, and in some cases dementia. Can delay clinical progression to.

Anti-Human Tau Antibody One way to neutralize tau is to use an antibody that binds to tau. In certain embodiments, these antibodies bind to an epitope within amino acids 1-25, 1-18, 9-18, 13-24, 15-44, or 15-24 of SEQ ID NO: 6. In one particular embodiment, the anti-tau antibody binds to an epitope within AGTYGLGDRK (SEQ ID NO: 11).

（配列番号：１２）
を有している。 An exemplary anti-human tau antibody is called "BIIB092". BIIB092 is a humanized IgG4 / κ antibody that recognizes human tau. The heavy chain variable domain of the BIIB092 antibody has the following amino acid sequence,

(SEQ ID NO: 12)
have.

（配列番号：１３）
を有している。 The light chain variable domain of the BIIB092 antibody has the following amino acid sequence,

(SEQ ID NO: 13)
have.

（配列番号：１４）
を有している。 The heavy chain of the BIIB092 antibody has the following amino acid sequence.

(SEQ ID NO: 14)
have.

（配列番号：１５）
を有している。 The light chain of the BIIB092 antibody has the following amino acid sequence,

(SEQ ID NO: 15)
have.

In some embodiments, the anti-human tau antibody or tau binding fragment thereof comprises three light chain variable domain CDRs of BIIB092. In some embodiments, the anti-human tau antibody or tau binding fragment thereof comprises three heavy chain variable domain CDRs of BIIB092. In yet other embodiments, the anti-human tau antibody or tau binding fragment thereof comprises three heavy chain variable domain CDRs of BIIB092 and three light chain variable domain CDRs. The CDRs may be based on any CDR definition in the art, such as Kabat, Chothia, Chothia from Abysis, enhanced Chothia / AbM, or may be based on a contact definition. The CDR sequences of BIIB092 are listed in Table 1 below.
Table 1: CDR sequence of BIIB092

In some embodiments, the anti-human tau antibody or tau binding fragment thereof comprises the amino acid sequence set forth in VH CDR1, SEQ ID NO: 17, comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 16. VH CDR2 comprising or consisting of VH CDR2 and VH CDR3 comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 18, and VL CDR1 comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 19. Includes VL CDR2 comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 20 and VL CDR3 comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 21.

In some embodiments, the anti-human tau antibody or tau binding fragment thereof comprises a VH comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 12. In some embodiments, the anti-human tau antibody or tau binding fragment thereof comprises a VL comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 13. In some embodiments, the anti-human tau antibody or tau binding fragment thereof comprises a VH comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 12, and the amino acid sequence set forth in SEQ ID NO: 13. Includes VL consisting of or consisting of it.

In some embodiments, the anti-human tau antibody or tau binding fragment thereof comprises a heavy chain comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 14. In some embodiments, the anti-human tau antibody or tau binding fragment thereof comprises a light chain comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 15. In some embodiments, the anti-human tau antibody or tau binding fragment thereof comprises a heavy chain comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 14, and the amino acid sequence set forth in SEQ ID NO: 15. Includes a light chain containing or consisting of it.

In certain embodiments, the anti-human tau antibody is an IgG antibody. In certain embodiments, the anti-human tau antibody has a heavy chain constant region selected from, for example, IgG1, IgG2, IgG3, IgG4, IgM, IgA1, IgA2, IgD, and IgE. In one embodiment, the anti-human tau antibody is an IgG4 isotype anti-human tau antibody. In another embodiment, the anti-human tau antibody is an IgG1 isotype anti-human tau antibody. In yet another embodiment, the anti-human tau antibody is an IgG2 isotype anti-human tau antibody. In another embodiment, the anti-human tau antibody is an IgG3 isotype anti-human tau antibody. In a further embodiment, the anti-human tau antibody has, for example, a light chain constant region selected from the human kappa light chain constant region or the human λ light chain constant region. In certain embodiments, the anti-human tau antibody is an IgG4 / human kappa light chain antibody.

In some embodiments, the anti-human tau antibody is a full-length (complete) antibody or near-full-length. The protein may comprise at least one, preferably two complete heavy chains, and at least one, preferably two complete light chains. In some embodiments, the anti-human tau antibody is a tau binding fragment. In some examples, the tau-binding antibody fragment is Fab, Fab', F (ab') 2, Facb, Fv, single chain Fv (scFv), sc (Fv) 2, or deerbody.

The heavy and light chains of the anti-human tau antibody disclosed herein may also contain a signal sequence. The signal sequence may be selected from signal sequences well known in the art, such as MDMRBPAQLLLGLLLWLFPGSRC (SEQ ID NO: 22) or MDMRBPAQLLLGLLLWLPGARC (SEQ ID NO: 23).

An antibody such as BIIB092 or a tau-binding fragment thereof comprises, for example, a gene encoding an enumerated amino acid sequence by preparing and expressing a synthetic gene encoding the enumerated amino acid sequence, or by mutating a human germ cell gene. By obtaining it, it can be produced. Furthermore, this antibody and other anti-human tau antibodies can be produced, for example, using one or more of the following methods.

Methods for Making Anti-Human Tau Antibodies Anti-human tau antibodies or tau binding fragments can be made using bacterial cells or eukaryotic cells. Some antibodies, such as Fab', are bacterial cells, such as E. coli. It can be produced using colli cells. Antibodies can also be made using eukaryotic cells, such as transformed cell lines (eg, CHO, 293E, COS). In addition, the antibody (eg, scFv's) can be a yeast cell, eg, Pichia (see, eg, Powers et al., J Immunol Methods. 251: 123-35 (2001)), Hanseula, or Saccharomyces. It can be expressed using such as. To produce the antibody of interest, one polynucleotide or multiple polynucleotides encoding the antibody is constructed, introduced into one expression vector or multiple expression vectors, and then in a suitable host cell. Express. To improve expression, the light chain and heavy chain genes are expressed without altering (or minimally altering, eg, removing the C-terminal residues of the heavy or light chain). The nucleotide sequence may be recoded. Candidate regions for recoding include regions involved in translation initiation, codon utilization, and possible unintended mRNA splicing. Those skilled in the art will readily imagine the polynucleotides encoding anti-human tau antibodies, including VH and / or VL, HC and / or LC of the tau antibodies described herein.

Recombinant expression vectors (s) are prepared using standard molecular biology techniques, transfected into host cells, transformed cells are selected, host cells are cultured, and anti-human tau antibodies are recovered. ..

When expressing an anti-human tau antibody or tau binding fragment in a bacterial cell (eg, E. coli), the expression vector needs to have properties such that the vector can be amplified in the bacterial cell. In addition, E. When a colli, such as JM109, DH5α, HB101, or XL1-Blue, is used as the host, the vector is a promoter, eg, E.I. The lacZ promoter (Ward et al., 341: 544-546 (1989)), the araB promoter (Better et al., Science, 240: 1041-1043 (1988)), or T7, which allows efficient expression in E. coli. Must have a promoter. Examples of such vectors include, for example, M13 series vectors, pUC series vectors, pBR322, pBluescript, pCR-Script, pGEX-5X-1 (Pharmacia), "QIAexpress system". (QIAGEN), pEGFP, and pET (when using this expression vector, the host is preferably BL21 expressing the T7 RNA polymerase). The expression vector contains a signal sequence for antibody secretion. The pelB signal sequence (Lei et al., J. Bacteriol., 169: 4379 (1987)) may be used as a signal sequence for antibody secretion to be produced within the periplasm of E. coli. In order to carry out expression by bacteria, the expression vector may be introduced into bacterial cells using the calcium chloride method or the electroporation method.

When the antibody is expressed in animal cells such as CHO, COS and NIH3T3 cells, the expression vector is a promoter required for expression in these cells, such as the SV40 promoter (Mulligan et al., Nature, 277: 108). (1979)) (eg, Simian virus 40 early promoter), MMLV-LTR promoter, EF1α promoter (Mizushima et al., Nuclear Acids Res., 18: 5322 (1990)), or CMV promoter (eg, human cytomegalovirus). Pre-early promoter) is included. In addition to the nucleic acid sequence encoding the immunoglobulin or its domain, the recombinant expression vector has another sequence, eg, a sequence that regulates the replication of the vector in the host cell (eg, the origin of replication), and a selectable marker gene. You may be doing it. The selectable marker gene facilitates the selection of host cells into which the vector has been introduced (see, eg, US Pat. Nos. 4,399,216, 4,634,665 and 5,179,017). For example, selectable marker genes typically confer resistance to drugs such as G418, hyglomycin or methotrexate to host cells into which the vector has been introduced. Examples of vectors with selectable markers include pMAM, pDR2, pBK-RSV, pBK-CMV, pOPRSV, and pOP13.

In one embodiment, the anti-human tau antibody is made using mammalian cells. Exemplary mammalian host cells for expressing the antibody include Chinese hamster ovary (CHO cells) (eg, with the DHFR selection marker described in Kaufman and Sharp (1982) Mol. Biol. 159: 601-621. Used, Urlaub and Chasin (1980) Proc. Natl. Acad. Sci. USA 77: 4216-4220, including dhfr-CHO cells, human fetal kidney 293 cells (eg, 293, 293E, 293T). ), COS cells, NIH3T3 cells, lymphocyte cell lines, such as NS0 myeloma cells and SP2 cells, and transgenic animals, such as cells derived from transgenic mammals. For example, the cell is a mammary epithelial cell. In one particular embodiment, the mammalian cell is a CHO-DG44I cell.

In an exemplary system for antibody expression, recombinant expression vectors encoding both the anti-human tau antibody heavy chain and the anti-human tau antibody light chain of an anti-human tau antibody (eg, BIIB092) are dhfr ^- CHO by calcium phosphate mediated transfection. Introduced into cells. In the recombinant expression vector, the antibody heavy chain gene and the antibody light chain gene are derived from enhancer / promoter regulatory elements (eg, SV40, CMV, adenovirus, etc., respectively, such as CMV enhancer / AdMLP promoter regulatory element, or, respectively. It is functionally linked to the SV40 enhancer / AdMLP promoter regulatory element, etc.) to drive high levels of gene transcription. The recombinant expression vector also has a DHFR gene, which allows selection of vector-transfected CHO cells using methotrexate selection / amplification. The selected transformed host cells are cultured to express the heavy and light chains of the antibody and the antibody is recovered from the medium.

Antibodies can also be made using transgenic animals. For example, US Pat. No. 5,849,992 describes a method for expressing an antibody in the mammary gland of a transgenic mammal. The transgene is constructed to include a milk-specific promoter, a nucleic acid encoding the antibody of interest, and a signal sequence for secretion. The milk produced by females of such transgenic mammals contains the antibody of interest secreted therein. The antibody may be purified from milk or the antibody may be used directly. Animals are also provided to include one or more of the nucleic acids described herein.

The antibodies of the present disclosure may be isolated from the inside or outside of the host cell (such as medium) and purified to a near pure and uniform antibody. Methods for isolation and purification commonly used for antibody purification may be used for antibody isolation and purification and are not limited to any particular method. For example, column chromatography, filtration, ultrafiltration, salting out, solvent precipitation, solvent extraction, distillation, immunoprecipitation, SDS polyacrylamide gel electrophoresis, isoelectric point electrophoresis, dialysis, and recrystallization are appropriately selected and combined. Thereby, the antibody may be isolated and purified. Chromatography includes, for example, affinity chromatography, ion exchange chromatography, hydrophobic chromatography, gel filtration, reverse phase chromatography, and adsorption chromatography. Daniel R. Marshak et al., Cold Spring Harbor Laboratory Press, 1996). Chromatography may be performed using liquid phase chromatography such as HPLC and FPLC. Examples of the column used for affinity chromatography include a protein A column and a protein G column. Examples of columns using a protein A column include Hyper D, POROS, and Sepharose FF (GE Healthcare Biosciences). The disclosure also includes antibodies that have been highly purified using these purification methods.

Anti-Human Tau Antibody Preparation Any of the anti-human tau antibodies described herein may be dispensed as a pharmaceutical composition. The pharmaceutical composition may contain 10 mg / mL, 60 mg / mL or 50 mg / mL of the anti-human tau antibodies described herein. In certain embodiments, the pharmaceutical composition comprises 50 mg / mL of the anti-human tau antibody described herein. In addition, the pharmaceutical composition comprises 20 mM histidine, 250 mM sucrose, and 0.05% (weight / volume) concentration of polysorbate 80. In certain examples, the pharmaceutical composition further comprises 50 μM diethylenetriamine pentaacetic acid (DTPA). The pharmaceutical composition has a pH of 6.0.

In some examples, the anti-human tau antibody of the pharmaceutical composition is an immunoglobulin heavy chain variable region (VH) containing VH complementarity determining regions (VH-CDRs), and an immunoglobulin light chain variable region containing VL-CDRs. Includes (VL). Whether VH-CDR1 contains or consists of the amino acid sequence of SEQ ID NO: 16, VH-CDR2 contains or consists of the amino acid sequence of SEQ ID NO: 17, and VH-CDR3 contains or consists of the amino acid sequence of SEQ ID NO: 18. Or it consists of it. Whether VL-CDR1 contains or consists of the amino acid sequence of SEQ ID NO: 19, VL-CDR2 contains or consists of the amino acid sequence of SEQ ID NO: 20, and VL-CDR3 contains the amino acid sequence of SEQ ID NO: 21. Or it consists of it.

In certain examples, the anti-human tau antibody of the pharmaceutical composition comprises VH comprising or consisting of SEQ ID NO: 12. In certain examples, the anti-human tau antibody of the pharmaceutical composition comprises VL comprising or consisting of SEQ ID NO: 13. In certain examples, the anti-human tau antibody of the pharmaceutical composition comprises a VH comprising or consisting of SEQ ID NO: 12, and a VL comprising or consisting of SEQ ID NO: 13.

In certain examples, the anti-human tau antibody of the pharmaceutical composition comprises a heavy chain comprising or consisting of SEQ ID NO: 14. In certain examples, the anti-human tau antibody of the pharmaceutical composition comprises a light chain comprising or consisting of SEQ ID NO: 15. In certain examples, the anti-human tau antibody of the pharmaceutical composition comprises a heavy chain comprising or consisting of SEQ ID NO: 14 and a light chain comprising or consisting of SEQ ID NO: 15.

Indications The anti-human tau antibodies described herein are expected to be useful in the treatment of tauopathy. Tauopathy is a class of neurodegenerative diseases associated with abnormal aggregation of tau protein in changes in neurofibrils or glial fibrils in the human brain.

Exemplary tauopathy include progressive supranuclear palsy, Alzheimer's disease, muscular atrophic lateral sclerosis / Parkinson's dementia complex, silver granule dementia, British amyloid angiopathy, cerebral amyloid angiopathy, and cortical basal nucleus. Degeneration, Kreuzfeld-Jakob disease, boxer dementia, diffuse neurofibrillary tangle with calcification, Down syndrome, frontotemporal dementia (FTD), frontotemporal dementia linked to chromosome 17 with parkinsonism Type dementia, frontotemporal degeneration, Gerstmannstroisler Scheinker's disease, Hallerfoldenspats' disease, inclusion myositis, polyline atrophy, muscle tonic dystrophy, type C Niemannpic's disease, neurofibrillary tangles Non-Guam type motor neuron disease with, Pick's disease, post-encephalitis Parkinson's syndrome, prion protein cerebral amyloid angiopathy, progressive subcortical gliosis, globular gliatauopathy, subacute sclerosing panencephalitis, neurofibrillary senile dementia , Multiple infarct dementia, stroke, chronic traumatic encephalopathy, traumatic brain injury, tremor, epilepsy, epilepsy, and acute lead encephalopathy.

In one embodiment, the anti-human tau antibodies described herein are used to treat progressive supranuclear palsy.

In another embodiment, the anti-human tau antibody described herein is used to treat Alzheimer's disease.

Therapeutic Methods The present disclosure features methods for treating human subjects with tauopathy (see above) using the anti-human tau antibodies disclosed herein or the pharmaceutical compositions disclosed herein. do. In one embodiment, tauopathy is a progressive supranuclear palsy. In another embodiment, tauopathy is Alzheimer's disease.

In certain embodiments, the method comprises tau (eg, serum or plasma) in an individual's extracellular fluid (eg, cerebrospinal fluid (CSF), interstitial fluid (ISF), blood, or blood fraction (eg, serum or plasma)). For example, it comprises administering it to a human subject in need of an anti-human tau antibody in an amount effective to significantly reduce the level of total tau and / or free tau). "Free tau" means a tau polypeptide that is not bound to an anti-human tau antibody. In one embodiment, the free tau is extracellular tau (e-tau). "Total tau" includes free tau and tau bound to anti-human tau antibodies. In one particular embodiment, the method comprises administering to a human subject in need of an anti-human tau antibody in an amount effective to significantly reduce the level of free e-tau. In some embodiments, the level of tau (eg, total tau and / or free tau) is significantly reduced within 36 hours of administration of the anti-human tau antibody. In some embodiments, the level of tau (eg, total tau and / or free tau) is significantly reduced within 24 hours of administration of the anti-human tau antibody. In some embodiments, the level of free e-tau is significantly reduced within 24 hours of administration of the anti-human tau antibody. In some cases, an effective amount of anti-human tau antibody is defined as within 48 hours, within 36 hours, within 24 hours, within 12 hours, within 8 hours, within 4 hours, within 2 hours, after administration of this anti-human tau antibody. An amount effective in significantly reducing the level of tau (eg, total tau and / or free tau) in extracellular fluid within 1 hour, 30 minutes, 15 minutes, or 5 minutes. be. For example, in some cases, an effective amount of anti-human tau antibody is 5 minutes to about 10 minutes, about 10 minutes to about 15 minutes, about 15 minutes to about 30 minutes, about 30 minutes to about 1 hour. Within, about 1 hour to about 2 hours, about 2 hours to about 4 hours, about 4 hours to about 8 hours, about 8 hours to about 12 hours, about 12 hours to about 24 hours, about 24 hours Significantly reduce the level of tau (eg, total tau and / or free tau) in extracellular fluid within ~ 36 hours, about 24 to about 48 hours, or about 36 hours to about 48 hours. It is an effective amount.

A significant reduction in the level of tau (eg, total tau and / or free tau) in an individual's extracellular fluid (eg, CSF, ISF, blood, or blood fraction (eg, serum or plasma)) is at least 30. % Reduction, at least 35% reduction, at least 40% reduction, at least 45% reduction, at least 50% reduction, at least 55% reduction, at least 60% reduction, at least 65% reduction, at least 70% reduction Reduction, at least 75% reduction, at least 80% reduction, at least 85% reduction, at least 90% reduction, at least 95% reduction, or more than 90% reduction. In some cases, a significant decrease is a statistically significant decrease. In some cases, a significant reduction is a clinically significant reduction. In some embodiments, the level of tau (eg, total tau and / or free tau) in extracellular fluid is reduced to normal control levels (eg, about 100 pg / ml). In some embodiments, the level of tau (eg, total tau and / or free tau) in extracellular fluid is reduced to undetectable levels. In some examples, the extracellular fluid is CSF. In another example, the extracellular fluid is ISF. In another example, the extracellular fluid is plasma. In another example, the extracellular fluid is whole blood. In another example, the extracellular fluid is serum.

In certain examples, the anti-human tau antibodies described herein are administered to a human subject at a fixed dose of 2000 mg. In certain examples, the anti-human tau antibody is administered to a human subject at a fixed dose of 2100 mg. In another example, the anti-human tau antibody is administered to a human subject at a fixed dose of 150 mg. In a further example, the anti-human tau antibody is administered to a human subject at a fixed dose of 4200 mg. In certain embodiments, the fixed doses of anti-human tau antibodies described herein are administered to a human subject once every four weeks.

In certain examples, the anti-human tau antibodies described herein are administered to a human subject as part of a pharmaceutical composition. In one embodiment, the pharmaceutical composition comprises 50 mg / mL of the anti-human tau antibody, 20 mM histidine, 250 mM sucrose, and 50 μM DTPA. The pharmaceutical composition has a pH of 6.0. In certain embodiments, the pharmaceutical composition is administered to a human subject in an amount sufficient to deliver a fixed dose of 2000 mg of anti-human tau antibody. In certain embodiments, the pharmaceutical composition is administered to a human subject in an amount sufficient to deliver a fixed dose of 2100 mg of anti-human tau antibody. In certain embodiments, the pharmaceutical composition is administered to a human subject in an amount sufficient to deliver 700 mg of a fixed dose of anti-human tau antibody. In certain embodiments, the pharmaceutical composition is administered to a human subject in an amount sufficient to deliver a fixed dose of 150 mg of anti-human tau antibody. In certain embodiments, the pharmaceutical composition is administered to a human subject in an amount sufficient to deliver a fixed dose of 210 mg of anti-human tau antibody. In certain embodiments, the pharmaceutical composition is administered to a human subject in an amount sufficient to deliver a fixed dose of 4200 mg of anti-human tau antibody.

In certain embodiments of these methods, the anti-human tau antibody is administered by an intravenous route to a human subject in need of the anti-human tau antibody.

In certain embodiments, the anti-human tau antibody comprises an immunoglobulin heavy chain variable region (VH) and an immunoglobulin light chain variable region (VL), where VH comprises VH complementarity determining regions (VH-CDR) and VH. -CDR1 contains or consists of the amino acid sequence of SEQ ID NO: 16, VH-CDR2 contains or consists of the amino acid sequence of SEQ ID NO: 17, and VH-CDR3 contains or consists of the amino acid sequence of SEQ ID NO: 18. VL-CDR1 comprises or comprises the amino acid sequence of SEQ ID NO: 19, VL-CDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 20 and VL-CDR3. Contains or consists of the amino acid sequence of SEQ ID NO: 21.

In certain embodiments, the VH of the anti-human tau antibody comprises or comprises SEQ ID NO: 12, and the VL comprises or comprises SEQ ID NO: 13.

In certain embodiments, the anti-human tau antibody comprises or comprises heavy and light chains, the heavy chain comprising or consisting of SEQ ID NO: 14, and the light chain comprising or consisting of SEQ ID NO: 15.

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

Example 1: Single escalating dose test of anti-human tau antibody BIIB092 BIIB092 is a humanized antibody that recognizes human extracellular tau (e-tau). The purpose of this study is to provide extracellular tau (e) in addition to the safety, tolerability and pharmacokinetics (PK) of BIIB092 after a single intravenous (IV) injection of BIIB092 into healthy human subjects. It was to evaluate the pharmacodynamic (PD) effect of BIIB092 on tau). In particular, BIIB092 was tested to measure the effect of BIIB092 on preventing the transmission of tau lesions by binding to and reducing free e-tau in human CSF.

The study was a randomized, double-blind, placebo-controlled, single-dose study. BIIB092 (6 subjects) or BIIB092 (6 subjects) in healthy subjects (ages: 21-65) in 6 incremental dose cohorts (21 mg, 70 mg, 210 mg, 700 mg, 2100 mg, and 4200 mg BIIB092), including 8 subjects per cohort. A single intravenous (IV) injection of placebo (2 subjects) was performed. See FIG. A safety assessment was performed over 12 weeks and serum and CSF samples (including 4 lumbar punctures) were collected. Pharmacokinetic parameters (in serum and CSF) and pharmacodynamic indicators (concentrations of free e-tau in CSF), as well as corresponding changes and rates of change from baseline were evaluated.

The increase in BIIB092 peak (Cmax) and exposure (AUC [INF]) in serum appeared to be dose-proportional. The elimination half-life of the terminal phase of BIIB092 was about 25 days. The CSF concentration of BIIB092 increased with dose and appeared to be dose-proportional. The CSF to serum ratio of BIIB092 was about 0.2%, similar between dose ranges. Most adverse events were mild. No serious adverse events or discontinuation due to adverse events were observed. The degree of e-tau suppression increased with dose, and the duration of e-tau suppression increased with dose. Suppression of e-tau in CSF on day 28 after a single dose of BIIB092 ranged from 65% to 96% at doses ranging from 70 mg to 4200 mg.

The ability of BIIB092 to strongly suppress CSF concentrations of free e-tau in this phase 1 study suggests that BIIB092 has demonstrated efficacy in the treatment of human tauopathy (eg, progressive supranuclear palsy). There is. A single dose of BIIB092 was safe at doses up to 4200 mg and was well tolerated.

Example 2: Bayesian Emax model An exposure-response model (Bayesian _Emax ) of CSF medium concentration vs. e-tau suppression was constructed (see FIG. 3). The Bayes E _max model captured the observed e-tau suppression reasonably well.

Example 3: Multiple doses of BIIB092, an anti-human tau antibody in patients with advanced nuclear palsy The purpose of this study is to inject BIIB092 intravenously (IV) into patients with advanced nuclear palsy (PSP). By assessing the safety, tolerability, pharmacokinetic (PK) and pharmacological (PD) effects of BIIB092 on free extracellular tau (e-tau) after injections are given every 4 weeks (Q4W). there were. Specifically, this study was designed to evaluate the safety profile of BIIB092 and the ability of BIIB092 to suppress free e-tau in CSF in patients with PSP.

The basic demographic characteristics for this study are shown in Figure 4. The study was a randomized, double-blind, placebo-controlled, multiple-dose study of 48 patients with PSP, of whom 12 (25%) received placebo. Three escalating dose panels (150 mg, 700 mg, and 2100 mg) containing 8 patients per panel were given IV injections of BIIB092 (6 patients) or placebo (2 patients) at Q4W for 12 weeks. Another 24 patients were treated with Q4W at a 2100 mg dose of BIIB092 (18 patients) or placebo (6 patients) for 12 weeks. See FIG. All patients were also offered the opportunity to participate in the open label expansion trial. A safety assessment was performed for 12 weeks and serum and CSF samples were collected. PK parameters (in serum and CSF) and PD indicators (concentration of free e-tau in CSF), as well as corresponding changes and rates of change from baseline were evaluated. Clinical efficacy indicators including PSP Racing Scale and Schwab and England Activities of Daily Living Scale were also adopted.

The average age of the patients was 67.4 ± 5.5 years, with 54.2% being female. Concentrations of BIIB092 in serum and CSF increased with dose. The percentage of patients who suffered an adverse event (AE) was similar (about 75%) in the BIIB092 and placebo groups. Most AEs were mild. See FIG. No death or discontinuation due to AE was observed. A list of serum PK parameters of BIIB092 is shown in FIG. The average suppression of free e-tau in CSF was about 90-96% (day 29) and 91-97% (day 85) at doses ranging from 150 mg to 2100 mg. Exposure to CSF and serum and suppression of free e-tau in CSF increased with the BIIB092 dose, whereas significant suppression of free e-tau in CSF was observed at all doses used in the study. See FIGS. 8 and 9.

Multiple doses of BIIB092 were safe and well tolerated at doses of up to 2100 mg in patients with PSP.

Example 4: Selection of BIIB092 dose based on simulated PK and e-tau suppression Using estimated PK-PD model parameters in PSP patients and variability around these parameters, 1000 profiles were simulated in serum. And the time course of PK in CSF and PD (e-tau) in CSF were obtained from two doses administered once every 4 weeks (Q4W), namely 2000 mg and 700 mg BIIB092. Before summarizing based on the simulation, the e-tau concentration was converted to the suppression rate based on the baseline value of each target. Table 1 below shows summary statistics of e-tau suppression rates relative to baseline levels at 4, 12, 24, and 48 weeks after the 2000 mg Q4W dosing regimen.
Table 1: Summary statistics of e-tau suppression rate based on baseline after 2000 mg Q4W (simulation)

Table 2 shows summary statistics of e-tau suppression rates relative to baseline levels at 4, 12, 24, and 48 weeks after the 700 mg Q4W dosing regimen.
Table 2: Summary statistics (simulation) of e-tau suppression rate based on baseline after 700 mg Q4W

Both the 2000 mg dose BIIB092 and the 700 mg dose BIIB092 administered in Q4W resulted in a strong suppression of e-tau in the trough. The 2000 mg dose of BIIB092 is involved in the trough with slightly higher inhibition (3-5%) compared to the 700 mg dose. It is expected that 90 percent of all subjects receiving the 2000 mg Q4W dose will exhibit an e-tau suppression rate of 96% or higher in the trough. It is expected that 90 percent of all subjects receiving the 700 mg Q4W dose will exhibit an e-tau suppression rate of 93% or higher in the trough.

Less than given the strong inhibition lasting up to 12 weeks after administration of doses of 210 mg and above tested, and the approximately 2 x higher CSF concentrations of BIIB092 observed in PSP patients compared to healthy subjects. BIIB092 may be administered on a frequency basis, eg, once every 12 weeks (Q12W). Simulations were performed using the same PK-PD model with a 2000 mg dose administered at Q12W. Table 3 summarizes e-tau suppression.
Table 3: Summary statistics (simulation) of e-tau suppression rate based on baseline after 2000 mg Q12W

It is expected that 90 percent of all subjects receiving the 2000 mg Q12W dose will exhibit an e-tau suppression rate of 86% or higher in the trough (ie, at the end of the 12-week dosing interval). However, subjects are expected to exhibit greater than 95% inhibition for the first month immediately following injection and slightly attenuated inhibition over the next two months. Overall, Q12W administration is also expected to be involved in the strong and sustained suppression of e-tau and may be favorable for patients and caregivers.

全ての製剤を、４０±２℃／７５±５％ＲＨで最長３ヶ月間にわたり、５±３℃及び２５±２℃／６０±５％ＲＨで最長１２ヶ月間にわたり保管した。初期の試料及び時点における試料について、外観によるＡＰＩ安定性、ｐＨ、Ａ_２８０、ＨＩＡＣ、ＳＥＣ、ＣＥＸ、及び力価ＥＬＩＳＡを評価した。初期、２ヶ月、及び１２ヶ月の時点にのみペプチドマッピング解析を実施した。３ヶ月の時点にのみマイクロフローイメージングを評価した。試験方法については表５に示しており、それぞれの時点における試験の一覧については表６に示している。
表５．ＢＩＩＢ０９２賦形剤最適化試験における解析検査

^１３×０．５ｍＬのランで検査を実施し、全３回のランに由来するデータを含めた
^２３ヶ月の時点にのみＭＦＩを試験し記録した
^３初期の時点、２ヶ月の時点、及び最終の時点にのみＥＬＩＳＡ検査及びペプチドマッピング検査を実施した
表６．ＢＩＩＢ０９２賦形剤最適化試験のプルスケジュール

Ａ＝ＭＦＩを除き表２に従った検査。
Ｂ＝ＥＬＩＳＡ、ＭＦＩ及びペプチドマップを除き表２に従った検査。
Ｃ＝ＥＬＩＳＡ及びペプチドマップを除き表２に従った検査。
Ｄ＝ＭＦＩ、ＥＬＩＳＡ及びペプチドマップを除き表２に従った製剤Ｅ１～Ｅ３、Ｅ７、及びＥ８の検査。
Ｅ＝ＭＦＩを除き表２に従った製剤Ｅ１～Ｅ３、Ｅ７、及びＥ８の検査。 Example 5: Optimization of Excipient Content in BIIB092 Formulation In this excipient optimization stabilization, either 0.05% PS-80 and 50 μM DTPA plus either 250 mM sucrose or 250 mM sorbitol. 10 mg / mL BIIB092 in 20 mM histidine buffer at pH 5.5, 6.0, and 6.5 was evaluated. In addition, 20 mg / mL and 50 mg / mL BIIB092 in 20 mM histidine at pH 6.0 containing 0.05% PS-80, 50 μM DTPA, and 250 mM sucrose were evaluated. A list of formulations is shown in Table 4.
Table 4. List of formulations for BIIB092 excipient optimization test

All formulations were stored at 40 ± 2 ° C./75 ± 5% RH for up to 3 months and at 5 ± 3 ° C. and 25 ± 2 ° C./60 ± 5% RH for up to 12 months. API stability, pH, A280, _HIAC , SEC, CEX, and titer ELISA by appearance were evaluated for early and current samples. Peptide mapping analysis was performed only at the initial, 2 and 12 month points. Microflow imaging was evaluated only at 3 months. The test methods are shown in Table 5, and the list of tests at each time point is shown in Table 6.
Table 5. Analytical inspection in BIIB092 excipient optimization test

Tests were performed on ¹³ x 0.5 mL runs and included data from all three runs.
MFI was tested and recorded ^only at 23 months
^3. The ELISA test and peptide mapping test were performed only at the initial time point, 2 months time point, and final time point. Table 6. BIIB092 Excipient Optimization Test Pull Schedule

Inspection according to Table 2 except A = MFI.
B = Testing according to Table 2 except for ELISA, MFI and peptide map.
Tests according to Table 2 except C = ELISA and peptide map.
Examination of formulations E1-E3, E7, and E8 according to Table 2 except for D = MFI, ELISA and peptide map.
Examination of formulations E1-E3, E7, and E8 according to Table 2 except E = MFI.

Appearance All samples were evaluated at all time points and conditions, except for one, which was a clear, colorless solution and contained no visible product-related particulates. It was confirmed that one sample (E3, 6 months, 25 ± 2 ° C / 60 ± 5% RH) contained a single large white particle, which was considered to be a contaminant. Further inspection of certain samples was stopped.

Measurement of pH All samples at all time points and conditions showed pH values within ± 0.1 pH units of the normal values for each pharmaceutical product. Therefore, all the differences observed in pH were within the variability of the method.

Protein content by UV / visible spectroscopy All samples at all time points and conditions showed protein concentrations not significantly different from their corresponding initial values. All the formulations (E1 to E6) targeting 10 mg / mL showed protein concentrations between 9.8 and 11.4 mg / mL. The measured concentration of the sorbitol preparation (E4 to E6) is higher than about 0.5 mg / mL compared to the concentration of the sucrose preparation (E1 to E3), but this simply reflects the slightly higher concentration due to sample preparation. Note that it is just doing. Formulation E7 (targeted at 20 mg / mL) ranges from 19.0 to 20.2 mg / mL, and E8 (targeted at 50 mg / mL) ranges from 50.0 to 53.7 mg / mL. rice field. The protein content by A280 showed that there was no significant trend in protein concentration over the time of stabilization of all the tested formulations.

Particle count (HIAC)
In all formulations, an increase in the number of larger particles (10 μm and 25 μm) was observed over the time of stabilization. However, there was no dependence on storage temperature in particle formation. The formulations E1 to E3 showed an overall increase in the count of large particles up to 12 months, and all differences among the three formulations were considered to be due to method variability. It should be noted that the absolute value of the count in the sample is considerable, although the tendency towards noise is not very strong. The increase in particles was particularly pronounced in the E7 and E8 formulations (20 mg / mL and 50 mg / mL APIs, respectively), with 10 μm particles in E7 being 9 months / 25 ° C./60% RH samples from the initial 12 counts. Particles of 10 μm in E8 increased from 18 counts at the initial time point to 1003 counts of 9 months / 25 ° C./60% RH sample. The particle counts of these two samples at other longer ranges of stabilization time points and conditions were also much higher than the initial particle counts. The difference between the storage temperatures is considered to be due to the variation in the method. For the formulations E4 to E6 tested only for 3 months, the particle count up to that point was equivalent to the particle count observed in E1 to E3 and less than the particle count observed in E7 to E8.

Size Exclusion Chromatography At each time point, a small but significant decrease in HMW percent was observed with all formulations as the stabilizing condition temperature increased. Generally, the temperature-dependent difference in HMW impurities was less in the 250 mM sucrose-prepared formulations (E1-E3) compared to the 250 mM sorbitol-prepared formulations (E4-E6). At 12 months, formulations E2 and E3 showed the highest main peak purity and showed the least aggregation and fragmentation of API in these formulations.

There is no convincing evidence that the HMW content is increased under any of the storage conditions in the time point data of each formulation. Problems in interpoint comparisons make further separate decisions difficult. However, if the performance of the reference standard in each sample queue is comparable, it is possible to make a constant comparison between time points. Two such time points were the initial and final (12 months) time points when all reference standard injections showed 1.5 ± 0.1% HMW. At these two time points, there was almost no difference in the HMW content of the test sample.

At each time point, the LMW percentage did not appear to change significantly (at certain formulations and time points / conditions there was an increase in the LMW percentage between 0.1 and 0.3% and no change in the LMW percentage was observed. Some are).

Cation exchange chromatography Acidic variant percent increased over time in all formulations, showing a greater increase with increasing temperature and appearing to decrease with lower pH. Conversely, the percentage of basic mutants increased over time and at higher stabilization temperatures, while showing fewer mutants as the pH increased. Either the acidic or basic variant percent in the sample formulated at higher concentrations (E7-20 mg / mL, E8-50 mg / mL) is 10 mg / mL at the same pH (6.0). There did not appear to be any significant difference compared to comparable samples.

Overall, the main peak percentages of the samples tested up to 12 months (E1 to E3, E7 to E8) are comparable to the main peak percentages of all concentrations of samples blended at pH 5.5 and 6.0. There seemed to be. The sample (E3) compounded at pH 6.5 showed a slightly lower main peak percentage compared to the main peak percentage of the lower pH sample (70 of E3 at 12 months / 25 ° C./60% RH). 6.6% vs. 76.9% of E2).

Titers by enzyme immunoassay The sample showed titers in the range of 82-126%. There did not appear to be any trend in the titer percentage for either the formulation type or the time of stabilization / condition.

Peptide mapping The peptide map was interpreted in light of the identification performed by LC / MS. Regarding the amide decomposition of asparagine at the light chain site N33 or N35, no significant difference was observed between the preparation stored at 5 ° C. and the frozen reference standard solution even up to 12 months. At room temperature (25 ° C./60% RH), trans-background amide degradation could be detected in the 2 month pH 6.5 formulations (E3 and E6). After 12 months, 25 ° C./60% RH, a significant pH-dependent mode of amide degradation (> 2% increase relative to the reference) was observed. Similar increases were observed after 2 months at 40 ° C./75% RH, with a clear dependence on pH, but the results for sorbitol and sucrose were comparable. It should be noted that the modification level of the reference standard solution measured by UV (6.1-8.3%) is reasonably similar to the modification level of the parent substance measured by LC / MS (3.0%). sea bream.

The amide degradation of asparagine at the heavy chain site N381 or N386 corresponded very similarly to the light chain amide degradation site. Even up to 12 months, no significant difference was observed between the pharmaceutical product stored at 5 ° C. and the frozen reference standard solution. At room temperature (25 ° C./60% RH), trans-background amide degradation could be detected in the 2 month pH 6.5 formulations (E3 and E6). After 12 months, 25 ° C./60% RH, a significant pH-dependent mode of amide degradation (> 2% increase relative to the reference) was observed. Similar increases were observed after 2 months at 40 ° C./75% RH, with a clear dependence on pH, but the results for sorbitol and sucrose were comparable. The modification level of the reference standard solution measured by UV (5.9 to 6.8%) was similar to the modification level of the parent substance measured by LC / MS (3.3%).

No significant difference was observed between the entire test and the frozen reference standard for amide degradation of asparagine at the heavy chain site N312. Even at 25 ° C./60% RH, 12 months and 40 ° C./75% RH, 2 months, the peak percentages were not significantly different from the frozen reference standard. This may indicate that this asparagine site is resistant to amide degradation, or that the presence of another species in the migration time of the peak identified as modified asparagine masks the amide degradation reaction. Supporting this theory, total N312 modifications were slightly higher in the reference standard solution by UV analysis (5.1-9.8%) compared to the parent material by LC / MS (2.8%). Since the succinimide morphology of the Asn / Asp intermediate is the most abundant in LC / MS data, the actual amide degradation at all time points may be simply low (<1%).

Hydroxylation of proline and lysine (although it was also quite abundant in the remixed sample) did not appear to change over time, regardless of storage temperature. Hydroxylation of P189 was extremely stable and unchanged at all time points. Interestingly, this proline hydroxylation was less in the frozen reference standard. The proline hydroxylation level (1.1-1.6%) of the reference standard solution measured by UV is reasonably similar to the proline hydroxylation level (1.0%) of the reference standard solution measured by LC / MS. rice field. The measurement of K121 hydroxylation produced a much larger difference in quantity. However, since no consistent trend could be observed, it was concluded that these differences indicate analytical noise. The lysine hydroxylation level (4.1-7.9%) of the reference standard solution measured by UV was reasonably similar to the lysine hydroxylation level (2.8%) of the parent substance measured by LC / MS. ..

Methionine oxidation at the heavy chain site M425 contributed significantly to the analysis. A slight but constant level of oxidation was observed in all formulations. Oxidation gradually increased as a function of time and temperature, but was unaffected by pH or the components of the pharmaceutical product. Even after 2 months at 40 ° C./75% RH, oxidation was only increased by about 0.5%. The modification level (0.2-0.6%) of the reference standard solution measured by UV was very similar to the modification level (0.3%) of the parent substance measured by LC / MS.

Particle count (microflow imaging)
Particle counting by MFI was performed only at 3 months. Overall, samples containing 250 mM sorbitol appeared to show lower particle counts for larger particle sizes (10 μm and 25 μm). Of the samples containing 250 mM sucrose, pharmaceutical E2 generally appeared to show a slightly lower particle count. There did not appear to be a significant difference in particle count between formulations with different API concentrations.

Other Embodiments Although the present invention has been described in relation to the embodiments for carrying out the invention, the above description is for illustration purposes only and does not limit the scope of the present invention. Is defined by the scope of the attached claims. Other aspects, advantages and modifications are within the scope of the following claims.

Claims (20)

A method for performing it in a human subject in need of treatment for tauopathy, said method comprising intravenously administering 2000 mg of a fixed dose of anti-human tau antibody to the human subject once every four weeks. , The anti-human tau antibody comprises an immunoglobulin heavy chain variable region (VH) and an immunoglobulin light chain variable region (VL).
(A) The VHs include VH complementarity determining regions (VH-CDRs).
VH-CDR1 consists of the amino acid sequence of SEQ ID NO: 16.
VH-CDR2 consists of the amino acid sequence of SEQ ID NO: 17.
VH-CDR3 consists of the amino acid sequence of SEQ ID NO: 18.
(B) The VL includes VL-CDR and contains.
VL-CDR1 consists of the amino acid sequence of SEQ ID NO: 19.
VL-CDR2 consists of the amino acid sequence of SEQ ID NO: 20.
VL-CDR3 consists of the amino acid sequence of SEQ ID NO: 21.
The method. The tauopathy includes Alzheimer's disease, muscular atrophic lateral sclerosis / Parkinson's dementia complex, silver granule dementia, British amyloid angiopathy, cerebral amyloid angiopathy, cortical basal nucleus degeneration, Kreuzfeld Jacob's disease, and boxer. Dementia, diffuse neurofibrillary tangle with calcification, Down's syndrome, frontotemporal dementia (FTD), frontotemporal dementia linked to chromosome 17 with parkinsonism, frontotemporal degeneration Disease, Gerstmannstroisler Scheinker's disease, Hallerfoldenspatz's disease, encapsulated myopathy, multilineage atrophy, muscle tonic dystrophy, C-type Niemannpic's disease, non-Guam-type motor neuron disease with neurofibrillary tangles, Pick's disease, post-encephalitis Parkinson's syndrome, prion protein cerebral amyloid angiopathy, progressive subcortical gliosis, progressive nuclear palsy, subacute sclerosing panencephalitis, neurofibrillary senile dementia, multiple infarct dementia, stroke, The method of claim 1, wherein the method is chronic traumatic encephalopathy, traumatic brain injury, tremor, epileptic attack, epilepsy, or acute lead encephalopathy. The method of claim 1, wherein the tauopathy is progressive supranuclear palsy. The method according to claim 1, wherein the tauopathy is Alzheimer's disease. The method according to any one of the preceding claims, wherein the VH comprises SEQ ID NO: 12 and the VL comprises SEQ ID NO: 13. The method according to any one of the preceding claims, wherein the anti-human tau antibody comprises a heavy chain and a light chain, wherein the heavy chain comprises SEQ ID NO: 14 and the light chain comprises SEQ ID NO: 15. (I) Anti-human tau antibody at a concentration of 50 mg / ml and
(Ii) With histidine at a concentration of 20 mM,
(Iii) With sucrose at a concentration of 250 mM,
(Iv) Polysorbate 80 at a concentration of 0.05% (weight / volume) and
(V) 50 μM diethylenetriamine pentaacetic acid (DTPA) and
A pharmaceutical composition containing
The anti-human tau antibody comprises an immunoglobulin heavy chain variable region (VH) and an immunoglobulin light chain variable region (VL).
(A) The VHs include VH complementarity determining regions (VH-CDRs).
VH-CDR1 consists of the amino acid sequence of SEQ ID NO: 16.
VH-CDR2 consists of the amino acid sequence of SEQ ID NO: 17.
VH-CDR3 consists of the amino acid sequence of SEQ ID NO: 18.
(B) The VL includes VL-CDR and contains.
VL-CDR1 consists of the amino acid sequence of SEQ ID NO: 19.
VL-CDR2 consists of the amino acid sequence of SEQ ID NO: 20.
VL-CDR3 consists of the amino acid sequence of SEQ ID NO: 21.
The composition has a pH of 6.0,
The pharmaceutical composition. The pharmaceutical composition according to claim 7, wherein the VH comprises SEQ ID NO: 12 and the VL comprises SEQ ID NO: 13. The pharmaceutical composition according to claim 7, wherein the anti-human tau antibody comprises a heavy chain and a light chain, the heavy chain comprises SEQ ID NO: 14, and the light chain comprises SEQ ID NO: 15. A method for performing this in a human subject in need of treatment for tauopathy, wherein the pharmaceutical composition according to any one of claims 7 to 9 is intravenously administered to the human subject. Said method comprising administering. The method of claim 10, wherein the anti-human tau antibody is administered at a fixed dose of 150 mg once every 4 weeks. The method of claim 10, wherein the anti-human tau antibody is administered at a fixed dose of 210 mg once every 4 weeks. The method of claim 10, wherein the anti-human tau antibody is administered at a fixed dose of 700 mg once every 4 weeks. The method of claim 10, wherein the anti-human tau antibody is administered at a fixed dose of 2000 mg once every 4 weeks. The method of claim 10, wherein the anti-human tau antibody is administered at a fixed dose of 2100 mg once every 4 weeks. The method of claim 10, wherein the anti-human tau antibody is administered at a fixed dose of 4200 mg once every 4 weeks. The method according to any one of claims 11 to 16, wherein the pharmaceutical composition is administered for at least 12 weeks. The tauopathy includes Alzheimer's disease, muscular atrophic lateral sclerosis / Parkinson's dementia complex, silver granule dementia, British amyloid angiopathy, cerebral amyloid angiopathy, cortical basal nucleus degeneration, Kreuzfeld Jacob's disease, and boxer. Dementia, diffuse neurofibrillary tangle with calcification, Down's syndrome, frontotemporal dementia (FTD), frontotemporal dementia linked to chromosome 17 with parkinsonism, frontotemporal degeneration Disease, Gerstmannstroisler Shineker's disease, Hallerfoldenspatz's disease, encapsulated myopathy, multilineage atrophy, muscle tonic dystrophy, C-type Niemannpic's disease, non-Guam-type motor neuron disease with neurofibrillary tangles, Pick's disease, post-encephalitis Parkinsonism, prion protein cerebral amyloid angiopathy, progressive subcortical gliosis, progressive nuclear palsy, subacute sclerosing panencephalitis, neurofibrillary senile dementia, multiple infarct dementia, stroke, The method according to any one of claims 10 to 17, wherein the method is chronic traumatic encephalopathy, traumatic brain injury, tremor, epileptic attack, epilepsy, or acute lead encephalopathy. The method according to any one of claims 10 to 17, wherein the tauopathy is progressive supranuclear palsy. The method according to any one of claims 10 to 17, wherein the tauopathy is Alzheimer's disease.

Images