Compound and method targeting interleukin-34

Abstract

To solve the problems described in the specifications (provided that there is no "Problem" section in the text).SOLUTION: The present disclosure relates to IL-34 antibodies, compositions comprising the same, and methods of using the antibodies and / or compositions thereof for treating immune-mediated diseases such as neurodegenerative diseases, for example, Alzheimer's disease or a tauopathy disease.SELECTED DRAWING: None

Description

Detailed Description of the Invention

[0001] The present disclosure relates to compounds, pharmaceutical compositions, and methods comprising antibodies against human interleukin-34 (IL-34), which are expected to be useful in the field of neuroinflammation and acute or chronic inflammatory diseases. In particular, embodiments are expected to be useful for therapeutic and / or diagnostic applications related to Alzheimer's disease, as well as other tauopathies.

[0002] Alzheimer's disease (AD), the leading cause of dementia, affects 1% of the population between the ages of 65 and 69, increasing to 40%-50% in those aged 95 and over. Patients with AD exhibit obvious clinical symptoms, including cognitive impairment and memory function deficits. In these patients, the presence of AD is confirmed by the severe senile plaque burden and neurofibrillary tangles (NFTs) found in the cerebral cortex on postmortem histopathological examination. Mature senile plaques are composed of extracellular β-amyloid peptides derived from enzymatic processing of amyloid precursor proteins, and intracellular neurofibrillary tangles (NFTs) derived from filaments of hyperphosphorylated tau protein. Aggregates of hyperphosphorylated tau, such as neurofibrillary tangles, are associated with the degree of cognitive impairment in Alzheimer's disease. In AD and various other tauopathies, tau aggregates appear in specific brain regions and patterns that are associated with disease risk, onset, and / or progression, and these regions and patterns are known to those skilled in the art.

[0003] Cytokines regulate normal homeostatic tissue function, and dysregulation of these cytokine networks is associated with pathological conditions. The central nervous system (CNS), with few circulating blood-derived immune cells, appears to be particularly vulnerable to dysregulated cytokine networks. In neurodegenerative diseases, CNS-resident cells are major producers of proinflammatory cytokines and may contribute to dysregulated cytokine networks and neuroinflammation. CNS injury may involve recruitment of circulating immune cells resulting in an innate immune response consisting of resident microglia, peripherally derived monocytes, macrophages, and dendritic cells. The activation state of microglia and macrophages is not strictly pro- or anti-inflammatory, but may instead have a range of functional states. Microglia and / or peripherally derived monocytes and macrophages may acquire an anti-inflammatory phenotype that clears necrotic debris and promotes regeneration and homeostasis. Neuronal dysfunction or injury may also activate microglia to produce proinflammatory cytokines and recruit leukocytes from the bloodstream. In neurodegenerative conditions such as Alzheimer's disease (AD), microglial activation is frequently observed, reflecting a tissue response to the accumulation of extracellular beta-amyloid plaques and hyperphosphorylated tau aggregates. Neuroinflammation is a key component of neurodegenerative diseases and is characterized by elevated production of proinflammatory cytokines by CNS cells (Becher, B., Spath, S. & Goverman, J. Cytokine networks in neuroinflammation. Nat Rev Immunol 17, 49-59 (2017)). Neuroinflammation and microgliosis are thought to be mechanisms underlying neurodegenerative diseases such as plaque accumulation in Alzheimer's disease and neuronal cell death and dysfunction in Parkinson's and Huntington's diseases.

[0004] Microgliosis involves abnormal proliferation and / or hypertrophy of microglia in response to inflammatory signals. Overall, IL-34 acts as a potent and pleiotropic cytokine in regulating inflammatory and immune processes and is a key regulatory cytokine for the proliferation of CNS-resident microglia in normal tissue homeostasis. IL-34 is expressed by neurons in the cortex, anterior olfactory nucleus, and hippocampus. IL-34 shows low sequence homology with CSF-1 but has a similar general structure, and both cytokines bind to a common receptor, CSF-1R, causing receptor autophosphorylation and dimerization, followed by activation of multiple signaling pathways (A.Freuchet,et al J Leukoc Biol 2021 Oct;110(4):771-796). IL-34 is a secreted homodimeric cytokine that acts as one of two activating ligands of CSF1R, causing autophosphorylation and dimerization of the receptor, followed by activation of multiple signaling pathways (see, for example, Structural basis of dual recognition of helical cytokines IL-34 and CSF-1 by CSF-1R, Structure 20, 676-687, and Felix J, De Munck S, Verstraete K, Meuris L, Callewaert N, Elegheert J. et al.). Human IL-34 polypeptide is disclosed, for example, in U.S. Patent No. 9,770,486, and consists of 242 amino acids with a leader sequence and 222 amino acids in the mature form (SEQ ID NO: 31).

[0005] Anti-IL-34 antibodies have been described in the art, for example WO2016 / 196679 details various anti-IL-34 antibodies and their potential uses, however, to date no antibody targeting IL-34 has been approved for therapeutic use.

[0006] Thus, there remains an unmet need for alternative and / or improved anti-IL-34 antibodies, pharmaceutical compositions thereof, and methods of using them for therapeutic and / or diagnostic applications related to immune-mediated diseases in which IL-34 is involved, and / or diseases treatable with anti-IL-34 antibodies, such as neuroinflammatory disorders, and / or Alzheimer's disease. Summary of the Invention

[0007] Embodiments of the present disclosure provide novel anti-human IL-34 antibodies. According to some embodiments, the disclosure provides antibodies comprising a light chain variable region (LCVR) and a heavy chain variable region (HCVR), wherein the LCVR comprises complementarity determining regions (CDRs), LCDR1, LCDR2, and LCDR3, and the HCVR comprises CDRs, HCDR1, HCDR2, and HCDR3, selected from the population of CDR combinations provided in Table 1. Sequence identifiers used herein are listed in Table 1 and throughout the specification, and sequences are set forth in the amino acid and nucleotide sequence tables provided herein. [Table 1]

[0008] Accordingly, embodiments of the present disclosure provide an antibody that binds to human IL-34, the antibody comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises heavy chain complementarity determining regions (HCDRs) HCDR1, HCDR2, and HCDR3, and the VL comprises light chain complementarity determining regions (LCDRs) LCDR1, LCDR2, and LCDR3, wherein HCDR1 comprises SEQ ID NO:5, HCDR2 comprises SEQ ID NO:6, HCDR3 comprises SEQ ID NO:7, LCDR1 comprises SEQ ID NO:8, LCDR2 comprises SEQ ID NO:9, and LCDR3 comprises SEQ ID NO:10.

[0009] Thus, embodiments of the present disclosure also provide an LCVR having the amino acid sequence of SEQ ID NO:4, and an HCVR having the amino acid sequence of SEQ ID NO:3.

[0010] Accordingly, embodiments of the present disclosure further provide an antibody that binds to human IL-34, wherein the antibody comprises a heavy chain (HC) comprising SEQ ID NO:1 and a light chain (LC) comprising SEQ ID NO:2.

[0011] According to other embodiments, the present disclosure also provides an antibody comprising an LCVR having the amino acid sequence of SEQ ID NO:4 and an HCVR having the amino acid sequence of SEQ ID NO:3, wherein the hinge and Fc regions are selected from SEQ ID NO:32 and SEQ ID NO:33.

[0012] As used herein, "Antibody 1" refers to an antibody having an HCDR1 amino acid sequence of SEQ ID NO:5, an HCDR2 amino acid sequence of SEQ ID NO:6, an HCDR3 amino acid sequence of SEQ ID NO:7, an LCDR1 amino acid sequence of SEQ ID NO:8, an LCDR2 amino acid sequence of SEQ ID NO:9, an LCDR3 amino acid sequence of SEQ ID NO:10, an HCVR amino acid sequence of SEQ ID NO:3, an LCVR amino acid sequence of SEQ ID NO:4, an HC amino acid sequence of SEQ ID NO:1, and an LC amino acid sequence of SEQ ID NO:2. Antibody 1 may be encoded by an HC DNA sequence of SEQ ID NO:11, and an LC DNA sequence of SEQ ID NO:12. The framework and CDR sequences in each of the antibodies whose sequences are presented herein have been annotated using annotation rules consistent with the method of North, et al.. J. Mol. Biol. 2011:406:228-256, unless otherwise indicated.

[0013] According to other embodiments, the present disclosure also provides an antibody comprising an LC having an amino acid sequence having at least 95% sequence identity to SEQ ID NO:2, and an HC having an amino acid sequence having at least 95% sequence identity to SEQ ID NO:1.

[0014] According to other embodiments, the present disclosure also provides an antibody comprising an LC having the amino acid sequence of SEQ ID NO:2 and an HC having the amino acid sequence of SEQ ID NO:35, further referred to herein as Antibody 2.

[0015] According to other embodiments, the present disclosure also provides an antibody comprising an LC having the amino acid sequence of SEQ ID NO:2 and an HC having the amino acid sequence of SEQ ID NO:36, further referred to herein as Antibody 3.

[0016] According to other embodiments, the present disclosure also provides an antibody comprising an LC having the amino acid sequence of SEQ ID NO:2 and an HC having the amino acid sequence of SEQ ID NO:37, further referred to herein as Antibody 4.

[0017] The carboxy-terminal portion of each HC defines a constant region primarily responsible for effector function, and in some embodiments of the disclosure, the antibody has one or more modifications in the constant region of each HC that reduce effector function. Preferably, embodiments of the disclosure are IgG4 antibodies and thus include an IgG4 Fc region, or an Fc region derived from human IgG4, such as a modified IgG4 Fc region.

[0018] According to some embodiments, modifications in the constant regions of both HCs that reduce effector function and amino acid substitutions are introduced into the IgG4 hinge and Fc regions. Thus, some embodiments have modifications in the constant regions of both HCs to include the amino acid alanine at both residues 230 and 231 (exemplified in the HC of Antibody 1 and SEQ ID NO: 33, respectively), and further modifications in the constant regions of both HCs to include the amino acid proline at residue 224 to promote stability (exemplified in the HC of Antibody 1 and for example in SEQ ID NO: 32), and deletion of the amino acid lysine at residue 443 (exemplified in the HC and SEQ ID NO: 1).

[0019] The antibodies of the present disclosure are believed to have a combination of particularly advantageous properties over prior art anti-IL-34 antibodies, including, but not limited to, one or more of the following properties: 1) desirable association and dissociation rates; 2) potency in neutralizing human IL-34 to achieve an anti-neuroinflammatory response and in vivo efficacy; 3) sufficient potency as a monotherapy for the treatment and / or prevention of immune-mediated and / or inflammatory disorders; 4) long duration of action; 5) sufficiently limited induction of undesirable cytokine release; 6) acceptably low immunogenicity (i.e., sufficiently non-immunogenic in humans); 7) avoidance of deleterious immune disorders; and / or 8) desirable in vivo stability, physical and chemical stability, including, but not limited to, thermal stability, solubility, low self-association, and acceptable pharmacokinetic properties for development and / or use in the treatment of inflammatory or neuroinflammatory disorders, such as AD. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0020] The present disclosure provides a significant advance over the prior art by providing compositions and methods useful for preventing, downregulating, or ameliorating inflammatory and / or neuroinflammatory related disorders through neutralization of IL-34 using pharmacologically advantageous anti-human IL-34 antibodies as provided in the embodiments described herein. The present disclosure's anti-human IL-34 antibodies can improve immune and / or inflammatory pathology or restore immune homeostasis, preferably through inhibition of the innate arm of the immune response and / or suppression of microgliosis or other monocytes. Directly modify the underlying disease pathology by activating and / or expanding cells of the macrophage lineage. Clinical use of such antibodies may lead to long-term persistence of the disease being treated.

[0021] Further, there is a need for diagnostic anti-human IL-34 antibodies that are specific for human IL-34, have improved binding affinity, and exhibit increased sensitivity in measuring human IL-34, as well as improved enzyme-linked immunosorbent assay (ELISA) assay conditions that result in minimal interference and broad dilution linearity. According to some aspects of the present disclosure, anti-human IL-34 antibodies, including human IL-34 neutralizing antibodies, that bind to human IL-34 as given by SEQ ID NO:31 are provided. Interleukin-34 (IL-34; also known as unidentified protein C16orf77) is secreted as a homodimer consisting of a 39 kDa monomer. It does not belong to any known cytokine family. Human IL-34 is synthesized as a 242 amino acid (AA) precursor that includes a 20 AA signal sequence, resulting in a 222 AA mature chain. As used herein, IL-34 refers to the mature chain. The mature chain contains one potential site for N-linked glycosylation. IL-34 is expressed in various tissues, such as heart, brain, liver, kidney, spleen, thymus, testis, ovary, small intestine, prostate, colon, and is most abundant in the spleen. "hIL-34" or "human IL-34", as used herein with respect to IL-34 polypeptide, refers to wild-type human IL-34, unless otherwise specified, and preferably has the amino acid sequence shown in SEQ ID NO: 31, which is mature IL-34 with the leader sequence removed. (See, e.g., Lin et.al., Science (2008) Vol. 320, Issue 5877, pp. 807-811).

[0022] An exemplary human IL-34 (SEQ ID NO:31) has the amino acid sequence: NEPLEMWPLTQNEECTVTGFLRDKLQYRSRLQYMKHYFPINYKISVPYEGVFRIANVTRLQRAQVSERELRYLWVLVSLSATESVQDVLLEGHPSWKYLQEVETLLLNVQQ GLTDVEVSPKVESVLSLLNAPGPNLKLVRPKALLDNCFRVMELLYCSCCKQSSVLNWQDCEVPSPQSCSPEPSLQYAATQLYPPPPWSPSSPPHSTGSVRPVRAQGEGLLP.

[0023] As used herein, "human anti-IL34 antibody" or "anti-human IL-34 antibody" refers to an antibody that binds to human IL-34. Preferably, the "human anti-IL34 antibody" or "anti-human IL-34 antibody" administered in vitro or in vivo results in IL-34 activity neutralization and / or response blocking, e.g., a desired reduction in IL-34 signaling, as evidenced by at least one significantly reduced desired activity, e.g., a change in an IL-34-responsive molecule or cellular endpoint. For example, the number, density, or phenotype of microglia in the central nervous system are examples of possible IL-34-responsive molecules or cellular effects. As used herein, the terms "signaling" and "signal transduction" and "IL-34-mediated" when related to IL-34, refer to cellular and / or intercellular responses resulting from the activity of IL-34.

[0024] As used herein, the term "antibody" refers to an immunoglobulin molecule that binds to an antigen. Antibody embodiments include monoclonal, polyclonal, human, humanized, chimeric, or conjugated antibodies. Antibodies may be of any class (e.g., IgG, IgE, IgM, IgD, IgA) and any subclass (e.g., IgG1, IgG2, IgG3, IgG4). An exemplary antibody is an immunoglobulin G (IgG) type antibody composed of four polypeptide chains: two heavy chains (HC) and two light chains (LC) cross-linked via interchain disulfide bonds. The LCs are classified as kappa or lambda, each of which is characterized by a specific constant region. Embodiments of the present disclosure may include IgG1, IgG2, or IgG4 antibodies and may further include a kappa or lambda light chain. Preferably, the antibodies of the present disclosure include a light chain constant region that is a kappa constant region.

[0025] HCs are classified as gamma, mu, alpha, delta, or epsilon, which define the antibody's isotype as IgG, IgM, IgA, IgD, or IgE, respectively. The amino-terminal portion of each of the four polypeptide chains contains a variable region of about 100-125 or more amino acids that is primarily responsible for antigen recognition. The carboxy-terminal portion of each of the four polypeptide chains contains a constant region that is primarily responsible for effector function. Each heavy chain is composed of a heavy chain variable region (VH) and a heavy chain constant region. The heavy chain constant region contains CH1, CH2, and CH3 domains. CH1 is after the HCVR, and CH1 and HCVR form the heavy chain portion of the antigen-binding (Fab) fragment, which is the part of the antibody that binds to the antigen. CH2 is after the hinge region and before CH3. CH3 is after CH2 and at the carboxy-terminus of the heavy chain. The light chain constant region contains one domain, CL. The CL is followed by the LCVR, and the CL and LCVR form the light chain portion of the Fab.

[0026] The antibodies of the present disclosure include an IgG HC, which can be further classified into subclasses, e.g., IgG1, IgG2, IgG3, IgG4, and embodiments of the present disclosure may include one or more modifications in the constant region of each HC, e.g., enhancing or reducing effector function. The term "Fc region" as used herein refers to the region of an antibody that includes the CH2 and CH3 domains of the antibody heavy chain. Optionally, the Fc region may include a portion of the hinge region or the entire hinge region of the antibody heavy chain. IgG1 is known to induce antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC), and the Fc mutations described herein may reduce aggregation, reduce or enhance ADCC or CDC activity (or other functions), and / or modify the pharmacokinetics of the antibody. Embodiments of the anti-human IL-34 antibodies described herein reduce binding to FcγR and C1q receptors, thereby reducing or eliminating cytotoxicity that can be induced by antibodies with wild-type IgG Fc regions. Thus, according to some embodiments, mutations are introduced into the Fc region at the positions described herein. By sufficiently reducing or eliminating effector function of such anti-human IL-34 antibodies comprising modified Fc regions, patient safety can be improved and, in combination with other properties described herein, can provide therapeutics with an improved profile of useful activity while avoiding undesirable activity.

[0027] When expressed in a particular biological system, an antibody is glycosylated in the Fc region. Typically, glycosylation occurs in the Fc region of an antibody at a highly conserved N-glycosylation site. N-glycans are typically linked to asparagine. Antibodies may also be glycosylated at other positions. The antibodies of the present disclosure are monoclonal antibodies. A monoclonal antibody is an antibody derived from a single copy or clone (including, for example, any eukaryotic, prokaryotic, or phage clone), not the method by which it is produced. Monoclonal antibodies can be produced, for example, by hybridoma technology, recombinant technology, phage display technology, synthetic technology, such as CDR grafting, or a combination of such techniques or other techniques known in the art. The present disclosure contemplates that the antibodies of the present disclosure are human or humanized antibodies. In the context of monoclonal antibodies, the terms "human" and "humanized" are familiar to those of skill in the art (Weiner LJ, J. Immunother. 2006;29:1-9; Mallbris L, et al., J. Clin. Aesthet. Dermatol. 2016;9:13-15). Exemplary embodiments of the antibodies of the present disclosure also include antibody or antigen-binding fragments that include at least a portion of an antibody that retains the ability to specifically interact with an antigen, such as Fab, Fab', F(ab')2, Fv fragments, scFv antibody fragments, disulfide-linked Fv (sdFv), Fd fragments, and linear antibodies.

[0028] The amino-terminal portion of each LC and HC contains a variable region of about 100-120 amino acids that is primarily responsible for antigen recognition via the CDRs contained therein. The VH and VL regions can be further subdivided into hypervariable regions, called complementarity determining regions (CDRs), interspersed with more conserved regions, called framework regions (FRs). The CDRs are exposed on the surface of the protein and are the critical regions of the antibody for antigen-binding specificity. Each VH and VL is composed of three CDRs and four FRs, arranged from the amino-terminus to the carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. Herein, the three CDRs of the heavy chain are referred to as "HCDR1, HCDR2, and HCDR3," and the three CDRs of the light chain are referred to as "LCDR1, LCDR2, and LCDR3." The CDRs contain the majority of the residues that form specific interactions with the antigen. The functional ability of an antibody to bind to a particular antigen is largely influenced by the six CDRs.The assignment of amino acid residues to CDRs has been reported by Kabat (Kabat et al., "Sequences of Proteins of Immunological Interest", National Institutes of Health, Bethesda, Md. (1991)), Chothia (Chothia et al., "Canonical structures for the hypervariable regions of immunoglobulins", Journal of Molecular Biology, 196, 901-917 (1987), Al-Lazikani et al., "Standard conformations for the canonical structures of immunoglobulins", Journal of Molecular Biology, 273, 927-948 (1997)), North (North et al., "A New Clustering of Antibody CDR Loop Conformations", Journal of Molecular Biology, 406, 228-256 (2011)), or IMGT (the international ImMunoGeneTics database available at www.imgt.org; see Lefranc et al., Nucleic Acids Res. 1999; 27:209-212).

[0029] For purposes of this disclosure, unless otherwise specified, the North CDR definitions are used for the assignment of amino acids to the CDR domains within the anti-IL-34 antibodies described herein and the LCVR and HCVR regions. Table 2 below provides the CDR sequences for Antibody 1 and / or antibodies of the present disclosure based on the North, Kabat, Chothia, and / or IMGT rules, respectively, generated using Benchling informatics software. [Table 2]

[0030] Antibody embodiments of the present disclosure have a combination of pharmacologically useful and important activities and properties, in some respects being able to bind human IL-34 with high affinity and high specificity, as well as other useful properties. As used herein, the term "bind" refers to the ability of a protein or molecule to form an attractive interaction with another protein or molecule, resulting in the proximity of the two proteins or molecules, as determined by common methods known in the art, unless otherwise indicated. The phrase "specifically binds" as used herein with respect to the affinity of an anti-IL-34 antibody for human IL-34, unless otherwise indicated, refers to a binding affinity of about 1×10, preferably about 1×10, as determined by common methods known in the art, including by the use of a surface plasmon resonance (SPR) biosensor, and / or solution equilibrium titration (SET) measured by a Meso Scale Discovery (MSD) instrument, essentially as described herein. -10 M or less, and even more preferably about 1×10 -10 M ~ approx. 1×10 -12 K of M D The phrase "specifically binds" is also intended to mean the relative affinity of an anti-IL-34 antibody for human IL-34 compared to other antigens, where affinity for human IL-34 results in specific recognition of human IL-34.

[0031] The antibody embodiments of the present disclosure can be expressed and produced by various techniques known in the art from constructs containing the sequences of the present embodiments. The terms "nucleic acid" or "polynucleotide", as used interchangeably herein, refer to polymers of nucleotides, including single-stranded and / or double-stranded nucleotide-containing molecules, such as DNA, cDNA, and RNA molecules, incorporating natural nucleotides, modified nucleotides, and / or nucleotide analogs. Polynucleotides of the present disclosure can also include substrates incorporated therein, for example, by DNA or RNA polymerase reactions or synthetic reactions. DNA molecules of the present disclosure are DNA molecules that include a non-naturally occurring polynucleotide sequence that encodes a polypeptide having the amino acid sequence of at least one of the polypeptides (e.g., heavy chain, light chain, variable heavy chain, and variable light chain) in the antibody of the present disclosure.

[0032] The isolated DNA encoding the HCVR or LCVR region can be converted into a full-length heavy chain gene by operably linking the DNA encoding the HCVR or LCVR to another DNA molecule encoding a heavy or light chain constant region, respectively, to form a heavy or light chain, respectively. The sequences of human and other mammalian heavy chain constant region genes are known in the art. DNA fragments encompassing these regions can be obtained, for example, by standard PCR amplification.

[0033] The polynucleotides of the present disclosure can be expressed in a host cell after the sequence is operably linked to an expression control sequence. Expression vectors are typically replicable in the host organism, either as episomes or as an integral part of the host chromosomal DNA. In general, expression vectors contain a selection marker, such as tetracycline, neomycin, and dihydrofolate reductase, to allow detection of those cells transformed with the desired DNA sequence. The vector containing the polynucleotide sequence of interest (e.g., a polynucleotide encoding an antibody polypeptide and an expression control sequence) can be introduced into the host cell by well-known methods that vary depending on the type of cellular host.

[0034] The antibodies of the present disclosure can be readily produced in mammalian cells, non-limiting examples of which include CHO, NS0, HEK293, or COS cells. The host cells are cultured using techniques well known in the art. Mammalian antibody expression typically results in glycosylation. Glycosylation of antibodies is usually either N-linked or O-linked. N-linked glycosylation refers to the attachment of a carbohydrate moiety to the side chain of an asparagine residue. O-linked glycosylation refers to the attachment of a sugar, such as N-acetylgalactosamine, galactose, or xylose, to a hydroxyamino acid. Typically, glycosylation occurs in the Fc region of an antibody at a highly conserved N-glycosylation site (e.g., position 297 in IgG1 according to the IMGT or EU index numbering). Glycosylation sites can be modified to alter glycosylation (e.g., to block or reduce glycosylation, or to change the amino acid sequence to generate additional or diverse glycosylation).

[0035] Expression of antibodies from the IgG subclass in a mammal can cause clipping of the C-terminal amino acids from one or both of the heavy chains, for example, in an IgG1 antibody, one or two C-terminal amino acids may be removed. In the case of an IgG1 antibody, the C-terminal lysine, if present, may be truncated or clipped from the heavy chain during expression. Additionally, the penultimate glycine may be similarly truncated or clipped from the heavy chain.

[0036] Expression of the antibody in a mammal may also result in modification of the N-terminal amino acid, for example, if the most N-terminal amino acid of the heavy or light chain is glutamine, it may be modified to pyroglutamic acid.

[0037] The antibody of the present disclosure, or pharmaceutical composition comprising the same, can be administered by parenteral route, non-limiting examples of which are subcutaneous administration and intravenous administration. The antibody of the present disclosure can be administered to a patient in a single dose or multiple doses together with a pharmaceutically acceptable carrier, diluent, or excipient. The pharmaceutical composition of the present disclosure can be prepared by a method well known in the art (e.g., Remington: The Science and Practice of Pharmacy, 22nd ed. (2012), A. Loyd et al., Pharmaceutical Press), and comprises an antibody disclosed herein and one or more pharmaceutically acceptable carriers, diluents, or excipients.

[0038] Uses of antibody embodiments of the invention: According to some embodiments, the anti-IL-34 antibodies of the present disclosure are useful for the treatment of immune-mediated diseases. As used herein, the terms "immune-mediated disease" or "inflammatory disease or disorder" are used interchangeably and refer to undesirable conditions resulting from inappropriate or excessive immune responses in which IL-34 inhibition results in more homeostasis and less pathological reactions. The terms "immune-mediated disease" or "inflammatory disorder" are meant to include such conditions, whether mediated by the cellular immune response of microglia or macrophages, or by the manifestation of similar tissue-resident cell types such as histiocytes, Kupffer cells, alveolar macrophages, intestinal macrophages, macrophage-like synoviocytes, or Langerhans cells. Exemplary diseases contemplated to be treated by the antibodies of the disclosure described herein include Alzheimer's disease, tauopathy, Sjogren's syndrome (SS), rheumatoid arthritis (RA), inflammatory bowel disease (IBD), atopic dermatitis, kidney disease, sepsis, amyotrophic lateral sclerosis (ALS), and / or non-alcoholic fatty liver disease (NAFLD).

[0039] In some more specific embodiments, the immune-mediated disease is Alzheimer's disease (AD). According to other embodiments of the present disclosure, the anti-IL-34 antibodies are useful for diagnostic applications of immune-mediated diseases. In some embodiments, the immune-mediated disease is at least one of AD, Sjogren's syndrome (SS), rheumatoid arthritis (RA), inflammatory bowel disease (IBD), atopic dermatitis, kidney disease, sepsis, and / or non-alcoholic fatty liver disease (NAFLD).

[0040] The present disclosure further provides a pharmaceutical composition comprising an anti-IL-34 antibody of the present disclosure and one or more pharma- ceutically acceptable carriers, diluents, or excipients.The present disclosure further provides a method of treating an immune-mediated disease, such as AD, Sjogren's syndrome (SS), rheumatoid arthritis (RA), inflammatory bowel disease (IBD), atopic dermatitis, renal disease, sepsis, and / or nonalcoholic fatty liver disease (NAFLD), comprising administering to a patient in need of treatment a pharmaceutical composition of the present disclosure.

[0041] In addition, the present disclosure provides methods of treating immune-mediated diseases, more specifically, the present disclosure provides methods of treating immune-mediated diseases, such as AD, Sjogren's syndrome (SS), rheumatoid arthritis (RA), inflammatory bowel disease (IBD), atopic dermatitis, kidney disease, sepsis, and / or non-alcoholic fatty liver disease (NAFLD), comprising administering to a patient in need of treatment an effective amount of an anti-IL-34 antibody of the present disclosure.

[0042] The present disclosure also provides an anti-IL-34 antibody of the present disclosure for use in therapy. More specifically, the present disclosure provides an anti-IL-34 antibody of the present disclosure for use in the treatment of immune-mediated diseases, including AD, Sjogren's syndrome (SS), rheumatoid arthritis (RA), inflammatory bowel disease (IBD), atopic dermatitis, kidney disease, sepsis, and / or non-alcoholic fatty liver disease (NAFLD).

[0043] In certain embodiments, the disclosure provides use of an anti-IL-34 antibody of the disclosure in the manufacture of a medicament for the treatment of one or more immune-mediated diseases, including AD, Sjogren's syndrome (SS), rheumatoid arthritis (RA), inflammatory bowel disease (IBD), atopic dermatitis, renal disease, sepsis, and / or non-alcoholic fatty liver disease (NAFLD).

[0044] The antibodies of the disclosure are useful for identifying immune-mediated disorders in which IL-34 may contribute to the pathogenesis of the disorder. In a further embodiment, the disclosure provides a method of treating an immune-mediated disease in a patient. Such a method includes contacting a patient sample with an IL-34 antibody and detecting binding between human IL-34 in the patient sample and the antibody, and diagnosing the patient as having, at risk for, in need of treatment for, and / or at risk for symptoms associated with an immune-mediated disease if the presence of IL-34 in the patient sample is detected above baseline levels observed in healthy individuals (see, e.g., Xie, HH, et al. Elevated Serum Interleukin-34 Level in Patients with Systemic Lupus Erythematosus Is Associated with Disease Activity. Sci Rep 2003, 144:1311-1323). 8,3462 (2018). According to some more specific embodiments of the therapeutic methods provided herein, such methods further include determining a reference value, further comprising contacting the control standard with a first antibody that binds to the same first epitope region of IL-34 as used in contacting the patient sample, contacting the control standard with a second antibody having a detectable label and that binds to the same second epitope region of IL-34 as used in contacting the patient sample, and detecting a signal provided by the detectable signal. In some specific embodiments, the anti-IL-34 antibody is an antibody that binds to the LC and HC as provided in Table 1. In further embodiments, the second antibody comprises a combination of LCVR and HCVR as provided in Table 1. According to some embodiments, the reference value is about 10-30 pg / mL, for example from a CNS tissue lysate. In certain embodiments, the immune-mediated disease is one of AD, Sjogren's syndrome (SS), rheumatoid arthritis (RA), inflammatory bowel disease (IBD), atopic dermatitis, renal disease, sepsis, and / or non-alcoholic fatty liver disease (NAFLD). In some embodiments, the patient sample is one of CSF, blood, serum, tissue lysate, or plasma.According to some embodiments, the method further comprises contacting the patient sample with a second anti-IL-34 antibody that binds to a second epitope region of IL-34 and has a detectable label, and detecting the signal provided by the detectable signal. In further embodiments, the second antibody comprises a combination of LC and HC CDRs provided in Table 1. In further embodiments, the second antibody comprises a combination of LCVR and HCVR provided in Table 1. According to certain embodiments, the first and second anti-IL-34 antibodies are not bottled together.

[0045] According to some embodiments, the present disclosure provides a method of detecting IL-34 in a patient sample, comprising contacting the patient sample with a first antibody that binds to a first epitope region of IL-34, contacting the patient sample with a second antibody that binds to a second epitope region of IL-34 and has a detectable label, and detecting a signal provided by the detectable label. In some embodiments, the patient sample is one of blood, serum, tissue lysate, or plasma. According to some more specific embodiments, the first epitope region of IL-34 overlaps with the second epitope region of IL-34. Moreover, in some embodiments, the steps of contacting with the first and second antibodies occur simultaneously. In some specific embodiments, the first antibody comprises a combination of LC and HC CDRs provided in Table 1. In further embodiments, the first antibody comprises a combination of LCVR and HCVR provided in Table 1.

[0046] According to some embodiments of the present disclosure, a method of quantifying IL-34 in a patient sample is provided. Such a method includes contacting the patient sample with a first antibody that binds to a first epitope region of IL-34, contacting the patient sample with a second antibody that binds to a second epitope region of IL-34 and has a detectable label, detecting a signal provided by the detectable label, contacting a control standard with a first antibody that binds to the same first epitope region of IL-34 (used to contact the patient sample), contacting the control standard with a second antibody that binds to the same second epitope region of IL-34 (used to contact the patient sample) and has a detectable label, and detecting a signal provided by the detectable label. In some embodiments, the patient sample is one of blood, serum or plasma, or tissue lysate. According to some more specific embodiments, the first epitope region of IL-34 overlaps with the second epitope region of IL-34. Further, in some embodiments, the steps of contacting with the first and second antibodies occur simultaneously. In some specific embodiments, the first antibody comprises a combination of LC and HC CDRs provided in Table 1. In further embodiments, the first antibody comprises a combination of LCVRs and HCVRs provided in Table 1. In some specific embodiments, the second antibody comprises a combination of LC and HC CDRs provided in Table 1 or herein. In further embodiments, the second antibody comprises a combination of LCVRs and HCVRs provided in Table 1.

[0047] According to some embodiments, a method for diagnosing an immune-mediated disease is provided. Such a method includes contacting a patient sample with an anti-IL-34 antibody and detecting binding between IL-34 in the patient sample and the antibody. According to some specific embodiments, the diagnostic method includes diagnosing the patient as having, at risk of, in need of treatment for, and / or at risk of symptoms associated with an immune-mediated disease if the presence of IL-34 in the patient sample is detected above the reference value. According to some more specific embodiments, such a method further includes determining the reference value, which further includes contacting the control standard with a first antibody that binds to a first epitope region of IL-34 as used in contacting the patient sample, contacting the control standard with a second antibody having a detectable label and that binds to a second epitope region of IL-34 as used in contacting the patient sample, and detecting a signal provided by the detectable signal. In some embodiments, the first antibody comprises a combination of LC and HC CDRs as provided in Table 1. According to some embodiments, the method further comprises contacting the patient sample with a second anti-IL-34 antibody that binds to a second epitope region of IL-34 and has a detectable label, and detecting the signal provided by the detectable signal. In some specific embodiments, the anti-IL-34 antibody comprises a combination of LC and HC CDRs as provided in Table 1. In further embodiments, the antibody comprises a combination of LCVR and HCVR as provided in Table 1. According to more specific embodiments, the first epitope region of IL-34 overlaps with the second epitope region of IL-34. According to certain embodiments, the first and second antibodies are not bottled together. According to further embodiments, the reference value is in the approximate range of about 10-30 pg / mL from CNS tissue lysates and / or as determined by one skilled in the art for the appropriate reference group and sample source.In further embodiments, the immune-mediated disease is one of AD, a tauopathy, Sjogren's syndrome (SS), rheumatoid arthritis (RA), inflammatory bowel disease (IBD), atopic dermatitis, renal disease, sepsis, and / or non-alcoholic fatty liver disease (NAFLD).

[0048] In one embodiment, the disclosure provides a method for determining human IL-34 levels in a body fluid, comprising: (a) contacting the body fluid with an anti-human IL-34 diagnostic monoclonal antibody or antigen-binding fragment thereof, which specifically binds to human IL-34, the antibody or antigen-binding fragment thereof having the amino acid sequence of SEQ ID NO:31 (comprising light chain complementarity determining regions LCDR1, LCDR2, and LCDR3 comprising amino acid sequences (SEQ ID NO:8), (SEQ ID NO:9), and (SEQ ID NO:10), respectively, and heavy chain complementarity determining regions HCDR1, HCDR2, and HCDR3 comprising amino acid sequences (SEQ ID NO:5), (SEQ ID NO:6), and (SEQ ID NO:7), respectively); (b) optionally removing non-specifically bound monoclonal antibody or antigen-binding fragment thereof; and (c) detecting and / or quantifying the amount of monoclonal antibody or antigen-binding fragment thereof specifically bound to human IL-34. Preferably, the body fluid is blood, serum or plasma, or cerebrospinal fluid, and the contacting occurs ex vivo.

[0049] Tauopathy diseases include, but are not limited to, Alzheimer's disease (AD), Pick's disease (PiD), progressive supranuclear palsy (PSP), corticobasal degeneration (CBD), argyrophilic grain dementia, Down's syndrome, chronic traumatic encephalopathy (CTE), traumatic brain injury (TBI), frontotemporal dementia with parkinsonism linked to chromosome 17 (FTDP-17), Parkinsonism-dementia complex of Guam, Niemann-Pick disease type C, and myotonic dystrophy (see Li, C., Gotz, J. Tau-based therapies in neurodegeneration: opportunities and challenges. Nat Rev Drug Discov 16, 863-883 (2017)).

[0050] In an embodiment of the present disclosure, the patient is a human diagnosed with a medical risk, condition, or disorder, such as one of the diseases or disorders described herein, that requires treatment with the antibody described herein. In cases where the disorder that can be treated by the method of the present disclosure is known by established and accepted classifications such as Alzheimer's disease, tauopathy disease, Sjogren's syndrome (SS), rheumatoid arthritis (RA), inflammatory bowel disease (IBD), atopic dermatitis, kidney disease, sepsis, and / or non-alcoholic fatty liver disease (NAFLD), those classifications are described in various well-known medical texts. For example, the Diagnostic and Statistical Manual of Mental Disorders, 5th Edition (DSM-5) currently provides diagnostic tools for identifying certain disorders described herein. Also, the International Classification of Diseases, 10th Edition (ICD-10) provides classifications for certain disorders described herein. Those of skill in the art will recognize that there are alternative nomenclatures, nosologies, and classification systems for the diseases and disorders described herein, including those described in DSM-5 and ICD-10, and that terminology and classification systems evolve as medical science advances.

[0051] The term "treating" (or "treat" or "treatment") refers to slowing, preventing, arresting, ameliorating, halting, reducing, or reversing the progression or severity of an existing symptom, disorder, condition, or disease in a subject. The term "subject" refers to a human. The terms "human subject" and "patient" are used interchangeably in this disclosure.

[0052] As used herein, "method of treatment" is equally applicable to the use of a composition to treat a disease or disorder as described herein, and / or the use of a composition in and / or for use in the manufacture of a medicament for treating a disease or disorder as described herein.

[0053] The terms "prevent" or "prevention" refer to the prophylactic administration of an antibody of the present disclosure to an asymptomatic subject or a subject suffering from preclinical Alzheimer's disease to prevent the onset or progression of Alzheimer's disease.

[0054] As used herein, the term "slow progression" means slowing or arresting the progression of a disease or a symptom thereof in a subject.

[0055] The term "disease characterized by deposition of Aβ" or "disease characterized by Aβ deposits" refers to a disease pathologically characterized by Aβ deposits in the brain or cerebral vasculature. This includes diseases such as Alzheimer's disease, Down's syndrome, cerebral amyloid angiopathy, etc. Clinical diagnosis, staging, or progression of Alzheimer's disease can be readily determined by the attending diagnostician or medical professional, such as one of ordinary skill in the art, by using known techniques and observing the results. This will generally involve brain plaque imaging, psychiatric or cognitive assessments (e.g., Clinical Dementia Rating-Summary of Boxes (CDR-SB), Mini-Mental State Examination (MMSE) or Alzheimer's Disease Assessment Scale-Cognitive (ADAS-Cog)), or functional assessments (e.g., Alzheimer's Disease Cooperative Study-Activities of Daily Living (ADCS-ADL). Cognitive and functional assessments can be used to determine changes in a patient's cognition (e.g., cognitive decline) and function (e.g., functional decline). Thus, a subject may be determined to have "slow" cognitive decline in accordance with the techniques described herein. In an exemplary embodiment, "slow" cognitive decline is when a subject's iADR decreases to about 20% or more over a predetermined period of time (e.g., 6, 12, 18 or 24 months). In another exemplary embodiment, "slow" cognitive decline can be identified by APOE-4 genotyping, where the subject is APOE-4 homozygous negative or APOE-4 heterozygous. In another exemplary embodiment, "slow" cognitive decline can be identified by MMSE, where the subject is determined to have an MMSE of about 27, or an MMSE decline of less than about 3 over a given period (e.g., 6, 12, 18, or 24 months). As used herein, "clinical Alzheimer's disease" is a diagnosed stage of Alzheimer's disease. This includes conditions diagnosed as prodromal Alzheimer's disease, mild Alzheimer's disease, moderate Alzheimer's disease, and severe Alzheimer's disease.The term "preclinical Alzheimer's disease" refers to the stage preceding clinical Alzheimer's disease, where measurable changes in biomarkers (such as CSF Aβ42 levels by amyloid PET or accumulated brain plaques) indicate the earliest signs of Alzheimer's disease patients progressing to clinical Alzheimer's disease. This is usually before symptoms such as memory loss and confusion become evident. Preclinical Alzheimer's disease includes presymptomatic autosomal dominant carriers as well as patients who are at high risk of developing AD because they carry one or two APOE e4 alleles.

[0056] The reduction or slowing of cognitive decline can be measured by cognitive assessments such as the Clinical Dementia Assessment-Summary of Boxes, Mini-Mental State Examination, or Alzheimer's Disease Assessment Scale-Cognition. The reduction or slowing of functional decline can be measured by functional assessments such as the ADCS-ADL.

[0057] As used herein, "mg / kg" refers to the amount of antibody or drug administered to a subject in milligrams based on body weight in kilograms. The dose is given at one time. For example, a 10 mg / kg dose of antibody to a subject weighing 70 kg is a single 700 mg dose of antibody administered in a single dose. Similarly, a 20 mg / kg dose of antibody to a subject weighing 70 kg is a single 1400 mg dose of antibody administered in a single dose.

[0058] As used herein, 18Using F-flortaucipir-based quantitative analysis, a human subject has a "very low tau" burden if the tau burden is less than 1.10 SUVr (<1.10 SUVr). Quantitative analysis refers to the calculation of SUVr, which represents counts within a specific target region of interest in the brain (Multiblock Centroid Discriminant Analysis or MUBADA, see Devous et al., "Test-Retest Reproducibility for the Tau PET Imaging Agent Flortaucipir F18," J. Nucl. Med. 59:937-943 (2018)) when compared to a reference region (Parametric Estimation of Reference Signal Intensity or PERSI, see Southekal et al., "Flortaucipir F 18 Quantitation Using Parametric Estimation of Reference Signal Intensity," J. Nucl. Med. 59:944-951 (2018)). As used herein, a human subject has a "very low to moderate tau" load if the tau load is 1.46 SUVr or less (i.e., 1.46 SUVr or less) using 18F-flortaucipir-based quantitative analysis. Quantitative analysis refers to the calculation of SUVr, which represents counts within a specific target region of interest in the brain when compared to a reference region (PERSI, Southekal et al., "Flortaucipir F 18 Quantitation Using Parametric Estimation of Reference Signal Intensity," J. Nucl. Med. 59:944-951 (2018)) (MUBADA, see Devous et al., "Test-Retest Reproducibility for the Tau PET Imaging Agent Flortaucipir F18," J. Nucl. Med. 59:937-943 (2018)).

[0059] As used herein, 18Using F-flortaucipir-based quantitative analysis, a human subject has a "low to moderate tau" load if the tau load is ≥1.10 - ≤1.46 (i.e., ≤1.10 SUVr - ≤1.46 SUVr). Quantitative analysis refers to the calculation of SUVr, which represents counts within a specific target region of interest in the brain when compared to a reference region (PERSI, see Southekal et al., "Flortaucipir F 18 Quantitation Using Parametric Estimation of Reference Signal Intensity," J. Nucl. Med. 59:944-951 (2018)) (MUBADA, see Devous et al., "Test-Retest Reproducibility for the Tau PET Imaging Agent Flortaucipir F18," J. Nucl. Med. 59:937-943 (2018)). Human subjects with a "low to moderate tau" load can also be referred to as having an "intermediate" tau load.

[0060] As used herein, 18 Using F-flortaucipir-based quantitative analysis, a human subject has a "high tau" burden if the tau burden is greater than 1.46 SUVr (i.e., >1.46 SUVr). Quantitative analysis refers to the calculation of SUVr, which represents counts within a specific target region of interest in the brain when compared to a reference region (PERSI, see Southekal et al., "Flortaucipir F 18 Quantitation Using Parametric Estimation of Reference Signal Intensity," J. Nucl. Med. 59:944-951 (2018)) (MUBADA, see Devous et al., "Test-Retest Reproducibility for the Tau PET Imaging Agent Flortaucipir F18," J. Nucl. Med. 59:937-943 (2018)).

[0061] As used herein, the term "about" means up to ±10%.

[0062] As used herein, the term "innate immunity" includes the arm of the immune response required to initiate and maintain the adaptive immune response (antibody and T cell responses), as opposed to the adaptive arm of the immune response.

[0063] By "effective amount" is meant an amount of an anti-human IL-34 antibody of the present disclosure, or a pharmaceutical composition comprising such an antibody, that will induce the biological or medical response of a tissue, system, or human being sought by a treating medical professional, or the desired therapeutic effect. As used herein, the term "effective response" of a patient, or the patient's responsiveness to a treatment, refers to the clinical or therapeutic benefit conferred upon the patient upon administration of an antibody of the present disclosure. An effective amount of an antibody may vary depending on factors such as the individual's medical condition, age, sex, and weight, and the ability of the antibody to induce a desired response in the individual. An effective amount is also an amount in which any toxic or detrimental effects of the antibody are outweighed by the therapeutically beneficial effects. Such benefits include any one or more of the following: reduced levels of inflammation or immune activation, stabilization of an immune-mediated disease or disorder, or amelioration of signs or symptoms of an immune-mediated disorder. Alternatively, such benefits include any one or more of the following: increased immune tolerance of a transplanted organ, stabilized autoimmune disease or disorder, or amelioration of signs or symptoms of an autoimmune disorder, and the like.

[0064] A potential advantage of the methods disclosed herein is the potential to provide significant and / or long-lasting relief in patients suffering from immune-mediated or neuroinflammatory disorders with acceptable safety profiles, including acceptable tolerability, toxicity, and / or adverse events, so that patients benefit from an overall therapeutic approach. The efficacy of the treatments of the present disclosure can be measured by a variety of endpoints commonly used in evaluating the treatment of various immune-mediated disorders. Other approaches to determining the efficacy of any particular therapy of the present disclosure can optionally be used, including, for example, immune cell activation markers, measures of inflammation, cell cycle-dependent biomarker measurements and visualization, and / or measurements of various inflammatory or immune-mediated responses, or tissue-specific biomarker assessments.

[0065] The effective amount can be easily determined by those skilled in the art by using known techniques and by observing results obtained under similar circumstances. An effective amount of the anti-human IL-34 antibody of the present disclosure can be administered in a single dose or multiple doses. Furthermore, an effective amount of the antibody of the present disclosure can be administered in multiple doses of an amount that is less than the effective amount if administered only once. In determining an effective amount for a patient, a number of factors are considered by the diagnostician in charge, including, but not limited to, the size (e.g., weight or mass), body surface area, age, and general health of the patient; the particular disease or disorder involved; the extent, or involvement, or severity of the disease or disorder; the response of the individual patient; the particular compound administered; the mode of administration; the bioavailability characteristics of the preparation administered; the selected dosing regimen; the use of concomitant pharmaceutical agents; and other relevant circumstances known to the diagnostician.

[0066] Weekly, biweekly, monthly, or quarterly parenteral (including but not limited to subcutaneous, intramuscular, and / or intravenous) doses can be from about 0.5 mg / kg to about 50 mg / kg. As used herein, "monthly" or derivatives thereof refers to a period of time that includes 28 to 31 consecutive days.

[0067] A potential advantage of the methods disclosed herein is the potential to provide significant and / or long-lasting relief in patients suffering from immune-mediated or neuroinflammatory disorders with acceptable safety profiles, including acceptable tolerability, toxicity, and / or adverse events, so that patients benefit from the overall treatment regimen; more specifically, the antibodies of the present disclosure provide effective treatment while avoiding clinically undesirable immune suppression and / or immune-related adverse events, such as "cytokine storm" or significant cytokine release. The antibodies of the present disclosure may be useful in treating cytokine storm, or harmful release of cytokines. As used herein, "significant cytokine release" refers to a significant increase in measurable cytokines that can be detected by methods known to those skilled in the art. For example, significant cytokine release can be detected in human blood samples by ELISA, where cytokine levels from unstimulated blood are compared to cytokine levels with blood incubated with the antibody. In some such tests, significant cytokine release can be detected, for example, when the levels of IL-6, or IL-8, or IFN-γ are at least three-fold higher in blood incubated with the antibody compared to levels in unstimulated blood. Preferably, treatment of an immune-mediated disorder as described in the embodiments herein occurs when the patient does not experience significant cytokine release.

[0068] Combination Uses of Antibodies of the Disclosure: The present disclosure further provides simultaneous, separate, or sequential combinations of the antibodies of the present disclosure, in particular antibody 1, and an anti-N3pGlu Aβ antibody, and methods of using the combination to treat diseases characterized by the deposition of amyloid beta (Aβ), such as AD. Some known anti-Aβ antibodies useful in the present combination include donanemab, bapineuzumab, gantenerumab, aducanumab, GSK933776, solanezumab, crenezumab, ponezumab, and lecanemab (BAN2401). The disclosure further provides simultaneous, separate, or sequential combinations of antibody 1 and donanemab (CAS No. 1931944-80-7, SEQ ID NOs: 38 and 39), and methods of using these combinations to treat diseases characterized by deposition of amyloid beta (Aβ), such as AD (Donanemab in Early Alzheimer's Disease, Mintun, MA et al, New England Journal of Medicine (2021), 384(18), 1691-1704). Preferably, the combination provides for the use of antibody 1 sequentially after a course of treatment with donanemab.

[0069] As used herein, "anti-N3pGlu Aβ antibody", "anti-N3pG antibody", or "anti-N3pE antibody" are used interchangeably and refer to an antibody that preferentially binds to N3pGlu Aβ over Aβ1-40 or Aβ1-42. Those skilled in the art will understand and appreciate that "anti-N3pGlu Aβ antibodies", as well as several specific antibodies, including "hE8L", "B12L" and "R17L", are identified and disclosed (along with methods of making and using) in U.S. Pat. No. 8,679,498 B2, which is incorporated herein by reference in its entirety. See, for example, Table 1 in U.S. Pat. No. 8,679,498 B2. Each of the antibodies disclosed in U.S. Pat. No. 8,679,498 B2, including the "hE8L", "B12L" and "R17L" antibodies, can be used as the anti-N3pGlu Aβ antibody of the present disclosure or in place of the anti-N3pGlu Aβ antibodies described in various aspects of the present invention. The anti-N3pGlu Aβ antibody of the present combination method is an antibody that comprises the HC and LC of SEQ ID NOs: 40 and 41, respectively.

[0070] Other representative types of anti-N3pGlu Aβ antibodies include, but are not limited to, those disclosed in U.S. Pat. No. 8,961,972, U.S. Pat. No. 10,647,759, U.S. Pat. No. 9,944,696, WO2010 / 009987A2, WO2011 / 151076A2, WO2012 / 136552A1 and equivalents thereof, such as antibodies disclosed under 35 U.S.C. § 112(f).

[0071] Those skilled in the art will understand and appreciate that "anti-N3pGlu Aβ antibodies," and several specific antibodies, are identified and disclosed (along with methods of making and using such antibodies) in U.S. Patent No. 8,961,972 (hereby incorporated by reference in its entirety), U.S. Patent No. 10,647,759 (hereby incorporated by reference in its entirety), and U.S. Patent No. 9,944,696 (hereby incorporated by reference in its entirety). Any of the anti-N3pGlu Aβ antibodies disclosed in U.S. Patent Nos. 8,961,972, 9,944,696, and 10,647,759 may be used as the anti-N3pGlu Aβ antibody of the present disclosure or in place of the anti-N3pGlu Aβ antibodies described in various aspects of the present invention.

[0072] Those skilled in the art will understand and appreciate that several specific antibodies, including "anti-N3pGlu Aβ antibodies," as well as "antibody VI," "antibody VII," "antibody VIII," and "antibody IX," have been identified and disclosed (along with methods of making and using such antibodies) in WO2010 / 009987A2, which is incorporated herein by reference in its entirety. Each of these four antibodies (e.g., "antibody VI," "antibody VII," "antibody VIII," and "antibody IX") can be used as or in place of the anti-N3pGlu Aβ antibodies of the present disclosure. N3pGlu Aβ antibodies are described in various aspects of the present invention.

[0073] Those skilled in the art will understand and appreciate that "anti-N3pGlu Aβ antibodies," as well as several specific antibodies, including "Antibody X" and "Antibody XI," have been identified and disclosed (along with methods of making and using such antibodies) in WO2011 / 151076A2, which is incorporated herein by reference in its entirety. Each of these two antibodies (e.g., "Antibody X" and "Antibody XI") may be used as the anti-N3pGlu Aβ antibody of the present disclosure or in place of the anti-N3pGlu Aβ antibodies described in various aspects of the present invention.

[0074] Those skilled in the art will understand and appreciate that several specific antibodies, including "anti-N3pGlu Aβ antibodies," as well as "Antibody XII" and "Antibody XIII," have been identified and disclosed (along with methods for making and using said antibodies) in WO2012 / 136552A1, which is incorporated herein by reference in its entirety. Each of these two antibodies (e.g., "Antibody XII" and "Antibody XIII") may be used as the anti-N3pGlu Aβ antibody of the present disclosure or in place of the anti-N3pGlu Aβ antibodies described in various aspects of the present disclosure.

[0075] Aspects of the present disclosure provide the use of a combination of an antibody of the present disclosure, particularly antibody 1, and an anti-N3pGlu Aβ antibody, particularly donanemab, for a method of treating a disease characterized by deposition of Aβ in a subject, the subject being selected based on i) tau levels / burden in the entire brain (global tau), ii) tau levels / burden in brain regions (e.g., different lobes of the brain), and / or the presence of one or two alleles of APOE e4 in the subject's genome. Diseases that can be treated or prevented using the combination methods disclosed herein include, for example, Alzheimer's disease (AD), Down's syndrome, and cerebral amyloid angiopathy (CAA). The present disclosure also relates to the use of the combination provided herein to slow disease progression in a subject with early symptomatic Alzheimer's disease (AD) in the presence of intermediate brain tau burden.

[0076] Antibodies against N3pGlu Aβ are known in the art. For example, U.S. Patent No. 8,679,498 (incorporated herein by reference in its entirety, including the anti-N3pGlu Aβ antibodies disclosed therein) discloses anti-N3pGlu Aβ antibodies and methods of treating diseases such as Alzheimer's disease with the antibodies. Passive immunization by long-term chronic administration of antibodies against Aβ, including N3pGlu Aβ, found in deposits, has been shown to disrupt Aβ aggregates in the brain in various animal models and promote plaque clearance. Donanemab (disclosed in U.S. Patent No. 8,679,498, see also CAS No. 1931944-80-7) is an antibody against a pyroglutamic acid modification of the third amino acid of the amyloid beta (N3pGlu Aβ) epitope, which is present only in amyloid plaques in the brain. The mechanism of action of donanemab is the targeting and removal of existing amyloid plaques, which are a key pathological feature of AD. The second neuropathological hallmark of AD is the presence of intracellular neurofibrillary tangles containing hyperphosphorylated tau protein. Aβ drives tau pathology, and more complex and synergistic interactions between Aβ and tau may emerge at later stages and promote disease progression (Busche et al., “Synergy Between Amyloid-β and Tau in Alzheimer's disease”, Nature Neuroscience 23:1183-93(2020)).

[0077] Administration of Aβ antibodies has resulted in adverse events in humans, including amyloid-related imaging abnormalities (ARIA), suggestive of vasogenic edema and crevicular effusion (ARIA-E), microhemorrhages and hemosiderin deposits (ARIA-H), injection site reactions, and the risk of immunogenicity. For example, Piazza and Winblad, “Amyloid-Related Imaging Abnormalities (ARIA) in Immunotherapy Trials for Alzheimer's Disease:Need for Prognostic Biomarkers?” Journal of Alzheimer's Disease, 52:417-420 (2016); Sperling, et al., “Amyloid-related Imaging Abnormalities in Patients with Alzheimer's Disease Treated with Bapineuzumab: A Retrospective Analysis,”The Lancet Neurology 11.3:241-249(2012);Brashear et al.,“Clinical Evaluation of Amyloid-related Imaging Abnormalities in Bapineuzumab Phase III Studies,”J.of Alzheimer's Disease 66.4:1409-1424(2018);Budd et al.,“Clinical Development of Aducanumab,an Anti-Aβ Human Monoclonal antibody See “Being Investigated for the Treatment of Early Alzheimer’s Disease,” The Journal of Prevention of Alzheimer’s Disease 4.4:255 (2017).

[0078] The disclosed combination treatment strategy for donanemab and antibody 1 involves targeting N3pGlu Aβ specific to amyloid plaques in a population of early symptomatic AD patients with existing brain amyloid burden, and targeting neuroinflammation in these patients. The rationale is based on the amyloid hypothesis of AD, which states that Aβ production and deposition are an early and necessary event in the pathogenesis of AD. See, e.g., Selkoe, “The Origins of Alzheimer Disease: A is for Amyloid,” JAMA 283:1615-1617 (2000). Clinical support for this hypothesis comes from the demonstration that parenchymal Aβ levels are elevated before AD symptoms appear, and is supported by genetic variants of AD that overproduce brain Aβ and genetic variants that prevent Aβ production. See, for example, Jonsson et al., “A Mutation in APP Protects Against Alzheimer's Disease and Age-related Cognitive Decline,” Nature 488(7409):96-99(2012) and Fleisher et al., “Associations Between Biomarkers and Age in the Presenilin 1 E280A Autosomal Dominant Alzheimer Disease Kindred: A Cross-sectional Study,” JAMA Neurol. 72:316-24(2015). Thus, there is a need for improved drug combinations to treat subjects without causing or increasing problematic adverse events. Neuroinflammation is a key component of neurodegenerative diseases and is characterized by increased production of proinflammatory cytokines by CNS cells. Neuroinflammation and microgliosis are believed to be mechanisms underlying Alzheimer's disease and / or neuronal cell death and dysfunction. Microgliosis involves abnormal proliferation and / or hypertrophy of microglia in response to inflammatory signals. IL-34 acts as a potent, pleiotropic cytokine in regulating inflammatory and immune processes and is expressed by neurons in the cortex, anterior olfactory nucleus, and hippocampus.Treatment with an N3pGlu Aβ antibody, particularly donanemab, followed by simultaneous, separate or preferably sequential treatment with Antibody 1 is believed to ameliorate the contribution of neuroinflammation and / or microgliosis to AD pathogenesis and slow or prevent the progression of the neurodegenerative process in these patients.

[0079] One aspect of the present disclosure is that Alzheimer's patients with low or moderate tau, very low to moderate tau, or no high tau respond to combination therapy with an anti-N3pGlu Aβ antibody, such as donanemab, and an antibody of the present disclosure, such as antibody 1. Another aspect of the present disclosure is based on the concept that Alzheimer's patients with one or two alleles of APOE e4 respond to treatment with an anti-N3pGlu Aβ antibody. Yet another aspect of the present disclosure is that Alzheimer's patients with one or two alleles of APOE e4 and low or moderate tau, very low to moderate tau, or no high tau respond to combination therapy with an anti-N3pGlu Aβ antibody, such as donanemab and an antibody of the present disclosure, such as antibody 1. Some aspects of the present disclosure are directed to diagnosing and treating patients based on their brain pathology. Selecting patients based on brain pathology not only provides a more homogenous population for clinical trials, but also allows for proper identification of the stage of AD and its progression. Proper identification of the stage of AD may allow, for example, timely referral to a memory clinic, accurate and early diagnosis of AD, initiation of symptomatic treatment, future planning, and even initiation of disease modifying treatment with combination therapy of an anti-N3pGlu Aβ antibody, such as donanemab, and an antibody of the disclosure, such as Antibody 1.

[0080] Some aspects of the present disclosure provide combination embodiments for treating a human subject suffering from a disease characterized by Aβ deposits in the brain, where the subject is first administered an anti-N3pGlu Aβ antibody, such as donanemab, administered in two stages in combination with simultaneous, separate, or sequential treatment with an antibody of the present disclosure, such as antibody 1. In the first stage, the human subject is administered one or more first doses of about 100 mg to about 700 mg of the anti-N3pGlu Aβ antibody, each first dose being administered about once every four weeks. About four weeks after the one or more administrations of the first doses, in a second step, the human subject is administered one or more second doses of greater than 700 mg to about 1400 mg, each second dose being administered about once every four weeks. Preferably, the anti-N3pGlu Aβ antibody is donanemab. Antibody 1 is administered simultaneously, separately, or sequentially after the course of treatment with donanemab. Preferably, antibody 1 is administered sequentially following a course of treatment with donanemab.

[0081] Some embodiments of the therapeutic combination methods relate to identifying the stage / progression of AD in a patient based on i) the overall or global tau burden in the brain of a human subject, or ii) the spread of tau in the subject's brain or regions or portions thereof.

[0082] In some embodiments, patients can be stratified / identified / selected / treated based on the amount of tau present in the subject's brain (e.g., whole brain or a portion of the brain). In some embodiments, patients can be stratified / identified / selected / treated based on the amount of tau present in the subject's brain (e.g., whole brain or a portion of the brain) and the presence of one or two alleles of APOE e4.

[0083] In other embodiments, patients are stratified / identified / selected / treated based on the stage of AD progression (e.g., based on the spread of tau in the brain). For example, in some stages, the tau burden in AD patients is isolated to regions of the frontal lobe or temporal lobe that does not include the posterior lateral temporal region (PLT). Another stage of AD is that the tau burden in AD patients is limited to the posterior lateral temporal (PLT) or occipital regions. Yet another stage of AD is that the tau burden in AD patients is present in the parietal or precuneus or frontal regions with tau burden in the PLT or occipital regions. In some embodiments, patients can be stratified / identified / selected / treated based on the stage of AD progression (e.g., based on the spread of tau in the brain) and based on the presence of one or two alleles of APOE e4.

[0084] Stratification of patients based on the amount of tau in the brain, the progression of AD in a portion of the brain, and / or the presence of one or two alleles of APOE e4 can be used to determine, for example, whether a patient will respond to a combination therapy of an anti-N3pGlu Aβ antibody, such as donanemab, and an antibody of the disclosure, such as antibody 1. Stratification / selection of patient populations based on the amount of tau in the brain, the progression of AD in a portion of the brain, and / or the presence of one or two alleles of APOE e4 also helps to solve the problems of patient heterogeneity and replicability faced during the design and execution of clinical trials in addition to treatment.

[0085] Another aspect of the disclosure provides for human subjects that respond to combination therapy or prevention of an anti-N3pGlu Aβ antibody, such as donanemab, and an antibody of the disclosure, such as antibody 1, for a disease characterized by amyloid beta (Aβ) deposits in the brain of the human subject. In some embodiments of this aspect of the disclosure, the responding human subjects include human subjects with low to moderate tau burden, very low to moderate tau burden, and / or one or two alleles of APOE e4. In some embodiments of this aspect of the disclosure, the responding human subjects exclude human subjects with high tau burden. In some embodiments of this aspect of the disclosure, the responding human subjects exclude human subjects with high tau burden and / or one or two alleles of APOE e4. In some embodiments, a combination of an anti-N3pGlu Aβ antibody, such as donanemab, and an antibody of the disclosure, such as antibody 1, is administered to the responding human subject for treatment or prevention of a disease characterized by amyloid beta (Aβ) deposits.

[0086] In one aspect, the disclosure relates to a simultaneous, separate or sequential combined treatment or prevention of a disease characterized by Aβ deposits in the brain of a human subject using an anti-N3pGlu Aβ antibody, particularly donanemab, and an antibody of the disclosure, particularly antibody 1, comprising: i) administering to the human subject one or more first doses of about 100 mg to about 700 mg of an anti-N3pGlu Aβ antibody, each first dose being administered about once every four weeks; and ii) about four weeks after administering the one or more first doses, administering to the human subject one or more second doses of greater than 700 mg to about 1400 mg of an anti-N3pGlu Aβ antibody, each second dose being administered about once every four weeks, the anti-N3pGlu Aβ antibody comprising donanemab, and administering to the human subject an antibody of the disclosure, particularly antibody 1. Preferably, antibody 1 is administered sequentially after a course of treatment with donanemab.

[0087] To date, the clinical focus of treatment with donanemab has been limited to early symptomatic AD patients with pre-existing cerebral amyloid burden. However, a second neuropathological hallmark of AD is the presence of intracellular neurofibrillary tangles containing hyperphosphorylated tau protein. Current disease models suggest that Aβ drives tau pathology and that more complex and synergistic interactions between Aβ and tau emerge at later stages, driving disease progression (Busche et al., “Synergy Between Amyloid-β and Tau in Alzheimer's disease”, Nature Neuroscience 23:1183-93(2020)).

[0088] Currently, there is no disease modifying treatment for AD. Thus, there is a need for improved methods of treating diseases, including AD, characterized by the deposition of Aβ in human subjects. Such methods should be useful for identifying patients based on whether they may have therapeutic benefit from such treatment. Furthermore, such treatments and methods should not be associated with increased cytotoxicity or other known adverse events. The present disclosure meets one or more of these needs.

[0089] Doody et al., “Phase 3 Trials of Solanezumab for Mild-to-Moderate Alzheimer's Disease,” NEJM, 370;4, 311-321 (2014) indicate that “no clear differential treatment effects on efficacy measures were observed between APOE ε4 carriers and non-carriers.” It is believed that administering an anti-N3pGlu Aβ antibody in combination with an antibody of the present disclosure to a human subject having one or two APOE e4 alleles (e.g., APOE e4 carriers) will provide unexpected efficacy when compared to non-carriers of one or more of those alleles. Thus, the present embodiments include administering an anti-N3pGlu Aβ antibody, particularly donanemab, in combination with an antibody of the present disclosure, particularly antibody 1, to patients having one or two APOE e4 alleles as a means of slowing cognitive decline in those patients.

[0090] According to certain embodiments, the present disclosure provides a method of treating or preventing a disease characterized by amyloid beta (Aβ) deposits in the brain of a human subject determined to have a high neurological tau burden, comprising administering, in simultaneous, separate, or sequential doses, a therapeutically effective amount of an anti-Aβ antibody, particularly donanemab, and a therapeutically effective amount of an antibody of the present disclosure, particularly antibody 1. Additionally, according to certain embodiments, the present disclosure provides a combination method of treating or preventing a disease characterized by Aβ deposits in the brain of a human subject determined to have a posterior lateral temporal lobe tau burden, comprising administering, in simultaneous, separate, or sequential doses, a therapeutically effective amount of an anti-Aβ antibody, particularly donanemab, and a therapeutically effective amount of an antibody of the present disclosure, particularly antibody 1.

[0091] According to certain embodiments, the present disclosure provides a combination method for treating or preventing a disease characterized by Aβ deposits in the brain of a human subject determined to have a high neurological tau burden and one or two alleles of the epsilon 4 allele of apolipoprotein E (referred to herein as APOE e4 or APOE4), comprising administering, in simultaneous, separate, or sequential doses, a therapeutically effective amount of an anti-Aβ antibody, particularly donanemab, and a therapeutically effective amount of an antibody of the present disclosure, particularly antibody 1. Additionally, according to certain embodiments, the present disclosure provides a method for treating or preventing a disease characterized by Aβ deposits in the brain of a human subject determined to have a posterior lateral temporal lobe tau burden, comprising administering, in simultaneous, separate, or sequential doses, a therapeutically effective amount of an anti-Aβ antibody, particularly donanemab, and a therapeutically effective amount of an antibody of the present disclosure, particularly antibody 1.

[0092] According to some embodiments, the present disclosure provides an anti-Aβ antibody, particularly donanemab, for simultaneous, separate or sequential use with an antibody of the present disclosure, particularly antibody 1, for the treatment or prevention of a disease characterized by Aβ deposits in the brain of a human subject determined to have a high neurological tau burden, comprising administering a therapeutically effective amount of an anti-Aβ antibody, particularly donanemab, and a therapeutically effective amount of an antibody of the present disclosure, particularly antibody 1, with simultaneous, separate or sequential doses of a therapeutically effective amount of anti-Aβ. In some embodiments, the human subject has been determined to have a high neurological tau burden and has one or two alleles of APOE e4.

[0093] In some embodiments, the present disclosure provides an anti-Aβ antibody, particularly donanemab, for simultaneous, separate, or sequential use with an antibody of the present disclosure, particularly antibody 1, for the treatment or prevention of a disease characterized by Aβ deposits in the brain of a human subject determined to have a posterior lateral temporal lobe tau burden. In some embodiments, the human subject has been determined to have a posterior lateral temporal lobe tau burden and to have one or two alleles of APOE e4.

[0094] Additionally, in some embodiments, the present disclosure provides anti-Aβ antibodies, particularly donanemab, for simultaneous, separate, or sequential use with an antibody of the present disclosure, particularly antibody 1, to treat, prevent, or slow the progression of Alzheimer's disease (AD). Additionally, in some embodiments, the present disclosure provides anti-Aβ antibodies, particularly donanemab, for simultaneous, separate, or sequential use with an antibody of the present disclosure, particularly antibody 1, to treat, prevent, or slow the progression of Alzheimer's disease (AD) in a human subject determined to have slowly progressing AD cognitive decline. Some embodiments of the present disclosure provide anti-Aβ antibodies, particularly donanemab, for simultaneous, separate, or sequential use with an antibody of the present disclosure, particularly antibody 1, to treat, prevent, or slow the progression of Alzheimer's disease (AD) in a human subject determined to have slowly progressing AD cognitive decline and to have one or two alleles of APOE e4.

[0095] Further, according to some embodiments, the present disclosure provides for the use of an anti-Aβ antibody, particularly donanemab, in combination simultaneously, separately, or sequentially with an antibody of the present disclosure, particularly antibody 1, in the manufacture of a medicament for the treatment or prevention of Alzheimer's disease. Further, according to some embodiments, the present disclosure provides for the use of an anti-Aβ antibody, particularly donanemab, in combination simultaneously, separately, or sequentially with an antibody of the present disclosure, particularly antibody 1, in the manufacture of a medicament for the treatment or prevention of a disease characterized by i) high neurological tau burden, or ii) high neurological tau burden and Aβ deposits in the brain of a human subject determined to have one or two alleles of APOE e4.

[0096] In some embodiments, the present disclosure provides for the use of an anti-Aβ antibody, particularly donanemab, in combination with, simultaneously, separately, or sequentially, an antibody of the present disclosure, particularly antibody 1, in the manufacture of a medicament for the treatment or prevention of a disease characterized by i) posterior lateral temporal lobe tau burden, or ii) posterior lateral temporal lobe tau burden and Aβ deposits in the brain of a human subject determined to have one or two alleles of APOE e4. In further embodiments, the present disclosure provides for the use of an anti-Aβ antibody, particularly donanemab, in combination with, simultaneously, separately, or sequentially, an antibody of the present disclosure, particularly antibody 1, in the manufacture of a medicament for treating, preventing, or slowing the progression of Alzheimer's Disease (AD) in a human subject determined to have i) a slowly progressing AD cognitive decline, or ii) a slowly progressing AD cognitive decline, having one or two alleles of APOE e4.

[0097] According to some embodiments provided herein, the human subject is determined to have a tau load in the posterior lateral temporal lobe and the occipital lobe. In some embodiments, the human subject is determined to have a tau load in the posterior lateral temporal lobe, the occipital lobe, and the parietal lobe. In some embodiments, the human subject is determined to have a tau load in the posterior lateral temporal lobe, the occipital lobe, the parietal lobe, and the frontal lobe. In some embodiments, the human subject is determined to have one or more of the tau loads in the posterior lateral temporal lobe, the occipital lobe, the parietal lobe, and / or the frontal lobe by neurological PET imaging. In some embodiments, one or more of the tau loads in the posterior lateral temporal lobe, the occipital lobe, the parietal lobe, and / or the frontal lobe correspond to a neurological tau load of more than 1.46 SUVr.

[0098] According to some embodiments provided herein, a human subject is determined to have one or two alleles of APOE e4 and a tau load in the posterior lateral temporal lobe and the occipital lobe. In some embodiments, a human subject is determined to have one or two alleles of APOE e4 and a tau load in the posterior lateral temporal lobe, the occipital lobe and the parietal lobe. In some embodiments, a human subject is determined to have one or two alleles of APOE e4 and a tau load in the posterior lateral temporal lobe, the occipital lobe, the parietal lobe and the frontal lobe. In some embodiments, a human subject is determined to have one or more of the posterior lateral temporal lobe, the occipital lobe, the parietal lobe and / or the frontal lobe tau load by neurological PET imaging and one or two alleles of APOE e4. In some embodiments, one or more of the tau load in the posterior lateral temporal lobe, the occipital lobe, the parietal lobe and / or the frontal lobe corresponds to a neurological tau load of more than 1.46 SUVr.

[0099] According to additional embodiments, the present disclosure provides a method of treating, preventing, or slowing the progression of Alzheimer's Disease (AD) in a human subject determined to have slowly progressing AD cognitive decline, comprising administering simultaneous, separate, or sequential doses of a therapeutically effective amount of an anti-Aβ antibody, particularly donanemab, and a therapeutically effective amount of an antibody of the present disclosure, particularly antibody 1. According to some embodiments, the human subject is determined to have a high neurological tau burden. According to some embodiments, the human subject is determined to have one or two alleles of APOE e4. In some embodiments, the human subject is determined to have a posterior lateral temporal lobe tau burden. In some embodiments, the human subject is determined to have a posterior lateral temporal lobe and occipital lobe tau burden. In some embodiments, the human subject is determined to have a posterior lateral temporal lobe, occipital lobe, and parietal lobe tau burden. In some embodiments, the human subject is determined to have a tau load in the posterior lateral temporal lobe, the occipital lobe, the parietal lobe, and the frontal lobe. In some embodiments, the human subject is determined to have a tau load in the posterior lateral temporal lobe and one or two alleles of APOE e4. In some embodiments, the human subject is determined to have one or two alleles of APOE e4 and a tau load in the posterior lateral temporal lobe and the occipital lobe. In some embodiments, the human subject is determined to have one or two alleles of APOE e4 and a tau load in the posterior lateral temporal lobe, the occipital lobe, and the parietal lobe. In some embodiments, the human subject is determined to have one or two alleles of APOE e4 and a tau load in the posterior lateral temporal lobe, the occipital lobe, and the parietal lobe.

[0100] According to embodiments of the disclosure provided herein, the human subject has been determined to have slowly progressing AD cognitive decline by one or more of ADAS-Cog, iADL, CDR-SB, MMSE, APOE-4 genotyping, and / or iADRS. In some embodiments, the human subject has been determined to have slowly progressing AD cognitive decline by iADRS. In some embodiments, the iADRS has been reduced by less than 20. In some embodiments, the iADRS has been reduced by less than 20 over 6 months. In some embodiments, the iADRS has been reduced by less than 20 over 12 months. In some embodiments, the iADRS has been reduced by less than 20 over 18 months. In some embodiments, the iADRS has been reduced by less than 20 over 24 months. In some embodiments, the human subject has been determined to have slowly progressing AD cognitive decline by APOE-4 genotyping. In some embodiments, the human subject has been determined to be APOE-4 heterozygote. In some embodiments, the human subject has been determined to be APOE-4 homozygote negative. In some embodiments, the human subject is determined to have slow AD cognitive decline by MMSE. In some embodiments, the human subject is determined to have an MMSE of greater than 27. In some embodiments, the MMSE has decreased by less than 3. In some embodiments, the MMSE has decreased by less than 3 over 6 months. In some embodiments, the MMSE has decreased by less than 3 over 12 months. In some embodiments, the MMSE has decreased by less than 3 over 18 months. In some embodiments, the MMSE has decreased by less than 3 over 24 months.

[0101] According to embodiments of the disclosure provided herein, a human subject has a high neurological tau burden determined by neurological PET imaging. In some embodiments, a human subject has a high neurological tau burden of greater than 1.46 SUVr determined by neurological PET imaging. In some embodiments, a human subject has a high neurological tau burden determined by quantification of human tau phosphorylated at threonine residue 217 ("hTau-pT217"). In some embodiments, hTau-pT217 is quantified in a biological sample of the human subject. In some embodiments, the biological sample is cerebrospinal fluid. In some embodiments, the biological sample is one of blood, plasma, or serum.

[0102] For purposes of the present invention, a human subject's tau level or burden (used interchangeably herein) can be determined, for example, using techniques or methods that detect or quantify i) neurological or brain tau deposits, ii) tau in blood, serum and / or plasma, or iii) tau in cerebrospinal fluid. In some embodiments, neurological tau burden (whether determined via PET or via blood, serum, plasma or cerebrospinal fluid assays) can be used to stratify subjects based on neurological tau burden (e.g., low, moderate, or high neurological tau burden).

[0103] Neurological tau burden can be determined using methods such as tau imaging with radiolabeled PET compounds (Leuzy et al., “Diagnostic Performance of RO948 F18 Tau Positron Emission Tomography in the Differentiation of Alzheimer Disease from Other Neurodegenerative Disorders,” JAMA Neurology 77.8:955-965(2020); Ossenkoppele et al., “Discriminative Accuracy of [ 18F]-flortaucipir Positron Emission Tomography for Alzheimer Disease vs Other Neurodegenerative Disorders,” JAMA 320, 1151-1162, doi:10.1001 / jama.2018.12917(2018), and PET ligand [ 18 F]-florbtaucipir), which are incorporated by reference in their entirety. PET tau images can be used to estimate SUVr (standardized uptake value) by published methods, for example (Pontecorvo et al., “A Multicentre Longitudinal Study of Flortaucipir (18F) in Normal Ageing, Mild Cognitive Impairment and Alzheimer's Disease Dementia,” Brain 142:1723-35 (2019); Devous et al., “Test-Retest Reproducibility for the Tau PET Imaging Agent Flortaucipir F18,” Journal of Nuclear Medicine 59:937-43 (2018); Southekal et al., “Flortaucipir F18 Quantitation Using Parametric Estimation of Reference Signal 59:944-51 (2018), which is incorporated by reference in its entirety), and / or can be quantitatively assessed to visually assess the patient, e.g., to determine whether the patient has an AD pattern (Fleisher et al., “Positron Emission Tomography Imaging With 18F]-flortaucipir and Postmortem Assessment of Alzheimer Disease Neuropathologic Changes,” JAMA Neurology 77:829-39 (2020), incorporated herein by reference in its entirety). Lower SUVr values ​​indicate lower tau burden, and higher SUVr values ​​indicate higher tau burden. In one embodiment, quantitative assessment of flortaucipir scans is achieved by an automated image processing pipeline as described below (Southekal et al., “Flortaucipir F18 Quantitation Using Parametric Estimation of Reference Signal Intensity,” J. Nucl. Med. 59:944-951 (2018), incorporated herein by reference in its entirety). In some embodiments, counts within specific target regions in the brain are assessed (e.g., using multiblock centroid discriminant analysis or MUBADA, Devous et al., “Test-Retest Reproducibility for the Tau PET Imaging Agent Flortaucipir F18," J. Nucl. Med. 59:937-943 (2018) (incorporated herein by reference in its entirety), compared to a reference region, e.g., whole Cerebellum (wholeCere), cerebellar GM (cereCrus), atlas-based white matter (atlasWM), subject-specific WM (ssWM, e.g., using Parametric Estimation of Reference Signal Intensity (PERSI)), see Southekal et al., "Flortaucipir F18 Quantitation Using Parametric Estimation of Reference Signal Intensity," J. Nucl. Med. 59:944-951 (2018) (incorporated herein by reference in its entirety). An exemplary method for determining tau burden is a quantitative analysis reported as standardized uptake value ratio (SUVr).This represents counts within a specific target region of interest in the brain (e.g., MUBADA) when compared to a reference region (e.g., using PERSI).

[0104] In some embodiments, phosphorylated tau (P-tau, phosphorylated at threonine 181 or 217, or a combination thereof) can be used to measure tau loading / burden for purposes of the present disclosure (Barthelemy et al., “Cerebrospinal Fluid Phospho-tau T217 Outperforms T181 as a Biomarker for the Differential Diagnosis of Alzheimer's Disease and PET Amyloid-positive Patient Identification,” Alzheimer's Res. Ther. 12, 26, doi:10.1186 / s13195-020-00596-4 (2020); Mattsson et al., “Aβ Deposition is Associated with Increases in Soluble and Phosphorylated Tau that Precede a Positive Tau PET in Alzheimer's Disease,” Science Advances 6, eaaz2387 (2020), which is incorporated herein by reference in its entirety). In certain embodiments, an antibody against human tau phosphorylated at threonine residue 217 can be used to measure tau load / burden in a subject (see International Patent Application Publication No. WO2020 / 242963, which is incorporated by reference in its entirety). The present disclosure includes, in some embodiments, measuring tau load / burden in a subject using an anti-tau antibody disclosed in WO2020 / 242963. The anti-tau antibody disclosed in WO2020 / 242963 is made against an isoform of human tau expressed in the CNS (e.g., recognizes an isoform expressed in the CNS and does not recognize an isoform of human tau expressed exclusively outside the CNS).

[0105] A subject is positive for amyloid deposits if amyloid is detected in the brain by methods such as amyloid imaging with radiolabeled PET compounds or by using diagnostic methods that detect Aβ or biomarkers of Aβ. Exemplary methods that can be used to measure brain amyloid load / burden include, for example, florbetapir (Carpenter, et al., "The Use of the Exploratory IND in the Evaluation and Development of 18 F-PET Radiopharmaceuticals for Amyloid Imaging in the Brain: A Review of One Company's Experience," The Quarterly Journal of Nuclear Medicine and Molecular Imaging 53.4:387 (2009), which is incorporated by reference in its entirety); florbetaben (Syed et al., "[ 18 F]Florbetaben: A Review in β-Amyloid PET Imaging in Cognitive Impairment,” CNS Drugs 29, 605-613 (2015), incorporated herein by reference in its entirety); and flutemetamol (Heurling et al., “Imaging β-amyloid Using [ 18 [F]Flutemetamol Positron Emission Tomography: From Dosimetry to Clinical Diagnosis,” European Journal of Nuclear Medicine and Molecular Imaging 43.2:362-373 (2016), the entire contents of which are incorporated herein by reference. 18 [F]-Florbetapir provides a qualitative and quantitative measurement of cerebral plaque burden in patients, including those with prodromal AD or mild AD dementia, and can also be used to assess the reduction of amyloid plaques from the brain.

[0106] Additionally, cerebrospinal fluid or plasma-based analysis of β-amyloid can be used to measure amyloid load / burden. For example, Aβ42 can be used to measure brain amyloid (Palmqvist, S. et al., “Accuracy of Brain Amyloid Detection in Clinical Practice Using Cerebrospinal Fluid Beta-amyloid 42: a Cross-validation Study Against Amyloid Positron Emission Tomography. JAMA Neurol 71, 1282-1289 (2014), incorporated herein by reference in its entirety). In some embodiments, the ratio of Aβ42 / Aβ40 or Aβ42 / Aβ38 can be used as amyloid beta biomarkers (Janelidze et al., “CSF Abeta42 / Abeta40 and Abeta42 / Abeta38 Ratios: Better Diagnostic Markers of Alzheimer Disease,” Ann Clin Transl Neurol 3, 154-165 (2016), which is incorporated herein by reference in its entirety. In some embodiments, cerebral amyloid plaques or Aβ deposited in CSF or plasma can be used to stratify subjects into groups based on amyloid load / burden.

[0107] Use of Antibody 1 for the Treatment or Prevention of ARIA: In some embodiments, the disclosure provides for the use of antibody 1 for the treatment or prevention of ARIA. Some therapeutic amyloid-targeting antibodies have shown a dose-response related increase in ARIA-E. See, e.g., Brashear et al., "Clinical Evaluation of Amyloid-related Imaging Abnormalities in Bapineuzumab Phase III Studies," J. of Alzheimer's Disease 66.4:1409-1424 (2018); Budd et al., "Clinical Development of Aducanumab, an Anti-Aβ Human Monoclonal Antibody Being Investigated for the Treatment of Early Alzheimer's Disease," The Journal of Prevention of Alzheimer's Disease 4.4:255 (2017).ARIA-E and ARIA-H are associated with amyloid plaque removal therapy. Sperling et al., “Amyloid-related imaging abnormalities in amyloid-modifying therapeutic trials: Recommendations from the Alzheimer's Association Research Roundtable Workgroup,” Alzheimer's & Dementia 7:367-85(2011); Sevigny et al., “The Antibody Aducanumab Reduces Aβ Plaques in Alzheimer's Disease,” Nature 537:50-6(2016); Ostrowitzki et al., “Mechanism of Amyloid Removal in Patients With Alzheimer Disease Treated With Gantenerumab,” Archives of Neurology 69:198-207(2012); Salloway et al., “Two Phase 3 Trials of Bapineuzumab in Mild-to-Moderate Alzheimer's Disease,” New England Journal of Medicine 370:322-33 (2014); Salloway et al., "A Phase 2 Multiple Ascending Dose Trial of Bapineuzumab in Mild to Moderate Alzheimer Disease," Neurology 73:2061-70 (2009) and Sperling et al., "Amyloid-related Imaging Abnormalities in Patients with Alzheimer's Disease Treated with Bapineuzumab: A Retrospective Analysis," Lancet Neurol. 11:241-9 (2012), each of which is incorporated by reference in its entirety.

[0108] As used herein, "amyloid-related imaging abnormalities" and "ARIA" are interchangeable and include vasogenic edema and cleft exudate (ARIA-E) and microhemorrhages and hemosiderin deposits (ARIA-H), and represent underlying pathological conditions recognized by those skilled in the art (see, e.g., Amyloid-Related Imaging Abnormalities and β-Amyloid-Targeting Antibodies, A Systematic Review, Massimo Filippi, MD; et al., JAMA Neurol. 2022; 79(3):291-304, and Amyloid-Related Imaging Abnormalities with Emerging Alzheimer Disease Therapeutics: Detection and Reporting Recommendations for Clinical Practice, PM Cogswell, et al., Am J Neuroradiol 43:E19-E35 Sep 2022). ARIA can be scored on a scale of 0 to 5. Although the exact cause of such adverse events is unknown, it is generally believed that anti-amyloid antibody treatment disrupts the blood-brain barrier through interaction with cerebrovascular amyloid, which leads to a leaky barrier and the development of edema in patients. Several possible mechanisms of action have been hypothesized, such as parenchymal plaque clearance or altered localization of AQP-4 in astrocyte end foot processes of the neurovascular unit, such as removal of amyloid from the vascular wall destabilizing the neurovascular unit, localizing inflammation / infiltration in the neurovascular unit, and increasing the level of cerebrovascular amyloid due to higher levels of interstitial soluble Aβ.

[0109] Amyloid deposition in the vessel wall (CAA) can lead to loss of vascular integrity and reduced perivascular clearance, which can spontaneously be associated with microhemorrhage. When anti-amyloid monoclonal antibody therapy is initiated, antibody-mediated degradation of amyloid plaques and mobilization of parenchymal and vascular Aβ increases the load of perivascular drainage. Overload of perivascular drainage pathways can lead to a transient increase in amyloid deposition in the arterial wall. Concurrently, antibody-mediated inflammation and amyloid degradation also occur within the vessel wall. These processes lead to further loss of vascular integrity and disruption of the blood-brain barrier. As a result, proteinaceous fluid and / or red blood cells leak into the parenchyma and / or pial space, causing edema / exudate (ARIA-E) or microhemorrhage / superficial siderosis (ARIA-H).

[0110] Methods for identifying patients in need of treatment or prevention of ARIA are known to those of skill in the art and are described, for example, in Amyloid-Related Imaging Abnormalities with Emerging Alzheimer Disease Therapeutics: Detection and Reporting Recommendations for Clinical Practice, P. M. Cogswell, et al., Am J Neuroradiol 43:E19-E35 Sep 2022, Detection and Management of Amyloid-Related Imaging Abnormalities in Patients with Alzheimer's Disease Treated with Anti-Amyloid Beta Therapy, J. Barakos et al., J Prev Alz Dis 2022;2(9):211-220 (incorporated herein by reference), and other prior references herein.

[0111] ARIA-E is most frequently detected on routine protocol-specified surveillance MRI in clinically asymptomatic patients. When ARIA-E is symptomatic, symptoms are most commonly nonlocalized, such as headache or confusion, but may additionally include visual disturbances, visuospatial impairment, or executive difficulties, given the relative predilection for posterior involvement in ARIA-E. The E in ARIA-E stands for edema, transudate, or exudate. Transudation of proteinaceous fluid into the parenchyma causes edema, the image appearance of which resembles vasogenic edema, best visualized on T2-FLAIR sequences. T2 hyperintense signals occur in white matter, gray matter, or both. Associated focal mass effect and gyral swelling may be present. Findings may be differentiated from cytotoxic edema by the absence of diffusion restriction, and the severe diffusion restriction associated with acute infarction is not a feature of ARIA. When leakage occurs in the leptomeningeal space, the result is a sulcal effusion or exudate that is only recognized on T2-FLAIR sequences because of T1 shortening associated with proteinaceous content. ARIA-E may manifest as either parenchymal edema or sulcal effusion, or both may occur together. In some antibody test analyses, sulcal effusion was the most common symptom of ARIA-E, and in others, parenchymal edema. ARIA-E most commonly affects the occipital lobe, followed by the parietal, frontal, and temporal lobes, and least frequently the cerebellum. The intensity and size of the signal abnormality can vary from subtle to small 1-2 cm zones of cortical-subcortical abnormality to multifocal to parahemispheric T2-hyperintense signal changes. These areas of signal abnormality generally have ill-defined margins, but in rare cases may have circumscribed margins and mimic neoplastic lesions. (See, e.g., Amyloid-Related Imaging Abnormalities with Emerging Alzheimer Disease Therapeutics: Detection and Reporting Recommendations for Clinical Practice, PM Cogswell, et al., Am J Neuroradiol 43:E19-E35 Sep 2022).

[0112] ARIA-H, hemorrhage includes microhemorrhage and superficial siderosis. Microhemorrhage occurs when leakage of heme products occurs in the parenchyma. Microhemorrhages are punctate, rounded, grossly hypointense foci in the brain parenchyma on T2 sequences, less than 10 mm in diameter. Leakage of heme products into the leptomeningeal or subpial space results in superficial siderosis, appearing as curvilinear hypointensities along the brain surface. Lobar macrohemorrhage (foci of hemorrhage identifiable on T1- or T2-weighted images, usually greater than 10 mm in diameter on gradient recalled echo [GRE]) rarely occurs with antiamyloid agents and, when it does occur, may be the result of an underlying disease process such as CAA. (See, e.g., Amyloid-Related Imaging Abnormalities with Emerging Alzheimer Disease Therapeutics: Detection and Reporting Recommendations for Clinical Practice, PM Cogswell, et al., Am J Neuroradiol 43:E19-E35 Sep 2022).

[0113] In some cases, patients with the epsilon 4 allele of apolipoprotein E (herein referred to as APOE e4 or APOE4) have a higher incidence of ARIA-E. Subjects with one or more copies of APOE4 may be at higher risk and in higher need of prevention and / or treatment. Monitoring the need for prevention or treatment includes genotyping, family history, and MRI or CT imaging as described above, and monitoring known symptoms consistent with ARIA. Patients with vascular or brain parenchymal amyloid disease may be at risk of ARIA and are subjects who require antibody 1 for prevention or treatment of ARIA.

[0114] Therefore, there is a need for improved methods of treating or preventing ARIA in patients, such as AD patients, treated with therapeutic amyloid-targeting antibodies. In particular, there is a need for simultaneous, separate, or sequential combinations of the antibodies of the present disclosure, particularly antibody 1, with one or more therapeutic amyloid-targeting antibodies, where antibody 1 is used to prevent or treat ARIA. Known anti-Aβ antibodies whose amyloid-targeting therapy may cause ARIA include donanemab, bapineuzumab, gantenerumab, aducanumab, GSK933776, solanezumab, crenezumab, ponezumab, and lecanemab (BAN2401), or anti-N3pGlu Aβ antibodies.

[0115] The disclosure further provides simultaneous, separate, or sequential combinations of Antibody 1 and one or more therapeutic amyloid-targeting antibodies for preventing or treating ARIA. In some embodiments, therapeutic amyloid-targeting antibodies whose treatment may be related to ARIA include donanemab, bapineuzumab, gantenerumab, aducanumab, GSK933776, solanezumab, crenezumab, ponezumab, and lecanemab (BAN2401), or anti-N3pGlu Aβ antibodies.

[0116] In these embodiments, "anti-N3pGlu Aβ antibody", "anti-N3pG antibody", or "anti-N3pE antibody" may be used interchangeably and refer to an antibody that preferentially binds to N3pGlu Aβ over Aβ1-40 or Aβ1-42. One of skill in the art will understand and appreciate that "anti-N3pGlu Aβ antibodies", as well as several specific antibodies, including "hE8L", "B12L" and "R17L", are identified and disclosed (along with methods of making and using) in U.S. Pat. No. 8,679,498 B2, which is incorporated herein by reference in its entirety. See, for example, Table 1 in U.S. Pat. No. 8,679,498 B2. Each of the antibodies disclosed in U.S. Pat. No. 8,679,498 B2, including the "hE8L", "B12L" and "R17L" antibodies, may be used as the anti-N3pGlu Aβ antibody of the present disclosure or in place of the anti-N3pGlu Aβ antibodies described in various aspects of the present invention. The anti-N3pGlu Aβ antibody of the present combination method is an antibody comprising the HC and LC of SEQ ID NOs: 40 and 41, respectively. Other representative species of anti-N3pGlu Aβ antibodies include, but are not limited to, those disclosed in U.S. Pat. Nos. 8,961,972, 10,647,759, 9,944,696, WO2010 / 009987A2, WO2011 / 151076A2, WO2012 / 136552A1 and equivalents thereof, such as antibodies disclosed under 35 U.S.C. § 112(f). One aspect of the present disclosure provides the use of antibody 1 for the prevention or treatment of ARIA observed in patients receiving an anti-N3pGlu Aβ antibody.

[0117] One aspect of the disclosure provides use of Antibody 1 for the prevention or treatment of ARIA, which is observed in patients receiving therapeutic antibodies that bind to deposited amyloid and is dose-limiting in some clinical development programs.

[0118] In one embodiment, the present disclosure provides a method of preventing ARIA comprising administering an effective amount of an anti-IL-34 antibody of the present disclosure to a patient in need thereof. In one embodiment, the present disclosure provides a method of preventing ARIA comprising administering an effective amount of antibody 1 to a patient in need thereof. In one embodiment, the present disclosure provides a method of treating ARIA comprising administering an effective amount of an antibody of the present disclosure to a patient in need thereof. In one embodiment, the present disclosure provides a method of treating ARIA comprising administering an effective amount of an antibody of the present disclosure to a patient in need thereof.

[0119] The present disclosure also provides an anti-IL-34 antibody of the present disclosure for use in the prevention or treatment of ARIA. The present disclosure also provides antibody 1 for use in the prevention or treatment of ARIA.

[0120] In certain embodiments, the disclosure provides for the use of an anti-IL-34 antibody of the disclosure in the manufacture of a medicament for the prevention or treatment of ARIA.

[0121] Further embodiments of combination uses and methods using the antibodies of the present disclosure are given below. The combination embodiments may refer to antibody 1, but the embodiments further include similar methods, uses, and all limitations described herein for the antibodies of the present disclosure described herein. The combination embodiments may refer to "anti-N3pG Aβ antibodies" which refers to each of the anti-N3pG Aβ antibodies described herein, but for clarity, these embodiments further include similar methods, uses, and all limitations described herein for each of the anti-N3pG Aβ antibodies individually, for example, preferably used in combination with donanemab. Further embodiments of the present disclosure are given below as numbered, including internal references to other numbered embodiments. For clarity, these embodiments should be read individually and / or collectively with the numbered embodiments to which they refer. The embodiments described below start with number 26. The term "course of treatment" refers to a particular patient or subject, the antibodies mentioned, the doses mentioned, the frequency and / or duration mentioned, the sequence mentioned, and other limitations to the extent described in each case.

[0122] Further combination embodiments of the present disclosure include the following:

[0123] 26. A method for treating or preventing a disease characterized by amyloid beta (Aβ) deposits in the brain of a human subject, comprising administering to a human subject in need of such treatment or prevention an effective amount of an anti-N3pG Aβ antibody in combination with an effective amount of antibody 1, either simultaneously, separately or sequentially.

[0124] 27. The method of embodiment 26, wherein the anti-N3pG Aβ antibody is donanemab.

[0125] 28. The method of embodiment 26, wherein the disease is Alzheimer's disease.

[0126] 29. The method of embodiment 26, wherein the anti-N3pG Aβ antibody is donanemab and the disease is Alzheimer's disease.

[0127] 30. The method of embodiment 29, wherein antibody 1 is administered sequentially after a course of treatment with donanemab.

[0128] 31. A method for treating or preventing a disease characterized by amyloid beta (Aβ) deposits in the brain of a human subject, comprising: i) administering to a human subject one or more first doses of about 100 mg to about 700 mg of an anti-N3pG Aβ antibody, wherein each first dose is administered once about every four weeks; ii) administering to the human subject one or more second doses of greater than 700 mg to about 1400 mg of an anti-N3pG Aβ antibody about four weeks after administering the one or more first doses, wherein each second dose is administered once about every four weeks; the anti-N3pGlu Aβ antibody is donanemab; iii) administering to a human subject an effective amount of antibody 1 simultaneously, separately or sequentially.

[0129] 32. The method of embodiment 31, wherein the human subject is administered one, two, or three doses of donanemab prior to administration of the second dose.

[0130] 33. The method of embodiment 31 or 32, wherein the human subject is administered a first dose of about 700 mg of donanemab.

[0131] 34. The method of any one of embodiments 31-33, wherein the human subject is administered one or more second doses of about 800 mg, about 900 mg, about 1000 mg, about 1100 mg, about 1200 mg, about 1300 mg, or about 1400 mg of donanemab.

[0132] 35. The method of any one of embodiments 31-34, wherein the human subject is administered one or more second doses of about 1400 mg of donanemab.

[0133] 36. The method of any one of embodiments 31-35, wherein the anti-N3pGlu Aβ antibody is administered to the human subject over a course of treatment for up to 72 weeks, or until normal levels of amyloid are achieved.

[0134] 37. The method of any one of embodiments 31-36, wherein the anti-N3pGlu Aβ antibody is administered to the human subject until the patient's amyloid plaque level is about 25 centiloids or less.

[0135] 38. The method of any one of embodiments 31-36, wherein the anti-N3pGlu Aβ antibody is administered to the human subject for a course of treatment until amyloid plaque levels in the human subject are about 25 centiloids or less in two consecutive PET imaging scans, optionally with two consecutive PET imaging scans spaced at least 6 months apart, or about 11 centiloids or less in one PET imaging scan.

[0136] 39. The method of any one of embodiments 31-36, wherein the human subject is administered three first doses of 700 mg of donanemab once every four weeks, followed by a second dose of 1400 mg once every four weeks, over a course of treatment of up to 72 weeks.

[0137] 40. The method of any one of embodiments 31-36, wherein a human subject is administered three first doses of 700 mg once every four weeks, followed by a second dose of 1400 mg once every four weeks until the subject's amyloid plaque level is about 25 centiloids or less.

[0138] 41. The method of any one of embodiments 31-36, wherein a human subject is administered three first doses of 700 mg of donanemab once every four weeks, followed by a second dose of 1400 mg once every four weeks, until the subject's amyloid plaque levels are about 25 centiloids or less in two consecutive PET imaging scans, optionally with two consecutive PET imaging scans separated by at least 6 months, or about 11 centiloids or less in one PET imaging scan.

[0139] 42. The method of any one of embodiments 31-41, wherein the human subject is administered a second dose of donanemab over a course of treatment sufficient to treat or prevent the disease.

[0140] 43. The method according to any one of embodiments 31 to 42, wherein the treatment or prevention of the disease results in i) a reduction in Aβ deposits in the brain of the human subject, and / or ii) a delay in cognitive or functional decline in the human subject.

[0141] 44. The method of embodiment 43, wherein the reduction in Aβ deposits in the brain of the human subject is determined by amyloid PET brain imaging or a diagnostic method that detects a biomarker of Aβ.

[0142] 45. The method of embodiment 43 or 44, wherein the second dose is administered to the human subject until Aβ deposits in the brain of the human subject are reduced by about 20-100%.

[0143] 46. ​​The method of embodiment 45, wherein Aβ deposits in the brain of the human subject are reduced by about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 75%, or about 100%.

[0144] 47. The method of any one of embodiments 31-44, wherein a second dose of donanemab is administered to the human subject until Aβ deposits in the brain of the human subject are reduced by i) about an average of about 25 to about 100 centiloids, ii) about an average of about 50 to about 100 centiloids, iii) about 100 centiloids, or iv) about 84 centiloids.

[0145] 48. The method of any one of embodiments 31 to 47, wherein the disease characterized by Aβ deposits in the brain of a human subject is selected from preclinical Alzheimer's disease (AD), clinical AD, prodromal AD, mild AD, moderate AD, severe AD, Down's syndrome, clinical cerebral amyloid angiopathy, or preclinical cerebral amyloid angiopathy.

[0146] 49. The method of any one of embodiments 31-48, wherein the human subject is an early symptomatic AD patient.

[0147] 50. The method of embodiment 49, wherein the human subject has prodromal AD and mild dementia due to AD.

[0148] 51. The method of any one of embodiments 26-50, wherein the human subject i) has a very low to moderate tau load or has been determined to have a very low to moderate tau load, ii) has a low to moderate tau load or has been determined to have a low to moderate tau load, iii) has a very low to moderate tau load or has a very low to moderate tau load and one or two alleles of APOE e4, iv) has a low to moderate tau load or has a low to moderate tau load and one or two alleles of APOE e4, or v) has one or two alleles of APOE e4.

[0149] 52. The method of embodiment 51, wherein the human subject has i) very low to moderate tau load if the tau load as measured by PET brain imaging is less than or equal to 1.46 SUVr, or ii) low to moderate tau load if the tau load as measured by PET brain imaging is between 1.10 SUVr and 1.46 SUVr.

[0150] 53. The method of any one of embodiments 26 to 50, wherein the human subject i) does not have or has been determined to not have a high tau burden, or ii) carries one or two alleles of APOE e4 and does not have or has been determined to not have a high tau burden.

[0151] 54. The method of embodiment 53, wherein the human subject has high tau burden if the tau burden as measured by PET brain imaging is greater than 1.46 SUVr.

[0152] 55. The method of embodiment 51 or 53, wherein the tau load of a human subject is determined using PET brain imaging or a diagnostic method that detects a biomarker of tau.

[0153] 56. Use of an anti-N3pGlu Aβ antibody in combination with antibody 1, simultaneously, separately or sequentially, in the manufacture of a medicament for the treatment or prevention of a disease characterized by Aβ deposits in the brain of a human subject, comprising one or more first doses of between about 100 mg and about 700 mg of an anti-N3pGlu Aβ antibody are administered, each first dose being administered once about every four weeks, followed by one or more second doses of greater than 700 mg to about 1400 mg administered four weeks after administration of the one or more first doses, each second dose of an anti-N3pGlu Aβ antibody being administered once about every four weeks; The anti-N3pGlu Aβ antibody is donanemab.

[0154] 57. The use according to embodiment 56, in which the human subject is administered one, two or three first doses of donanemab prior to administration of a second dose of donanemab.

[0155] 58. The use of embodiment 56 or 57, wherein the human subject is administered three first doses of about 700 mg of donanemab.

[0156] 59. The use of any one of embodiments 56-58, wherein a human subject is administered one or more second doses of about 800 mg, about 900 mg, about 1000 mg, about 1100 mg, about 1200 mg, about 1300 mg, or about 1400 mg of donanemab.

[0157] 60. The use according to any one of embodiments 56 to 59, in which the human subject is administered one or more second doses of about 1400 mg of donanemab.

[0158] 61. The use according to any one of embodiments 56 to 60, wherein the anti-N3pGlu Aβ antibody is administered to the human subject over a course of treatment for up to 72 weeks or until normal levels of amyloid are achieved.

[0159] 62. The use according to any one of embodiments 56 to 61, wherein the anti-N3pGlu Aβ antibody is administered to a human subject until the amyloid plaque level in the patient is about 25 centiloids or less.

[0160] 63. The use of any one of embodiments 56-61, wherein the anti-N3pGlu Aβ antibody is administered to a human subject until amyloid plaque levels in the patient are about 25 centiloids or less in two consecutive PET imaging scans, optionally with two consecutive PET imaging scans spaced at least 6 months apart, or about 11 centiloids or less in one PET imaging scan.

[0161] 64. The use of any one of embodiments 56-61, wherein a human subject is administered three first doses of 700 mg of donanemab once every four weeks, followed by a second dose of 1400 mg of donanemab once every four weeks, for a period of up to 72 weeks.

[0162] 65. The use of any one of embodiments 56-61, wherein a human subject is administered three first doses of 700 mg of donanemab once every four weeks, followed by a second dose of 1400 mg of donanemab once every four weeks until the amyloid plaque level in the patient is about 25 centiloids or less.

[0163] 66. The use of any one of embodiments 56-61, wherein a human subject is administered three first doses of 700 mg of donanemab once every four weeks, followed by a second dose of 1400 mg of donanemab once every four weeks, until amyloid plaque levels in the patient are about 25 centiloids or less in two consecutive PET imaging scans, where appropriate, the two consecutive PET imaging scans being separated by at least 6 months, or about 11 centiloids or less in one PET imaging scan.

[0164] 67. The use according to any one of embodiments 56 to 66, wherein the human subject is administered a second dose of donanemab over a course of treatment sufficient to treat or prevent the disease.

[0165] 68. The use according to any one of embodiments 56 to 67, wherein the treatment or prevention of the disease results in i) a reduction in Aβ deposits in the brain of a human subject, and / or ii) a delay in cognitive or functional decline in a human subject.

[0166] 69. The use according to embodiment 68, wherein the reduction of Aβ deposits in the brain of a human subject is determined by amyloid PET brain imaging or a diagnostic method detecting a biomarker of Aβ.

[0167] 70. The use of embodiment 68 or 69, wherein a second dose of donanemab is administered to the human subject until Aβ deposits in the brain of the human subject are reduced by about 20 to 100%.

[0168] 71. The use according to embodiment 70, wherein Aβ deposits in the brain of a human subject are reduced by about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 75%, or about 100%.

[0169] 72. The use according to embodiment 70 or 71, in which Aβ deposits in the brain are reduced by 100% in the patient.

[0170] 73. The use of any one of embodiments 56-72, wherein a second dose of donanemab is administered to the human subject until Aβ deposits in the brain of the human subject are reduced by i) about an average of about 25 centiloids to about 100 centiloids, ii) about an average of about 50 centiloids to about 100 centiloids, iii) about 100 centiloids, or iv) about 84 centiloids.

[0171] 74. The use according to any one of embodiments 56 to 73, wherein the disease characterized by Aβ deposits in the brain of a human subject is selected from preclinical Alzheimer's disease, clinical AD, prodromal AD, mild AD, moderate AD, severe AD, Down's syndrome, clinical cerebral amyloid angiopathy, or preclinical cerebral amyloid angiopathy.

[0172] 75. The use according to any one of embodiments 56 to 74, wherein the human subject is an early symptomatic AD patient, or the human subject has prodromal AD or mild dementia due to AD.

[0173] 76. The use according to any one of embodiments 56 to 75, wherein the human subject i) has a very low to moderate tau load or has been determined to have a very low to moderate tau load, ii) has a low to moderate tau load or has been determined to have a low to moderate tau load, iii) has a very low to moderate tau load or has a very low to moderate tau load and one or two alleles of APOE e4, iv) has a low to moderate tau load or has a low to moderate tau load and one or two alleles of APOE e4, or v) has one or two alleles of APOE e4.

[0174] 77. The use of embodiment 76, wherein the human subject has i) very low to moderate tau load if the tau load as measured by PET brain imaging is below 1.46 SUVr, or ii) low to moderate tau load if the tau load as measured by PET brain imaging is between 1.10 SUVr and 1.46 SUVr.

[0175] 78. The use according to any one of embodiments 56 to 75, wherein the human subject i) does not have a high tau burden or has been determined to not have a high tau burden, or ii) carries one or two alleles of APOE e4 and does not have a high tau burden or has been determined to not have a high tau burden.

[0176] 79. The use of embodiment 78, wherein the human subject has a high tau load if the tau load as measured by PET brain imaging is greater than 1.46 SUVr.

[0177] 80. The use according to embodiment 76 or 78, wherein the tau load of a human subject is determined using tau PET brain imaging or a diagnostic method detecting a biomarker of tau.

[0178] 81. A method of treating or preventing a disease characterized by brain amyloid beta (Aβ) deposits in a human subject who has been determined to have ii) a very low to moderate tau burden, or a low to moderate tau burden, or ii) a very low to moderate tau burden, or a low to moderate tau burden and one or two alleles of APOE e4, comprising: i) administering to a human subject one or more first doses of about 100 mg to about 700 mg of donanemab, wherein each first dose of donanemab is administered once about every 4 weeks; ii) four weeks after administering the one or more first doses, administering to the human subject one or more second doses of greater than 700 mg to about 1400 mg of donanemab, wherein each second dose is administered once about every four weeks; A method of combining with an effective amount of antibody 1 simultaneously, separately or sequentially.

[0179] 82. A method for treating or preventing a disease characterized by amyloid beta (Aβ) deposits in the brain of a human subject, comprising: If the human subject has tau burden in the temporal, occipital, parietal, or frontal lobes, determining whether the human subject has tau burden in the temporal, occipital, parietal, or frontal lobes of the brain, and then i) administering to a human subject one or more first doses of about 100 mg to about 700 mg of an anti-N3pGlu Aβ antibody, wherein each first dose is administered once about every four weeks; ii) about 4 weeks after administering the one or more first doses, administering to the human subject one or more second doses of greater than 700 mg to about 1400 mg of an anti-N3pGlu Aβ antibody, wherein each second dose is administered once about every 4 weeks; A method of combining with an effective amount of antibody 1 simultaneously, separately or sequentially.

[0180] 83. The method of embodiment 82, wherein the human subject has tau burden in the posterior lateral temporal lobe or temporal lobe of the brain.

[0181] 84. The method of embodiment 82, wherein the human subject has tau burden in the occipital lobe of the brain.

[0182] 85. The method of embodiment 82, wherein the human subject has tau burden in the parietal lobe of the brain.

[0183] 86. The method of embodiment 82, wherein the human subject has tau burden in the frontal lobe of the brain.

[0184] 87. The method of embodiment 82, wherein the human subject has tau burden in the posterior lateral temporal lobe (PLT) and / or occipital lobe of the brain.

[0185] 88. The method of any one of embodiments 82-87, wherein the human subject has tau burden in the PLT or occipital regions of the brain, as well as i) parietal or precuneus regions, or ii) frontal regions.

[0186] 89. The method of any one of embodiments 82-86, wherein the human subject has i) tau burden isolated to the frontal lobe, or ii) tau burden in regions of the temporal lobe that do not include the posterolateral temporal region (PLT) of the brain.

[0187] 90. The method of any one of embodiments 82-88, wherein the human subject has tau burden in the posterior lateral temporal, occipital, and parietal lobes of the brain.

[0188] 91. The method of any one of embodiments 82-88, wherein the human subject has tau burden in the posterior lateral temporal lobe, occipital lobe, parietal lobe, and frontal lobe of the brain.

[0189] 92. The method of any one of embodiments 82-88, wherein the human subject has tau burden in the posterior lateral temporal lobe, occipital lobe, parietal lobe and / or frontal lobe of the brain.

[0190] 93. The method of any one of embodiments 82-92, wherein the human subject is administered one, two, or three first doses prior to administration of the second dose.

[0191] 94. The method of any one of embodiments 82-93, wherein the human subject is administered a first dose of about 700 mg.

[0192] 95. The method of any one of embodiments 82-94, wherein the human subject is administered one or more second doses of about 800 mg, about 900 mg, about 1000 mg, about 1100 mg, about 1200 mg, about 1300 mg, or about 1400 mg.

[0193] 96. The method of any one of embodiments 82-95, wherein the human subject is administered one or more second doses of about 1400 mg.

[0194] 97. The method of any one of embodiments 82-96, wherein the anti-N3pGlu Aβ antibody is administered to the human subject for a period of up to 72 weeks or until normal levels of amyloid are achieved.

[0195] 98. The method of any one of embodiments 82-97, wherein the anti-N3pGlu Aβ antibody is administered to the human subject until the patient's amyloid plaque level is about 25 centiloids or less.

[0196] 99. The method of any one of embodiments 82-98, wherein the anti-N3pGlu Aβ antibody is administered to the human subject until amyloid plaque levels in the human subject are about 25 centiloids or less in two consecutive PET imaging scans, optionally with two consecutive PET imaging scans spaced at least 6 months apart, or about 11 centiloids or less in one PET imaging scan.

[0197] 100. The method of any one of embodiments 82-99, wherein a human subject is administered a first dose of 700 mg once every 4 weeks for three doses, followed by a second dose of 1400 mg once every 4 weeks for up to 72 weeks.

[0198] 101. The method of any one of embodiments 82-100, wherein a human subject is administered three first doses of 700 mg once every four weeks, followed by a second dose of 1400 mg once every four weeks, until the amyloid plaque level in the subject is about 25 centiloids or less.

[0199] 102. The method of any one of embodiments 82-101, wherein a human subject is administered three first doses of 700 mg once every four weeks, and then a second dose of 1400 mg once every four weeks, until the amyloid plaque level in the subject is about 25 centiloids in two consecutive PET imaging scans, optionally wherein the two consecutive PET imaging scans are separated by at least six months, or is about 11 centiloids or less in one PET imaging scan.

[0200] 103. The method of any one of embodiments 82-102, wherein the second dose is administered to the human subject for a period sufficient to treat or prevent the disease.

[0201] 104. The method according to any one of embodiments 82 to 103, wherein the treatment or prevention of the disease results in i) a reduction in Aβ deposits in the brain of the human subject, and / or ii) a delay in cognitive or functional decline in the human subject.

[0202] 105. The method of embodiment 97, wherein the reduction of Aβ deposits in the brain of the human subject is determined by amyloid PET brain imaging or a diagnostic method that detects a biomarker of Aβ.

[0203] 106. The method of embodiment 97 or 98, wherein the second dose is administered to the human subject until Aβ deposits in the brain of the human subject are reduced by about 20-100%.

[0204] 107. The method of embodiment 106, wherein Aβ deposits in the brain of a human subject are reduced by about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 75%, or about 100%.

[0205] 108. The method of any one of embodiments 82-107, wherein the second dose is administered to the human subject until Aβ deposits in the human subject's brain are reduced by i) about an average of about 25 centiloids to about 100 centiloids, ii) about an average of about 50 centiloids to about 100 centiloids, iii) about 100 centiloids, or iv) about 84 centiloids.

[0206] 109. The method of any one of embodiments 82 to 108, wherein the disease characterized by Aβ deposits in the brain of a human subject is selected from preclinical Alzheimer's disease (AD), clinical AD, prodromal AD, mild AD, moderate AD, severe AD, Down's syndrome, clinical cerebral amyloid angiopathy, or preclinical cerebral amyloid angiopathy.

[0207] 110. The method of any one of embodiments 82-109, wherein the human subject is an early symptomatic AD patient.

[0208] 111. The method of embodiment 109, wherein the human subject has prodromal AD and mild dementia due to AD.

[0209] 112. The method of any one of embodiments 82 to 111, wherein the human subject i) has a very low to moderate tau load or has been determined to have a very low to moderate tau load, or ii) has a low to moderate tau load or has been determined to have a low to moderate tau load.

[0210] 113. The method of embodiment 112, wherein the human subject has i) very low to moderate tau load if the tau load as measured by PET brain imaging is less than or equal to 1.46 SUVr, or ii) low to moderate tau load if the tau load as measured by PET brain imaging is between 1.10 SUVr and 1.46 SUVr.

[0211] 114. The method of any one of embodiments 82-113, wherein the human subject does not have a high tau burden or has been determined to not have a high tau burden.

[0212] 115. The method of embodiment 114, wherein the human subject has high tau load if the tau load as measured by PET brain imaging is greater than 1.46 SUVr.

[0213] 116. The method of embodiment 114 or 115, wherein the tau load of a human subject is determined using PET brain imaging or a diagnostic method that detects a biomarker of tau.

[0214] 117. The method of any one of embodiments 82-116, wherein the anti-N3pGlu Aβ antibody comprises donanemab.

[0215] 118. The method of any one of embodiments 82 to 117, wherein the patient has one or two alleles of APOE e4.

[0216] 119. A method for reducing / preventing further increase in tau burden or slowing the rate of tau accumulation in the temporal, occipital, parietal or frontal lobes of the human brain, comprising administering to a human subject an anti-N3pGlu Aβ antibody in simultaneous, separate or sequential combination with an effective amount of antibody 1.

[0217] 120. A method for treating ARIA in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of antibody 1 or a pharmaceutical composition thereof.

[0218] 121. A method for preventing ARIA in a subject in need thereof, comprising administering a therapeutically effective amount of antibody 1 or a pharmaceutical composition thereof to the subject. [Brief description of the drawings]

[0219] [Figure 1] 1 shows Antibody 1 neutralization of human IL-34-induced luciferase reporter activity in hCSF1R-expressing 293SRE cells. [Diagram 2] Figure 1 shows the ability of Antibody 1 to inhibit ERK phosphorylation in NIH-3T3 / CSF1R cells. Triangles represent cells treated with isotype control antibody, circles represent cells treated with Antibody 1, and asterisks represent no addition of IL-34 to the assay (assay baseline). EXAMPLES

[0220] The following examples are provided to illustrate, but not limit, the claimed invention. The results of the following assays demonstrate that the exemplified monoclonal antibodies, such as antibody 1 of the present disclosure, bind and / or neutralize IL-34 and therefore may be used to treat immune-mediated and inflammatory diseases as described herein.

[0221] Example 1: Antibody generation, expression and purification A panel of human anti-IL-34 antibodies is obtained using a fully human yeast display library and screened to identify reagents that may be effective human IL-34 neutralizing antibodies. To isolate clones with improved affinity, mutations are systematically introduced into individual complementarity determining regions (CDRs) of each antibody, and the resulting library is subjected to multiple rounds of selection with decreasing antigen concentration and / or increasing dissociation time. Individual variants are sequenced and used to construct combinatorial libraries that are subjected to additional rounds of selection with increasing stringency to identify additive or synergistic pairings of mutations between individual CDR regions. Individual combinatorial clones are sequenced and binding characteristics are determined. To further increase affinity for IL-34, these combinatorial clones may be subjected to additional rounds of single and combinatorial mutagenesis. This screening can be performed against human or cynomolgus IL-34 to increase affinity for the species of choice. Selected antibodies can also be mutagenized to repair post-translational modifications such as isomerization while maintaining binding affinity to IL-34. Additionally, framework (FW) or CDR substitutions can be made to the antibodies to revert sequences to their germline state to reduce potential immunogenicity risks.

[0222] For example, a modified and / or optimized anti-IL-34 antibody, referred to herein as Antibody 1, has been obtained having heavy and light chain variable region amino acid sequences, complete heavy and light chain amino acid sequences, and nucleotide sequences encoding the same as those listed in the section entitled "Amino Acid and Nucleotide Sequence Listing" below. The SEQ ID NOs corresponding to these sequences, as well as the light and heavy chain CDR amino acid sequences, are shown in Table 1.

[0223] Exemplary anti-IL-34 antibodies of the present disclosure can be expressed and purified essentially as follows: Suitable host cells, such as HEK293, NS0 or CHO, can be transiently or stably transfected with an expression system for secretion of the antibody using an optimal predetermined HC:LC vector ratio (such as 1:3 or 1:2 or 1:1) or a single vector system encoding both the HC and LC.

[0224] The expression plasmid contains, for example, DNA encoding the LC and HC of Antibody 1 (the DNA sequence of SEQ ID NO: 11 encoding the HC of exemplified Antibody 1, and the DNA sequence of SEQ ID NO: 12 encoding the LC amino acid sequence of exemplified Antibody 1), and is expressed from a construct commonly used and suitable for this purpose. The clone-derived cell line is grown and screened for Antibody 1 production, and the clone-derived cell line is selected and established. The cell line is generated without the use of any material containing animal components and is used for production.

[0225] The clarified medium into which the antibody is secreted may be purified by conventional techniques such as mixed-mode methods of ion exchange and hydrophobic interaction chromatography. For example, the medium may be applied to and eluted from a Protein A or Protein G column using conventional methods, and mixed-mode methods of ion exchange and hydrophobic interaction chromatography may also be used. Soluble aggregates and multimers may be effectively removed by common techniques including size exclusion, hydrophobic interaction, ion exchange, or hydroxyapatite chromatography. The exemplified IL-34 antibodies of the present disclosure are concentrated and / or sterile filtered using common techniques. The purity of the exemplified antibodies after these chromatography steps is greater than 95%. The exemplified anti-IL-34 antibodies of the present disclosure may be immediately frozen at -70°C or stored at 4°C for several months.

[0226] Example 2: Characterization of anti-IL-34 antibodies Binding affinity to human and cynomolgus monkey IL-34 The binding affinity of the anti-IL-34 monoclonal antibodies of the present disclosure to human and / or cynomolgus monkey (cyno) IL-34 can be determined by methods known in the art. Briefly, the binding affinity and kinetics of the antibodies are assessed by surface plasmon resonance using a BIAcore™ 8K (Cytiva) at 37° C. The binding affinity is measured by immobilizing the anti-IL-34 antibodies on a BIAcore™ Sensor Chip Protein A (Cytiva) and flowing human or cynomolgus monkey IL-34 starting at 25 nM or 12.5 nM in two-fold serial dilutions in HBS-EP+ buffer (Teknova). In each cycle, 200 μL of IL-34 is flowed over the immobilized antibody at 100 μL / min, followed by dissociation for 20 minutes. The chip surface is regenerated with 50 μL of glycine buffer at pH 1.5 at a flow rate of 100 μL / min. The data is fitted to a 1:1 Langmiur binding model to derive k, k, and calculate K. Table 3 shows the average of at least three experiments for exemplified Antibody 1 for human and cynomolgus IL-34. [Table 3]

[0227] Example 3: In vitro functional characterization of anti-human IL-34 antibodies Antibodies of the disclosure are tested for their ability to neutralize IL-34 binding and / or activity. Neutralization of IL-34 binding and / or activity by antibodies of the disclosure can be assessed, for example, by one or more IL-34 / CSF1R receptor binding assay formats, as well as IL-34 cell-based activity assays, as described below.

[0228] Ability of Antibody 1 to displace IL-34 from CSF1R Assays for neutralizing antibodies of IL-34 / CSF1R binding can be performed using enzyme assays. Such assays can use recombinantly expressed CSF1R extracellular domain proteins capable of binding IL-34. These proteins can be bound to ELISA plates to capture soluble IL-34. IL-34 can then be detected by either biotinylation of the antigen and detection with streptavidin / neutravidin-conjugated peroxidase or phosphatase enzymes. Such neutralization assays include pre-incubation (e.g., 1 hour) of the antibody to be evaluated with labeled IL-34 (as well as a control sample that does not involve an antibody targeting IL-34) before adding to the binding assay.

[0229] CSF1R extracellular domain protein (hCSF1R_Fc, commercially available from R&D, catalog number 329-MR, cynomolgus monkey CSF1R ECD-Fc (AAA is the linker between CSF1R extracellular domain and Fc) (SEQ ID NO: 34)) can be bound to ELISA plates at a concentration of 30 nM to capture soluble biotinylated IL-34 and allowed to bind for 1 hour. After washing and blocking the plate, biotinylated IL-34 can be added and then detected with streptavidin-conjugated peroxidase. To determine the concentration of antibody required to displace IL-34 from CSF1R, the 80% binding level (EC 80 A concentration of labeled IL-34 close to IL-34 (3.7 nM) can be used in combination with various antibody concentrations (0-100 nM). After 1 hour of incubation, IL-34 bound to CSF1R is detected via streptavidin-conjugated peroxidase. Antibodies are assayed (n=2) and the mean and standard deviation at each concentration are calculated. The potency of the antibody to displace IL-34 from CSF1R is expressed as IC 50 Reported as (nM) and calculated confidence intervals (CI) are shown in Tables 4 and 5. [Table 4] [Table 5]

[0230] IL-34 binds to human CSF1R with an affinity of approximately 50-100 pM, therefore, a high affinity antibody is required to effectively neutralize this cytokine in the CNS. The results in Table 4 show that Antibody 1 has high affinity for human IL-34, with an IC of 0.07882 nM. 50 The results in Table 4 show that antibody 1 has high affinity for human IL-34, and in particular antibody 1 exhibits affinity for human IL-34 comparable to hCSF1R, thus possessing binding properties that allow them to effectively neutralize IL-34 in vivo. Blocking IL-34 is believed to provide a useful means for disease modification while avoiding the safety concerns associated with some existing immunomodulatory therapies. Thus, neutralization of IL-34-mediated signaling represents a therapeutic approach for the management of neuroinflammation, microgliosis, and neurodegenerative diseases such as Alzheimer's disease, and other tauopathies and inflammatory diseases. (See, for example, Lelios, I. et al. Emerging role of IL-34 in health and disease, J Exp Med (2020) 217 ​​(3): e20190290).

[0231] Ability of Antibody 1 to neutralize CSF1R dimerization in the PathHunter® eXpress Dimerization Assay: Human IL-34 neutralization can be further assessed by seeding U2OS CSF1R / CSF1R cells (Path Hunter® eXpress Dimerization Assay, DiscoverX) in 96-well plates to assess the ability of anti-IL-34 antibodies to inhibit CSF1R dimerization. These assays utilize Enzyme Fragment Complementation (EFC) technology, in which the b-galactosidase (b-gal) enzyme is split into two fragments, ProLink (PK) and Enzyme Acceptor (EA). Independently, these fragments have no b-gal activity, but when forced to complement by protein-protein interactions, they form the active b-gal enzyme. The PathHunter® eXpress Dimerization Assay detects ligand-induced dimerization of the two subunits of the CSF1R receptor-dimer pair. Cells are engineered to co-express one CSF1R receptor subunit fused to an enzyme donor (ED) and a second dimer partner fused to an enzyme acceptor (EA). Binding of human IL-34 to one of the receptor subunits induces an interaction with its dimer partner, forcing the complementation of the two enzyme fragments. This results in the formation of a functional enzyme that hydrolyzes a substrate and generates a chemiluminescent signal. The reduction in relative fluorescence units (RFU) shown in Table 6 reflects the ability of Antibody 1 to neutralize human IL-34 and reduce chemiluminescence. The half maximal inhibitory concentration (IC 50 ) value is 1.035 nM. Human CSF1R-Fc was used as a positive control in this assay and had an IC of 1.025 nM. 50 The data in Table 6 support the ability of Antibody 1 to block the interaction of human IL-34 with CSF1R, thereby inhibiting CSF1R dimerization in this assay. This data supports the use of the antibodies of the present disclosure to neutralize human IL-34. [Table 6]

[0232] Inhibition of IL-34-induced responses in vitro Neutralization of IL-34 activity by the antibodies of the present disclosure can be assessed by one or more IL-34 cell-based assays, for example as described below. The ability of the antibodies of the present disclosure to neutralize human IL-34-inducible luciferase reporter activity can be assessed in 293 hCSF1R SRE cells transfected with cDNA expressing human CSF1R (Accession: NP_001275634.1). For example, 293 / SRE cells stably overexpressing human CSF1R (hCSF1R) are dissociated in 0.05% trypsin-PBS and seeded at 70,000 cells per 100 ul in tissue culture-treated 96-well plates. The next day, growth medium is removed and cells are starved with DMEM-F12 (Dulbecco's Modified Eagle Medium: Nutrient Mixture F-12) supplemented with 1% heat-inactivated FBS (fetal bovine serum). After 24 hours of starvation, cells are treated with 100ng / ml human IL-34 and multiple concentrations of either hCSF1R-Fc or Antibody 1 for 6 hours. After incubation, cells are lysed with 50ul of Promega™ Glo™ Lysis Buffer (Promega™ E266A) for 5 minutes with gentle agitation. 50ml of BrightGlo™ Luminescence Reagent (Promega™ E2620) is added and incubated on the lysed cells for 2 minutes. Luminescence is read on a Perkin Elmer Wallac 1420 Victor2™ microplate reader. The reduction in relative fluorescence units (RFU) shown in Table 7 and Figure 1 reflects the ability of Antibody 1 to neutralize human IL-34-induced luciferase activity. The half-maximal inhibitory concentration (IC) of Antibody 1 for neutralization of hIL-34 was calculated. 50 The IC value is 0.05037ug / ml. Human CSF1R-Fc was used as a positive control in this assay and had an IC of 0.09603ug / ml. 50 Inhibits luciferase activity. [Table 7]

[0233] Ability of anti-IL-34 antibodies to inhibit ERK phosphorylation in NIH-3T3 / CSF1R cells: Neutralization of IL-34 may be determined by assessing the ability of anti-IL-34 antibodies to inhibit extracellular signal-regulated kinase (ERK) phosphorylation in NIH-3T3 / CSF1R. In this assay, on day 1, cells are seeded in DMEM supplemented with 10% FBS and incubated overnight at 37°C. On day 2, the medium is removed, cells are washed with serum-free DMEM, and further incubated for 24 hours. On day 3, the medium is replaced with serum-free DMEM containing anti-IL-34 antibodies. Human or cynomolgus IL-34 is added for 5 minutes to a final concentration of 1ug / ml. Either human or cynomolgus IL-34 and an isotype control antibody serve as positive and negative controls, respectively. Phospho / total ERK1 / 2 levels are assessed by measuring electrochemiluminescence signal using Whole Cell Lysate Kit (Meso Scale Discovery, Catalog No. K15107D). Data are calculated as the ratio of electrochemiluminescence signal of phospho-ERK1 / 2 to total ERK1 / 2 protein. The reduction in the signal ratio shown in Table 8 and / or Figure 2 reflects the ability of Antibody 1 to neutralize IL-34 activity. The half-maximal inhibitory concentration (IC) of Antibody 1 against human IL-34 was 50 ) values ​​are 26 nM and 53 nM for cynomolgus IL-34. [Table 8] [Table 9]

[0234] Ability of anti-IL34 antibodies to inhibit IL-34-induced expression of CD163 on human monocytes by flow cytometry: Neutralization of IL-34 can also be assessed by measuring the expression of the cell surface antigen CD163 on human monocytes after treatment with IL-34 by flow cytometry (see, for example, Boulakirba, S., et al., IL-34 and CSF-1 exhibit comparable macrophage differentiation capacity but different polarization potentials. Sci Rep 8, 256 (2018)). CD14 positive monocytes are treated with IL-34 for 6 days and stained with antibodies for CD163, followed by evaluation of CD163 expression by flow cytometry. In the experiment, the change in the number of cells expressing CD163 indicates that IL-34 treatment increases the expression of this antigen on monocytes. Addition of antibody 1 suppresses the increased expression of CD163. In this experiment, an isotype-matched IgG4 antibody is used as a negative control. The results are shown in Table 10.

[0235] CD14+ human monocytes can be differentiated into macrophages by the addition of IL-34 (100ng / ml). The macrophage marker CD163 can be used to monitor the degree of differentiation. This differentiation into macrophages can be inhibited by the addition of anti-IL-34 antibodies. CD14+ human monocytes are seeded into 6-well plates with or without IL-34. Cells are treated with anti-IL-34 antibodies, such as antibody 1, or 15ug / ml IgG4 PAA for a total of 6 days, refreshing the treatment on day 3. On day 6, the cells are removed from the plate with non-enzymatic cell dissociation buffer to collect the multicellular cells and washed with FACS buffer (PBS+2%FBS+0.1% sodium azide+2%EDTA). Cells are blocked with TruStain FcX (cat. no. 422302) for 30 minutes, following the manufacturer's recommendations. After blocking, cells are washed with FACS buffer and stained with anti-CD163-PE or IgGk isotype control-PE for 1 h at 4 C. At the end of the incubation, cells are washed and flow analysis is performed on Accuri using a minimum of 10,000 events. Median-PE-A levels are collected per treatment. [Table 10]

[0236] Inhibition of CD163 expression on human monocytes in response to IL-34 by Antibody 1 demonstrates the ability of the disclosed antibodies to modulate monocyte / macrophage numbers and / or phenotypic differentiation responses to IL-34 and supports the use of the antibodies to treat immune-mediated diseases such as neuroinflammation and other inflammatory conditions (see, e.g., Lelios, I. et al. Emerging role of IL-34 in health and disease, J Exp Med (2020) 217(3):e20190290).

[0237] Example 4: Characterization of the immunogenic potential of Antibody 1 Dendritic cell (DC) internalization assay Monocyte-derived DC culture (MDDC) CD14+ monocytes are isolated from peripheral blood mononuclear cells (PBMCs), cultured, and differentiated into DCs according to standard protocols. Briefly, PBMCs are isolated from LRS-WBCs using density gradient centrifugation with Ficoll (#17-1440-02, GE Healthcare) and Sepmate 50 (#15450, STEMCELL Technologies). CD14+ monocytes are isolated using a positive control with CD14+ microbead kit (#130-050-201, Miltenyi Biotec) according to the manufacturer's manual. The cells are then cultured at 1 million / ml for 6 days with 1000 units / ml GM-CSF and 600 units / ml IL-4 to induce immature dendritic cells (MDDCs) in RPMI medium (hereafter referred to as complete RPMI medium or medium purchased from Life Technologies) supplemented with L-glutamine and 25 mM HEPES, 10% FBS, 1 mM sodium pyruvate, 1× penicillin-streptomycin, 1× non-essential amino acids, and 55 μM 2-mercaptoethanol. The medium is changed twice, on days 2 and 5. On day 6, the cells are gently collected with a cell scraper and used for experiments. MDDCs are visually characterized for dendritic morphology by microscopy and expression of CD14, CD11c, and HLA-DR by flow cytometry. The ability to respond to LPS treatment is confirmed by measuring the increase in CD80, CD83, and CD86 using flow cytometry.

[0238] Binding of Fab-TAMRA-QSY7 F(ab')2 fragment goat anti-human IgG (Jackson ImmunoResearch) is dual-labeled with QSY7-NHS and TAMRA-SE (Molecular Probes) to obtain Fab-TAMRA-QSY7, which is used as a universal probe to follow the internalization of test articles. Each vial of F(ab')2 (approximately 1 ml at 1.3 mg / ml) is concentrated to approximately 2 mg / ml by centrifugation at 14,000 rcf for 2 min using an Amico Ultra-0.5 centrifugal filter device (#UFC501096, Millipore). The pH is adjusted to basic (>pH 8) with 10% (v / v) 1 M sodium bicarbonate and 6.8 μl of 10 mM stock solution of QSY-NHS in DMSO is added and mixed. The reaction vial is kept in the dark at room temperature for 30 min. The intermediate product, Fab-QSY7, is purified using a Zeba Spin desalting column (#89890, Thermo Scientific) by centrifugation at 1000 relative centrifugal force (RCF) for 2 minutes. The concentration and degree of labeling (DOL) are calculated by measuring the absorbance at 280 nm and 560 nm with a NanoDrop (ThermoFisher). Fab-QSY7 is then concentrated to approximately 2 mg / ml by centrifugation again at 14,000 rcf for 2 minutes using an Amico Ultra-0.5 centrifugal filter device. After adjusting the pH with 10% (v / v) 1 M sodium bicarbonate, 4.3 μl of 15 mM TAMRA-SE stock solution in DMSO is added and mixed. After 30 minutes at room temperature in the dark, the final product, Fab-TAMRA-QSY7, is purified and collected using a Zeba Spin desalting column by centrifugation at 1000 rcf for 2 minutes. Concentration and DOL are again quantified by reading absorbance at 280 nm, 555 nm, and 560 nm on a NanoDrop spectrophotometer. Using this protocol, approximately 300 μl of Fab-TAMRA-QSY7 at approximately 1.5 mg / ml is obtained, containing approximately 2 QSY7 and 2 TAMRA per F(ab')2.

[0239] Standardized internalization assay by FACS Individual test molecules are normalized to 1 mg / ml in PBS and then further diluted to 8 μg / ml in complete RPMI medium. Fab-TAMRA-QSY7 is diluted to 5.33 μg / ml in complete RPMI medium. Equal volumes of antibody and Fab-TAMRA-QSY7 are mixed and incubated for 30 min at 4°C in the dark for complex formation. MDDCs are resuspended at 4 million / ml in complete RPMI medium and plated at 50 μl per well in a 96-well round-bottom plate with 50 μl of antibody / probe complex. Cells are incubated for 24 hours at 37°C in a CO2 incubator. Cells are washed with 2% FBS PBS and resuspended in 100 μl of 2% FBS PBS with Cytox Green live / dead dye. Data are collected on a BD LSR Fortessa X-20 and analyzed with FlowJo. Live single cells are gated and the percentage of TAMRA fluorescent positive cells is recorded as readout.

[0240] Data presentation and statistical analysis Molecules are tested in duplicate or triplicate in three or more donors. For each donor, the percentage of the TAMRA positive population is considered. To allow comparison of molecules with data generated from different donors, a normalized internalization index (NII) is used. The internalization signal is normalized to the IgG1 isotype (NII=0) and the internal positive control PC (NII=100) using the following formula:

number

[0241] MAPPs assay (MHC-associated peptide proteomics) Primary human dendritic cells from 10 normal human donors were prepared from buffy coats by isolation of CD-14 positive cells as described below and differentiated into immature dendritic cells by incubation with 20ng / ml IL-4 and 40ng / ml GM-CSF in complete RPMI medium containing 5% serum replacement (Thermo Fisher Scientific, Cat. No. A2596101) at 37°C and 5% CO2 for 3 days (Knierman et al., “The Human Leukocyte Antigen Class II Immunopeptidome of the SARS-CoV-2 Spike Glycoprotein”, Cell Reports, 33, 108454 (2020)). On day 4, 3 micromolar test antibodies were added to approximately 5 × 10 6 Fresh medium containing 5 μg / ml LPS is added to the cells and replaced after 5 hours of incubation to transform the cells into mature dendritic cells. The next day, mature cells are lysed in 1 ml of RIPA buffer containing protease inhibitors and DNAse. Lysates are stored at -80°C until sample analysis.

[0242] HLA-II molecules are isolated from thawed lysates using a biotinylated anti-pan HLA class II antibody (clone Tu39) using an automated liquid handling system. Bound receptor-peptide complexes are eluted with 5% acetic acid, 0.1% TFA. Eluted MHC-II peptides are passed through a pre-washed 10k MWCO filter to remove high molecular weight proteins. Separated MHC-II peptides are analyzed by nano LC / MS using a Thermo easy 1200 nLC-HPLC system equipped with a Thermo LUMOS mass spectrometer. The separation uses a 75 μm × 7 cm YMC-ODS C18 column with a flow rate of 250 nL / min and a 65 min gradient of 0.1% formic acid in water as solvent A and 80% acetonitrile with 0.1% formic acid as solvent B. Mass spectrometry was performed in full scan mode at a resolution of 240,000, followed by a 3 second data-dependent MS / MS cycle consisting of a rapid scan in the ion trap with HCD and EThcD fragmentation.

[0243] Peptide identification is generated by an internal proteomics pipeline (Higgs et al., “Label-free LC-MS method for the identification of biomarkers”, Methods in Molecular Biology, 428, 209-230 (2008)) using multiple search algorithms without enzymatic search parameters against bovine / human databases containing the test antibody sequences. The KNIME workflow is used to process the sample identification files. Peptides identified from the test articles are aligned against the parent sequence. A summary of all donors is created annotating the percentage of donors displaying non-germline residues, the number of different regions displaying peptides containing non-germline residues, and the depth of peptide display in each region containing non-germline residues. An increasing degree of non-germline peptide display is associated with an increased risk of immunogenicity. Results for Antibody 1 are shown in Table 12. [Table 12]

[0244] T cell proliferation assay This assay evaluates the ability of the test candidate or peptide clusters derived from the MAPPs of the test candidate to activate CD4+ T cells by inducing cell proliferation as described below (Walsh et al., “Post-hoc assessment of the immunogenicity of three antibodies reveals distinct immune stimulatory mechanisms”, mAbs, 12, 1764829 (2020)). Cryopreserved PBMCs from 10 healthy donors were used, and PBMCs were depleted of CD8+ T cells and labeled with 1 μM carboxyfluorescein diacetate succinimidyl ester (CFSE). PBMCs were cultured at 4 × 10 in AIM-V medium (Life Technologies, Catalog No. 12055-083) containing 5% CTS™ Immune Cell SR (Gibco, Catalog No. A2596101). 6 Cells were seeded at 1000 cells / ml / well and tested in triplicate in 2.0 mL containing various test substances, DMSO control, medium control, and keyhole limpet hemocyanin (KLH; positive control). Cells were cultured and incubated at 37°C with 5% CO2 for 7 days. On day 7, samples were stained with the following cell surface markers for viability detection by flow cytometry using a BD LSRFortessa™ equipped with a high throughput sampler (HTS). Data were analyzed and mitotic index (CDI) was calculated using FlowJo® software (FlowJo, LLC, TreeStar). Briefly, the CDI for each test molecule was calculated as the ratio of the number of CFSE-positive cells growing in stimulated wells to the number of CFSE-positive cells growing in stimulated wells. dim The percentage of CD4+ T cells proliferating in unstimulated wells was compared with that of CFSE dim The CD4+ T cell percentage was calculated by dividing by the percentage of CD4+ T cells. A CDI of 2.5 or greater was considered to represent a positive response. Donor frequency percentages across all donors were assessed. The results for Antibody 1 are shown in Table 13. [Table 13]

[0245] Example 5: Antibody pharmacokinetics in cynomolgus monkeys Cynomolgus monkeys are administered a single intravenous (IV) dose of 3 mg / kg of Antibody 1 in PBS (pH 7.4) at a volume of 1 mL / kg. For pharmacokinetic characterization, blood is collected from two animals / time points at 1, 3, 6, 24, 48, 72, 96, 120, 168, 240, 336, 408, 504, and 672 hours post-dose and processed to serum. Serum concentrations of Antibody 1 are determined by a qualified immunoaffinity liquid chromatography mass spectrometry method. Antibody 1 and a human antibody internal standard (stable isotope-labeled human IgG) are extracted from 100% cynomolgus monkey serum using a biotinylated goat anti-human IgG antibody, followed by quantification of tryptic surrogate peptides using a Q-Exactive™ Orbitrap® mass spectrometer. Pharmacokinetic parameters are calculated for each animal (N=2) using noncompartmental analysis (NCA) and parameters are summarized by mean values. NCA and summary statistical calculations are performed using Phoenix. As shown in Table 14, Antibody 1 exhibits an enhanced pharmacokinetic profile in cynomolgus monkeys. [Table 14]

[0246] Amino acid and nucleotide sequences Heavy chain of Antibody 1 (SEQ ID NO:1) EVQLLESGGGLVQPGGSLRLSCAASGFAFSNYAMSWVRQAPGKGLEWVSAISASGGKTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKRGYLWHAFDHWGR GTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYG PPCPCPPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEK TISKAKGQPREPQVYTLPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG Light chain of antibody 1 (SEQ ID NO:2) EIVLTQSPGTLSLSPGERATLSCRASQSVSSLYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQVVGSSPPFTFGGGTKVEI KRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC HCVR of Antibody 1 (SEQ ID NO:3) EVQLLESGGGLVQPGGSLRLSCAASGFAFSNYAMSWVRQAPGKGLEWVSAISASGGKTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKRGYLWHAFDH LCVR of Antibody 1 (SEQ ID NO: 4) EIVLTQSPGTLSLSPGERATLSCRASQSVSSLYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQVVGSSPPFT HCDR1 of Antibody 1 (SEQ ID NO:5) AASGFAFSNYAMS HCDR2 of Antibody 1 (SEQ ID NO:6) AISASGGKTY HCDR3 of Antibody 1 (SEQ ID NO:7) AKRGYLWHAFDH LCDR1 of Antibody 1 (SEQ ID NO:8) RASQSVSSLYLA LCDR2 of Antibody 1 (SEQ ID NO: 9) YGASS RAT LCDR3 of Antibody 1 (SEQ ID NO: 10) QVVGSSPPFT DNA encoding the heavy chain of Antibody 1 (SEQ ID NO:11) DNA encoding the light chain of Antibody 1 (SEQ ID NO: 12) gaaatagttctcactcagtcccctgggacactctccctgagtccaggagaacgtgcaacactcagttgccgtgcaagccagtccgtctcatccttgtatcttgcttggtaccaacaaaaacctggacaggccccccgtcttcttatctatggtgcctccagt cgcgcaactggtattcccgaccggttcagcggcagtgggtccggcactgacttcaccctgactataagtcggttggagccagaggactttgccgtgtactattgccaagtggtgggaagctcccctcccttcacttttcggcggagggaccaaggtagaaatc aaaagaactgtggcggcgccatctgtcttcatcttcccgccatctgatgagcagttgaaatccggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccag gagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgc HCDR1 of Antibody 1 (Kabat) (SEQ ID NO: 13) NYAMS HCDR2 of Antibody 1 (Kabat) (SEQ ID NO: 14) AISASGGKTYYADSVKG HCDR3 of Antibody 1 (Kabat) (SEQ ID NO: 15) RGYLWHAFDH LCDR1 (SEQ ID NO: 16) of Antibody 1 (Kabat) RASQSVSSLYLA LCDR2 of Antibody 1 (Kabat) (SEQ ID NO: 17) GASSRAT LCDR3 of Antibody 1 (Kabat) (SEQ ID NO: 18) QVVGSSPPFT HCDR1 of Antibody 1 (Chothia) (SEQ ID NO: 19) GFAFSNY HCDR2 of Antibody 1 (Chothia) (SEQ ID NO: 20) SASGGK HCDR3 of Antibody 1 (Chothia) (SEQ ID NO: 21) RGYLWHAFDH LCDR1 (SEQ ID NO: 22) of Antibody 1 (Chothia) RASQSVSSLYLA LCDR2 of Antibody 1 (Chothia) (SEQ ID NO: 23) GASSRAT LCDR3 of Antibody 1 (Chothia) (SEQ ID NO: 24) QVVGSSPPFT HCDR1 (SEQ ID NO:25) of Antibody 1 (IMGT) GFAFSNYA HCDR2 of Antibody 1 (IMGT) (SEQ ID NO:26) ISASGGKT HCDR3 of Antibody 1 (IMGT) (SEQ ID NO: 27) AKRGYLWHAFDH LCDR1 (SEQ ID NO: 28) of Antibody 1 (IMGT) QSVSSLY LCDR2 (SEQ ID NO:29) of Antibody 1 (IMGT) GAS LCDR3 (SEQ ID NO: 30) of Antibody 1 (IMGT) QVVGSSPPFT Human IL-34 (SEQ ID NO:31) NEPLEMWPLTQNEECTVTGFLRDKLQYRSRLQYMKHYFPINYKISVPYEGVFRIANVTRLQRAQVSERELRYLWVLVSLSATESVQDVLLEGHPSWKYLQEVETLLLNVQQ GLTDVEVSPKVESVLSLLNAPGPNLKLVRPKALLDNCFRVMELLYCSCCKQSSVLNWQDCEVPSPQSCSPEPSLQYAATQLYPPPPWSPSSPPHSTGSVRPVRAQGEGLLP IgG4 PAA hinge region (SEQ ID NO: 32) ESKYGPPCPPCP IgG4PAA Fc region (SEQ ID NO:33) APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISK AKGQPREPQVYTLPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG Cynomolgus CSF1R ECD-Fc sequence (SEQ ID NO: 34) IPVIEPSGPELVVKPGETVTLRCVGNGSVEWDGPISPHWTLYSDGPSSVLTTNNATFQNTRTYRCTEPGDPLGGSAAIHLYVKDPARPWNVLAKEVVVFEDQDALLPCLLTDPVLEAGVSLVRLRGRPLLRHTNYSFSPWHGFIIHRAKFIQGQDYQCSALMGGRKVMSISIRLKVQKVIP GPPALTLVPAELVRIRGEAAQIVCSASNIDVDFDVFLQHNTTKLAIPQRSDFHDNRYQKVLTLSLGQVDFQHAGNYSCVASNVQGKHSTSMFRVVESAYLDLSSEQNLIQEVTVGEGLNLKVMVEAYPGLQGFNWTYLGPFSDHQPEPKLANATTKDTYRHTFTLSLPRLKPSEAGRYSFL ARNPGGWRALTFELTLRYPPEVSVIWTSINGSGTLLCAASGYPQPNVTWLQCAGHTDRCDEAQVLQVWVDPHPEVLSQEPFQKVTVQSLLTAETLEHNQTYECRAHNSVGSGSWAFIPISAGARTHPPDEAAAEPKSSDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVV VDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP Heavy chain of antibody 2 (SEQ ID NO:35) EVQLLESGGGLVQPGGSLRLSCAASGFAFSNYAMSWVRQAPGKGLEWVSAISASGGKTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKRGYLWHAFDHWGRG TLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDK THTCPPCPAPEAEGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPSSIEK TISKAKGQPREPQVYTLPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK Heavy chain of antibody 3 (SEQ ID NO:36) EVQLLESGGGLVQPGGSLRLSCAASGFAFSNYAMSWVRQAPGKGLEWVSAISASGGKTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKRGYLWHAFDHWGRG TLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDK THTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEK TISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG Heavy chain of antibody 4 (SEQ ID NO:37) EVQLLESGGGLVQPGGSLRLSCAASGFAFSNYAMSWVRQAPGKGLEWVSAISASGGKTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKRGYLWHAFDHWGR GTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCC VECPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKT ISKTKGQPREPQVYTLPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG Heavy chain of donanemab (SEQ ID NO:38) QVQLVQSGAEVKKPGSSVKVSCKASGYDFTRYYINWVRQAPGQGLEWMGWINPGSGNTKYNEKFKGRVTITADESTSTAYMELSSLRSEDTAVYYCAREGITVYWGQGTTV TVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTH TCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKT ISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG Donanemab light chain (SEQ ID NO:39) DIVMTQTPLSLSVTPGQPASISCKSSQSLLYSRGKTYLNWLLQKPGQSPQLLIYAVSKLDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCVQGTHYPFTFGQGTKL EIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC Heavy chain of anti-N3pG antibody (SEQ ID NO: 40) EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYPMSWVRQAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAREGGSGSYYNGFDYW GQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC DKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIE KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG Light chain of anti-N3pG antibody (SEQ ID NO: 41) DIQMTQSPSTLSASVGDRVTITCRASQSLGNWLAWYQQKPGKAPKLLIYQASTLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQHYKGSFWTFGQGTKVEIK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

Claims

1. An antibody that binds to human IL-34, the antibody comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises heavy chain complementarity determining regions (HCDRs) HCDR1, HCDR2, and HCDR3, and the VL comprises light chain complementarity determining regions (LCDRs) LCDR1, LCDR2, and LCDR3; the HCDR1 comprises SEQ ID NO: 5; said HCDR2 comprising SEQ ID NO: 6; the HCDR3 comprises SEQ ID NO: 7; the LCDR1 comprises SEQ ID NO: 8; the LCDR2 comprises SEQ ID NO: 9, and An antibody wherein the LCDR3 comprises SEQ ID NO:

10.

2. The antibody of claim 1, wherein the VH comprises SEQ ID NO: 3 and the VL comprises SEQ ID NO:

4.

3. The antibody of claim 1, wherein the antibody comprises a heavy chain (HC) comprising an amino acid sequence having at least 95% sequence identity with SEQ ID NO: 1 and a light chain (LC) comprising an amino acid sequence having at least 95% sequence identity with SEQ ID NO:

2.

4. A pharmaceutical composition comprising the antibody of any one of claims 1 to 3 and a pharmaceutically acceptable excipient, diluent, or carrier.

5. A therapeutic agent for immune-mediated diseases, comprising the antibody according to any one of claims 1 to 3.

6. 6. The therapeutic agent of claim 5, wherein the immune-mediated disease is selected from the group consisting of Alzheimer's disease, tauopathy, Sjogren's syndrome (SS), rheumatoid arthritis (RA), inflammatory bowel disease (IBD), atopic dermatitis, kidney disease, sepsis, and / or non-alcoholic fatty liver disease (NAFLD).

7. The therapeutic agent according to claim 6, wherein the immune-mediated disease is Alzheimer's disease.

8. Use of an antibody according to any one of claims 1 to 3 in the manufacture of a medicament for the treatment of an immune-mediated disease.

9. 9. The use of claim 8, wherein the immune-mediated disease is selected from the group consisting of Alzheimer's disease, tauopathy, Sjogren's syndrome (SS), rheumatoid arthritis (RA), inflammatory bowel disease (IBD), atopic dermatitis, kidney disease, sepsis, and / or non-alcoholic fatty liver disease (NAFLD).

10. 9. The use according to claim 8, wherein the immune-mediated disease is Alzheimer's disease.

11. An agent for treating or preventing a disease characterized by amyloid beta (Aβ) deposition in the brain of a human subject, comprising an anti-N3pG Aβ antibody, characterized in that the anti-N3pG Aβ antibody is administered simultaneously, separately, or sequentially in combination with an effective amount of the antibody according to any one of claims 1 to 3 to the human subject in need of said treatment or prevention.

12. The agent according to claim 11 , wherein the anti-N3pG Aβ antibody is donanemab.

13. The agent according to claim 11, wherein the antibody according to any one of claims 1 to 3 is antibody 1.

14. The agent according to claim 13 , wherein antibody 1 is administered sequentially after a course of treatment with an anti-N3pG Aβ antibody, and the anti-N3pG Aβ antibody is donanemab.

15. The agent according to claim 11, i) administering to the human subject one or more first doses of 100 mg to 700 mg of an anti-N3pG Aβ antibody, wherein each first dose is administered once every four weeks; ii) four weeks after administering the one or more first doses, administering to the human subject one or more second doses of greater than 700 mg to 1400 mg of the anti-N3pG Aβ antibody, wherein each second dose is administered once every four weeks; the anti-N3pGlu Aβ antibody is donanemab; iii) A method for treating a human subject comprising administering to said subject simultaneously, separately or sequentially an effective amount of an antibody according to any one of claims 1 to 3, wherein said antibody according to any one of claims 1 to 3 is used as antibody 1.

16. 16. The method of claim 15, wherein the human subject is administered the first dose of donanemab once, twice, or three times prior to administering the second dose.

17. 16. The method of claim 15, wherein the human subject is administered a first dose of 700 mg of donanemab.

18. 16. The method of claim 15, wherein the human subject is administered one or more second doses of 800 mg, 900 mg, 1000 mg, 1100 mg, 1200 mg, 1300 mg, or 1400 mg of donanemab.

19. 16. The method of claim 15, wherein the human subject is administered one or more second doses of 1400 mg of donanemab.

20. 16. The method of claim 15, wherein the anti-N3pGlu Aβ antibody is administered to the human subject over a course of treatment of up to 72 weeks, or until normal levels of amyloid are achieved.

21. 16. The method of claim 15, wherein the anti-N3pGlu Aβ antibody is administered to the human subject until the amyloid plaque level in the patient is 25 centiloids or less.

22. 16. The agent of claim 15, wherein the anti-N3pGlu Aβ antibody is administered for a course of treatment to the human subject until amyloid plaque levels in the human subject are 25 centiloids or less in two consecutive PET imaging scans, optionally wherein the two consecutive PET imaging scans are separated by at least six months, or 11 centiloids or less in one PET imaging scan.

23. 16. The method of claim 15, wherein the human subject is administered three 700 mg first doses of donanemab once every four weeks, followed by a 1400 mg second dose once every four weeks for a course of treatment of up to 72 weeks.

24. 16. The method of claim 15, wherein the human subject is administered three 700 mg first doses once every four weeks, followed by a 1400 mg second dose once every four weeks until the amyloid plaque level in the subject is 25 centiloids or less.

25. 16. The method of claim 15, wherein the human subject is administered three 700 mg first doses of donanemab once every four weeks, followed by a 1400 mg second dose once every four weeks, until amyloid plaque levels in the subject are 25 centiloids or less in two consecutive PET imaging scans, optionally wherein the two consecutive PET imaging scans are separated by at least six months, or 11 centiloids or less in one PET imaging scan.

26. 16. The method of claim 15, wherein the human subject is administered the second dose of donanemab over a course of treatment sufficient to treat or prevent the disease.

27. The agent according to claim 15, wherein the treatment or prevention of the disease results in i) a reduction in Aβ deposits in the brain of the human subject, and / or ii) a delay in cognitive or functional decline in the human subject.

28. 28. The agent of claim 27, wherein the reduction in Aβ deposits in the brain of the human subject is determined by amyloid PET brain imaging or a diagnostic method that detects a biomarker of Aβ.

29. The method of claim 27, wherein the second dose is administered to the human subject until Aβ deposits in the brain of the human subject are reduced by 20 to 100%.

30. 30. The method of claim 29, wherein Aβ deposits in the brain of the human subject are reduced by 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 100%.

31. 16. The agent of claim 15, wherein the second dose of donanemab is administered to the human subject until Aβ deposits in the brain of the human subject are reduced by i) an average of 25 to 100 centiloids, ii) an average of 50 to 100 centiloids, iii) 100 centiloids, or iv) 84 centiloids.

32. 16. The agent according to claim 15, wherein the disease characterized by Aβ deposition in the brain of the human subject is selected from preclinical Alzheimer's disease (AD), clinical AD, prodromal AD, mild AD, moderate AD, severe AD, Down's syndrome, clinical cerebral amyloid angiopathy, or preclinical cerebral amyloid angiopathy.

33. The method of claim 15, wherein the human subject is an early-symptomatic AD patient.

34. The method of claim 33, wherein the human subject has prodromal AD and mild dementia due to AD.

35. 16. The agent of claim 15, wherein the human subject i) has been determined to have a very low to moderate tau burden or has a very low to moderate tau burden, ii) has been determined to have a low to moderate tau burden or has a low to moderate tau burden, iii) has been determined to have a very low to moderate tau burden or has a very low to moderate tau burden and one or two alleles of APOE e4, iv) has been determined to have a low to moderate tau burden or has a low to moderate tau load and one or two alleles of APOE e4, or v) has one or two alleles of APOE e4.

36. 36. The agent of claim 35, wherein the human subject i) has a very low to moderate tau burden when the tau burden as measured by PET brain imaging is 1.46 SUVr or less, or ii) has a low to moderate tau burden when the tau burden as measured by PET brain imaging is 1.10 SUVr to 1.46 SUVr.

37. 16. The agent of claim 15, wherein the human subject i) does not have a high tau burden or has been determined to not have a high tau burden, or ii) carries one or two alleles of APOE e4 and does not have a high tau burden or has been determined to not have a high tau burden.

38. 38. The agent of claim 37, wherein the human subject has a high tau burden if the tau burden as measured by PET brain imaging is greater than 1.46 SUVr.

39. 36. The agent of claim 35, wherein the tau burden of the human subject is determined using PET brain imaging or a diagnostic method that detects a tau biomarker.

40. 38. The agent of claim 37, wherein the tau burden of the human subject is determined using PET brain imaging or a diagnostic method that detects a tau biomarker.

41. An agent for treating or preventing a disease characterized by amyloid beta (Aβ) deposits in the brain of a human subject, comprising an anti-N3pGlu Aβ antibody, i) administering to the human subject one or more first doses of 100 mg to 700 mg of an anti-N3pGlu Aβ antibody, wherein each first dose is administered once every four weeks; ii) four weeks after administering the one or more first doses, one or more second doses of greater than 700 mg to 1400 mg of an anti-N3pGlu Aβ antibody are administered to the human subject, wherein each second dose is administered once every four weeks; combined simultaneously, separately or sequentially with an effective amount of antibody 1; if the human subject has tau burden in the temporal lobe, occipital lobe, parietal lobe, or frontal lobe of the brain prior to administration of the one or more first doses of an anti-N3pGlu Aβ antibody, determining whether the human subject has tau burden in the temporal lobe, occipital lobe, parietal lobe, or frontal lobe of the brain; The agent, wherein the antibody 1 is an antibody according to any one of claims 1 to 3.

42. 42. The method of claim 41, wherein the human subject has tau burden in the posterior lateral temporal lobe or temporal lobe of the brain.

43. 42. The method of claim 41, wherein the human subject has tau burden in the occipital lobe of the brain.

44. 42. The method of claim 41, wherein the human subject has tau burden in the parietal lobe of the brain.

45. 42. The agent of claim 41, wherein the human subject has tau burden in the frontal lobe of the brain.

46. 42. The method of claim 41, wherein the human subject has tau burden in the posterolateral temporal lobe (PLT) and / or occipital lobe of the brain.

47. 42. The agent of claim 41, wherein the human subject has tau burden in i) the parietal or precuneus region, or ii) the frontal region, along with tau burden in the PLT or occipital region of the brain.

48. 42. The agent of claim 41, wherein the human subject has tau burden isolated to i) the frontal lobe of the brain, or ii) tau burden in regions of the temporal lobe that do not include the posterolateral temporal region (PLT).

49. 42. The method of claim 41, wherein the human subject has tau burden in the posterior lateral temporal, occipital, and parietal lobes of the brain.

50. 42. The agent of claim 41, wherein the human subject has tau burden in the posterolateral temporal lobe, occipital lobe, parietal lobe, and frontal lobe of the brain.

51. 42. The method of claim 41, wherein the human subject has tau burden in the posterolateral temporal lobe, occipital lobe, parietal lobe and / or frontal lobe of the brain.

52. 42. The method of claim 41, wherein the human subject is administered the first dose once, twice, or three times before receiving the second dose.

53. 53. The method of claim 52, wherein the human subject is administered a first dose of 700 mg.

54. 53. The method of claim 52, wherein the human subject is administered one or more second doses of 800 mg, 900 mg, 1000 mg, 1100 mg, about 1200 mg, 1300 mg, or 1400 mg.

55. 53. The method of claim 52, wherein the human subject is administered one or more second doses of 1400 mg.

56. 53. The method of claim 52, wherein the anti-N3pGlu Aβ antibody is administered to the human subject for a period of up to 72 weeks or until normal levels of amyloid are achieved.

57. 53. The method of claim 52, wherein the anti-N3pGlu Aβ antibody is administered to the human subject until amyloid plaque levels in the patient are 25 centiloids or less.

58. 53. The method of claim 52, wherein the anti-N3pGlu Aβ antibody is administered to the human subject until amyloid plaque levels in the human subject are 25 centiloids or less in two consecutive PET imaging scans, optionally wherein the two consecutive PET imaging scans are separated by at least six months, or 11 centiloids or less in one PET imaging scan.

59. 53. The method of claim 52, wherein the human subject receives three 700 mg first doses once every four weeks, followed by a 1400 mg second dose once every four weeks for up to 72 weeks.

60. 53. The method of claim 52, wherein the human subject is administered three 700 mg first doses once every four weeks, followed by a 1400 mg second dose once every four weeks, until the amyloid plaque level in the subject is 25 centiloids or less.

61. 53. The method of claim 52, wherein the human subject is administered three 700 mg first doses once every four weeks, followed by a 1400 mg second dose once every four weeks, until the amyloid plaque level in the subject is 25 centiloids or less in two consecutive PET imaging scans, optionally wherein the two consecutive PET imaging scans are separated by at least six months, or 11 centiloids or less in one PET imaging scan.

62. 53. The method of claim 52, wherein the human subject is administered the second dose for a period of time sufficient to treat or prevent the disease.

63. 53. The method of claim 52, wherein treating or preventing a disease results in i) a reduction in Aβ deposits in the brain of the human subject, and / or ii) a delay in cognitive or functional decline in the human subject.

64. 53. The agent of claim 52, wherein the reduction in Aβ deposits in the brain of the human subject is determined by amyloid PET brain imaging or a diagnostic method that detects a biomarker of Aβ.

65. 53. The method of claim 52, wherein the second dose is administered to the human subject until Aβ deposits in the brain of the human subject are reduced by 20-100%.

66. 53. The agent of claim 52, wherein Aβ deposits in the brain of the human subject are reduced by 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 100%.

67. 53. The agent of claim 52, wherein the second dose is administered to the human subject until Aβ deposits in the brain of the human subject are reduced by i) an average of 25 to 100 centiloids, ii) an average of 50 to 100 centiloids, iii) 100 centiloids, or iv) 84 centiloids.

68. 53. The agent of claim 52, wherein the disease characterized by Aβ deposits in the brain of the human subject is selected from preclinical Alzheimer's disease (AD), clinical AD, prodromal AD, mild AD, moderate AD, severe AD, Down's syndrome, clinical cerebral amyloid angiopathy, or preclinical cerebral amyloid angiopathy.

69. 53. The method of claim 52, wherein the human subject is an early-symptomatic AD patient.

70. 53. The method of claim 52, wherein the human subject has prodromal AD and mild dementia due to AD.

71. 53. The agent of claim 52, wherein the human subject i) has a very low to moderate tau burden or has been determined to have a very low to moderate tau burden, or ii) has a low to moderate tau burden or has been determined to have a low to moderate tau burden.

72. 53. The agent of claim 52, wherein the human subject i) has a very low to moderate tau burden when the tau burden as measured by PET brain imaging is 1.46 SUVr or less, or ii) has a low to moderate tau burden when the tau burden as measured by PET brain imaging is 1.10 SUVr to 1.46 SUVr.

73. 53. The agent of claim 52, wherein the human subject does not have, or has been determined to not have, a high tau burden.

74. 53. The agent of claim 52, wherein the human subject has a high tau burden if the tau burden as measured by PET brain imaging is greater than 1.46 SUVr.

75. 53. The agent of claim 52, wherein the tau burden of the human subject is determined using PET brain imaging or a diagnostic method that detects a tau biomarker.

76. The agent of claim 52, wherein the anti-N3pGlu Aβ antibody comprises donanemab.

77. 53. The method of claim 52, wherein the patient has one or two alleles of APOE e4.

78. An agent for reducing / preventing further increase in tau burden or slowing the rate of tau accumulation in the temporal lobe, occipital lobe, parietal lobe, or frontal lobe of the human brain, comprising an anti-N3pGlu Aβ antibody, characterized in that the agent is used by administering to a human subject the anti-N3pGlu Aβ antibody and an effective amount of antibody 1 simultaneously, separately, or sequentially in combination, wherein antibody 1 is the antibody according to any one of claims 1 to 3.

79. A therapeutic agent for ARIA, comprising the antibody according to any one of claims 1 to 3.

80. A preventive agent for ARIA, comprising the antibody according to any one of claims 1 to 3.

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