Combinatorial therapy for alzheimer's disease

Abstract

The present invention provides novel compositions and methods for treating diseases and conditions associated with amyloid beta in a subject, said method comprising administering to the subject (i) an anti-amyloid beta antibody or an antigen binding fragment thereof and (ii) a regulatory T cell (Treg) inducing or activating agent.

Description

[0001] COMBINATORIAL THERAPY FOR ALZHEIMER’S DISEASE

[0002] By Howard E. Gendelman R. Lee Mosley Pravin Yeapuri Rana Kadry Shaurav Bhattarai Emma Foster

[0003] This application claims priority under 35 U.S.C. §119(e) to U.S. Provisional Patent Application No. 63 / 680,162, filed August 7, 2024. The foregoing application is incorporated by reference herein.

[0004] This invention was made with government support under Grant Nos. P01 DA028555; R01 NS036126; P01 NS031492; P01 MH064570; P01 NS043985; P30 MH062261; R01 AG043540; and R01 NS034239 awarded by the National Institutes of Health. The government has certain rights in the invention.

[0005] FIELD OF THE INVENTION

[0006] The present invention relates generally to amyloid beta related diseases and conditions. More specifically, the present invention provides compositions and methods for treating, inhibiting, and / or preventing a disease or condition associated with aberrant amyloid beta.

[0007] BACKGROUND OF THE INVENTION

[0008] Amyloid plaques are a hallmark of Alzheimer’s disease. In patients with Alzheimer’s disease, abnormal levels of amyloid beta clump together to form plaques that disrupt normal cell function and lead to neuronal cell death. Amyloid plaques have been a popular target for developing potential therapies for Alzheimer’s disease. Recently, humanized antibodies that target aggregated amyloid beta have been approved by the FDA. However, these therapies have had limited clinical success and are also associated with adverse events. Some patients that receive amyloid beta targeted antibody therapy develop what are known as amyloid related imaging abnormalities (ARIA) such as ARIA-edema / effusion and ARIA-hemosiderosis / microhemorrhages. Furthermore, it has been shown that some of these antibodies that target amyloid beta induce microglia activation with a shift to a pro-inflammatory phenotype and consequent peripheral transformation to an effector T cell (Teff) response which exacerbates neuroinflammation and neuronal loss. Accordingly, compositions and methods for improved treatment of Alzheimer’s disease are needed.

[0009] SUMMARY OF THE INVENTION

[0010] In accordance with the instant invention, methods of inhibiting, treating, and / or preventing a disease or condition associated with amyloid beta, particularly oligomeric or aggregated amyloid beta, are provided. In certain embodiments, the method comprises administering to a subject at least one anti-amyloid beta antibody or an antigen binding fragment thereof and at least one Treg inducing / activating agent. In certain embodiments, the anti-amyloid beta antibody or an antigen binding fragment thereof comprises one, two, or all three complementarity determining regions (CDRs) of SEQ ID NO: 6 and one, two, or all three CDRs of SEQ ID NO: 10. In certain embodiments, the anti-amyloid beta antibody or an antigen binding fragment thereof comprises a heavy chain complementarity determining region 1 (CDR-H1), a heavy chain complementarity determining region 2 (CDR-H2), and a heavy chain complementarity determining region 3 (CDR-H3) comprising amino acid sequences SEQ ID NOs: 7, 8, and 9, respectively, and a light chain complementarity determining region 1 (CDR-L1), a light chain complementarity determining region 2 (CDR-L2) and a light chain complementarity determining region 3 (CDR-L3) comprising amino acid sequences SEQ ID NOs: 11, 12, and 13, respectively. In certain embodiments, the anti-amyloid beta antibody or an antigen binding fragment thereof is selected from the group consisting of 2CD3M, lecanemab, donanemab, aducanumab, Remternetug, Gantenerumab, Trontinemab, ABBV916, ACU193, SHR-1707, PMN310, and PRX012. In certain embodiments, the Treg inducing / activating agent is selected from the group consisting of interleukin-2 (IL-2), low-dose IL-2, granulocyte-macrophage colony stimulating factor (GM-CSF), GM-CSF analogs, TGF-P, IL-10, IL-2, glatiramer acetate, anti- CD3, anti-CD28, bee venom phospholipase A2, rapamycin, histone deacetylase inhibitors, vasoactive intestinal peptide (VIP), VIP analogs and VIP receptor-2 agonists, and combinations thereof. In certain embodiments, the Treg inducing / activating agent is IL-2 or low-dose IL-2. In certain embodiments, the anti-amyloid beta antibody or an antigen binding fragment thereof and the Treg inducing / activating agent are administered separately and / or at different times. In certain embodiments, the anti-amyloid beta antibody or an antigen binding fragment thereof and the Treg inducing / activating agent are administered together or simultaneously. In certain embodiments, the disease or condition is a neurodegenerative disease. In certain embodiments, the disease or condition is Alzheimer’s disease. In certain embodiments, anti-amyloid beta antibody or an antigen binding fragment thereof comprises one, two, or all three complementarity determining regions (CDRs) of SEQ ID NO: 6 and one, two, or all three CDRs of SEQ ID NO: 10, the Treg inducing / activating agent is low-dose IL-2, and the disease or condition is Alzheimer’s disease.

[0011] In accordance with another aspect of the instant invention, compositions and kits are provided. In certain embodiments, the compositions comprise (i) an antiamyloid beta antibody or an antigen binding fragment thereof; (ii) a Treg inducing / activating agent; and (iii) a pharmaceutically acceptable carrier.

[0012] BRIEF DESCRIPTIONS OF THE DRAWING

[0013] Figure 1 A provides a schematic of a study for treating APP / PS1 mice with mAb (2CD3M), interleukin-2 (IL-2), or both. Figure IB provides results of surface plasmon resonance assays showing the binding of amyloid beta monomers (top row) and amyloid beta oligomers (bottom row) with aducanumab (left column), lecanemab (middle column), or 2CD3M (right column).

[0014] Figure 2 provides graphs showing the number of errors by the indicated mice in a radial arm water maze test. BM mice were used as a control. APP / PS1 mice were untreated, treated with mAb (2CD3M), and treated with mAb (2CD3M) and IL-2. Mice are assessed over a 9 day period. Each graph represents one 3 day block for days 1-3 (left graph), days 4-6 (middle graph), and days 7-9 (right graph). Each block shows learning trials 1 and 4 (T1 and T4), and retention trial (T5). * p < 0.05. • BM; ■ APP / PS1; ▲ mAb; ▼ mAb + IL-2; ♦ IL-2.

[0015] Figures 3 A And 3B provide graphs of the %Treg and %Teff in the blood (Fig. 3 A) and brain (Fig. 3B) of BM mice (control) and APP / PS1 mice which were untreated or treated with mAb (2CD3M) or with mAb (2CD3M) and IL-2.

[0016] Figure 4A provides images of the hippocampus and cortex of mice receiving the indicated treatments. PBS treatment serves as a control. Figure 4B provides graphs of the amount of amyloid beta in the hippocampus (top) and cortex (bottom) of mice receiving the indicated treatments. DETAILED DESCRIPTION OF THE INVENTION

[0017] The present invention describes a novel combination therapy that helps induce an anti-inflammatory T cell response by inducing regulatory T cells (Tregs). Immune tolerance is activated by Tregs. Tregs signal the immune system to dampen pro-inflammatory responses and become tolerant to self- or modified self-antigens that stimulate pro-inflammatory responses. In the case of neurodegenerative disorders, Tregs mitigate the neuroinflammation that drives neuronal loss in Alzheimer’s disease. Therefore, optimal induction of Treg levels or activity would lead to control of disease-initiating inflammatory activities and mitigate disease progression.

[0018] The present invention describes a combination therapy and methods for treating amyloid beta associated diseases and conditions such as Alzheimer’s disease. The combination therapy comprises an anti-amyloid beta antibody and a Treg inducing / activating agent. In certain embodiments, the anti-amyloid beta antibody is an antibody that binds to amyloid beta, including both monomeric and oligomeric forms of amyloid beta. In certain embodiments, the anti-amyloid beta antibody preferentially binds to oligomeric forms over monomeric forms of amyloid beta. In certain embodiments, the anti-amyloid beta antibody comprises a humanized antibody, an antibody fragment, or a bispecific antibody.

[0019] Amyloid beta precursor amino acid and nucleotide sequences are provided, e.g., in GenBank Gene ID: 351 and GenBank Accession Nos. NM_000484.4 and NP 000475.1. Mature amyloid beta can be a peptide of about 36-43 amino acids in length. In certain embodiments, amyloid beta comprises the amino acid sequence DAEFRHDSGYEVHHQKLVFFAEDVGSNKGAIIGLMVGGVVIA (SEQ ID NO: 1). In certain embodiments, amyloid beta comprises the amino acid sequence DAEFRHDSGYEVHHQKLVFFAEDVGSNKGAIIGLMVGGVV (SEQ ID NO: 2).

[0020] In certain embodiment, the anti-amyloid beta antibody or antigen-binding fragment thereof specifically binds (e.g., is immunologically specific for) to human amyloid beta. In certain embodiments, the anti-amyloid beta antibody or antigenbinding fragment thereof specifically binds to oligomeric amyloid beta and / or fibrillar amyloid beta. In certain embodiments, the anti-amyloid beta antibody or antigen-binding fragment thereof specifically binds aggregated forms of amyloid beta, such as oligomeric and fibril forms. In certain embodiments, the anti-amyloid beta antibody or antigen-binding fragment thereof specifically binds to oligomeric amyloid beta and fibrillar amyloid beta. In certain embodiments, the anti-amyloid beta antibody or antigen-binding fragment thereof does not bind or does not bind significantly to monomeric amyloid beta.

[0021] Examples of anti-amyloid beta antibodies include, without limitation: donanemab (Kisunla™), lecanemab (Leqembi®), aducanumab (Aduhelm™), Remternetug, Gantenerumab, Trontinemab, ABBV916, ACU193, SHR-1707, PMN3 10, PRX012, and 2CD3M (WO 2025 / 111402). In certain embodiments, the anti-amyloid beta antibody is lecanemab. In certain embodiments, the anti-amyloid beta antibody is an antibody described in WO 2025 / 111402, incorporated herein by reference.

[0022] In certain embodiments, the anti-amyloid beta antibody or antigen-binding fragment thereof binds to or specifically binds an epitope or region of amyloid beta that comprises residues 22 to 26 of amyloid beta. In certain embodiments, the antiamyloid beta antibody or antigen-binding fragment thereof binds to or specifically binds to an epitope or region of amyloid beta that comprises the amino acid sequence EDVGS (SEQ ID NO: 3). In certain embodiments, the anti-amyloid beta antibody or antigen-binding fragment thereof binds to or specifically binds to an epitope or region of amyloid beta that comprises the amino acid sequence KLVFFAEDVGSNKGA (SEQ ID NO: 4). In certain embodiments, the antiamyloid beta antibody or antigen-binding fragment thereof binds to or specifically binds to the amino acid sequence CGEDVGSG (SEQ ID NO: 5). The above epitopes or amino acid sequences may be longer or shorter than the above identified sequences by 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more amino acids, particularly 1, 2, 3, 4, or 5 amino acids, at the N-terminus and / or C-terminus of the sequence. In certain embodiments, the above epitopes or amino acid sequences have at least 90%, 95%, 97%, 99%, or 100% homology or identity with SEQ ID NO: 1 or 2. Antibodies which bind the same epitope as an antibody provided herein are also encompassed by the instant invention.

[0023] In certain embodiments, the anti-amyloid beta antibody or antigen-binding fragment thereof comprises a heavy chain (e.g., a heavy chain variable region; VH region) comprising:

[0024] QVQLQQSGAELARPGASVKLSCKASGYTFTSYGISWVKQRTGQGLEWIGEI YPRSGNTYYNEKFKGKATLTADKSSSTAYMELRSLTSEDSAVYFCAREAIR IGRRDAMDYWGQGTSVTVSS (SEQ ID NO: 6). In certain embodiments, the anti-amyloid beta antibody or antigen-binding fragment thereof comprises one, two, or all three CDRs of SEQ ID NO: 6 (e.g., as determined by IMGT, Chothia, Kabat, Martin (e.g., enhanced Chothia or AbM), or Honneger's numbering scheme (Aho)). In certain embodiments, the anti-amyloid beta antibody or antigen-binding fragment thereof comprises one, two, or all three CDRs of SEQ ID NO: 6 as determined by Kabat. In certain embodiments, the anti-amyloid beta antibody or antigen-binding fragment thereof comprises a heavy chain (e.g., VH domain) comprising one, two, or all three CDRs of SEQ ID NO: 6. In certain embodiments, the anti-amyloid beta antibody or antigen-binding fragment thereof comprises one, two, or all three of: SYGIS (SEQ ID NO: 7), EIYPRSGNTYYNEKFKG (SEQ ID NO: 8), and EAIRIGRRDAMDY (SEQ ID NO: 9). In certain embodiments, the anti-amyloid beta antibody or antigen-binding fragment thereof comprises a CDR-H1 which comprises the amino acid sequence SYGIS (SEQ ID NO: 7) or an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology or identity with SEQ ID NO: 7. In certain embodiments, the anti-amyloid beta antibody or antigen-binding fragment thereof comprises a CDR-H2 which comprises the amino acid sequence EIYPRSGNTYYNEKFKG (SEQ ID NO: 8) or an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology or identity with SEQ ID NO: 8. In certain embodiments, the anti-amyloid beta antibody or antigenbinding fragment thereof comprises a CDR-H3 which comprises the amino acid sequence EAIRIGRRDAMDY (SEQ ID NO: 9) or an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology or identity with SEQ ID NO: 9. In certain embodiments, the anti-amyloid beta antibody or antigen-binding fragment thereof comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology or identity with SEQ ID NO: 6. In certain embodiments, the anti-amyloid beta antibody or antigen-binding fragment thereof comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology or identity with SEQ ID NO: 7, SEQ ID NO: 8, and / or SEQ ID NO: 9. In certain embodiments, the anti-amyloid beta antibody or antigen-binding fragment thereof comprises a light chain (e.g., a light chain variable region; VL region) comprising:

[0025] DIVLTQSPATLSVTPGDSVSLSCRASQS ISNNLHWYQQKSHESPRLLIKYA SQSISGIPSRFSGSGSGTDFTLS INSVETEDFGMYFCQQSNSWPHTFGSGT KLEIK (SEQ ID NO: 10).

[0026] In certain embodiments, the anti-amyloid beta antibody or antigen-binding fragment thereof comprises one, two, or all three CDRs of SEQ ID NO: 10 (e.g., as determined by IMGT, Chothia, Kabat, Martin (e.g., enhanced Chothia or AbM), or Honneger's numbering scheme (Aho)). In certain embodiments, the anti-amyloid beta antibody or antigen-binding fragment thereof comprises one, two, or all three CDRs of SEQ ID NO: 10 as determined by Kabat. In certain embodiments, the antiamyloid beta antibody or antigen-binding fragment thereof comprises a light chain (e.g., VL domain) comprising one, two, or all three CDRs of SEQ ID NO: 10. In certain embodiments, the anti-amyloid beta antibody or antigen-binding fragment thereof comprises one, two, or all three of: RASQSISNNLH (SEQ ID NO: 11), YASQSIS (SEQ ID NO: 12), and QQSNSWPHT (SEQ ID NO: 13). In certain embodiments, the anti-amyloid beta antibody or antigen-binding fragment thereof comprises a CDR-L1 which comprises the amino acid sequence RASQSISNNLH (SEQ ID NO: 11) an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology or identity with SEQ ID NO: 11. In certain embodiments, the anti-amyloid beta antibody or antigen-binding fragment thereof comprises a CDR-L2 which comprises the amino acid sequence YASQSIS (SEQ ID NO: 12) or an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology or identity with SEQ ID NO: 12. In certain embodiments, the anti-amyloid beta antibody or antigen-binding fragment thereof comprises a CDR-L3 which comprises the amino acid sequence QQSNSWPHT (SEQ ID NO: 13) an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology or identity with SEQ ID NO: 13. In certain embodiments, the anti-amyloid beta antibody or antigen-binding fragment thereof comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology or identity with SEQ ID NO: 10. In certain embodiments, the anti-amyloid beta antibody or antigen-binding fragment thereof comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology or identity with SEQ ID NO: 11, SEQ ID NO: 12, and / or SEQ ID NO: 13.

[0027] In certain embodiments, the anti-amyloid beta antibody or antigen-binding fragment thereof comprises one, two, or all three CDRs of SEQ ID NO: 6 and one, two, or all three CDRs of SEQ ID NO: 10. In certain embodiments, the antiamyloid beta antibody or antigen-binding fragment thereof comprises a VH region comprising a heavy chain complementarity determining region 1 (CDR-H1), a heavy chain complementarity determining region 2 (CDR-H2), and a heavy chain complementarity determining region 3 (CDR-H3) comprising amino acid sequences SEQ ID NOs: 7, 8, and 9, respectively, and a VL region comprising a light chain complementarity determining region 1 (CDR-L1), a light chain complementarity determining region 2 (CDR-L2) and a light chain complementarity determining region 3 (CDR-L3) comprising amino acid sequences SEQ ID NOs: 11, 12, and 13, respectively. In certain embodiments, the anti-amyloid beta antibody or antigenbinding fragment thereof comprises a VH region comprising SEQ ID NO: 6 and a VL region comprising SEQ ID NO: 10.

[0028] In certain embodiments, the anti-amyloid beta antibody or antigen-binding fragment thereof is a humanized. In certain embodiments, the anti-amyloid beta antibody or antigen-binding fragment thereof is recombinant. In certain embodiments, the anti-amyloid beta antibody or antigen-binding fragment thereof is a chimeric antibody. In certain embodiments, the anti-amyloid beta antibody or antigen-binding fragment thereof is monoclonal. In certain embodiments, the antiamyloid beta antibody or antigen-binding fragment thereof is an antibody of any class or sub-class, including, without limitation: IgG, IgM, IgE, IgA, and IgD. In certain embodiments, the anti-amyloid beta antibody or antigen-binding fragment thereof is an IgM antibody. In certain embodiments, the anti-amyloid beta antibody or antigen-binding fragment thereof is an IgG or subclass thereof. In certain embodiments, the anti-amyloid beta antibody or antigen-binding fragment thereof is an IgGl antibody. In certain embodiments, the anti-amyloid beta antibody or antigen-binding fragment thereof is an IgG2a antibody.

[0029] In certain embodiments, the anti -amyloid beta antibody is an antigen-binding fragment thereof. Examples of antigen-binding fragments include, without limitation: Fab fragments, F(ab’)2 fragments, Fab’ fragments, Fv fragments, an scFv, SCFV2, SCFV-FC, or a single domain antibody. In certain embodiments, the antiamyloid beta antibody or antigen-binding fragment thereof is a single-chain fragment. In certain embodiments, the single-chain fragment comprises the VH and VL regions joined by a linker (e.g., peptide linker), optionally a flexible immunoglobulin linker. In certain embodiments, the anti-amyloid beta antibody or antigen-binding fragment thereof comprises an scFv comprising a VH region and a VL region, particularly as described herein.

[0030] In certain embodiments, the anti -amyloid beta antibody or an antigenbinding fragment thereof is a bispecific antibody. In certain embodiments, the bispecific antibody comprises a first antigen-binding domain that specifically binds to amyloid beta and a second antigen-binding domain that specifically binds to a second antigen which is not amyloid beta (e.g., tau (e.g., human tau)). In certain embodiments, the first antigen-binding domain comprises a first heavy chain variable (VH) region and a first light chain variable (VL) region as described herein.

[0031] In certain embodiments, the anti-amyloid beta antibody or an antigenbinding fragment thereof is a conjugate. In certain embodiments, the anti-amyloid beta antibody or an antigen-binding fragment thereof is conjugated (e.g., directly or via a linker (e.g., peptide linker)) to a therapeutic agent, an imaging agent (e.g., a radionuclide or radioisotope), and / or a detectable moiety (e.g., radionuclide, enzyme, fluorescent moiety or protein, or luminescent moiety). In certain embodiments, the therapeutic agent is an anti-inflammatory agent.

[0032] As described herein, the instant invention also involves the use of an agent which activates and / or induces regulatory T cells (Tregs). Examples of Treg activating / inducing agent include, without limitation: IL-2 (e.g., low-dose IL-2), GM-CSF (e.g., human recombinant GM-CSF, molgramostim, sargramostim), GM- CSF analogs, TGF-P, IL-10, IL-2, glatiramer acetate, anti-CD3 (e.g., anti-CD3 antibodies), anti-CD28 (e.g., anti-CD28 antibodies), bee venom phospholipase A2 (bvPLA2), rapamycin, histone deacetylase inhibitors, vasoactive intestinal peptide (VIP), VIP analogs and VIP receptor-2 agonists (e.g., LBT-3627), or combinations thereof. In certain embodiments, the Treg activating / inducing agent is IL-2 or low dose IL-2. In certain embodiments, the Treg activating / inducing agent is low dose IL-2. The Treg activating / inducing agent may be of the species of the subject being treated (e.g., human). In certain embodiments, the low dose IL-2 is COYA 301. In certain embodiments, a low dose IL-2 is less than 5 million international units per day (MIU / day), less than 4 MIU / day, less than 3 MIU / day, less than 2 MIU / day, or less than 1 MIU / day. In certain embodiments, a low dose IL-2 is about 0.1 to about 5 MIU / day, about 0.5 to about 5 MIU / day, about 0.5 to about 3 MIU / day, or about 1 to about 3 MIU / day.

[0033] Treg activating / inducing agents may be proteins, peptides, small molecules, or nucleic acids (e.g., RNA (e.g., mRNA), DNA, inhibitory nucleic acid molecules (e.g., antisense, siRNA, or shRNA), or nucleic acids encoding a protein or peptide). For Treg activating / inducing agents that are proteins or peptides, these may be administered to a patient in the form of a protein or peptide. In certain embodiments, they may be administered as an oligonucleotide-based construct that is capable of being expressed as a protein or peptide once inside a cell. The Treg activating / inducing agents may be administered to a patient in a variety of ways. In certain embodiments, the Treg activating / inducing agents may be administered directly to a patient. In certain embodiments, formulations such as liposomes, dendrimers, microparticles, and / or nanoparticles, containing the Treg activating / inducing agents may be administered to a patient. In certain embodiments, the formulation or delivery vehicle in a slow-release formulation or delivery vehicle. In certain embodiments, expression constructs (such as vectors containing nucleic acids that encode the Treg activating / inducing agent) may be administered to a patient. Examples of vectors for expressing the molecules of the invention include, without limitation, plasmids and viral vectors (e.g., adeno- associated viruses (AAVs), retroviruses, and lentiviruses). The expression vectors of the instant invention may employ a strong promoter, a constitutive promoter, and / or a regulated promoter. In certain embodiments, the promoter is cell-type specific. In certain embodiments, the nucleic acid molecules are expressed transiently. In certain embodiments, the Treg activating / inducing agents may be administered ex vivo to cells which can then be administered to the patient as a cellbased therapy. If the Treg activating / inducing agents are being administered ex vivo to cells, the cells may be cells obtained from the patient or they may be cells obtained from a donor. The cells may be Tregs that are activated ex vivo or they may be precursor cells or other cells (e.g., immune cells, T cells, Tregs, precursor cells, pre-T cells, bone marrow cells, peripheral blood mononuclear cells (PBMCs)) that are transformed into Tregs ex vivo before being administered to a patient. In certain embodiments, the cells are autologous. In certain embodiments, the method further comprises obtaining the cells from the subject or a donor prior to treatment of the cells with the Treg activating / inducing agent.

[0034] In certain embodiments, more than one Treg activating / inducing agent is used. In certain embodiments, the more than one Treg activating / inducing agent act synergistically. Synergy allows for reduced doses and lowered toxicities. In certain embodiments, two or more Treg activating / inducing agents are administered to a patient. The two or more Treg activating / inducing agent may be administered at the same time and / or they may be administered at different times. In certain embodiments, the two or more Treg activating / inducing agents comprise GM-CSF and IL-2 (or low dose IL-2).

[0035] Compositions comprising (i) an anti-amyloid beta antibody or an antigenbinding fragment thereof of the instant invention and / or at least one Treg activating / inducing agent and (ii) a carrier such as a pharmaceutically acceptable carrier are also encompassed herein. In certain embodiments, the composition comprises at least one anti-amyloid beta antibody or an antigen-binding fragment thereof, at least one Treg activating / inducing agent, and at least one carrier (e.g., a pharmaceutically acceptable carrier). The anti-amyloid beta antibody or an antigenbinding fragment thereof and Treg activating / inducing agent may be formulated together or individually.

[0036] The instant invention also encompasses kits comprising (i) an anti-amyloid beta antibody or an antigen-binding fragment thereof and (ii) at least one Treg activating / inducing agent. As used herein, a “kit” is a packaged combination of agents for use together for a purpose (e.g., therapeutic use (e.g., treating Alzheimer’s disease)). A kit may be a delivery system or enclosure (e.g., package or box) for delivering materials such as a combination of components, compositions, and / or packaging. For example, the kit may be a packaged product which packaging includes two or more independent containers or bottles, each containing a different composition. The kit may further comprise additional agents and / or instructions for use of the combination of agents. In certain embodiments, the kit comprises a first composition comprising an anti-amyloid beta antibody or an antigen-binding fragment thereof and a carrier (e.g., a pharmaceutically acceptable carrier) and a second composition comprising a Treg activating / inducing agent and a carrier (e.g., a pharmaceutically acceptable carrier). In certain embodiments, the kit comprises the carriers (e.g., a pharmaceutically acceptable carriers) or diluents (e.g., sterilized water or saline solutions) in separate containers or vials (e.g., for reconstitution (e.g., of lyophilized agents)). In certain embodiments, the agents or compositions are in separate containers or vials.

[0037] In accordance with another aspect of the instant invention, methods for the inhibition, treatment, and / or prevention of a disease or condition associated with amyloid beta are provided. In certain embodiment, the methods comprise administering (i) at least one anti-amyloid beta antibody or an antigen-binding fragment thereof and (ii) at least one Treg activating / inducing agent to a subject (e.g., human) in need thereof. The anti-amyloid beta antibody or an antigen-binding fragment thereof may be administered in a composition further comprising a pharmaceutically acceptable carrier. The Treg activating / inducing agent may be administered in a composition further comprising a pharmaceutically acceptable carrier. The anti -amyloid beta antibody or an antigen-binding fragment thereof and Treg activating / inducing agent may be administered at the same time (e.g., simultaneously) and / or at different times. The anti-amyloid beta antibody or an antigen-binding fragment thereof may be administered before, after, and / or at the same time as the Treg activating / inducing agent. In certain embodiments, the antiamyloid beta antibody or an antigen-binding fragment thereof and Treg activating / inducing agent are administered in the same composition when administered at the same time. In certain embodiments, the anti-amyloid beta antibody or an antigen-binding fragment thereof and Treg activating / inducing agent are administered in separate compositions when administered at the same time. In certain embodiments, the anti-amyloid beta antibody or an antigen-binding fragment thereof and Treg activating / inducing agent are administered at least at different times. In certain embodiments, the anti-amyloid beta antibody or an antigen-binding fragment thereof and Treg activating / inducing agent are administered at least partially at the same time (e.g., their administration time frames overlap at least partially). In certain embodiments, the Treg activating / inducing agent is administered at least before the anti-amyloid beta antibody or an antigen-binding fragment thereof (e.g., in order to increase Treg numbers prior to administration of the anti-amyloid beta antibody or an antigen-binding fragment thereof).

[0038] In certain embodiments, the method comprises administering an anti-amyloid beta antibody or an antigen-binding fragment thereof to the subject for a first period of time (e.g., from the first to last administration of the anti-amyloid beta antibody or an antigen-binding fragment thereof) and a Treg activating / inducing agent to the subject for a second period of time (e.g., from the first to last administration of the Treg activating / inducing agent). In certain embodiments, the first period of time at least partially or completely overlaps with the second period of time. In certain embodiments, the first period of time does not overlap with the second period of time. In certain embodiments, the second period of time is before or at least begins before the first period of time. In certain embodiments, the second period of time is before or prior to the first period of time and does not overlap with the first period of time. When the first period of time does not overlap with the second period of time, the time in between the first period of time and the second period of time may be less than 2 weeks, less than one week, less than 6 days, less than 5 days, less than 4 days, less than 3 days, less than 2 days, or less than one day. In certain embodiments, the first period of time is for more than 1 month, more than 2 months, more than 3 months, more than 4 months, more than 5 months, more than 6 months, more than 7 months, more than 8 months, more than 9 months, more than 10 months, more than 11 months, more than 12 months, or more. In certain embodiments, the second period of time is less than 6 months, less than 5 months, less than 4 months, less than 10 weeks, less than 9 weeks, less than 8 weeks, less than 7 weeks, less than 6 weeks, less than 5 weeks, less than 4 weeks, less than 3 weeks, less than 2 weeks, or less than 1 week.

[0039] In certain embodiments, the anti-amyloid beta antibody or an antigenbinding fragment thereof is administered approximately once every two weeks (e.g., after the first administration of the Treg activating / inducing agent). In certain embodiments, the anti-amyloid beta antibody or an antigen-binding fragment thereof is administered at 0.1 mg / kg, 0.5 mg / kg, 1 mg / kg, 5 mg / kg, 10 mg / kg, or more.

[0040] In certain embodiments, the disease or condition is associated with aggregates of amyloid beta. In certain embodiments, the aggregates of amyloid beta comprise oligomeric amyloid beta, fibrillar amyloid beta, and / or amyloid beta plaques. In certain embodiments, the aggregates of amyloid beta comprise oligomeric amyloid beta. In certain embodiments, the aggregates of amyloid beta comprise fibrillar amyloid beta. In certain embodiments, the aggregates of amyloid beta comprise amyloid beta plaques. In certain embodiments, the aggregates of amyloid beta comprise oligomeric amyloid beta, fibrillar amyloid beta, and amyloid beta plaques. In certain embodiments, the disease or condition is associated with amyloid beta plaque formation. In certain embodiments, the disease or condition is associated with deposition of amyloid beta or amyloid beta plaques and / or increased deposition of amyloid beta or amyloid beta plaques as compared to a subject without the disease or condition. In certain embodiments, the disease or condition is associated with the presence of amyloid beta and / or the increased presence of amyloid beta as compared to a subject without the disease or condition. In certain embodiments, the disease or condition is associated with the presence of oligomeric amyloid beta and / or the increased presence of oligomeric amyloid beta as compared to a subject without the disease or condition.

[0041] In certain embodiments, the disease or condition is neurological. In certain embodiments, the disease or condition is a neurodegenerative disease. In certain embodiments, the disease or condition is Alzheimer’s disease (optionally including Alzheimer’s disease related dementias), mild cognitive impairment, Lewy body dementia (e.g., diffuse Lewy body disease), hereditary cerebral hemorrhage with amyloidosis, Down’s syndrome (optionally including neurodegeneration), vascular dementia, subcortical vascular dementia, cerebral amyloid angiopathy, inflammatory cerebral amyloid angiopathy, cerebral amyloidoma, Parkinson’s disease (e.g., Parkinson’s disease with dementia), amyloidosis, or post-ischemic stroke.

[0042] In certain embodiments, the disease or condition is Alzheimer’s Disease. The Alzheimer’s disease may be associated with amyloid beta. The Alzheimer’s disease may be associated with aggregates of amyloid beta. The Alzheimer’s disease may be associated with deposition of amyloid beta or amyloid beta plaques. The aggregates of amyloid beta may be oligomeric amyloid beta, fibrillar amyloid beta, and / or amyloid beta plaques. In certain embodiments, the Alzheimer’s disease is associated with amyloid beta plaque formation. In certain embodiments, the Alzheimer’s disease is associated with increased deposition of amyloid beta or amyloid beta plaques as compared to a subject without Alzheimer’s disease; and / or increased presence of amyloid beta as compared to a subject without Alzheimer’s disease; and / or increased presence of oligomeric amyloid beta as compared to a subject without Alzheimer’s disease.

[0043] In certain embodiments, the methods of the instant invention further comprise administering at least one other therapeutic agent to the subject. Examples of further therapeutic agents include, without limitation: a cholinesterase inhibitor (e.g., rivastigmine, galantamine, or donepezil), an N-methyl-D-aspartate (NMD A) antagonist, granulocyte-macrophage colony stimulating factor (GM-CSF), a casein kinase 1 delta (CK1-5) inhibitor, a vasopressin receptor 1 A (VI AR) antagonist, a sigma-2 receptor antagonist, or an O-GlcNAcase (OGA) inhibitor.

[0044] The compounds of the instant invention will generally be administered to a patient as a pharmaceutical preparation. The term “patient” as used herein refers to human or animal subjects. These compounds may be employed therapeutically, under the guidance of a physician for the treatment of cancer.

[0045] The pharmaceutical preparation comprising the compounds of the invention may be conveniently formulated for administration with an acceptable medium such as water, buffered saline, ethanol, polyol (for example, glycerol, propylene glycol, liquid polyethylene glycol and the like), dimethyl sulfoxide (DMSO), oils, detergents, suspending agents or suitable mixtures thereof. The concentration of the compounds in the chosen medium may be varied and the medium may be chosen based on the desired route of administration of the pharmaceutical preparation. Except insofar as any conventional media or agent is incompatible with the compounds to be administered, its use in the pharmaceutical preparation is contemplated.

[0046] The dose and dosage regimen of the compounds according to the invention that is suitable for administration to a particular patient may be determined by a physician considering the patient's age, sex, weight, general medical condition, and the specific condition and severity thereof for which the compound is being administered. The physician may also consider the route of administration of the compound, the pharmaceutical carrier with which the compounds may be combined, and the compounds’ biological activity.

[0047] Selection of a suitable pharmaceutical composition will also depend upon the mode of administration chosen. For example, the compositions of the invention may be administered by direct injection or intravenously. In this instance, a pharmaceutical composition comprises the components dispersed in a medium that is compatible with the site of injection.

[0048] Compositions of the instant invention may be administered by any method. For example, the compositions of the instant invention can be administered, without limitation parenterally, subcutaneously, orally, topically, pulmonarily, rectally, vaginally, intravenously, intraperitoneally, intrathecally, intracerbrally, epidurally, intramuscularly, intradermally, or intracarotidly. In certain embodiments, the methods of administration include but are not limited to: parenteral, subcutaneous, oral, topical, inhalation or intranasal, pulmonary, rectal, vaginal, intravenous, intraperitoneal, intrathecal, intracerebral, epidural, intramuscular, intradermal, or intracarotid administration. In certain embodiments, the compositions are administered intramuscularly, subcutaneously, or to the bloodstream (e.g., intravenously). In certain embodiments, the Treg activating / inducing agent is administered subcutaneously. In certain embodiments, the anti-amyloid beta antibody or antigen binding fragment thereof is administered intravenously or to the bloodstream.

[0049] Pharmaceutical compositions for injection are known in the art. If injection is selected as a method for administering the composition, steps must be taken to ensure that sufficient amounts of the components reach their target cells to exert a biological effect. Dosage forms for oral administration include, without limitation, tablets (e.g., coated and uncoated, chewable), gelatin capsules (e.g., soft or hard), lozenges, troches, solutions, emulsions, suspensions, syrups, elixirs, powders / granules (e.g., reconstitutable or dispersible) gums, and effervescent tablets. Dosage forms for parenteral administration include, without limitation, solutions, emulsions, suspensions, dispersions and powders / granules for reconstitution. Dosage forms for topical administration include, without limitation, creams, gels, ointments, salves, patches and transdermal delivery systems.

[0050] Pharmaceutical compositions containing compounds of the present invention as the active ingredient in intimate admixture with a pharmaceutical carrier can be prepared according to conventional pharmaceutical compounding techniques. The carrier may take a wide variety of forms depending on the form of preparation desired for administration. In preparing the compound in oral dosage form, any of the usual pharmaceutical media may be employed, such as, for example, water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents and the like in the case of oral liquid preparations (such as, for example, suspensions, elixirs and solutions); or carriers such as starches, sugars, diluents, granulating agents, lubricants, binders, disintegrating agents and the like in the case of oral solid preparations (such as, for example, powders, capsules and tablets). Because of their ease in administration, tablets and capsules represent the most advantageous oral dosage unit form in which case solid pharmaceutical carriers are obviously employed. If desired, tablets may be sugar-coated or enteric-coated by standard techniques. For parenterals, the carrier will usually comprise sterile water, though other ingredients, for example, to aid solubility or for preservative purposes, may be included. Injectable suspensions may also be prepared, in which case appropriate liquid carriers, suspending agents and the like may be employed.

[0051] Compositions of the invention may be formulated in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form, as used herein, refers to a physically discrete unit of the pharmaceutical composition appropriate for the patient undergoing treatment. Each dosage should contain a quantity of active ingredient calculated to produce the desired effect in association with the selected pharmaceutical carrier. Procedures for determining the appropriate dosage unit are well known to those skilled in the art. For example, the appropriate dosage unit for the administration of the composition may be determined by evaluating the toxicity of the composition in animal models. Various concentrations of the components in composition may be administered to mice or other mammals, and the minimal and maximal dosages may be determined based on the beneficial results and side effects observed as a result of the treatment.

[0052] A pharmaceutical preparation of the invention may be formulated in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form, as used herein, refers to a physically discrete unit of the pharmaceutical preparation appropriate for the patient undergoing treatment. Each dosage should contain a quantity of active ingredient calculated to produce the desired effect in association with the selected pharmaceutical carrier. Procedures for determining the appropriate dosage unit are well known to those skilled in the art. Dosage units may be proportionately increased or decreased based on the weight of the patient. Appropriate concentrations for alleviation of a particular pathological condition may be determined by dosage concentration curve calculations, as known in the art.

[0053] In accordance with the present invention, the appropriate dosage unit for the administration of the compounds of the invention may be determined by evaluating the toxicity of the compounds in animal models. Various concentrations of the compounds of the instant invention may be administered to mice with transplanted human tumors, and the minimal and maximal dosages may be determined based on the results of significant reduction of tumor size and side effects as a result of the treatment. Appropriate dosage unit may also be determined by assessing the efficacy of the compounds in combination with other standard anti-cancer drugs. The dosage units of the compounds may be determined individually or in combination with each anti-cancer treatment according to greater shrinkage and / or reduced growth rate of tumors.

[0054] The compositions comprising the compounds of the instant invention may be administered at appropriate intervals, for example, at least twice a day or more until the pathological symptoms are reduced or alleviated, after which the dosage may be reduced to a maintenance level. The appropriate interval in a particular case would normally depend on the condition of the patient.

[0055] Definitions

[0056] The following definitions are provided to facilitate an understanding of the present invention.

[0057] The singular forms “a,” “an,” and “the” include plural referents unless the context clearly dictates otherwise.

[0058] “Pharmaceutically acceptable” indicates approval by a regulatory agency of the Federal or a state government or listed in the U.S. Pharmacopeia or other generally recognized pharmacopeia for use in animals, and more particularly in humans.

[0059] A “carrier” refers to, for example, a diluent, adjuvant, preservative (e.g., Thimersol, benzyl alcohol), anti-oxidant (e.g., ascorbic acid, sodium metabisulfite), solubilizer (e.g., polysorbate 80), emulsifier, buffer (e.g., Tris HC1, acetate, phosphate), antimicrobial, bulking substance (e.g., lactose, mannitol), excipient, auxiliary agent or vehicle with which an active agent of the present invention is administered. Pharmaceutically acceptable carriers can be sterile liquids, such as water and oils, including those of petroleum, animal, vegetable or synthetic origin. Water or aqueous saline solutions and aqueous dextrose and glycerol solutions are preferably employed as carriers, particularly for injectable solutions. Suitable pharmaceutical carriers are described in “Remington's Pharmaceutical Sciences” by E.W. Martin (Mack Publishing Co., Easton, PA); Gennaro, A. R., Remington: The Science and Practice of Pharmacy, (Lippincott, Williams and Wilkins); Liberman, et al., Eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, N.Y.; and Kibbe, et al., Eds., Handbook of Pharmaceutical Excipients, American Pharmaceutical Association, Washington. The term “treat” as used herein refers to any type of treatment that imparts a benefit to a patient afflicted with a disease, including improvement in the condition of the patient (e.g., in one or more symptoms), delay in the progression of the condition, etc.

[0060] As used herein, the term “prevent” refers to the prophylactic treatment of a subject who is at risk of developing a condition (e.g., neurodegenerative disease) resulting in a decrease in the probability that the subject will develop the condition.

[0061] A “therapeutically effective amount” of a compound or a pharmaceutical composition refers to an amount effective to prevent, inhibit, treat, or lessen the symptoms of a particular disorder or disease. The treatment of a neurodegenerative disease herein may refer to curing, relieving, and / or preventing the neurodegenerative disease, the symptom(s) of it, or the predisposition towards it.

[0062] As used herein, the term “subject” refers to an animal, particularly a mammal, particularly a human.

[0063] As used herein, a “biological sample” refers to a sample of biological material obtained from a subject, preferably a human subject, including a tissue, a tissue sample, a cell sample, a tumor sample, and a biological fluid (e.g., blood, urine, or amniotic fluid).

[0064] An “antibody” or “antibody molecule” is any immunoglobulin, including antibodies and fragments thereof, that binds to a specific antigen. Antibody fragments include, without limitation, antigen-binding immunoglobulin fragments including, without limitation: single domain (dAb; e.g., single variable light or heavy chain domain), Fab, Fab', F(ab')2, and F(v); and fusions (e.g., via a linker) of these immunoglobulin fragments including, without limitation: scFv, scFv2, scFv- Fc, minibody, diabody, triabody, and tetrabody.

[0065] With respect to antibodies, the term “immunologically specific” refers to antibodies that bind to one or more epitopes of a protein or compound of interest, but which do not substantially recognize and bind other molecules in a sample containing a mixed population of antigenic biological molecules.

[0066] The term “vector” refers to a carrier nucleic acid molecule (e.g., RNA or DNA) into which a nucleic acid sequence can be inserted, e.g., for introduction into a host cell where it may be expressed and / or replicated. An “expression vector” is a specialized vector that contains a gene or nucleic acid sequence with the necessary operably linked regulatory regions needed for expression in a host cell. The term “operably linked” means that the regulatory sequences necessary for expression of a coding sequence are placed in the DNA molecule in the appropriate positions relative to the coding sequence so as to effect expression of the coding sequence. This same definition is sometimes applied to the arrangement of coding sequences and transcription control elements (e.g. promoters, enhancers, and termination elements) in an expression vector.

[0067] As used herein, the term “therapeutic agent” refers to a chemical compound or biological molecule including, without limitation, nucleic acids, peptides, proteins, and antibodies that can be used to treat a condition, disease, or disorder or reduce the symptoms of the condition, disease, or disorder.

[0068] As used herein, the term “small molecule” refers to a substance or compound that has a relatively low molecular weight (e.g., less than 2,000). Typically, small molecules are organic, but are not proteins, polypeptides, or nucleic acids.

[0069] The following example provides illustrative methods of practicing the instant invention, and is not intended to limit the scope of the invention in any way.

[0070] EXAMPLE

[0071] Tregs can transform pro-inflammatory, neurotoxic environments to more anti-inflammatory and neurotrophic environments, wherein dopaminergic neurons are rescued or spared from a degenerative fate (Reynolds, et al. (2010) J. Immunol., 184:2261-71; Gendelman et al. (2017) NPJ Parkinsons Dis., 3: 10; Olson, et al. (2021) EBioMedicine 67: 103380; Mosley, et al. (2019) Front. Cell Neurosci., 13:421; Olson, et al. (2021) Biomaterials 272: 120786; Olson, et al. (2020) Neurotherapeutics 17: 1861-77; Potter, et al. (2021) Alzheimer’s Dement., 7:el2158). Indeed, immunomodulatory agents such as granulocyte-macrophage colony stimulating factor (GM-CSF) increase Treg numbers and activity with parallel neuroprotective activities in PD and Alzheimer’s disease (Reynolds, et al. (2010) J. Immunol., 184:2261-71; Gendelman et al. (2017) NPJ Parkinsons Dis., 3: 10; Olson, et al. (2021) EBioMedicine 67: 103380; Thome, et al. (2021) NPJ Parkinson’s Dis 7:41; Olson, et al. (2021) Biomaterials 272: 120786; Olson, et al. (2020) Neurotherapeutics 17: 1861-77; Potter, et al. (2021) Alzheimer’s Dement., 7:el2158; Kiyota, et al. (2018) J. Neuroimmunol., 319:80-92; Kosloski, et al. (2013) J. Neuroimmunol., 265: 1-10). IL-2 has essential defined roles and direct interaction in survival and expansion of new or existing Tregs. IL-2 upregulates forkhead box P3 (FoxP3) expression during Treg development in the thymus and is involved in Treg differentiation, lineage stability, proliferation, and function (Zorn, et al. (2006) Blood 108:1571-9). Tregs constitutively express the high-affinity receptor for IL-2, while its expression by other subsets of T cells is induced after activation (Chinen, et al. (2016) Nat. Immunol., 17: 1322-33). IL-2 signaling is also a key component by which CD4+CD25+FoxP3+ iTregs exhibit functional dominance and can outgrow other T-cell types that typically express lower levels of the IL-2 receptor a chain (CD25) (Hotta-Iwamura, et al. (2018) J. Autoimmun., 90:39-48; Dong, et al. (2021) JCI Insight 6:el47474; MacMillan, et al. (2021) (2021) Blood Adv., 5: 1425-36; Hui, et al. (2021) Front. Immunol., 12:619932). Therefore, in the steady state, functional iTregs respond better to IL-2 than other T cells and do so in a direct manner, rather than via intermediary cell interactions (Abba, et al. (2018) Sci. Immunol., 3:eaatl482). This may account for the ability of low doses of IL-2 to preferentially increase Treg numbers without global immune activation of other T cell types (Grinberg-Bleyer, et al. (2010) J. Exp. Med., 207:1871-8; Tang, et al. (2008) Immunity 28:687-97; Wang, et al. (2021) Int. Immunopharmacol., 97: 107663). Therapies such as low-dose IL-2 or IL-2 / anti-IL-2 antibody complexes may preferentially expand Treg populations as a treatment for chronic inflammatory autoimmune diseases (Abbas, A.K. (2020) Am. J. Pathol., 190: 1776-81). Some neuroprotective effects of low-dose IL-2 treatment have been found in Alzheimer’s disease mice (Alves et al. (2017) Brain 140:826-42) and in amyotrophic lateral sclerosis (ALS) patients (Giovannelli, et al. (2021) Brain Commun., 3:fcabl41).

[0072] Misfolded amyloid beta monomers can aggregate into soluble oligomers which can further lead to the formation of insoluble fibril sand amyloid plaques. The oligomers, fibrils and plaques are neurotoxic amyloid beta isoforms. The poor specificity of certain amyloid beta antibodies to the pathological or neurotoxic amyloid beta forms leads to peripheral clearance from binding to monomeric forms.

[0073] Combinational therapy of monoclonal antibody specific to the oligomeric amyloid beta and anti-inflammatory Treg stimulating agent will increase amyloid clearance and decrease antibody induced neuroinflammatory Teff responses.

[0074] Figure 1 A provides a schematic of a study design. Briefly, an Alzheimer’s disease mouse model - APP / PS1 - was used. APP / PS1 mice are double transgenic mice expressing a transgene coding for the 695 amino acid isoform that harbors the Swedish mutation of human amyloid precursor protein (APPsw) and human presenilin 1 (PSI) carrying the M146V mutation. This mouse model expresses high concentrations of human amyloid beta protein and amyloid beta peptides via proteolytic processing (e.g. via P-secretases and y-secretases) and develops human Alzheimer’s disease-like amyloid pathology due to genetic mutations within the human amyloid precursor protein (APP) and PSEN1 genes. 7-8 month old APP / PS1 mice were administered mAb (2CD3M) (i.p. 30 mg / kg), IL-2 (i.p.), or both mAb and IL-2. Mock treated APP / PS1 or BM mice were used as Alzheimer’s disease control and wild-type control, respectively. Five treatment groups were utilized in the studies (n = 5-8 mice / group): 1. Non-Tg (BM), 2. APP / PS1, 3. APP / PS1 + 2CD3M, 4. APP / PS1 + 2CD3M + low-dose IL-2, and 5. APP / PS1 + low-dose IL-2.

[0075] Figure IB provides surface plasmon resonance (SPR) data which confirms the preferential binding of 2CD3M to neurotoxic amyloid beta isoforms. SPR is described in Hearty, et al. (Measuring Antibody-Antigen Binding Kinetics Using Surface Plasmon Resonance. In: Nevoltris, D., Chames, P. (eds) Antibody Engineering. Methods in Molecular Biology, (2018) vol 1827. Humana Press, New York, NY. doi.org / 10.1007 / 978-l-4939-8648-4_22). As seen in Figure IB, 2CD3M has undetectable binding to monomer amyloid beta.

[0076] As seen in Figure 2, the combinatorial therapy of mAb and IL-2 significantly improves cognitive function. In a radial arm water maze test, APP / PS1 mice treated with mAb and IL-2 demonstrated significantly improved cognitive function compared to mAb alone.

[0077] Briefly, mice from masked cages were introduced into a circular water filled tank (diameter-110 cm and height-91 cm, San Diego Instruments) with triangular inserts that produce six swim paths radiating from the center. Spatial cues are fixed on the tank wall to guide mouse orientation. At the end of any one arm, a circular plexiglass hidden platform (diameter- 10 cm) is placed submerged 1 cm beneath the water level. The platform was placed in the same arm for four consecutive acquisition trials (T1-T4), and retention trial (T5), but in a different arm on different experimental days. For T1-T4, the mouse started the task from a randomly chosen arm without a platform. After four trials, the mouse was returned to its cage for 30 minutes and reintroduced into the T4 arm, for the delayed retention trial (T5). Each trial lasted 1 min, and an error was scored when mouse entered the wrong arm; entered the arm with the platform, but did not climb on it; or did not make a choice for 20 seconds. The trial ended when the mouse climbed and stay on the platform for at least 10 seconds. The mouse was allowed to rest on the platform for 20 seconds between trials. If the mouse did not climb the platform, after 60 seconds it was gently guided to the submerged platform. The Tl, T4 and T5 trial errors over 9- day test were divided into three blocks (block-1 days 1-3, block-2 days 4-6, block-3 days 7-9), and the errors in each block were averaged for statistical analysis.

[0078] Figures 3 A And 3B provide graphs of the %Treg and %Teff in the blood (Fig. 3 A) and brain (Fig. 3B). The combinatorial therapy of mAb and IL-2 dramatically rescues the brain from antibody mediated T-effector (Teff; pro- inflammatory) stimulation.

[0079] Figure 4A provides images of the hippocampus and cortex of mice receiving the indicated treatments. Figure 4B provides graphs of the amount of amyloid beta in the hippocampus and cortex of mice receiving the indicated treatments. The combinatorial therapy significantly decreases the hippocampal amyloid load and a decrease in cortex amyloid load.

[0080] Combinatorial treatment with anti-amyloid beta monoclonal antibody (2CD3M) and low-dose IL-2 significantly improves cognitive performance in APP / PS1 mice compared to antibody monotherapy. Cognitive improvement is accompanied by a marked reduction in hippocampal amyloid burden, consistent with the known role of this region in memory and learning. A trend toward reduced cortical amyloid was also observed, indicating broader efficacy, particularly with prolonged treatment. These findings support the therapeutic use of a dual-modality approach combining targeted antibody -mediated clearance of neurotoxic amyloid beta oligomers with Treg-inducing agents to mitigate neuroinflammation, highlighting its use as a next-generation immunotherapy strategy for Alzheimer’s disease.

[0081] A number of publications and patent documents are cited throughout the foregoing specification in order to describe the state of the art to which this invention pertains. The entire disclosure of each of these citations is incorporated by reference herein.

[0082] While certain of the preferred embodiments of the present invention have been described and specifically exemplified above, it is not intended that the invention be limited to such embodiments. Various modifications may be made thereto without departing from the scope and spirit of the present invention, as set forth in the following claims.

Claims

What is claimed is:

1. A method of inhibiting, treating, and / or preventing a disease or condition associated with amyloid beta in a subject, said method comprising administering to the subject (i) an anti -amyloid beta antibody or an antigen binding fragment thereof and (ii) a regulatory T cell (Treg) inducing or activating agent.

2. The method of claim 1, wherein anti-amyloid beta antibody or an antigen binding fragment thereof comprises one, two, or all three complementarity determining regions (CDRs) of SEQ ID NO: 6 and one, two, or all three CDRs of SEQ ID NO: 10.

3. The method of claim 1, wherein anti-amyloid beta antibody or an antigen binding fragment thereof comprises a heavy chain complementarity determining region 1 (CDR-H1), a heavy chain complementarity determining region 2 (CDR- H2), and a heavy chain complementarity determining region 3 (CDR-H3) comprising amino acid sequences SEQ ID NOs: 7, 8, and 9, respectively, and a light chain complementarity determining region 1 (CDR-L1), a light chain complementarity determining region 2 (CDR-L2) and a light chain complementarity determining region 3 (CDR-L3) comprising amino acid sequences SEQ ID NOs: 11, 12, and 13, respectively.

4. The method of claim 1, wherein anti-amyloid beta antibody or an antigen binding fragment thereof is selected from the group consisting of 2CD3M, lecanemab, donanemab, aducanumab, Remtemetug, Gantenerumab, Trontinemab, ABBV916, ACU193, SHR-1707, PMN310, and PRX012.

5. The method of any one of claims 1-4, wherein the Treg inducing or activating agent is selected from the group consisting of interleukin-2 (IL-2), low- dose IL-2, granulocyte-macrophage colony stimulating factor (GM-CSF), GM-CSF analogs, TGF-P, IL-10, IL-2, glatiramer acetate, anti-CD3, anti-CD28, bee venom phospholipase A2, rapamycin, histone deacetylase inhibitors, vasoactive intestinal peptide (VIP), VIP analogs and VIP receptor-2 agonists, and combinations thereof.

6. The method of any one of claims 1-5, wherein the Treg inducing or activating agent is IL-2 or low-dose IL-2.

7. The method of any one of claims 1-5, wherein the Treg inducing or activating agent is low-dose IL-2.

8. The method of claim 7, wherein low-dose IL-2 is less than about 5 million international units per day.

9. The method of any one of claims 1-8, wherein said anti-amyloid beta antibody or an antigen binding fragment thereof and said Treg inducing or activating agent are administered separately.

10. The method of any one of claims 1-8, wherein said anti-amyloid beta antibody or an antigen binding fragment thereof and said Treg inducing or activating agent are administered together.

11. The method of any one of claims 1-10, wherein said disease or condition is a neurodegenerative disease.

12. The method of any one of claims 1-10, wherein said disease or condition is Alzheimer’s disease.

13. The method of claim 2, wherein the Treg inducing or activating agent is low- dose IL-2 and wherein said disease or condition is Alzheimer’s disease.

14. A composition comprising (i) an anti-amyloid beta antibody or an antigen binding fragment thereof; (ii) a regulatory T cell (Treg) inducing or activating agent; and (iii) a pharmaceutically acceptable carrier.

Images