A combination of IL-4 / IL-13 pathway inhibitors and plasma cell ablation used to treat allergies.

Abstract

This application provides a method for treating allergies, comprising selecting a patient with an allergy and administering a therapeutically effective amount of an IL-4 / IL-13 pathway inhibitor (e.g., an anti-IL-4 receptor antibody or its antigen-binding fragment) combined with a therapeutically effective amount of an agent that depletes plasma cells (e.g., an anti-BCMA / anti-CD3 bispecific antibody). In some embodiments, the plasma cell ablative, such as the anti-BCMA / anti-CD3 bispecific antibody, ablates plasma cells, including IgE+ plasma cells, while the IL-4 / IL-13 pathway inhibitor prevents the generation of new IgE+ plasma cells, thereby eliminating allergen-specific IgE from the patient.

Description

[0001] Cross-references to related applications

[0002] This application was filed as a PCT international patent application on March 20, 2020, and claims priority to U.S. Provisional Patent Application No. 62 / 822,022, filed March 21, 2019, and U.S. Provisional Patent Application No. 62 / 957,550, filed January 6, 2020, the entire contents of which are incorporated herein by reference. Invention Field

[0003] This application relates to a method for treating allergies, comprising administering to an individual in need a therapeutically effective amount of an IL-4 / IL-13 pathway inhibitor and a therapeutically effective amount of a plasma cell ablating agent. Background Technology

[0004] Allergies and allergic diseases are serious medical conditions with consequences ranging from non-life-threatening reactions that may resolve over time to life-threatening outcomes, such as anaphylactic reactions. Causes of anaphylactic reactions can include contact with or exposure to a wide variety of products, such as certain foods, insect venom, plant-derived substances (e.g., pollen), chemicals, pharmaceutical / drug treatments, and animal dander. The pathophysiology of allergies is influenced by complex interactions between immunoglobulin E (IgE)-mediated sensitization, the immune system, and environmental factors. Current options for treating allergies include avoidance, pharmacological symptomatic treatment, and prevention with allergen-specific immunotherapy (SIT). Unfortunately, these current treatment strategies are often inadequate, expensive, impractical, or carry significant risks. For example, allergen avoidance is not always possible but can negatively impact the quality of life for both patients and caregivers. On the other hand, immunotherapy involves the intentional administration of allergens to susceptible individuals, thus carrying inherent risks and the potential for undesirable severe anaphylactic reactions or allergic reactions. Therefore, there is an unmet need in the field for new treatment methods that prevent or treat allergies or anaphylactic reactions and reduce the risk of developing anaphylactic reactions.

[0005] Overview

[0006] In one aspect, this application provides methods for treating allergies, allergic reactions, or allergic disorders in an individual, preventing or reducing the severity of allergic reactions, or reducing or eliminating allergen-specific serum IgE. In some embodiments, the method comprises administering an IL-4 / IL-13 pathway inhibitor and a plasma cell ablation agent to an individual (e.g., an individual suffering from allergies, allergic disorders, mast cell activation disorders, or mast cell hyperplasia).

[0007] In some embodiments, this application includes methods for treating allergies or preventing or reducing the severity of allergic reactions, comprising: (a) selecting an individual suffering from an allergic disease or condition, mast cell activation disorder, or mast cell hyperplasia; and (b) administering to the individual in need a therapeutically effective amount of an IL-4 / IL-13 pathway inhibitor and a therapeutically effective amount of a plasma cell ablation agent.

[0008] In some embodiments, this application includes methods for treating allergies or preventing or reducing the severity of allergic reactions, comprising: (a) selecting an individual suffering from an allergic disease or condition, mast cell activation disorder, or mast cell hyperplasia; and (b) administering to the individual in need a therapeutically effective amount of an IL-4 / IL-13 pathway inhibitor and a plasma cell ablation agent.

[0009] In some embodiments, this application includes a method for treating an allergy or preventing or reducing the severity of an allergic reaction, comprising: (a) selecting an individual suffering from an allergic disease or condition, mast cell activation disorder, or mast cell hyperplasia, wherein the individual is undergoing a background therapy regimen comprising one or more doses of an IL-4 / IL-13 pathway inhibitor; and (b) administering at least one dose of a plasma cell ablation agent.

[0010] In one implementation, administration of an IL-4 / IL-13 pathway inhibitor prevents the generation of new IgE+ plasma cells, and administration of a plasma cell ablation agent leads to the elimination of bone marrow-resident IgE+ plasma cells, thereby eliminating allergen-specific serum IgE.

[0011] In some embodiments, this application includes methods for increasing the efficacy and / or tolerability of immunotherapy regimens in individuals with allergies. In some embodiments, the method comprises administering an IL-4 / IL-13 pathway inhibitor and a plasma cell ablation agent to the individual before or concurrently with the immunotherapy regimen. In some embodiments, the immunotherapy regimen is an oral immunotherapy (OIT) regimen. In some embodiments, the immunotherapy regimen is a subcutaneous immunotherapy (SCIT) regimen. In some embodiments, the immunotherapy is an allergen-specific immunotherapy regimen against a food allergen (e.g., peanut allergen). In some embodiments, the immunotherapy is an allergen-specific immunotherapy regimen against an environmental allergen.

[0012] In one embodiment of the method disclosed herein, the allergic disease or condition is selected from allergic asthma, hay fever, chronic urticaria, food allergy, pollen allergy, and allergies caused by environmental allergens. In one embodiment, the individual is at risk of an allergic reaction caused by an allergen. In one embodiment, the individual has seasonal allergies. In one embodiment, the individual has severe allergies. In one embodiment, the individual suffers from an allergy caused by one or more allergens selected from milk, dairy products, eggs, celery, sesame, wheat, meat, fruit, soy, fish, shellfish, sugar, peanuts, beans, tree nuts, dust, dust mites, pollen, insect venom, mold, animal fur, animal dander, wool, latex, metals, household cleaners, detergents, pharmaceuticals, cosmetics, fragrances, drugs (e.g., penicillin, sulfonamides, salicylates, therapeutic monoclonal antibodies (e.g., cetuximab), ragweed, grass, and birch). In one embodiment, The allergen is contained in food products selected from milk, dairy products, poultry eggs, celery, sesame, wheat, meat, fruit, soybeans, fish, shellfish, sugar, peanuts, legumes, and tree nuts. In one embodiment, the allergen is a non-food allergen selected from: dust, dust mites, pollen, insect venom, mold, animal fur, animal dander, wool, latex, metals, household cleaners, detergents, pharmaceuticals, cosmetics, fragrances, pharmaceuticals such as penicillin, sulfonamides or salicylates, therapeutic monoclonal antibodies (e.g., cetuximab), ragweed, grass, and birch.

[0013] In one embodiment of the method disclosed herein, the IL-4 / IL-13 pathway inhibitor is selected from anti-IL-4 antibodies, anti-IL-13 antibodies, anti-IL-4 / IL-13 bispecific antibodies, IL-4 receptor (IL-4R) inhibitors, IL-4 traps, IL-13 traps, and anti-IL-4R antibodies. In one embodiment, the IL-4 / IL-13 pathway inhibitor is an anti-IL-4 antibody (e.g., pascolizumab). In another embodiment, the IL-4 / IL-13 pathway inhibitor is an anti-IL-13 antibody (e.g., tralokinumab, lebrikizumab, dectrekumab, GSK679586, or MEDI7836). In yet another embodiment, the IL-4 / IL-13 pathway inhibitor is an anti-IL-4 / IL-13 bispecific antibody (e.g., romilkimab). In another embodiment, the IL-4 / IL-13 pathway inhibitor is an IL-4R inhibitor (e.g., an IL-4 mutant protein, such as pitrakinra, or an anti-IL-4R antibody). In another embodiment, the IL-4 / IL-13 pathway inhibitor is an anti-IL-4R antibody. In yet another embodiment, the IL-4 / IL-13 pathway inhibitor is an IL-4Trap or an IL-13Trap.

[0014] In one embodiment, the IL-4 / IL-13 pathway inhibitor is an anti-IL-4R antibody or its antigen-binding fragment. In one embodiment, the anti-IL-4R antibody comprises a heavy chain variable region (HCVR) including three heavy chain complementarity-determining regions (CDRs) (HCDR1, HCDR2, and HCDR3) and a light chain variable region (LCVR) including three light chain CDRs (LCDR1, LCDR2, and LCDR3), wherein: HCDR1 has the amino acid sequence of SEQ ID NO:3; HCDR2 has the amino acid sequence of SEQ ID NO:4; HCDR3 has the amino acid sequence of SEQ ID NO:5; LCDR1 has the amino acid sequence of SEQ ID NO:6; LCDR2 has the amino acid sequence of SEQ ID NO:7; and LCDR3 has the amino acid sequence of SEQ ID NO:8. In another embodiment, the anti-IL-4R antibody comprises a heavy chain variable region (HCVR) including the amino acid sequence of SEQ ID NO:1 and a light chain variable region (LCVR) including the amino acid sequence of SEQ ID NO:2. In another embodiment, the anti-IL-4R antibody comprises a heavy chain and a light chain, wherein the heavy chain has the amino acid sequence of SEQ ID NO:9. In another embodiment, the anti-IL-4R antibody comprises a heavy chain and a light chain, wherein the light chain has the amino acid sequence of SEQ ID NO:10. In another embodiment, the anti-IL-4R antibody comprises a heavy chain and a light chain, wherein the heavy chain has the amino acid sequence of SEQ ID NO:9 and the light chain has the amino acid sequence of SEQ ID NO:10. In another embodiment, the IL-4 / IL-13 pathway inhibitor is dupilumab or its bioequivalence. In another implementation, the IL-4 / IL-13 pathway inhibitor is selected from dupilumab, pacozumab, AMG317, MEDI2045, MEDI9314, troluuzumab, lerezumab, anrukinzumab, detrocurumab, GSK679586, MEDI7836, roximateb, IL-4Trap, IL-13Trap, AER-003, and picotrigin.

[0015] In one embodiment of the method disclosed herein, the plasma cell ablation agent is selected from B-cell maturation antigen (BCMA) targets, proteasome inhibitors, histone deacetylase inhibitors, B-cell activating factor (BAFF) inhibitors, and A proliferation-inducing ligand (APRIL; CD256) inhibitors. In one embodiment, the BCMA target is selected from anti-BCMA / anti-CD3 bispecific antibodies, chimeric antigen receptors targeting BCMA, and anti-BCMA antibodies conjugated to cytotoxic drugs.

[0016] In one embodiment, the plasma cell ablation agent is an anti-BCMA / anti-CD3 bispecific antibody or an antigen-binding fragment thereof, comprising: (a) a first antigen-binding domain specifically binding to BCMA; and (b) a second antigen-binding domain specifically binding to CD3. In one embodiment, the first antigen-binding domain comprises: three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) contained in a heavy chain variable region (HCVR) containing the amino acid sequence of SEQ ID NO:12; and three light chains (LCDR1, LCDR2, and LCDR3) contained in a light chain variable region (LCVR) containing the amino acid sequence of SEQ ID NO:20. In another embodiment, HCDR1 has the amino acid sequence of SEQ ID NO:14; HCDR2 has the amino acid sequence of SEQ ID NO:16; HCDR3 has the amino acid sequence of SEQ ID NO:18; LCDR1 has the amino acid sequence of SEQ ID NO:22; LCDR2 has the amino acid sequence of SEQ ID NO:24; and LCDR3 has the amino acid sequence of SEQ ID NO:26. In another embodiment, the second antigen-binding domain comprises: three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) contained in a heavy chain variable region (HCVR) comprising an amino acid sequence selected from SEQ ID NO: 28 and 36; and three light chains (LCDR1, LCDR2, and LCDR3) contained in a light chain variable region (LCVR) comprising an amino acid sequence selected from SEQ ID NO: 20. In another embodiment, HCDR1 has an amino acid sequence of SEQ ID NO: 30 or 38; HCDR2 has an amino acid sequence of SEQ ID NO: 32 or 40; HCDR3 has an amino acid sequence of SEQ ID NO: 34 or 42; LCDR1 has an amino acid sequence of SEQ ID NO: 22; LCDR2 has an amino acid sequence of SEQ ID NO: 24; and LCDR3 has an amino acid sequence of SEQ ID NO: 26.In one embodiment, the anti-BCMA / anti-CD3 bispecific antibody comprises: (a) a first antigen-binding domain that specifically binds to BCMA and comprises three heavy chain CDRs and three light chain CDRs, wherein the six CDRs HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 comprise the amino acid sequence of SEQ ID NO: 14-16-18-22-24-26; and (b) a second antigen-binding domain that specifically binds to CD3 and comprises three heavy chain CDRs and three light chain CDRs, wherein the six CDRs HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 comprise the amino acid sequence of SEQ ID NO: 30-32-34-22-24-26. In one embodiment, the anti-BCMA / anti-CD3 bispecific antibody comprises: (a) a first antigen-binding domain that specifically binds to BCMA and comprises three heavy chain CDRs and three light chain CDRs, wherein the six CDRs HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 comprise the amino acid sequence of SEQ ID NO: 14-16-18-22-24-26; and (b) a second antigen-binding domain that specifically binds to CD3 and comprises three heavy chain CDRs and three light chain CDRs, wherein the six CDRs HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 comprise the amino acid sequence of SEQ ID NO: 38-40-42-22-24-26. In one embodiment, the anti-BCMA / anti-CD3 bispecific antibody comprises: (a) a first antigen-binding domain comprising the HCVR of SEQ ID NO:12 and the LCVR of SEQ ID NO:20; and (b) a second antigen-binding domain comprising the HCVR of SEQ ID NO:28 and the LCVR of SEQ ID NO:20. In another embodiment, the anti-BCMA / anti-CD3 bispecific antibody comprises: (a) a first antigen-binding domain comprising the HCVR of SEQ ID NO:12 and the LCVR of SEQ ID NO:20; and (b) a second antigen-binding domain comprising the HCVR of SEQ ID NO:36 and the LCVR of SEQ ID NO:20.

[0017] In one embodiment, an IL-4 / IL-13 pathway inhibitor is administered prior to the plasma cell ablation agent. In another embodiment, the IL-4 / IL-13 pathway inhibitor is administered after the plasma cell ablation agent. In one embodiment, compared to individuals treated with a therapeutic agent as a monotherapy, administration of the combination of an IL-4 / IL-13 pathway inhibitor and a plasma cell ablation agent blocks IgE production and eliminates allergen-specific IgE from the patient's serum.

[0018] In another embodiment, one or more doses of an IL-4 / IL-13 pathway inhibitor are administered in combination with one or more doses of a plasma cell ablation agent. In another embodiment, at least one dose of the IL-4 / IL-13 pathway inhibitor comprises about 0.1 to about 50 mg / kg body weight. In another embodiment, at least one dose of the IL-4 / IL-13 pathway inhibitor comprises about 0.05 to about 600 mg of the inhibitor. In another embodiment, at least one dose of the plasma cell ablation agent comprises about 0.1 mg / kg to about 20 mg / kg body weight. In another embodiment, at least one dose of the plasma cell ablation agent comprises about 0.05 to about 500 mg of the active agent.

[0019] In another embodiment, the method further includes administering at least one other therapeutic agent or therapy. In another embodiment, the other therapeutic agent or therapy includes IgE antagonists, antihistamines, anti-inflammatory drugs, corticosteroids, leukotriene antagonists, mast cell inhibitors, bronchodilators, decongestants, adrenaline, IL-1 antagonists, IL-5 antagonists, IL-31 antagonists, IL-33 antagonists, IL-25 antagonists, interferon-gamma, TNF antagonists, and / or TSLP antagonists.

[0020] In another aspect, this application provides pharmaceutical compositions and combinations thereof for treating allergies, anaphylactic reactions, or allergic disorders in an individual, preventing or reducing the severity of anaphylactic reactions, reducing or eliminating allergen-specific serum IgE, or increasing the efficacy and / or tolerability of immunotherapy in an individual suffering from an allergy. In some embodiments, the pharmaceutical composition or combination comprises an IL-4 / IL-13 pathway inhibitor (e.g., an anti-IL-4Rα antibody) and a plasma cell ablation agent (e.g., a BCMA-targeting agent). In some embodiments, the pharmaceutical composition or combination comprises a therapeutically effective amount of an IL-4 / IL-13 pathway inhibitor (e.g., an anti-IL-4Rα antibody) and a therapeutically effective amount of a plasma cell ablation agent (e.g., a BCMA-targeting agent). In some embodiments, the pharmaceutical composition or combination comprises a subtherapeutic dose of an IL-4 / IL-13 pathway inhibitor (e.g., an anti-IL-4Rα antibody) and / or a plasma cell ablation agent (e.g., a BCMA-targeting agent).

[0021] In another aspect, this application provides the use of IL-4 / IL-13 pathway inhibitors (e.g., anti-IL-4Rα antibodies) and plasma cell ablation agents (e.g., BCMA-targeting agents) in the preparation of medicaments for treating allergies, anaphylactic reactions, or allergic disorders in an individual, preventing or reducing the severity of anaphylactic reactions, reducing or eliminating allergen-specific serum IgE, or increasing the efficacy and / or tolerability of immunotherapy in an individual with an allergy. In some embodiments, one or both of the IL-4 / IL-13 pathway inhibitors and plasma cell ablation agents are used in a therapeutically effective amount. In some embodiments, one or both of the IL-4 / IL-13 pathway inhibitors and plasma cell ablation agents are used in a subtherapeutic dose. Brief description of the attached diagram

[0023] Figure 1 This is an illustration of the house dust mite (HDM) exposure and antibody treatment regimen studied according to Example 1.

[0024] Figure 2A This shows the serum IgE levels of mice treated with saline, antibody-free, isotype control antibody, REGN5459 (anti-BCMA x anti-CD3 bispecific antibody), REGN1103 (anti-IL-4R antibody), or a combination of REGN5459 and REGN1103, after exposure to HDM for 11 weeks followed by a 1-week rest period, as described in Example 1. An asterisk (*) indicates statistical significance relative to the isotype control (IgG).

[0025] Figure 2B This shows the serum IgE levels of mice treated with saline, antibody-free, isotype control antibody, REGN5459 (anti-BCMA x anti-CD3 bispecific antibody), REGN1103 (anti-IL-4R antibody), or a combination of REGN5459 and REGN1103, after exposure to HDM for 11 weeks followed by a 6-week rest period, as described in Example 1. An asterisk (*) indicates statistical significance relative to the isotype control (IgG).

[0026] Figure 3 This is a diagram illustrating the HDM exposure and antibody treatment regimen studied according to Example 2.

[0027] Figure 4AThis shows the HDM-specific serum IgE levels in mice 1 week after continuous exposure to HDM and treatment with either anti-BCMA x anti-CD3 bispecific antibody (REGN5459) or an isotype control (REGN4460), as described in Example 2. An asterisk (*) indicates statistical significance relative to the isotype control (IgG). LLOQ = lower limit of quantitation.

[0028] Figure 4B This shows the HDM-specific serum IgE levels in mice that were continuously exposed to HDM and treated for 3 weeks with anti-BCMA x anti-CD3 bispecific antibody (REGN5459) and isotype control (REGN4460) as described in Example 2. An asterisk (*) indicates the degree of statistical significance relative to the isotype control (IgG).

[0029] Figure 4C This shows the HDM-specific serum IgE levels in mice that were continuously exposed to HDM and treated for 5 weeks with anti-BCMA x anti-CD3 bispecific antibody (REGN5459) and isotype control (REGN4460) as described in Example 2. An asterisk (*) indicates the degree of statistical significance relative to the isotype control (IgG).

[0030] Figure 5 This diagram illustrates the effects of anti-BCMA x anti-CD3 bispecific antibody and anti-IL-4Rα antibody, alone or in combination, on IgE bone marrow plasma cells 5 weeks after administration of anti-BCMA x anti-CD3 bispecific antibody. An asterisk (*) indicates statistical significance relative to the isotype control (IgG); *p≤0.05; **p≤0.01; *****p≤0.001; *****p≤0.0001.

[0031] Figure 6 This is a diagram illustrating the HDM exposure and antibody treatment regimen studied according to Example 4.

[0032] Figure 7This study demonstrates the effects of anti-BCMA x anti-CD3 bispecific antibody and anti-IL-4Rα antibody, alone or in combination, on serum HDM-specific IgE levels. Treatment groups were as described in Example 4 and Table 7, and were: Group A (saline), Group B (HDM 12 weeks, no antibody), Group C (HDM 15 weeks, no antibody), Group D (HDM 15 weeks, isotype control antibody), Group E (HDM 15 weeks, anti-BCMA x anti-CD3 bispecific antibody), Group F (HDM 15 weeks, anti-IL-4Rα antibody), and Group G (HDM 15 weeks, IL-4Rα antibody and anti-BCMA x anti-CD3 bispecific antibody). Statistical significance was compared between treatment groups as shown in the inset. ns = no statistical significance; *p≤0.05; **p≤0.01; **p≤0.001; **p≤0.0001. LLOQ = lower limit of quantitation.

[0033] Figures 8A-8D The anti-BCMA x anti-CD3 bispecific antibody, alone or in combination with the anti-IL-4Rα antibody, showed efficacy against total bone marrow plasma cells ( Figure 8A ), IgE bone marrow plasma cells ( Figure 8B ), total spleen plasma cells ( Figure 8C ) and IgE spleen plasma cells ( Figure 8D The treatment groups were as described in Example 4 and Table 7, and were: Group A (saline), Group B (HDM 12 weeks, no antibody), Group C (HDM 15 weeks, no antibody), Group D (HDM 15 weeks, allotype control antibody), Group E (HDM 15 weeks, anti-BCMA x anti-CD3 bispecific antibody), Group F (HDM 15 weeks, anti-IL-4Rα antibody), and Group G (HDM 15 weeks, IL-4Rα antibody and anti-BCMA x anti-CD3 bispecific antibody). Statistical significance was compared between the treatment groups as shown in the illustration. ns = no statistical significance; *p≤0.05; **p≤0.01; *****p≤0.001; *****p≤0.0001.

[0034] Detailed description

[0035] It should be understood that this application is not limited to the specific methods and experimental conditions described herein, as such methods and conditions can vary. It should also be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting, and the scope of this application will be defined only by the appended claims.

[0036] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application pertains. As used herein, the term “about” when applied to a particular specified numerical value indicates that the value may differ from the specified value by no more than 1%. For example, as used herein, the expression “about 100” includes 99 and 101 and all values ​​in between (e.g., 99.1, 99.2, 99.3, 99.4, etc.).

[0037] Although any methods and materials similar to or equivalent to those described herein may be used in the implementation of this application, preferred methods and materials are now described.

[0038] Foreword

[0039] Allergic symptoms, including allergic reactions, are driven by allergen-induced cross-linking, where allergen-specific IgE binds to FcεR on effector cells (mast cells and basophils) that can induce mast cell degranulation. In allergic individuals, circulating IgE originates from antibody-secreting cells and B cells in the bone marrow, which undergo class switching to produce new IgE-producing cells. Even in the absence of an allergen, antibody-secreting cells that accumulate in the bone marrow can survive for extended periods and are a source of allergen-specific IgE. Furthermore, allergen-specific IgE can survive long-term in individuals, as evidenced by at least the following: (1) IgE is maintained in atopic patients in the absence of allergens (Luger et al., Allergol Int 2010, 59:1-8); (2) allergies can be transferred from atopic patients to non-atopic individuals, i.e., during bone marrow transplantation from the former to the latter (Garzorz et al., J Eur Acad Dermatol Venereol 2016, 30:1136-1139; Hallstrand et al., Blood 2004, 104:3086-3090); and (3) serum IgE is not eliminated in patients who undergo IgE+B cell ablation (Gauvreau et al., Sci Transl Med 2014, 6:243ra85).

[0040] In their initial research, the inventors of this application observed that treatment with anti-IL-4R antibodies in a mouse model of allergen-induced lung inflammation prevented the class conversion and differentiation of B cells into IgE-producing plasma cells, but did not affect IgE+ plasma cells in the bone marrow during prolonged allergen exposure. The inventors therefore hypothesized that combined treatment with an IL-4 / IL-13 pathway inhibitor and targeted ablation of long-lived plasma cells (including IgE+ plasma cells) could potentially block IgE production from newly generated IgE+ plasma cells as well as from IgE+ plasma cells in the bone marrow. As shown herein, this treatment combination significantly reduces or completely blocks the production of allergen-specific IgE in an animal model of allergen-induced type 2 lung inflammation (HDM). Therefore, this combination of therapeutic agents could also be used to treat allergic diseases in atopic individuals.

[0041] Methods of treating allergies

[0042] In one aspect, this application relates to the inventors’ surprising results in which the administration of a combination of an IL-4 / IL-13 pathway inhibitor (e.g., an anti-IL-4R antibody) and a plasma cell ablative (e.g., an anti-BCMA / anti-CD3 bispecific antibody) resulted in the complete elimination of allergen-specific IgE in the serum of individuals in a chronic allergen-induced lung inflammation model.

[0043] Therefore, in some embodiments, this application provides methods for treating, improving, or reducing the severity of at least one symptom or indication of an individual's allergy. In some embodiments, this application provides methods for preventing or reducing the severity of an individual's allergic reaction. In one aspect, the disclosed method includes selecting an individual suffering from an allergic disease or condition, mast cell activation disorder, or mast cell hyperplasia; and administering to the individual in need a therapeutically effective amount of an IL-4 / IL-13 pathway inhibitor (e.g., anti-IL-4 antibody, anti-IL-13 antibody, anti-IL-4 / IL-13 bispecific antibody, IL-4 receptor (IL-4R) inhibitor, anti-IL-4R antibody, or any other "IL-4 / IL-13 pathway inhibitor" as described herein) and a therapeutically effective amount of a plasma cell ablation agent (e.g., anti-BCMA / anti-CD3 bispecific antibody or any other "plasma cell ablation agent" as described herein).

[0044] References are made in this application to any particular anti-IL-4R antibody and / or any particular plasma cell ablation agent to illustrate representative IL-4 / IL-13 pathway inhibitors and representative plasma cell ablation agents, respectively, but this does not limit the scope of this application, as other combinations of IL-4 / IL-13 pathway inhibitors with plasma cell ablation agents may also be used.

[0045] As used herein, the term "treatment" refers to relieving an individual's allergic symptoms, temporarily or permanently eliminating the cause of their allergic symptoms, or preventing or slowing the onset of their allergic symptoms. As used herein, the term also includes reducing or eliminating allergen-specific serum IgE to prevent allergic reactions. In some embodiments, the term refers to reducing serum allergen-specific IgE levels by at least 50%, 60%, 70%, 80%, or more compared to baseline when administering a combination of an IL-4 / IL-13 pathway inhibitor and a plasma cell ablation agent as provided in this application. In some embodiments, the term refers to eliminating serum allergen-specific IgE levels compared to baseline when administering a combination of an IL-4 / IL-13 pathway inhibitor and a plasma cell ablation agent as provided in this application.

[0046] As used herein, the term "individual in need" refers to a human or non-human animal that exhibits one or more symptoms or indications of anaphylaxis or atopy and / or has been diagnosed with allergy to an allergen. The terms "individual" and "patient" are used interchangeably herein. In some embodiments, the term "individual in need" includes an individual at increased risk of developing an allergic reaction or anaphylactic response to an allergen. In some embodiments, the term includes an individual showing allergen sensitization to one or more allergens. In some embodiments, the method of this application can be used to treat an individual showing elevated levels of one or more serum biomarkers, including but not limited to total IgE, allergen-specific IgE, thymus and activated chemokine (TARC), lung and activated chemokine (PARC), lactate dehydrogenase (LDH), and / or periostin. For example, in some embodiments, the method of this application comprises administering an IL-4 / IL-13 pathway inhibitor with a plasma cell ablation agent to a patient with elevated levels of allergen-specific serum IgE.

[0047] The term "individual with this need" also includes individuals suffering from an allergic disease or condition selected from allergic asthma, hay fever, chronic urticaria, food allergy, pollen allergy, and allergies caused by environmental (non-food) allergens. The term also includes individuals suffering from severe allergies due to one or more allergens. For example, in some embodiments, an individual is considered to have a "severe" allergy if they exhibit one or more severe symptoms of an allergic reaction, such as symptoms of an allergic reaction (e.g., difficulty breathing / noisy breathing, swollen tongue, swollen / tight throat, difficulty speaking and / or hoarseness, wheezing or persistent cough, nausea / vomiting, persistent dizziness, collapse, or loss of consciousness).

[0048] In some embodiments, the term "individual with this need" includes individuals who are susceptible to allergic reactions or who are at increased risk of developing an allergic reaction to an allergen. For example, this term includes individuals at risk of an allergic reaction due to an allergen, such as peanuts or penicillin. In some embodiments, an individual may be at increased risk of developing an allergy or allergic reaction to an allergen due to sensitization. For example, this term includes individuals exhibiting elevated serum IgE levels specific to one or more allergens ("allergen sensitization"), such as one or more food allergens and / or environmental allergens. In some embodiments, an individual has an allergen-specific IgE level of at least about 0.35 kU / L (e.g., for one or more allergens disclosed herein, such as food allergens or environmental allergens, or allergens selected from: milk, dairy products, eggs, celery, sesame, wheat, meat, fruit, soybeans, fish, shellfish, sugar, peanuts, beans, tree nuts, dust, dust mites, pollen, insect venom, mold, animal fur, animal dander, wool, latex, metals, household cleaners, detergents, pharmaceuticals, cosmetics, fragrances, pharmaceuticals such as penicillin, sulfonamides or salicylates, therapeutic monoclonal antibodies, ragweed, grass, and birch). In the context of this application, the term "individual with this need" also includes individuals suffering from atopic diseases and individuals suffering from diseases or disorders selected from atopic dermatitis, asthma, allergic rhinitis, eosinophilic esophagitis, and food allergies. The term "individual" also includes individuals with elevated levels of serum total IgE and allergen-specific IgE or serum chemokines (e.g., CCL17 or CCL27), who may have an increased risk of developing allergic reactions. In one aspect, this application provides a method for reducing the risk of allergies or allergic reactions in susceptible individuals.

[0049] As used herein, the terms "allergic reaction," "allergic symptoms," etc., include one or more signs or symptoms selected from urticaria (e.g., hives), angioedema, rhinitis, asthma, vomiting, sneezing, runny nose, sinusitis, lacrimation, wheezing, bronchospasm, decreased peak expiratory flow (PEF), gastrointestinal upset, flushing, swollen lips, swollen tongue, decreased blood pressure, anaphylaxis, and organ dysfunction / failure. "Allergic reaction," "allergic symptoms," etc., also include immune responses and immune reactions, such as increased IgE production and / or increased production of allergen-specific immunoglobulins.

[0050] As used herein, the term "allergen" includes any substance, chemical, particle, or composition capable of stimulating an allergic reaction in a susceptible individual. Allergens contained in or derived from food include, for example, dairy products (e.g., milk), eggs, celery, sesame seeds, wheat, meat, soy, fish, shellfish, sugars (e.g., sugars present in meat, such as α-galactose), peanuts, other legumes (e.g., beans, peas, soybeans, etc.)), and tree nuts; allergens contained in or derived from food are referred to herein as "food allergens." Alternatively, allergens can be contained in or derived from non-food substances, such as indoor or outdoor environmental allergens, such as dust (e.g., containing dust mites), pollen, insect venom (e.g., venom from bees, wasps, mosquitoes, fire ants, etc.), mold, animal fur, animal dander, wool, latex, metals (e.g., nickel), household cleaners, detergents, medications, cosmetics (e.g., fragrances), drugs (e.g., penicillin, sulfonamides, salicylates, etc.), therapeutic monoclonal antibodies (e.g., cetuximab), ragweed, grass, and birch. Exemplary pollen allergens include, for example, tree pollen, such as birch pollen, cedar pollen, oak pollen, alder pollen, hornbeam pollen, horse chestnut pollen, willow pollen, poplar pollen, sycamore pollen, linden pollen, oleanum pollen, ashe juniper pollen, and alstonia scholaris pollen. Other examples of allergens can be found in other parts of this article.

[0051] This application includes methods for treating allergies, including severe allergies, or preventing or reducing the severity of allergic reactions, comprising administering to an individual in need a therapeutically effective amount of an IL-4 / IL-13 pathway inhibitor (e.g., the anti-IL-4R antibody described herein) and a therapeutically effective amount of a plasma cell ablation agent (e.g., the anti-BCMA x anti-CD3 bispecific antibody described herein). In some embodiments, the disclosed methods target allergic diseases or conditions, mast cell activation disorders, or mast cell proliferation. In one embodiment, the allergic disease or condition is selected from allergic asthma, hay fever, chronic urticaria, food allergies, pollen allergies, and allergies caused by environmental (non-food) allergens. In some embodiments, the allergic disease is a food allergy, such as peanut allergy. In some embodiments, the allergic disease is a severe food allergy.

[0052] According to certain embodiments, this application includes a method for treating allergies or preventing or reducing the severity of allergic reactions, the method comprising: (a) selecting a patient suffering from an allergic disease or condition, mast cell activation disorder, or mast cell hyperplasia, wherein the patient is undergoing a background therapy regimen comprising one or more doses of an IL-4 / IL-13 pathway inhibitor; and (b) administering at least one dose of a plasma cell ablation agent. In this respect, the method enhances the therapeutic efficacy of the IL-4 / IL-13 pathway inhibitor in reducing serum levels of allergen-specific IgE. In some embodiments, the patient is undergoing a treatment regimen comprising one or more doses of the IL-4 / IL-13 pathway, and one or more doses of a plasma cell ablation agent are administered, thereby enhancing the anti-allergic effect of the IL-4 / IL-13 pathway inhibitor.

[0053] In some embodiments, the disclosed methods include administering a therapeutically effective amount of an IL-4 / IL-13 pathway inhibitor and a therapeutically effective amount of a plasma cell ablation agent along with other therapeutic agents or therapies (e.g., protocols or methods). Other therapeutic agents or therapies may be administered to increase anti-allergic efficacy, reduce the toxicity of one or more therapies, and / or lower the dosage of one or more therapies. In various embodiments, the other therapeutic agents or therapies may include one or more of IgE antagonists, antihistamines, anti-inflammatory drugs, corticosteroids, leukotriene antagonists, mast cell inhibitors, bronchodilators, decongestants, adrenaline, IL-1 antagonists, IL-5 antagonists, IL-31 antagonists, IL-33 antagonists, IL-25 antagonists, interferon-γ, TNF antagonists, and TSLP antagonists.

[0054] The other therapeutic agents may be, for example, another IL-4R antagonist, IL-1 antagonist (including, for example, IL-1 antagonists as described in US 6,927,044), IL-6 antagonist, IL-6R antagonist (including, for example, anti-IL-6R antibodies as described in US 7,582,298), IL-13 antagonist, tumor necrosis factor (TNF) antagonist, IL-8 antagonist, IL-9 antagonist, IL-17 antagonist, IL-5 antagonist, IgE antagonist (e.g., anti-IgE antibodies, such as omazomab), CD48 antagonist, IL-31 antagonist (including, for example, those described in US 7,531,637), thymic interstitial lymphopoietin (TSLP) antagonist (including, for example, those described in US 7,531,637), and thymic interstitial lymphopoietin (TSLP) antagonist (including, for example, those described in US 7,927,044). The following are included in 2011 / 027468: interferon-γ (IFNγ), antibiotics, topical corticosteroids, tacrolimus, pimecrolimus, cyclosporine, azathioprine, methotrexate, sodium cromoglycate, protease inhibitors, systemic corticosteroids, systemic immunotherapy, antihistamines, or combinations thereof.

[0055] In some embodiments, the disclosed method of administering a therapeutically effective amount of an IL-4 / IL-13 pathway inhibitor and a therapeutically effective amount of a plasma cell ablation agent results in a reduction or elimination of one or more symptoms or indications of allergy compared to untreated individuals or individuals treated with one of the inhibitors as a monotherapy.

[0056] In some embodiments, the disclosed method results in a reduction, preferably complete elimination, of allergen-specific IgE in the treated individual. For example, the disclosed method of administering a therapeutically effective amount of an IL-4 / IL-13 pathway inhibitor and a therapeutically effective amount of a plasma cell ablation agent promotes a reduction in serum allergen-specific IgE levels of the treated individual by at least about 50%, about 60%, about 70%, or more than about 80% compared to untreated individuals or individuals treated with one of the inhibitors as monotherapy. In some embodiments, the disclosed method results in complete elimination of allergen-specific serum IgE in the treated individual compared to individuals treated with one of the active agents as monotherapy.

[0057] According to certain implementations, after administration of one or more doses of an IL-4 / IL-13 pathway inhibitor (e.g., anti-IL-4R antibody) and one or more doses of a plasma cell ablation agent (e.g., anti-BCMA / anti-CD3 bispecific antibody), an individual may exhibit a decrease in serum IgE levels specific to one or more allergens. For example, according to this application, after administration of one or more doses of an anti-IL-4R antibody (e.g., dupilumab) with a plasma cell ablation agent, individuals may exhibit a reduction in allergen-specific IgE of approximately 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or more from baseline (wherein “baseline” is defined as the individual’s level of allergen-specific IgE prior to the first administration) on approximately day 8, day 15, day 22, day 25, day 29, day 36, day 43, day 50, day 57, day 64, day 71, day 85 or day 112.

[0058] Methods for detecting and / or quantifying allergen-specific IgE or total IgE in serum are well known in the art; kits for measuring them are available from various commercial sources; and various commercial diagnostic laboratories also offer services to obtain measurements of said levels.

[0059] For example, Phadiatop TMThis is a commercially available variant of a serum-specific or antigen-specific IgE assay, introduced for screening for allergic sensitization (Merrett et al. 1987, Allergy 17:409-416). The test simultaneously measures serum-specific IgE for a mixture of common inhaled allergens. The test provides a qualitative result, positive or negative, based on the observed fluorescence response. A positive result is indicated when a patient sample shows a fluorescence response equal to or higher than the reference value. A negative result is indicated when a patient sample shows a lower fluorescence response. This application includes a method comprising selecting individuals with a positive test result and administering to the individual a therapeutically effective amount of a combination of an IL-4 / IL-13 pathway inhibitor and a plasma cell ablation agent.

[0060] In some implementations, the combination of administered therapeutic agents is safe and well tolerated by the individual, such that there is no increase in adverse side effects compared to individuals treated with one of the therapeutic agents as a monotherapy.

[0061] IL-4 / IL-13 pathway inhibitors

[0062] The methods disclosed herein include administering a therapeutically effective amount of an IL-4 / IL-13 pathway inhibitor to an individual in need. As used herein, an "IL-4 / IL-13 pathway inhibitor" (also referred to herein as an "IL-4 / IL-13 pathway antagonist," "IL-4 / IL-13 pathway blocker," etc.) is any active agent that inhibits or attenuates at least one of the following effects: (i) binding of IL-4 and / or IL-13 to their respective receptors; (ii) signaling and / or activity of IL-4 and / or IL-13; and / or (iii) downstream signaling / activity resulting from the binding of IL-4 and / or IL-13 to their respective receptors. Exemplary IL-4 / IL-13 pathway inhibitors include, but are not limited to, anti-IL-4 antibodies (e.g., those disclosed in U.S. Patent 7,740,843 and U.S. Patent Application Publications 2010 / 0297110 and 2016 / 0207995), and anti-IL-13 antibodies (e.g., those disclosed in U.S. Patents 7,501,121, 7,674,459, 7,807,788, 7,910,708, 7,915,388, 7,935,343, 8,088,618, 8,691,233, and 9,605,065, U.S. Patents... Antibodies disclosed in application publications 2006 / 0073148 and 2008 / 0044420 and EP2627673B1, bispecific antibodies binding to IL-4 and IL-13 (e.g., antibodies disclosed in U.S. Patent 8,388,965 and U.S. Patent Application Publications 2011 / 0008345, 2013 / 0251718 and 2016 / 0207995), and IL-4 receptor (IL-4R) inhibitors (described below), portions of publications identifying inhibitors of the IL-4 / IL-13 pathway cited herein are incorporated herein by reference.

[0063] In some embodiments, the IL-4 / IL-13 pathway inhibitor may be an antibody, a small molecule compound, a nucleic acid, a peptide, or a functional fragment or variant thereof. Non-limiting examples of suitable IL-4 / IL-13 pathway inhibitor antibodies include anti-IL-4 antibodies, anti-IL-13 antibodies and anti-IL-4 / IL-13 bispecific antibodies, anti-IL-4R antibodies and any antigen-binding fragment thereof. Other non-limiting examples of suitable IL-4 / IL-13 pathway inhibitors include: RNAi molecules, such as anti-IL-4 RNAi molecules and anti-IL-13 RNAi; antisense molecules, such as anti-IL-4 antisense RNA and anti-IL-13 antisense RNA; and dominant / negative proteins, such as dominant / negative IL-4 protein and dominant / negative L-13 protein.

[0064] As used herein, "IL-4R inhibitor" (also referred to herein as "IL-4 / IL-13 pathway inhibitor," "IL-4Rα antagonist," "IL-4R blocker," "IL-4Rα blocker," etc.) is any active agent that binds to or interacts with IL-4Rα or IL-4R ligands and inhibits or attenuates the normal biological signal transduction function of type 1 and / or type 2 IL-4 receptors. Type 1 IL-4 receptors are dimerized receptors containing IL-4Rα and γc chains. Type 2 IL-4 receptors are dimerized receptors containing IL-4Rα and IL-13Rα1 chains. Type 1 IL-4 receptors interact with and are stimulated by IL-4, while type 2 IL-4 receptors interact with and are stimulated by both IL-4 and IL-13. Therefore, the IL-4R inhibitors that can be used in the methods of this application can act by blocking IL-4-mediated signal transduction, IL-13-mediated signal transduction, or IL-4- and IL-13-mediated signal transduction. The IL-4R inhibitors of this application can thus prevent IL-4 and / or IL-13 from interacting with type 1 or type 2 receptors.

[0065] Non-limiting examples of the IL-4R inhibitor class include IL-4 mutant proteins (e.g., picotriol), small molecule IL-4R inhibitors, anti-IL-4R aptamers, peptide-based IL-4R inhibitors (e.g., "peptibody" molecules), "receptor-body" molecules (e.g., modified molecules containing ligand-binding domains of IL-4R components), and antigen-binding fragments of antibodies or antibodies that specifically bind human IL-4Rα. As used herein, IL-4R inhibitors also include antigen-binding proteins that specifically bind IL-4 and / or IL-13.

[0066] Other non-limiting examples of suitable IL-4 / IL-13 pathway inhibitors that may be used in the context of this application include, for example, picoplanin (AER-001; BAY-16-9996), aeroderm (AER-003), and antibodies known and involved in the art, such as dupilumab, pacocillin, AMG-317, MILR1444A, CAT-354, QAX576, tranexamic acid (IMA-638), ISIS-369645 (AIR-645), IMA-026, APG-201, CNTO-607, MK-6105, MEDI9314, MEDI2045, trorocalumab, lerezumab, roximateb, and DOM-0910.

[0067] Anti-IL-4Rα antibody and its antigen-binding fragment

[0068] According to certain exemplary embodiments of this application, the IL-4 / IL-13 pathway inhibitor is an anti-IL-4Rα antibody or its antigen-binding fragment. The term "antibody" as used throughout this application includes immunoglobulin molecules and their multimers (e.g., IgM) comprising four polypeptide chains linked by disulfide bonds, namely two heavy (H) chains and two light (L) chains. In a typical antibody, each heavy chain contains a heavy chain variable region (abbreviated herein as HCVR or V). H The heavy-chain constant region contains three structural domains C. H 1. C H 2 and C H 3. Each light chain contains a light chain variable region (abbreviated as LCVR or V in this article). L The light chain constant region contains one structural domain (C) and a light chain constant region. L 1). V H and V L The region can be further divided into highly variable regions called complementary determinant regions (CDRs), within which more conservatively spaced regions are called frame regions (FRs). Each V H and V L It consists of 3 CDRs and 4 FRs, arranged in the following order from the amino terminus to the carboxyl terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. In different embodiments of this application, the FRs of the antibody (or its antigen-binding portion) may be identical to those of a human lineage or may be natural or artificially modified. A common amino acid sequence can be defined based on the alignment analysis of two or more CDRs.

[0069] The term "antibody" as used throughout this application includes its antigen-binding fragment, i.e., the antigen-binding fragment of the complete antibody molecule. The terms "antigen-binding portion" and "antigen-binding fragment" as used throughout this application include any naturally occurring, enzymatically obtained, synthetic, or genetically modified polypeptide or glycoprotein that specifically binds to an antigen to form a complex. The antigen-binding fragment of an antibody can be derived from the complete antibody molecule, for example, using any suitable standard technique, such as proteolytic digestion or recombinant genetic engineering, involving the manipulation and expression of DNA encoding variable and optionally constant domains of the antibody. Such DNA is known and / or readily available from, for example, commercial sources, DNA libraries (including, for example, phage-antibody libraries), or can be synthesized. For example, chemical or molecular biological techniques can be used to sequence and manipulate the DNA to arrange one or more variable and / or constant domains into a suitable conformation, or to introduce codons, generate cysteine ​​residues, modify, add, or delete amino acids, etc.

[0070] Non-limiting examples of antigen-binding fragments include: (i) Fab fragments; (ii) F(ab')2 fragments; (iii) Fd fragments; (iv) Fv fragments; (v) single-chain Fv (scFv) molecules; (vi) dAb fragments; and (vii) the smallest recognition unit consisting of amino acid residues of a hypervariable region of a mimic antibody (e.g., a separated complementarity-determining region (CDR), such as a CDR3 peptide) or a bound FR3-CDR3-FR4 peptide. Other modified molecules, such as domain-specific antibodies, single-domain antibodies, domain-deficient antibodies, chimeric antibodies, CDR-transplanted antibodies, biantibodies, triantibodies, tetraantibodies, small bodies, nanobodies (e.g., monovalent nanobodies, bivalent nanobodies, etc.), small motif immunopharmaceuticals (SMIPs), and shark variable IgNAR domains, are also included within the term "antigen-binding fragment" as used throughout this application.

[0071] Antibody antigen-binding fragments typically contain at least one variable domain. Variable domains can have any size or amino acid composition and generally contain at least one CDR adjacent to or in frame one or more frames. L V with domain combination H In the antigen-binding fragment of the domain, V H and V L Domains can be positioned relative to each other in any suitable arrangement. For example, variable regions can be dimerized and contain V. H -V H V H -V L or V L -V L Dimer. Alternatively, the antigen-binding fragment of the antibody may contain monomer V. H or V L Structural domain.

[0072] In some embodiments, the antigen-binding fragment of the antibody may include at least one variable domain covalently linked to at least one constant domain. Non-limiting exemplary configurations of the variable and constant domains that may be found within the antigen-binding fragment of the antibody of this application include: (i) V H -C H 1; (ii)V H -C H 2; (iii)V H -C H 3; (iv)V H -C H 1-C H 2; (v)V H -C H 1-C H 2-C H3;(vi)V H -C H 2-C H 3;(vii)V H -C L (viii)V L -C H 1; (ix)V L -C H 2; (x)V L -C H 3;(xi)V L -C H 1-C H 2;(xii)V L -C H 1-C H 2-C H 3; (xiii)V L -C H 2-C H 3; and (xiv)V L -C L In any configuration of the variable and constant domains, including any of the exemplary configurations listed above, the variable and constant domains may be directly connected to each other or may be connected via complete or partial hinge or connector regions. The hinge region may consist of at least two (e.g., 5, 10, 15, 20, 40, 60, or more) amino acids, creating a flexible or semi-flexible connection between adjacent variable and / or constant domains in a single polypeptide molecule. Furthermore, the antigen-binding fragment of the antibody of this application may comprise the domains listed above that are non-covalently linked to each other and / or have one or more monomeric V... H or V L Homodimers or heterodimers (or other polymers) of any variable and constant domain configurations of the domain (e.g., via disulfide bonds).

[0073] The term "antibody" as used throughout this application also includes multispecific (e.g., bispecific) antibodies. Multispecific antibodies or antigen-binding fragments of antibodies typically comprise at least two distinct variable domains, each capable of specifically binding to a single antigen or different epitopes on the same antigen. Conventional techniques available in the art can be applied to make any form of multispecific antibody suitable for the context of the antibodies or antigen-binding fragments of antibodies of this application. For example, this application includes a method comprising using a bispecific antibody, wherein one arm of an immunoglobulin is specific for IL-4Rα or a fragment thereof, while the other arm is specific for or conjugated to a second therapeutic target. Exemplary bispecific forms that may be used in the context of this application include, but are not limited to, scFv-based biantibody-specific forms, IgG-scFv fusions, dual variable domain (DVD)-Ig, quadroma, knocks-in-holes, universal light chains (e.g., universal light chains with knocks), CrossMab, CrossFab, (SEED) bodies, leucine zippers, Duobody, IgG1 / IgG2, dual-action Fab (DAF)-IgG, and Mab. 2 Bispecific forms (for a review of the above forms, see, for example, Klein et al. 2012, mAbs 4(6):653-663 and the references cited therein). Bispecific antibodies can also be constructed using peptide / nucleic acid conjugations, for example, in which non-natural amino acids with orthogonal chemical reactivity are used to generate site-specific antibody-oligonucleotide conjugations, which then self-assemble into polymerized complexes with defined composition, titer, and geometry. (See, for example, Kazane et al., J.Am.Chem.Soc., 2013, 135(1):340-46).

[0074] The antibodies used in the methods of this application may be human antibodies. The term "human antibody" as used throughout this application is intended to include antibodies having variable and constant regions derived from human germline immunoglobulin sequences. However, the human antibodies of this application may include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced through random or site-specific mutagenesis in vitro or through somatic mutation in vivo), such as in CDRs, particularly CDR3. However, the term "human antibody" as used throughout this application is not intended to include antibodies in which a CDR sequence derived from another mammalian species, such as a mouse, has been grafted onto a human frame sequence.

[0075] The antibody used in the methods of this application may be a recombinant human antibody. As used throughout this application, the term "recombinant human antibody" is intended to include all human antibodies prepared, expressed, generated, or isolated in a recombinant manner, such as antibodies expressed using a recombinant expression vector transfected into host cells (further described below), antibodies isolated from recombinant, combined human antibody libraries (further described below), antibodies isolated from animals (e.g., mice) transgenic for human immunoglobulin genes (see, for example, Taylor et al. (1992) Nucl. Acids Res., 20:6287-6295), or antibodies prepared, expressed, generated, or isolated by any other method involving splicing a human immunoglobulin gene sequence to another DNA sequence. Such recombinant human antibodies have variable and constant regions derived from human germline immunoglobulin sequences. However, in some embodiments, such recombinant human antibodies may be mutagenized in vitro (or, when using animals transgenic for human Ig sequences, in vivo somatic cell mutagenization), thus the V of the recombinant antibody... H and V L The amino acid sequence of the region is such that, although it originates from human lineage V, it is not a sequence that can be easily replicated. H and V L The sequence is associated with it, but it may not be naturally present in the human antibody germline library.

[0076] According to certain embodiments, the antibodies used in the methods of this application specifically bind IL-4Rα. The term "specific binding," etc., refers to the formation of a complex between an antibody or its antigen-binding fragment and an antigen, which is relatively stable under physiological conditions. Methods for determining whether an antibody specifically binds to an antigen are well known in the art and include, for example, equilibrium dialysis, surface plasmon resonance assays, etc. For example, in some embodiments, antibodies that "specifically bind" IL-4Rα as used in the context of this application include, as measured in surface plasmon resonance assays, at concentrations of less than about 500 nM, less than about 300 nM, less than about 200 nM, less than about 100 nM, less than about 90 nM, less than about 80 nM, less than about 70 nM, less than about 60 nM, less than about 50 nM, less than about 40 nM, less than about 30 nM, less than about 20 nM, less than about 10 nM, less than about 5 nM, less than about 4 nM, less than about 3 nM, less than about 2 nM, less than about 1 nM, or less than about 0.5 nM. D Antibodies that bind to IL-4Rα or a fraction thereof. However, isolated antibodies that specifically bind to human IL-4Rα may be cross-reactive with other antigens, such as IL-4Rα molecules from other (non-human) species.

[0077] According to certain exemplary embodiments of this application, the IL-4 / IL-13 pathway inhibitor is an anti-IL-4Rα antibody or its antigen-binding fragment, comprising: a heavy chain variable region (HCVR), a light chain variable region (LCVR), and / or a complementarity-determining region (CDR) comprising any amino acid sequence of an anti-IL-4R antibody as described in U.S. Patent No. 7,608,693, which is incorporated herein by reference. In certain exemplary embodiments, the anti-IL-4Rα antibody or its antigen-binding fragment that can be used in the method context of this application comprises: a heavy chain complementarity-determining region (HCDR) containing the heavy chain variable region (HCVR) of the amino acid sequence of SEQ ID NO:1 and a light chain complementarity-determining region (LCDR) containing the light chain variable region (LCVR) of the amino acid sequence of SEQ ID NO:2. According to some embodiments, the anti-IL-4Rα antibody or its antigen-binding fragment comprises three HCDRs (HCDR1, HCDR2, and HCDR3) and three LCDRs (LCDR1, LCDR2, and LCDR3), wherein HCDR1 comprises the amino acid sequence of SEQ ID NO:3; HCDR2 comprises the amino acid sequence of SEQ ID NO:4; HCDR3 comprises the amino acid sequence of SEQ ID NO:5; LCDR1 comprises the amino acid sequence of SEQ ID NO:6; LCDR2 comprises the amino acid sequence of SEQ ID NO:7; and LCDR3 comprises the amino acid sequence of SEQ ID NO:8. In another embodiment, the anti-IL-4R antibody or its antigen-binding fragment comprises: an HCVR containing SEQ ID NO:1 and an LCVR containing SEQ ID NO:2. In some embodiments, the method of this application comprises using an anti-IL-4R antibody, wherein the antibody comprises a heavy chain containing the amino acid sequence of SEQ ID NO:9. In some embodiments, the anti-IL-4R antibody comprises a light chain containing the amino acid sequence of SEQ ID NO:10. An exemplary antibody comprising a heavy chain containing the amino acid sequence of SEQ ID NO:9 and a light chain containing the amino acid sequence of SEQ ID NO:10 is a complete human anti-IL-4R antibody, referred to as dupilumab (DUPIXENT™). According to certain exemplary embodiments, the method of this application includes the use of dupilumab or a bioequivalence thereof. The term "bioequivalence" in relation to dupilumab refers to an anti-IL-4R antibody or its IL-4R-binding protein or fragment, which is a pharmaceutical equivalent or substitute, whose rate and / or extent of absorption does not show significant difference from dupilumab when administered at the same molar dose (whether single or multiple doses) under similar experimental conditions. In the context of this application, the term refers to an antigen-binding protein that binds to IL-4R, which has no clinically significant difference from dupilumab in terms of safety, purity, and / or efficacy.

[0078] According to certain embodiments of this application, the anti-human IL-4R antibody or its antigen-binding fragment comprises an HCVR having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with SEQ ID NO:1. In some embodiments, the anti-human IL-4R antibody or its antigen-binding fragment comprises three HCDRs (HCDR1, HCDR2, and HCDR3) containing the amino acid sequences of SEQ ID NO:3, 4, and 5, respectively, and comprises an HCVR having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:1.

[0079] According to certain embodiments of this application, the anti-human IL-4R antibody or its antigen-binding fragment comprises an LCVR having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with SEQ ID NO:2. In some embodiments, the anti-human IL-4R antibody or its antigen-binding fragment comprises three LCDRs (LCDR1, LCDR2, and LCDR3) each containing the amino acid sequences of SEQ ID NO:6, 7, and 8, and comprises an LCVR having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:2.

[0080] According to certain embodiments of this application, an anti-human IL-4R antibody or its antigen-binding fragment comprises an HCVR having an amino acid sequence of SEQ ID NO:1 having no more than 5 amino acid substitutions. According to certain embodiments of this application, an anti-human IL-4R antibody or its antigen-binding fragment comprises an LCVR having an amino acid sequence of SEQ ID NO:2 having no more than 2 amino acid substitutions.

[0081] In some embodiments, the anti-human IL-4R antibody or its antigen-binding fragment comprises an HCVR having 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with SEQ ID NO:1 and an LCVR having 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with SEQ ID NO:2. In some embodiments, the anti-human IL-4R antibody or its antigen-binding fragment comprises: (a) HCDR1, HCDR2, and HCDR3 comprising the amino acid sequences of SEQ ID NO:3, 4, and 5, respectively, and HCVR having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:1; and (b) LCDR1, LCDR2, and LCDR3 comprising the amino acid sequences of SEQ ID NO:6, 7, and 8, respectively, and LCVR having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:2.

[0082] Sequence identity can be determined using methods known in the art (e.g., GAP, BESTFIT, and BLAST).

[0083] This application also includes the use of anti-IL-4R antibodies in methods of treating an individual's allergy or eliminating their allergen-specific IgE, wherein said anti-IL-4R antibody comprises a variant of any HCVR, LCVR, and / or CDR amino acid sequence disclosed herein having one or more conserved amino acid substitutions. For example, this application includes the use of anti-IL-4R antibodies having HCVR, LCVR, and / or CDR amino acid sequences having, for example, 10 or fewer, 8 or fewer, 6 or fewer, 4 or fewer, etc., conserved amino acid substitutions relative to any HCVR, LCVR, and / or CDR amino acid sequence disclosed herein. In some embodiments, this application includes the use of anti-IL-4R antibodies having HCVR, LCVR, and / or CDR amino acid sequences having 1, 2, 3, or 4 conserved amino acid substitutions relative to any HCVR, LCVR, and / or CDR amino acid sequence disclosed herein.

[0084] Other anti-IL-4Rα antibodies that may be used in the context of the methods described in this application include, for example, antibodies known and relevant to the art, such as AMG317 (Corren et al., 2010, Am J Respir Crit Care Med., 181(8):788-796) or MEDI 9314 or any of the anti-IL-4Rα antibodies described in the following documents: U.S. Patent Nos. 7,186,809, 7,605,237, 7,638,606, 8,092,804, 8,679,487, or 8,877,189. Representations of publications identifying anti-IL-4Rα antibodies cited herein are incorporated herein by reference.

[0085] Anti-IL-4Rα antibodies used in the context of the methods of this application may have pH-dependent binding characteristics. For example, anti-IL-4Rα antibodies used in the methods of this application may exhibit reduced binding to IL-4Rα at acidic pH compared to neutral pH. Alternatively, anti-IL-4Rα antibodies of this application may exhibit enhanced binding to their antigen at acidic pH compared to neutral pH. The expression "acidic pH" includes pH values ​​less than about 6.2, such as about 6.0, 5.95, 5.9, 5.85, 5.8, 5.75, 5.7, 5.65, 5.6, 5.55, 5.5, 5.45, 5.4, 5.35, 5.3, 5.25, 5.2, 5.15, 5.1, 5.05, 5.0 or lower. As used throughout this application, the expression "neutral pH" refers to a pH of about 7.0 to about 7.4. The term "neutral pH" includes pH values ​​of approximately 7.0, 7.05, 7.1, 7.15, 7.2, 7.25, 7.3, 7.35, and 7.4.

[0086] In some cases, "reduced binding to IL-4Rα at acidic pH compared to neutral pH" is expressed as the Kc of the antibody binding to IL-4Rα at acidic pH. D The K value of the antibody that binds to IL-4Rα at neutral pH D The ratio of values ​​(or vice versa). For example, for the purposes of this application, if the antibody or its antigen-binding fragment exhibits acidic / neutral K... D A ratio of approximately 3.0 or greater can be considered as exhibiting "reduced binding to IL-4Rα at acidic pH compared to neutral pH". In some exemplary embodiments, the acidic / neutral pH of the antibody or antigen-binding fragment of this application... DThe ratio can be approximately 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10.0, 10.5, 11.0, 11.5, 12.0, 12.5, 13.0, 13.5, 14.0, 14.5, 15.0, 20.0, 25.0, 30.0, 40.0, 50.0, 60.0, 70.0, 100.0 or higher.

[0087] Antibodies exhibiting pH-dependent binding characteristics can be obtained, for example, by screening for a population of antibodies that bind less (or more) to a specific antigen at acidic pH compared to neutral pH. Furthermore, modifications to the antigen-binding domain at the amino acid level can also produce antibodies with pH-dependent characteristics. For example, by substituting one or more amino acids of the antigen-binding domain (e.g., within the CDR) with histidine residues, antibodies exhibiting reduced antigen binding at acidic pH relative to neutral pH can be obtained. As used throughout this application, the term "acidic pH" refers to a pH of 6.0 or below.

[0088] Plasma cell ablation agent

[0089] The methods disclosed herein involve administering a therapeutically effective amount of a plasma cell ablative to an individual in need. As used herein, "plasma cell ablative" refers to any molecule capable of specifically binding to surface antigens on plasma cells and killing or ablating said plasma cells. In some embodiments, the plasma cell ablative may be an antibody, a small molecule compound, a nucleic acid, a peptide, or a functional fragment or variant thereof. In the context of this application, the plasma cell ablative is used in combination with an IL-4 / IL-13 pathway inhibitor from the disclosed methods.

[0090] Non-limiting examples of suitable plasma cell ablation agents include BCMA-targeting agents (as described elsewhere in this document), proteasome inhibitors [e.g., bortezomib (Velcade), carfilzomib (Kyprolis), ixazomib (Ninlaro)], histone deacetylase inhibitors [e.g., pabistat (Farydak)], B-cell activating factor (BAFF; also known as BLyS, TALL-1, or CD257) inhibitors (e.g., anti-BAFF antibodies, such as belimumab, tabalumab, AMG570; or anti-BAFF receptor antibodies, such as ianalumab)), and proliferation-inducing ligand (APRIL; also known as TNFSF13 or CD256) inhibitors (e.g., anti-APRIL antibodies, such as BION-1301 or VIS624).

[0091] BCMA-targeted drugs

[0092] According to certain exemplary embodiments, the plasma cell ablation agent used in the methods disclosed herein is a BCMA-targeting drug.

[0093] As used herein, the term "BCMA-targeting agent" refers to any molecule capable of specifically binding to BCMA expressed on the surface of individual cells, thereby targeting and destroying said cells. BCMA is expressed only in B-cell lineage cells, particularly in the interfollicular region of germinal centers and on plasmablasts and differentiated plasma cells. BCMA is selectively induced during plasma cell differentiation and is essential for the optimal survival of long-lived plasma cells in the bone marrow. Therefore, BCMA-targeting agents bind to BCMA expressed on the surface of plasma cells and mediate the killing or ablation (plasma cell ablation) of BCMA-expressing cells. In the context of this application, in some embodiments, a BCMA-targeting agent comprises a binding portion (antigen-binding portion or antigen-binding fragment thereof) that binds to BCMA expressed on the surface of plasma cells and a portion that promotes the killing of said plasma cells. In some embodiments, the BCMA portion expressed on the surface of plasma cells is an antibody or antigen-binding fragment thereof that specifically binds to BCMA. Such BCMA-binding portions are linked (e.g., covalently bound) to portions that help kill or destroy target plasma cells. The portion that helps target and kill the bound plasma cells can be a molecule that directly kills the target cells (e.g., a cytotoxic agent), or it can be a protein or fragment thereof that mediates the killing of target cells by immune cells such as T cells. In the context of this application, the term "BCMA-targeting agent" includes, but is not limited to, anti-BCMA antibodies ("BCMA ADC" or "anti-BCMA ADC") conjugated to therapeutic agents such as cytotoxic drugs, chimeric antigen receptors (CARs) that specifically bind to BCMA ("BCMA CAR" or "anti-BCMA CAR"), and anti-BCMA / anti-CD3 bispecific antibodies.

[0094] According to certain embodiments, the BCMA-targeting agent used in the disclosed method context is an antibody-drug conjugate (ADC) comprising an anti-BCMA antibody and a cytotoxic agent. In some embodiments, the anti-BCMA antibody or its antigen-binding fragment and the cytotoxic agent are covalently linked via a linker. Generally, an ADC comprises: A-[LP] yWhere A is an antigen-binding molecule, such as an anti-BCMA antibody or a fragment thereof, L is a linker, P is a payload or therapeutic agent portion (e.g., a cytotoxic agent), and y is an integer from 1 to 30. Examples of suitable cytotoxic agents and chemotherapeutic agents for forming an ADC are well known in the art. Non-limiting examples of suitable cytotoxic agents that can be conjugated to anti-BCMA antibodies for the disclosed methods are auristatin, such as monomethylauristatin E (MMAE) or monomethylauristatin F (MMAF), tubulysin, such as TUB-OH or TUB-OMOM, thromycin derivatives, dolalastatin derivatives, or maytansinoids, such as DM1 or DM4. According to certain exemplary embodiments, this application includes the use of an anti-BCMA ADC in a method of treating allergies, wherein the anti-BCMA ADC comprises any of the HCVR, LCVR, and / or CDR amino acid sequences disclosed in other parts of this document.

[0095] This application also includes the use of anti-BCMA ADCs in methods of treating allergic reactions, wherein the antibody comprises a variant of any of the disclosed HCVR, LCVR, and / or CDR amino acid sequences having one or more conserved amino acid substitutions. For example, this application includes the use of antibodies having HCVR, LCVR, and / or CDR amino acid sequences having, for example, 10 or fewer, 8 or fewer, 6 or fewer, 4 or fewer, etc., conserved amino acid substitutions relative to any of the disclosed HCVR, LCVR, and / or CDR amino acid sequences. In some embodiments, this application includes the use of antibodies having HCVR, LCVR, and / or CDR amino acid sequences having 1, 2, 3, or 4 conserved amino acid substitutions relative to any of the disclosed HCVR, LCVR, and / or CDR amino acid sequences.

[0096] Other anti-BCMA ADCs that may be used in the context of the methods described in this application include, for example, ADCs known and relevant to the art, such as belantamoxifen (mofodotin), GSK2857916, AMG224, HDP-101, MEDI2228, and TBL-CLN1, or any anti-BCMA ADC described, for example, in the following patent publications: WO2011 / 108008, WO2014 / 089335, WO2017 / 093942, WO2017 / 143069, and WO2019 / 025983. Representations of publications identifying anti-BCMA ADCs cited herein are incorporated herein by reference.

[0097] According to certain embodiments, the BCMA-targeting agent used in the context of the disclosed methods is a chimeric antigen receptor (CAR) that specifically binds to BCMA ("BCMA CAR"). The term "chimeric antigen receptor" (CAR) refers to a molecule that combines a binding domain of a component present on a target cell, such as an antibody specific for a target antigen (e.g., BCMA on plasma cells), with an intracellular domain of an activated T-cell receptor to produce a chimeric protein exhibiting specific anti-target cell immune activity. Typically, a CAR includes an extracellular single-chain antibody-binding domain (scFv) fused to an intracellular signaling domain of the ζ chain of the T-cell antigen receptor complex, and when expressed in T cells, possesses the ability to redirect antigen recognition based on monoclonal antibody specificity. In some embodiments, the BCMA CAR or its antigen-binding fragment comprises a heavy chain variable region (HCVR), a light chain variable region (LCVR), and / or a complementarity-determining region (CDR) comprising the amino acid sequence of any antibody described in U.S. Provisional Patent Application No. 62 / 700,615, filed July 19, 2018, or International Patent Application No. PCT / US2019 / 042452, which are incorporated herein by reference in their entirety. According to some exemplary embodiments, this application includes the use of an anti-BCMA CAR in a method of treating allergies, wherein the anti-BCMA CAR comprises any of the HCVR, LCVR, and / or CDR amino acid sequences disclosed in a further portion thereof.

[0098] This application also includes the use of an anti-BCMA CAR in a method of treating allergies, wherein the CAR comprises a variant of any of the disclosed HCVR, LCVR, and / or CDR amino acid sequences having one or more conserved amino acid substitutions. For example, this application includes the use of an anti-BCMA CAR having an HCVR, LCVR, and / or CDR amino acid sequence having, for example, 10 or fewer, 8 or fewer, 6 or fewer, 4 or fewer, etc., conserved amino acid substitutions relative to any of the disclosed HCVR, LCVR, and / or CDR amino acid sequences. In some embodiments, this application includes the use of an anti-BCMA CAR having an HCVR, LCVR, and / or CDR amino acid sequence having 1, 2, 3, or 4 conserved amino acid substitutions relative to any of the disclosed HCVR, LCVR, and / or CDR amino acid sequences.

[0099] Other anti-BCMA CARs that may be used in the context of the method of this application include, for example, CARs that are known and relevant to the art, such as bb2121, LCAR-B38M and 4C8A, or anti-BCMA CARs such as those in the following patent publications. Any of the following CARs: WO2015 / 052538, WO2015 / 052536, WO2016 / 094304, WO2016 / 166630, WO2016 / 151315, WO2016 / 130598, WO2017 / 183418, WO2017 / 173256, WO2017211900, WO2017 / 130223, WO2018 / 229492, WO2018 / 085690, WO2018 / 151836, WO2018 / 028647, WO2019 / 006072. Representations of publications identifying anti-BCMACAR cited herein are incorporated herein by reference.

[0100] According to certain exemplary embodiments, the BCMA-targeting agent used in the context of the disclosed methods is an anti-BCMA / anti-CD3 bispecific antibody (also referred to herein as "anti-BCMA x anti-CD3 bispecific antibody"). The anti-BCMA / anti-CD3 bispecific antibody is used for specific targeting of cells expressing BCMA and T-cell-mediated killing. The terms "antibody," "antigen-binding fragment," "human antibody," "recombinant antibody," and other related terms are as defined above. In the context of anti-BCMA / anti-CD3 antibodies and their antigen-binding fragments, this application includes the use of a bispecific antibody, wherein one arm of the immunoglobulin is specific for BCMA or a fragment thereof, while the other arm of the immunoglobulin is specific for a second therapeutic target (e.g., CD3 on T cells). Exemplary bispecific forms that may be used in the context of this application include, but are not limited to, scFv-based or biantibody-specific forms, IgG-scFv fusions, dual variable domain (DVD)-Ig, hybridoma cells, mortises, universal light chains (e.g., universal light chains with mortises), CrossMab, CrossFab, (SEED) bodies, leucine zippers, Duobody, IgG1 / IgG2, dual-action Fab (DAF)-IgG and Mab 2Bispecific forms (for a review of the above forms, see, for example, Klein et al. 2012, mAbs 4(6):653-663 and the references cited therein). Bispecific antibodies can also be constructed using peptide / nucleic acid conjugations, for example, in which non-natural amino acids with orthogonal chemical reactivity are used to generate site-specific antibody-oligonucleotide conjugates, which then self-assemble into polymerized complexes with defined composition, titer, and geometry. (See, for example, Kazane et al., J.Am.Chem.Soc., 2013, 135(1):340-46).

[0101] The term "specific binding" refers to the formation of a relatively stable complex between an antibody or its antigen-binding fragment and an antigen under physiological conditions. Methods for determining whether an antibody specifically binds to an antigen are well known in the art and include, for example, equilibrium dialysis and surface plasmon resonance assays. For example, antibodies that "specifically bind" to BCMA as used in the context of this application include, as measured in surface plasmon resonance assays, antibodies with K+ concentrations less than about 100 nM, less than about 50 nM, less than about 20 nM, less than about 10 nM, less than about 5 nM, less than about 1 nM, less than about 500 pM, less than about 200 pM, less than about 100 pM, or less than about 50 pM. D Antibodies that bind to BCMA or its fractions. However, isolated antibodies that specifically bind to human BCMA may be cross-reactive with other antigens, such as BCMA molecules from other (non-human) species.

[0102] According to certain exemplary embodiments, an anti-BCMA / anti-CD3 bispecific antibody or an antigen-binding fragment thereof comprises a heavy chain variable region (HCVR), a light chain variable region (LCVR), and / or a complementarity-determining region (CDR) containing the amino acid sequence of any antibody described in U.S. Provisional Patent Application USSN 62 / 793,645, filed January 17, 2019, or International Patent Application PCT / US2019 / 042447, which are incorporated herein by reference in their entirety. In certain exemplary embodiments, an anti-BCMA / anti-CD3 bispecific antibody or an antigen-binding fragment thereof that may be used in the context of this application comprises: (a) a first antigen-binding domain that specifically binds BCMA; and (b) a second antigen-binding domain that specifically binds CD3. In one embodiment, the first antigen-binding domain comprises: a heavy chain complementarity-determining region (HCDR) of a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO:12; and a light chain complementarity-determining region (LCDR) of a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:20. According to certain embodiments, the first antigen-binding domain comprises three HCDRs (HCDR1, HCDR2, and HCDR3) and three LCDRs (LCDR1, LCDR2, and LCDR3), wherein HCDR1 comprises the amino acid sequence of SEQ ID NO:14; HCDR2 comprises the amino acid sequence of SEQ ID NO:16; HCDR3 comprises the amino acid sequence of SEQ ID NO:18; LCDR1 comprises the amino acid sequence of SEQ ID NO:22; LCDR2 comprises the amino acid sequence of SEQ ID NO:24; and LCDR3 comprises the amino acid sequence of SEQ ID NO:26. In one embodiment, the second antigen-binding domain comprises: a heavy chain complementarity-determining region (HCDR) of a heavy chain variable region (HCVR) containing the amino acid sequence of SEQ ID NO:28 or SEQ ID NO:36; and a light chain complementarity-determining region (LCDR) of a light chain variable region (LCVR) containing the amino acid sequence of SEQ ID NO:20. In one embodiment, the second antigen-binding domain comprises three HCDRs (HCDR1, HCDR2, and HCDR3) and three LCDRs (LCDR1, LCDR2, and LCDR3), wherein HCDR1 contains the amino acid sequence of SEQ ID NO:30 or 38; HCDR2 contains the amino acid sequence of SEQ ID NO:32 or 40; HCDR3 contains the amino acid sequence of SEQ ID NO:34 or 42; LCDR1 contains the amino acid sequence of SEQ ID NO:22; LCDR2 contains the amino acid sequence of SEQ ID NO:24; and LCDR3 contains the amino acid sequence of SEQ ID NO:26.

[0103] In one embodiment, the anti-BCMA / anti-CD3 bispecific antibody or its antigen-binding fragment comprises: (a) a first antigen-binding domain comprising HCDR1, HCDR2, and HCDR3 domains, which respectively comprise the amino acid sequences of SEQ ID NO:14, 16, and 18; and LCDR1, LCDR2, and LCDR3 domains, which respectively comprise the amino acid sequences of SEQ ID NO:22, 24, and 26; and (b) a second antigen-binding domain comprising HCDR1, HCDR2, and HCDR3 domains, which respectively comprise the amino acid sequences of SEQ ID NO:30, 32, and 34; and LCDR1, LCDR2, and LCDR3 domains, which respectively comprise the amino acid sequences of SEQ ID NO:22, 24, and 26. In one embodiment, the anti-BCMA / anti-CD3 bispecific antibody or its antigen-binding fragment comprises: (a) a first antigen-binding domain comprising an HCVR containing the amino acid sequence of SEQ ID NO:12 and an LCVR containing the amino acid sequence of SEQ ID NO:20; and (b) a second antigen-binding domain comprising an HCVR containing the amino acid sequence of SEQ ID NO:28 and an LCVR containing the amino acid sequence of SEQ ID NO:20.

[0104] In one embodiment, the anti-BCMA / anti-CD3 bispecific antibody or its antigen-binding fragment comprises: (a) a first antigen-binding domain comprising HCDR1, HCDR2, and HCDR3 domains, which respectively comprise the amino acid sequences of SEQ ID NO:14, 16, and 18; and LCDR1, LCDR2, and LCDR3 domains, which respectively comprise the amino acid sequences of SEQ ID NO:22, 24, and 26; and (b) a second antigen-binding domain comprising HCDR1, HCDR2, and HCDR3 domains, which respectively comprise the amino acid sequences of SEQ ID NO:38, 40, and 42; and LCDR1, LCDR2, and LCDR3 domains, which respectively comprise the amino acid sequences of SEQ ID NO:22, 24, and 26. In one embodiment, the anti-BCMA / anti-CD3 bispecific antibody or its antigen-binding fragment comprises: (a) a first antigen-binding domain comprising an HCVR containing the amino acid sequence of SEQ ID NO:12 and an LCVR containing the amino acid sequence of SEQ ID NO:20; and (b) a second antigen-binding domain comprising an HCVR containing the amino acid sequence of SEQ ID NO:36 and an LCVR containing the amino acid sequence of SEQ ID NO:20.

[0105] Exemplary anti-BCMA / anti-CD3 bispecific antibodies include complete human bispecific antibodies referred to as REGN5458 and REGN5459. According to certain exemplary embodiments, the methods of this application include the use of REGN5458 or REGN5459 or their bioequivalents. As used herein, the term "bioequivalent" in relation to anti-BCMA / anti-CD3 antibodies refers to an antibody or its BCMA / CD3-binding protein or fragment that is a pharmaceutical equivalent or substitute that, when administered at the same molar dose in single or multiple doses under similar experimental conditions, shows no significant difference in absorption rate and / or extent compared to a reference antibody (e.g., REGN5458 or REGN5459); the term "bioequivalent" also includes an antigen-binding protein that binds to BCMA / CD3 and is not clinically significantly different from a reference antibody (e.g., REGN5458 or REGN5459) in terms of safety, purity, and / or efficacy.

[0106] In some embodiments, the anti-BCMA / anti-CD3 bispecific antibody or its antigen-binding fragment comprises: (a) a first antigen-binding domain comprising an HCVR having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:12 and an LCVR having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:20; and (b) a second antigen-binding domain comprising an HCVR having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:20 and an LCVR having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:20; and (c) a second antigen-binding domain comprising an HCVR having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:28 and an LCVR having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:20; and (d) a second antigen-binding domain comprising an HCVR having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the amino acid The amino acid sequence of NO:20 has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the LCVR. In some embodiments, the anti-BCMA / anti-CD3 bispecific antibody or its antigen-binding fragment comprises: (a) a first antigen-binding domain comprising three HCDRs (HCDR1, HCDR2, and HCDR3) comprising the amino acid sequences of SEQ ID NO:14, 16, and 18, respectively; and an HCVR having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:12; and three LCDRs (LCDR1, LCDR2, and LCDR3) comprising the amino acid sequences of SEQ ID NO:22, 24, and 26, respectively; and an LCVR having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:12 ... The amino acid sequence of NO:20 has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the LCVR; and (b) a second antigen-binding domain comprising three HCDRs (HCDR1, HCDR2, and HCDR3), which respectively comprise the amino acid sequences of SEQ ID NO:30, 32, and 34, and an HCVR having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:28; and comprising three LCDRs (LCDR1, LCDR2, and LCDR3), which respectively comprise the amino acid sequences of SEQ ID NO:22, 24, and 26, and an LCVR having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:28; and comprising three LCDRs (LCDR1, LCDR2, and LCDR3), which respectively comprise the amino acid sequences of SEQ ID NO:22, 24, and 26, and an LCVR having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:28. LCVRs with at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity of the amino acid sequence of NO:20.

[0107] In some embodiments, the anti-BCMA / anti-CD3 bispecific antibody or its antigen-binding fragment comprises: (a) a first antigen-binding domain comprising an HCVR having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:12 and an LCVR having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:20; and (b) a second antigen-binding domain comprising an HCVR having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:20 and an LCVR having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:20; and (b) a second antigen-binding domain comprising an HCVR having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:36 and an LCVR having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:20; and (c) an LCVR having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:2 The amino acid sequence of NO:20 has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the LCVR. In some embodiments, the anti-BCMA / anti-CD3 bispecific antibody or its antigen-binding fragment comprises: (a) a first antigen-binding domain comprising three HCDRs (HCDR1, HCDR2, and HCDR3) comprising the amino acid sequences of SEQ ID NO:14, 16, and 18, respectively; and an HCVR having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:12; and three LCDRs (LCDR1, LCDR2, and LCDR3) comprising the amino acid sequences of SEQ ID NO:22, 24, and 26, respectively; and an LCVR having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:12 ... The amino acid sequence of NO:20 has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the LCVR; and (b) a second antigen-binding domain comprising three HCDRs (HCDR1, HCDR2, and HCDR3), each comprising the amino acid sequences of SEQ ID NO:38, 40, and 42, respectively, and an HCVR having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:36; and comprising three LCDRs (LCDR1, LCDR2, and LCDR3), each comprising the amino acid sequences of SEQ ID NO:22, 24, and 26, respectively, and an LC ... an LCVR having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:36; and an LCVR having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97 LCVRs with at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity of the amino acid sequence of NO:20.

[0108] This application also includes the use of anti-BCMA / anti-CD3 antibodies in methods of treating allergies, wherein said antibody comprises a variant of any of the HCVR, LCVR, and / or CDR amino acid sequences disclosed herein having one or more conserved amino acid substitutions. For example, this application includes the use of anti-BCMA / anti-CD3 antibodies having HCVR, LCVR, and / or CDR amino acid sequences having conserved amino acid substitutions relative to any of the HCVR, LCVR, and / or CDR amino acid sequences disclosed herein, for example, 10 or fewer, 8 or fewer, 6 or fewer, 4 or fewer, etc. In some embodiments, this application includes the use of anti-BCMA / anti-CD3 antibodies having HCVR, LCVR, and / or CDR amino acid sequences having 1, 2, 3, or 4 conserved amino acid substitutions relative to any of the HCVR, LCVR, and / or CDR amino acid sequences disclosed herein.

[0109] Other anti-BCMA / anti-CD3 antibodies that may be used in the context of the methods described in this application include, for example, antibodies known and understood in the art, such as AMG420, AMG701, CC-93269, EM801, JNJ-64007957, and TNB384B, or, for example, anti-BCMA / anti-CD3 antibodies described in the following patent publications: WO2013 / 072415, WO2014 / 140248, WO2014 / 122144, WO2016 / 166629, WO2016 / 079177, WO2016 / 020332, WO2017031104, WO2017 / 223111, WO2017 / 134134, WO2018 / 083204, and WO2018 / 201051. The portions of publications cited in this article that identify anti-BCMA / anti-CD3 antibodies are incorporated herein by reference.

[0110] Combination of IgE depletion and allergen immunotherapy

[0111] This application also provides methods for enhancing the efficacy and / or tolerability of immunotherapy regimens (e.g., allergen-specific immunotherapy regimens) in individuals with allergies. In some embodiments, the method includes administering an IL-4 / IL-13 pathway inhibitor (e.g., anti-IL-4R antibody) and a plasma cell ablation agent (e.g., anti-BCMA / anti-CD3 antibody) to the individual with allergies before or simultaneously with the immunotherapy regimen.

[0112] In some embodiments, the treated individual suffers from a food allergy. For example, in some embodiments, the individual suffers from an allergy to milk, dairy products, eggs, celery, sesame, wheat, meat, fruit, soy, fish, shellfish, sugar, peanuts, beans, tree nuts, or combinations thereof; in some embodiments, the individual suffers from a peanut allergy. In some embodiments, the treated individual suffers from a non-food allergy (e.g., an allergy to environmental allergens). For example, in some embodiments, the treated individual suffers from an allergy to a non-food allergen selected from dust, dust mites, pollen, insect venom, mold, animal dander, animal dander, wool, latex, metals, household cleaners, detergents, pharmaceuticals, cosmetics, fragrances, pharmaceuticals such as penicillin, sulfonamides or salicylates, therapeutic monoclonal antibodies (e.g., cetuximab), ragweed, grass, and birch. In some embodiments, the treated individual suffers from a severe allergy (e.g., a severe food allergy or a severe non-food allergy).

[0113] As used herein, "allergen-specific immunotherapy" refers to the repeated administration of an allergen (such as the allergens disclosed herein) to an individual over time as a means of treating or preventing allergies and allergic reactions, or reducing or eliminating allergic reactions. In some embodiments, an allergen-specific immunotherapy regimen includes oral immunotherapy. In some embodiments, an allergen-specific immunotherapy regimen includes subcutaneous immunotherapy. In some embodiments, an allergen-specific immunotherapy regimen includes sublingual immunotherapy. Typically, immunotherapy regimens can be "conventional" immunotherapy regimens and "accelerated" immunotherapy regimens. Typically, in a conventional immunotherapy regimen, an increasing dose of allergen (also referred to as "escalation") is administered to the patient every week for several weeks to months (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 months or longer) under close medical supervision, followed by a maintenance dose, which typically consists of one or more doses of allergen administered at the highest dose administered during the escalation regimen. In accelerated immunotherapy regimens, the escalation process is accelerated compared to conventional immunotherapy. Examples of accelerated immunotherapy include "rush" immunotherapy and "cluster" immunotherapy. In rush immunotherapy, an escalating dose of the allergen is typically administered daily over several consecutive days (e.g., for 2, 3, 4, 5, 6 days, or 1 week) until the maximum tolerated dose is reached. In cluster immunotherapy, several escalating doses of the allergen are typically administered daily over discontinuous days until the maximum tolerated dose is reached, usually within 4–8 weeks.

[0114] In some embodiments, an IL-4 / IL-13 pathway inhibitor and a plasma cell ablation agent are administered before or concurrently with an allergen-specific immunotherapy regimen as disclosed herein (e.g., oral, sublingual, or subcutaneous immunotherapy, which may be conventional or accelerated immunotherapy). In some embodiments, a plasma cell ablation agent (e.g., 1, 2, 3, 4, 5, or more doses) is administered before initiating the immunotherapy regimen. In some embodiments, an IL-4 / IL-13 pathway inhibitor (e.g., 1, 2, 3, 4, 5, or more doses) is administered before initiating the immunotherapy regimen. In some embodiments, at least one dose of both a plasma cell ablation agent and an IL-4 / IL-13 pathway inhibitor is administered before initiating the immunotherapy regimen. In some embodiments, an IL-4 / IL-13 pathway inhibitor is administered concurrently with the immunotherapy regimen. In some embodiments, a plasma cell ablation agent is administered concurrently with the immunotherapy regimen.

[0115] Pharmaceutical composition and administration

[0116] The disclosed method involves administering a combination of an IL-4 / IL-13 pathway inhibitor and a plasma cell ablation agent to an individual in need, wherein the inhibitor is contained in a separate pharmaceutical composition or a combined (single) pharmaceutical composition. The pharmaceutical compositions of this application can be formulated with pharmaceutically acceptable carriers, excipients, and other agents that provide suitable transfer, delivery, tolerability, etc. A variety of suitable formulations can be found in pharmacist-known formulation sets: Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, Pa. These formulations include, for example, powders, pastes, ointments, gels, waxes, oils, lipids, vesicles containing lipids (cationic or anionic) (e.g., LIPOFECTINTM), DNA conjugates, anhydrous absorbent pastes, oil-in-water and water-in-oil emulsions, polyethylene glycol emulsions (polyethylene glycol of varying molecular weights), semi-solid gels containing carbon waxes, and semi-solid mixtures. See also Powell et al., 1998, J Pharm Sci Technol, 52:238-311.

[0117] In some embodiments, the pharmaceutical composition of this application comprises a therapeutically effective amount of an IL-4 / IL-13 pathway inhibitor (e.g., anti-IL-4R antibody) and / or a therapeutically effective amount of a plasma cell ablation agent (e.g., anti-BCMA / anti-CD3 antibody) and a pharmaceutically acceptable carrier. In some embodiments, the disclosed pharmaceutical composition is formulated for administration by injection, e.g., intravenous injection.

[0118] Various delivery systems are known and can be used to administer the pharmaceutical compositions of this application, such as encapsulation in liposomes, microparticles, microcapsules, recombinant cells capable of expressing mutant viruses, and receptor-mediated endocytosis (see, for example, Wu et al., 1987, J. Biol. Chem. 262:4429-4432). Administration methods include, but are not limited to, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, and oral routes. The compositions can be administered via any convenient route, such as by infusion or rapid bolus injection, absorption through the epithelial or mucosal lining (e.g., oral mucosa, rectal and intestinal mucosa), and can be administered together with other bioactive agents. In some embodiments, IL-4 / IL-13 pathway inhibitors and / or plasma cell ablation agents are administered intravenously. In some embodiments, IL-4 / IL-13 pathway inhibitors and / or plasma cell ablation agents are administered subcutaneously.

[0119] In some embodiments, the pharmaceutical composition of this application is contained within a container. Therefore, in another aspect, containers containing pharmaceutical compositions as disclosed herein are provided. For example, in some embodiments, the pharmaceutical composition is contained within a container selected from glass vials, syringes, pen-type delivery devices, and autoinjectors.

[0120] In some embodiments, the pharmaceutical composition of this application is delivered subcutaneously or intravenously using a standard needle and syringe. In some embodiments, the syringe is a pre-filled syringe. Furthermore, for subcutaneous delivery, pen-type delivery devices or auto-injectors are readily applicable in delivering the pharmaceutical composition of this application. Pen-type delivery devices can be reusable or disposable. Typically, reusable pen-type delivery devices utilize a replaceable cartridge containing the pharmaceutical composition. Once the entire pharmaceutical composition in the cartridge has been administered and the cartridge is empty, the empty cartridge is discarded and replaced with a new cartridge containing the pharmaceutical composition. The pen-type delivery device can then be reused. In disposable pen-type delivery devices, there is no replaceable cartridge. Instead, the disposable pen-type delivery device is pre-filled with the pharmaceutical composition held in a reservoir within the device. Once the reservoir is emptied of the pharmaceutical composition, the entire device is discarded.

[0121] Examples of suitable pen types and autoinjector delivery devices include, but are not limited to, AUTOPENTM (Owen Mumford, Inc., WOodstock, UK), DISETRONICTM pen (Disetronic Medical Systems, Bergdorf, Switzerland), HUMALOG MIX 75 / 25TM pen, HUMALOGTM pen, HUMALIN 70 / 30TM pen (Eli Lillyand Co., Indianapolis, IN), NOVOPENTM, II and III (Novo Nordisk, Copenhagen, Denmark), NOVOPEN JUNIORTM (Novo Nordisk, Copenhagen, Denmark), BDTM pen (Becton Dickinson, Franklin Lakes, NJ), OPTIPENTM, OPTIPEN PROTM, OPTIPEN STARLETTM and OPTICLIKTM (sanofi-aventis, Frankfurt, Germany). Examples of disposable pen-type delivery devices used in the subcutaneous delivery of the pharmaceutical compositions of this application include, but are not limited to, SOLOSTARTM pen (sanofi-aventis), FLEXPENTM (NovoNordisk), KWIKPENTM (Eli Lilly), and SURECLICK. TM Automatic injectors (Amgen, Thousand Oaks, CA), PENLET TM (Haselmeier, Stuttgart, Germany), EPIPEN (Dey, LP) and HUMIRA TM Pen (Abbott Labs, Abbott Park IL).

[0122] In some cases, one or two drug compositions can be delivered using a controlled-release system. In one embodiment, a pump can be used. In another embodiment, a polymeric material can be used. See Medical Applications of Controlled Release, Langer and Wise (eds.), 1974, CRC Pres., Boca Raton, Fla. In another embodiment, the controlled-release system can be placed near the target of the composition, thereby requiring only a portion of the systemic dose (see, for example, Goodson, 1984, M). EDICAL A PPLICATIONS OF C ONTROLLED R ELEASE(Volume 2, pp. 115-138). Other controlled release systems are discussed in Langer, 1990, Science 249: 1527-1533.

[0123] Suitable injectable formulations may include dosage forms for intravenous, subcutaneous, intradermal, and intramuscular injection, infusion, etc. These injectable formulations can be prepared by known methods. For example, injectable formulations can be prepared by dissolving, suspending, or emulsifying the aforementioned antibodies or their salts in a sterile aqueous or oily medium commonly used for injections. Aqueous media for injection include, for example, physiological saline, isotonic solutions containing glucose and other adjuvants, which can be used in combination with suitable solubilizers, such as alcohols (e.g., ethanol), polyols (e.g., propylene glycol, polyethylene glycol), nonionic surfactants [e.g., polysorbate 80, HCO-50 (a polyoxyethylene (50 mol) adduct of hydrogenated castor oil)], etc. Oily media can be used, for example, sesame oil, soybean oil, etc., which can be used in combination with solubilizers, such as benzyl benzoate, benzyl alcohol, etc. It is preferable to fill the injectable formulation into a suitable ampoule.

[0124] In some embodiments, the pharmaceutical composition intended for oral or parenteral use is formulated into a unit dose dosage form suitable for assembling a dose of the active ingredient. Such unit dose dosage forms include, for example, tablets, pills, capsules, injections (ampoules), cartridges, suppositories, etc.

[0125] Injectable formulations of pharmaceutical compositions can be prepared by known methods. For example, injections can be prepared, for instance, by dissolving, suspending, or emulsifying an inhibitor (e.g., an anti-IL-4R antibody) or a salt thereof in a sterile aqueous or oily medium commonly used for injections. Aqueous media for injection include, for example, physiological saline, isotonic solutions containing glucose and other adjuvants, which can be used in combination with suitable solubilizers, such as alcohols (e.g., ethanol), polyols (e.g., propylene glycol, polyethylene glycol), nonionic surfactants [e.g., polysorbate 80, HCO-50 (a polyoxyethylene (50 mol) adduct of hydrogenated castor oil)], etc. Oily media can be used, for example, sesame oil, soybean oil, etc., which can be used in combination with solubilizers, such as benzyl benzoate, benzyl alcohol, etc. Preferably, the injection thus prepared is filled into a suitable injection ampoule. In some embodiments, the injection may include a concentration of an inhibitor (e.g., an anti-IL-4R antibody) and one or more pharmaceutically acceptable solvents (e.g., distilled water, saline, etc.).

[0126] Exemplary pharmaceutical compositions comprising anti-IL-4R antibodies that may be used in the context of this application are disclosed, for example, in U.S. Patent No. 8,945,559, wherein a portion identifying a pharmaceutical composition comprising anti-IL-4R antibodies is incorporated herein by reference.

[0127] medicine box

[0128] In some embodiments, this application provides drug combinations and kits comprising an IL-4 / IL-13 pathway inhibitor as disclosed herein and a plasma cell ablation agent as disclosed herein. In some embodiments, the drug combination and kit comprise an anti-IL-4R antibody as disclosed herein (e.g., an anti-IL-4R antibody or its antigen-binding fragment comprising HCDR1, HCDR2, and HCDR3 containing amino acid sequences of SEQ ID NO: 3, 4, and 5, respectively, and LCDR1, LCDR2, and LCDR3 containing amino acid sequences of SEQ ID NO: 6, 7, and 8, respectively) and a plasma cell ablation agent as disclosed herein (e.g., an anti-BCMA / anti-CD3 bispecific antibody comprising a first antigen-binding domain specifically binding to BCMA; and a second antigen-binding domain specifically binding to CD3, wherein the first antigen-binding domain comprises HCDR1, HCDR2, and HCDR3 containing amino acid sequences of SEQ ID NO: 14, 16, and 18, respectively, and LCDR1, LCDR2, and LCDR3 containing amino acid sequences of SEQ ID NO: 22, 24, and 26, respectively; and wherein the second antigen-binding domain comprises HCDR1 containing amino acid sequences of SEQ ID NO: 30 or 38, and LCDR1 containing amino acid sequences of SEQ ID NO: 30 or 38, respectively). HCDR2 containing the amino acid sequence NO:32 or 40, HCDR3 containing the amino acid sequence SEQ ID NO:34 or 42, and LCDR1, LCDR2, and LCDR3 containing the amino acid sequences SEQ ID NO:22, 24, and 26, respectively.

[0129] In some embodiments, a combination or kit comprising an IL-4 / IL-13 pathway inhibitor and a plasma cell ablation agent is used in the methods disclosed herein. In some embodiments, the combination or kit is used to treat allergies or allergic disorders or to reduce or eliminate allergen-specific serum IgE in an individual. In some embodiments, the combination or kit further comprises one or more other therapeutic agents as disclosed herein.

[0130] In some embodiments, a combination or kit comprising an IL-4 / IL-13 pathway inhibitor and a plasma cell ablation agent is used to increase the efficacy and / or tolerability of immunotherapy regimens in individuals with allergies. Therefore, in some embodiments, the combination or kit further comprises one or more agents for use in immunotherapy regimens.

[0131] In some embodiments, the kit as disclosed herein further includes instructions for use. In some embodiments, the kit for use as disclosed herein includes one or more containers containing an IL-4 / IL-13 pathway inhibitor and a plasma cell ablation agent. In some embodiments, the kit includes a first container containing an IL-4 / IL-13 pathway inhibitor and a second container containing a plasma cell ablation agent.

[0132] Application plan

[0133] In some embodiments, the disclosed method includes sequentially administering a therapeutically effective amount of an IL-4 / IL-13 pathway inhibitor and a therapeutically effective amount of a plasma cell ablation agent to an individual in need, wherein each therapeutic agent is administered to the individual in one or more doses, for example as part of a specific treatment dosing regimen. In some embodiments, the method of this application includes administering an inhibitor with additive or synergistic activity to treat an allergic disease or condition, mast cell activation disorder, or mast cell enlargement.

[0134] As used herein, "sequential administration" means administering each dose of the inhibitor to an individual at different time points, such as on different days separated by predetermined intervals (e.g., hours, days, weeks, or months). In some embodiments, the disclosed method includes sequentially administering an initial single dose of an IL-4 / IL-13 pathway inhibitor to an individual, followed by one or more subsequent doses of the IL-4 / IL-13 pathway inhibitor. In some embodiments, the method also includes sequentially administering an initial single dose of a plasma cell ablation agent to an individual, followed by one or more subsequent doses of the plasma cell ablation agent.

[0135] In some embodiments, the treatment dosing regimen comprises administering one or more doses of an IL-4 / IL-13 pathway inhibitor in combination with one or more doses of a plasma cell ablation agent. In some embodiments, one or more doses of the IL-4 / IL-13 pathway inhibitor and / or one or more doses of the plasma cell ablation agent are administered to the individual at the following frequencies: approximately once daily, once every two days, once every three days, once every four days, once every five days, once every six days; once weekly, once every two weeks, once every three weeks, once every four weeks, once monthly, once every two months, once every three months, once every four months, or at lower frequencies.

[0136] As used herein, the phrase "in combination with" means the administration of an IL-4 / IL-13 pathway inhibitor before, after, or concurrently with a plasma cell ablation agent. The term "in combination with" also includes the sequential or concurrent administration of an IL-4 / IL-13 pathway inhibitor and a plasma cell ablation agent.

[0137] For example, when an IL-4 / IL-13 pathway inhibitor is administered "before" the plasma cell ablation agent, it can be administered more than 150 hours, approximately 150 hours, approximately 100 hours, approximately 72 hours, approximately 60 hours, approximately 48 hours, approximately 36 hours, approximately 24 hours, approximately 12 hours, approximately 10 hours, approximately 8 hours, approximately 6 hours, approximately 4 hours, approximately 2 hours, approximately 1 hour, approximately 30 minutes, or approximately 15 minutes before the plasma cell ablation agent is administered. When an IL-4 / IL-13 pathway inhibitor is administered "after" the plasma cell ablation agent, it can be administered approximately 15 minutes, approximately 30 minutes, approximately 1 hour, approximately 2 hours, approximately 4 hours, approximately 6 hours, approximately 8 hours, approximately 10 hours, approximately 12 hours, approximately 24 hours, approximately 36 hours, approximately 48 hours, approximately 60 hours, approximately 72 hours, or more than 72 hours after the plasma cell ablation agent is administered. "Simultaneous" administration of an IL-4 / IL-13 pathway inhibitor with a plasma cell ablation agent means administering the IL-4 / IL-13 pathway inhibitor in a separate formulation to an individual within 10 minutes of the plasma cell ablation agent administration (before, after, or simultaneously), or administering a single combination formulation containing both an IL-4 / IL-13 pathway inhibitor and a plasma cell ablation agent to an individual.

[0138] As used herein, "initial dose" refers to the dose administered at the start of a treatment regimen (also known as the "baseline dose"). One or more subsequent doses administered after the initial dose may all contain the same amount of an IL-4 / IL-13 pathway inhibitor or plasma cell ablation agent. However, in some embodiments, the amounts contained in the initial and subsequent doses differ from each other during treatment (e.g., adjusted up or down appropriately). In some embodiments, one or more (e.g., 1, 2, 3, 4, or 5) doses are administered at the start of a treatment regimen as a "loading dose," followed by subsequent doses administered at a lower frequency (e.g., "maintenance dose"). For example, an IL-4 / IL-13 pathway inhibitor or plasma cell ablation agent may be administered to a patient with an allergic disease with a loading dose of approximately 1 mg / kg to approximately 20 mg / kg, followed by one or more maintenance doses of approximately 0.1 mg / kg to approximately 10 mg / kg of the patient's body weight.

[0139] In one exemplary embodiment of this application, each subsequent dose is administered for 1 / 2 to 14 weeks or more (e.g., 1 / 2, 1 / 2, 2 / 2, 3, 3 / 2, 4, 5, 5 / 2, 6, 6 / 2, 7, 7 / 2, 8, 8 / 2, 9, 9 / 2, 10, 10 / 2 or more weeks) following an immediate preceding dose. As used herein, the term “previous dose” refers to the dose of each inhibitor administered to an individual in a sequence of multiple administrations, without any interval between doses prior to the next administration.

[0140] dose

[0141] In some embodiments, at least one dose of the IL-4 / IL-13 pathway inhibitor comprises about 0.1-50 mg / kg, for example, about 0.1-10 mg / kg of individual body weight. For example, at least one dose may comprise about 0.1, 1, 0.3, 3, 4, 5, 6, 7, 8, 9, or 10 mg / kg of individual body weight. In some embodiments, at least one dose of the IL-4 / IL-13 pathway inhibitor comprises about 0.05-600 mg of the IL-4 / IL-13 pathway inhibitor, for example, about 5-600 mg, about 10-300 mg, about 50-600 mg, or about 50-300 mg, for example, about 5, 10, 15, 20, 25, 40, 45, 50, 60, 70, 80, 90, 100, 200, 300, 400 mg, 500 mg, 600 mg, or more of the IL-4 / IL-13 pathway inhibitor. In one implementation, the IL-4 / IL-13 pathway inhibitor is REGN668 (dupilumab).

[0142] In some embodiments, at least one dose of the plasma cell ablation agent comprises about 0.1-20 mg / kg body weight, such as about 0.1, 1, 0.3, 3, 4, 5, 6, 7, 8, 9, or 10 mg / kg body weight. In some embodiments, at least one dose of the plasma cell ablation agent comprises about 0.05-500 mg of plasma cell ablation agent, such as about 5, 10, 15, 20, 25, 40, 45, 50, 60, 70, 80, 90, 100 mg or more of plasma cell ablation agent. In one embodiment, the plasma cell ablation agent is an anti-BCMA / anti-CD3 bispecific antibody (e.g., REGN5459). In one embodiment, the plasma cell ablation agent is a proteasome inhibitor, such as bortezomib.

[0143] The amount of IL-4 / IL-13 pathway inhibitor and plasma cell ablation agent administered to an individual according to the methods disclosed herein is a therapeutically effective amount. As used herein, the term "therapeuticly effective amount" refers to the amount of each therapeutic agent that results in one or more of the following outcomes: (a) anaphylaxis, such as a reduction in the severity or duration of symptoms or indications of an anaphylactic reaction; (b) a decrease in serum allergen-specific IgE levels; (c) elimination of serum IgE in the individual; (d) reduced allergen sensitization; (e) reduced susceptibility to anaphylactic reactions; and / or (f) a reduction in the use or need for conventional anti-allergy therapy (e.g., reduced or eliminated use of corticosteroids) compared to untreated individuals or individuals treated with one of the therapeutic agents as a monotherapy.

[0144] In the case of IL-4 / IL-13 pathway inhibitors (e.g., anti-IL-4R antibodies), the therapeutically effective dose can be approximately 0.05 mg to approximately 600 mg, for example, approximately 0.05 mg, approximately 0.1 mg, approximately 1.0 mg, approximately 1.5 mg, approximately 2.0 mg, approximately 10 mg, approximately 20 mg, approximately 30 mg, approximately 40 mg, approximately 50 mg, approximately 60 mg, approximately 70 mg, approximately 80 mg, approximately 90 mg, approximately 100 mg, approximately 110 mg, approximately 120 mg, approximately 130 mg, approximately 140 mg, approximately 150 mg, approximately 160 mg, approximately 170 mg, approximately 180 mg, approximately 190 mg, approximately 200 mg, approximately 210 mg, approximately 220 mg, approximately 230 mg, approximately 240 mg, approximately 250 mg, etc. IL-4 / IL-13 pathway inhibitors in doses of approximately 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, 350 mg, 360 mg, 370 mg, 380 mg, 390 mg, 400 mg, 410 mg, 420 mg, 430 mg, 440 mg, 450 mg, 460 mg, 470 mg, 480 mg, 490 mg, 500 mg, 510 mg, 520 mg, 530 mg, 540 mg, 550 mg, 560 mg, 570 mg, 580 mg, 590 mg, or 600 mg. In some implementations, an IL-4 / IL-13 pathway inhibitor is administered to an individual at doses of 10 mg, 25 mg, 50 mg, 75 mg, 150 mg, or 300 mg.

[0145] In the case of plasma cell ablation agents (such as anti-BCMA / anti-CD3 bispecific antibodies), the therapeutically effective dose can be about 0.05 mg to about 500 mg, about 1 mg to about 500 mg, about 10 mg to about 450 mg, about 50 mg to about 400 mg, about 75 mg to about 350 mg, or about 100 mg to about 300 mg of antibody. For example, in different embodiments, the amount of plasma cell ablation agent is about 0.05 mg, about 0.1 mg, about 1.0 mg, about 1.5 mg, about 2.0 mg, about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 30 mg, about 40 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 110 mg, about 120 mg, about 130 mg, about 140 mg, about 150 mg, about 160 mg, about 170 mg, about 180 mg, about 190 mg, about 200 mg, about 210 mg, and so on. Plasma cell ablation agents in doses of 220 mg, approximately 230 mg, approximately 240 mg, approximately 250 mg, approximately 260 mg, approximately 270 mg, approximately 280 mg, approximately 290 mg, approximately 300 mg, approximately 310 mg, approximately 320 mg, approximately 330 mg, approximately 340 mg, approximately 350 mg, approximately 360 mg, approximately 370 mg, approximately 380 mg, approximately 390 mg, approximately 400 mg, approximately 410 mg, approximately 420 mg, approximately 430 mg, approximately 440 mg, approximately 450 mg, approximately 460 mg, approximately 470 mg, approximately 480 mg, approximately 490 mg, or approximately 500 mg.

[0146] In some embodiments, the single dose of an IL-4 / IL-13 pathway inhibitor (e.g., anti-IL-4R antibody) and / or plasma cell ablation agent (e.g., anti-BCMA / anti-CD3 antibody) administered to an individual may be below the therapeutically effective amount, i.e., a subtherapeutic dose. For example, if the therapeutically effective amount of the inhibitor is 3 mg / kg, the subtherapeutic dose includes amounts below 3 mg / kg, such as 2 mg / kg, 1.5 mg / kg, 1 mg / kg, 0.5 mg / kg, or 0.3 mg / kg. As defined herein, a "subtherapeutic dose" refers to an amount of inhibitor that does not produce a therapeutic effect on its own. However, in some embodiments, multiple subtherapeutic doses of the inhibitor may be administered to achieve a therapeutic effect collectively in an individual. Example

[0147] The disclosed technology is described below through the following embodiments. The use of these and other embodiments anywhere in this specification is exemplary only and does not limit the scope and meaning of this application or any example form in any way. Similarly, this application is not limited to any particular preferred embodiment described herein. In fact, variations and modifications of this application will be apparent to those skilled in the art upon reading this specification, and such variations and modifications can be made without departing from its spirit and scope. Therefore, this application is limited only by the terminology of the claims and the full scope of the equivalents conferred by the claims. Furthermore, although attempts have been made to ensure the accuracy of the numerical values ​​used (e.g., quantities, temperatures, etc.), some experimental errors and deviations should be taken into account. Unless otherwise stated, parts are parts by weight, molecular weights are average molecular weights, temperatures are degrees Celsius, and pressures are atmospheric pressure or close to atmospheric pressure.

[0148] Example 1: The effect of combining anti-IL-4R antibody with anti-BCMA / anti-CD3 bispecific antibody

[0149] This embodiment relates to a study that demonstrated the enhanced efficacy of a combination of an IL-4 / IL-13 pathway inhibitor and a plasma cell ablation agent in blocking IgE production in a mouse model of chronic allergen-driven lung inflammation.

[0150] The IL-4 / IL-13 pathway inhibitor used in this embodiment is a mouse anti-IL-4R antibody identified as REGN1103, which is a mouse alternative antibody to the human monoclonal antibody against human IL-4R identified as REGN668 (also known as dupilumab). REGN1103 contains the HCVR / LCVR amino acid sequence pair of SEQ ID NO:43 / 44 and has an affinity for mouse IL-4R similar to that of dupilumab for human IL-4R. Furthermore, REGN1103 inhibits IL-4 and IL-13-dependent proliferation of cell lines, with IC50 values ​​of 1.9 nM and 11 pM, respectively.

[0151] The plasma cell ablation agent used in this embodiment is the anti-BCMA / anti-CD3 bispecific antibody REGN5459, which contains an anti-BCMA binding domain of HCVR (SEQ ID NO:12) and LCVR (SEQ ID NO:20); and an anti-CD3 binding domain of HCVR (SEQ ID NO:36) and LCVR (SEQ ID NO:20).

[0152] Materials and methods

[0153] To determine the effects of the anti-IL-4Rα and anti-BCMA x anti-CD3 combination on IgE production in a relevant in vivo model, a study of chronic house dust mite (HDM)-driven lung inflammation was conducted in homozygous mice where human BCMA and human CD3 were replaced with human CD3. Chronic lung inflammation and persistent IgE production were induced by intranasal exposure of mice to 25 μg HDM (Greer, catalog number XPB70D3A25) diluted in 20 μL saline (Sigma, catalog number S8776) or 20 μL saline (control group), three times a week for 11 weeks. This model induced a class shift of B cells to IgE-producing plasma cells in secondary lymphoid organs and drove the accumulation of IgE-producing plasma cells in the bone marrow. At week 8 following the first administration of HDM, a subgroup of mice began receiving subcutaneous injections of 25 mg / kg REGN1103 (anti-IL-4Rα) or 25 mg / kg isotype control until the end of the experiment. At week 11, mice received two subcutaneous injections of REGN5459 (anti-BCMA x anti-CD3) or two doses of isotype control antibody, and were allowed to rest for 9 weeks without intranasal HDM administration. Table 1 and Figure 1 The document provides a detailed overview of HDM exposure and antibody treatment regimens.

[0154] Table 1: HDM exposure and antibody treatment regimens in mice

[0155]

[0156]

[0157] One and six weeks after the last dose of HDM, ~100 μL of blood was collected from all groups of mice via retro-orbital pleophoresis and transferred to micro-collection tubes (BD, catalog #365967) for serum separation. Total IgE concentrations in serum were determined using the OptEIA™ ELISA kit (BD Biosciences, #555248) according to the manufacturer's instructions. Briefly, ELISA plates were coated with IgE capture antibodies diluted in coating buffer (carbonate-bicarbonate buffer, Sigma; catalog #C3041, diluted in 100 mL distilled water, Gibco; catalog #15230-270) and incubated overnight at 4°C. The coated plates were washed four times in washing buffer (0.05% Tween 20, Sigma; catalog #P1379, diluted in DPBS, GE; catalog #SH3001304). Block the plate for 1 hour at room temperature (RT) with assay diluent (BD; catalog #555213). Dilute 100 μL of serum sample at an initial concentration of 1:50 or 1:100 and further serially dilute 3-fold. Dilute IgE standard at an initial concentration of 100 ng / mL and further serially dilute 2-fold. Add the solutions to the plates and incubate at room temperature for 2 hours. Wash the plates four times with wash buffer and incubate at room temperature for 1 hour with 100 μL of working detector (detection antibody with sAv-HRP). Wash the plates seven times, soak in wash buffer for 5 minutes, and then wash four more times to remove unbound detection antibody. Add 100 μL of TMB substrate solution (BD; catalog #555214) to each sample, incubate the plates in the dark for 30 minutes, and then add 50 μL of stop solution (2N sulfuric acid, BDH VWR analytical type; catalog #BDH7500). Absorbance was measured at 450 nm, and IgE concentration was calculated based on a standard curve. Serum IgE values ​​are displayed as ng / mL. Statistical significance was determined using the Kronoswiss test and Dunn's post-hoc multiple comparison test in GraphPad Prism.

[0158] result

[0159] In a chronic HDM model, anti-IL4RA therapy can reduce, but does not eliminate, circulating IgE. Figure 2A and 2B (Table 2).

[0160] Table 2: Effects of anti-IL-4Rα and anti-BCMA x anti-CD3 antibody therapy on serum IgE in a chronic HDM model

[0161]

[0162] Anti-BCMA x anti-CD3 therapy alone temporarily reduces IgE production, but serum IgE levels recover after 6 weeks of rest. Figure 2A and 2B Table 2). Combination therapy with anti-IL-4Rα and anti-BCMA x anti-CD3 eliminated serum IgE (which was undetectable by IgE ELISA). Figure 2A and 2B (Table 2) This demonstrates the effectiveness of blocking IL-4RA and using anti-BCMA x anti-CD3 to deplete plasma cells as a successful strategy to block IgE production.

[0163] Example 2: The treatment combination of BCMAxCD3 bispecific antibody and anti-IL-4Rα antibody completely blocked HDM-specific IgE production during continuous HDM exposure.

[0164] This example demonstrates the efficacy of blocking IL-4R and depleting plasma cells in blocking allergen-specific IgE production, even in the presence of continuous allergen exposure.

[0165] In this embodiment, the IL-4 / IL-13 pathway inhibitor used is the mouse anti-IL-4R antibody REGN1103, which is a mouse alternative antibody to a human monoclonal antibody, REGN668 (also known as dupilumab), identified as targeting human IL-4R. REGN1103 is described in Example 1 above. The plasma cell ablation agent used is the anti-BCMA x anti-CD3 bispecific antibody REGN5459 described in Example 1 above. Mouse IgG1 antibody (REGN1094) and human IgG4 x anti-CD3 antibody (REGN4460) were used as isotype controls.

[0166] Materials and methods

[0167] To determine the effects of the anti-IL-4Rα and anti-BCMA x anti-CD3 combination on IgE production in a relevant in vivo model, a chronic house dust mite (HDM)-driven lung inflammation study was conducted in mice homozygous for human BCMA and CD3 instead of mouse BCMA and CD3. Chronic lung inflammation and sustained IgE production were induced in mice by intranasal exposure to 25 μg HDM (Greer, catalog #XPB70D3A25) diluted in 20 μL saline (Sigma, catalog #S8776) or 20 μL saline (control group) three times a week for 19 weeks. This model induced a class shift of B cells to IgE-producing plasma cells in secondary lymphoid organs and drove the accumulation of IgE-producing plasma cells in the bone marrow. At week 12 following the first HDM administration, a subgroup of mice began subcutaneous administration of either 25 mg / kg REGN1103 (anti-IL-4Rα) or 25 mg / kg REGN1094 (isotype control) until the end of the experiment. At week 15, mice were subcutaneously administered two doses of REGN5459 (anti-BCMA x anti-CD3) or two doses of REGN4460 (isotype control), and were then re-exposed to HDM for 4 weeks. Details of the HDM exposure and antibody treatment regimens are shown in Table 3 below. Figure 3 As shown in the image.

[0168] Table 3: HDM exposure and antibody treatment regimens in mice in a chronic (19-week) HDM model

[0169]

[0170] At 1, 3, and 5 weeks post-REGN5459 administration, ~100 μL of blood was collected from all mouse groups via retroorbital pleophoresis and transferred to micro-blood collection tubes (BD, catalog #365967) for serum separation. The concentration of HDM-specific IgE in serum was determined using the Mouse Serum Anti-HDM IgE Antibody Detection Kit (Chondrex catalog #3037) according to the manufacturer's instructions. Briefly, 100 μL of serum sample diluted 1:20 or 1:60 and HDM-IgE standards diluted to a starting concentration of 50 ng / mL and further serially diluted 2-fold were added to a pre-coated plate provided with the kit and incubated overnight at 4°C. The plate was then washed three times with wash buffer and incubated with 100 μL of biotinylated HDM provided with the kit. The plate was then washed four times with wash buffer and incubated with 100 μL of streptavidin peroxidase (provided with the kit) at room temperature for 30 min. The plate was washed 7 times, and 100 μL of TMB substrate solution (provided with the kit) was added to each sample. The plate was incubated in the dark for 25 min, followed by the addition of 50 μL of stop solution (2N sulfuric acid, provided with the kit). The absorbance was measured at 450 nm, and the HDM-IgE concentration was calculated based on the standard curve. Serum IgE values ​​are displayed as ng / mL. The limit of quantitation (LLOQ) for HDM-specific IgE ELISA was 15.62 ng / mL. Statistical significance was determined using the Kronoswiss test and Dunn's post-hoc multiple comparison test in GraphPad Prism.

[0171] result

[0172] In the chronic HDM model, anti-IL4RA treatment showed a trend toward decreasing serum HDM-specific IgE levels, but this did not reach statistical significance, and this trend was maintained at all three time points examined (see [reference needed]). Figure 4A , 4B (and 4C and Table 4). Compared to the isotype control group, one week after REGN5459 administration, anti-BCMA x anti-CD3 monotherapy resulted in a significant reduction in circulating HDM-specific IgE ( Figure 4A However, this effect is transient, and at 3 or 5 weeks after bispecific antibody administration, the level of HDM-specific IgE was not significantly different from that of the isotype control group. Figure 4B and 4C (and Table 4). The combination of anti-IL-4Rα and anti-BCMA x anti-CD3 therapy eliminated serum HDM-specific IgE (not detectable by ELISA). This effect was maintained during this experiment (see Table 4). Figure 4A , 4B(and 4C and Table 4) demonstrate that blocking IL-4RA and using anti-BCMA x anti-CD3 to deplete plasma cells is a successful strategy to block the production of allergen-specific IgE even in the presence of continuous allergen exposure.

[0173] Table 4: HDM-specific IgE levels at weeks 1, 3, and 5 after anti-BCMAxCD3 treatment

[0174]

[0175] Example 3: Effect of combined therapy with BCMAxCD3 bispecific antibody and anti-IL-4Rα antibody on IgE bone marrow plasma cells

[0176] IgE-mediated bone marrow plasma cell analysis was also performed on the mice described in Example 2. See Table 3 and... Figure 3 Following the HDM exposure and antibody treatment regimen described in the original text, mice were euthanized and femurs were collected. Bone marrow was extracted from the femurs by cutting both ends of each bone and placing each bone into a single well of a 96-well PCR plate (with an opening at the bottom of each well). The PCR plate was then placed on top of a 96-well 2mL deep-well collection plate and centrifuged at 500g for 4 minutes. The bone marrow was resuspended in 0.5mL RBC lysis buffer and incubated at room temperature for 3 minutes, followed by inactivation with 1-2mL PBS. The cells were centrifuged at 400g for 4 minutes, the supernatant was decanted, and the pellet was resuspended in 1mL DPBS and filtered through a Millipore plate filter (100μm) into a 2mL deep-well plate. The cells were then centrifuged and resuspended in 200μL PBS. Bone marrow cells were then seeded in 96-well plates, stained with live / dead cell markers, and then stained with antibodies B220, CD138, IgM, IgG1, IgA, IgD, IgE (extracellular blockade), and “dump” (including TCRβ, CD200R3, Ly6G, CD49b, and CD11b).

[0177] After staining, cells were washed twice with MACS buffer, fixed for 15 minutes with BD Cytofix (catalog #554655) diluted 1:4 in PBS, then resuspended in MACS buffer and stored at 4°C. On the day of collection, cells were washed, incubated for 10 minutes in BDPerm / wash buffer (catalog #554723), and stained with intracellular antibodies for light chain κ, IgG1, and Intra IgE. Cells were then acquired in an LSL Ortessa instrument and analyzed using FlowJo software. Mature IgE-positive bone marrow plasma cells were identified as viable Dump-B220-light chain κ+IgE+. The percentage reduction in plasma cells in each antibody-administered mouse was calculated using the following formula: 100 – (100 x plasma cell percentage / mean plasma cell percentage in the isotype group), where the plasma cell percentage is calculated relative to total viable cells.

[0178] result

[0179] In a chronic HDM model with continuous HDM exposure following antibody therapy, neither anti-IL-4RA nor BCMA x CD3 alone had a significant effect on IgE bone marrow plasma cells at collection time (5 weeks after BCMA x CD3 administration). However, anti-IL-4RA treatment showed a trend toward a decrease in IgE bone marrow plasma cells (see [link to relevant documentation]). Figure 5 (and Table 5). Conversely, the combination of continuous anti-IL-4Ra administration with transient anti-BCMA x anti-CD3 administration resulted in a significant reduction in IgE bone marrow plasma cells relative to untreated and allotype controls (see Table 5). Figure 5 (and Table 5).

[0180] Table 5: IgE bone marrow plasma cells (BMPC) after antibody treatment

[0181]

[0182] Example 4: Effects of combined treatment with BCMAxCD3 bispecific antibody and anti-IL-4Rα antibody on IgE-producing and IgE-generating cells.

[0183] To determine the effects of combined anti-IL-4Rα and anti-BCMA x anti-CD3 therapy on IgE production and IgE-producing cells in a relevant in vivo model, an HDM-driven lung inflammation study was conducted in mice, as described in Examples 1-2 above, where human BCMA and human CD3 were homozygous for mouse BCMA and CD3. Chronic lung inflammation and sustained IgE production were induced in mice by intranasal exposure to 25 μg HDM diluted in 20 μL saline or 20 μL saline (control group) three times a week for 15 weeks. This model induced a class shift of B cells to IgE-producing plasma cells in secondary lymphoid organs and drove the accumulation of IgE-producing plasma cells in the bone marrow. At week 12 following the first HDM administration, a subgroup of mice began receiving subcutaneous injections of 25 mg / kg REGN1103 (anti-IL-4Rα) or 25 mg / kg REGN1094 (isotype control) until the end of the experiment. At week 15, mice were administered two doses of REGN5459 (anti-BCMA x anti-CD3) or two doses of REGN4460 (isotype control) subcutaneously, and then allowed to rest for two weeks without further HDM administration. (See below) Figure 6 Table 6 provides a detailed overview of HDM exposure and antibody treatment regimens.

[0184] Table 6: Mouse HPD exposure and antibody treatment regimens to address the effects of anti-IL-4Rα and anti-BMCA x anti-CD3 antibody therapy on bone marrow plasma cell population in a chronic (15-week) HPD model.

[0185]

[0186] Following HDM exposure and antibody treatment, mice were euthanized and blood, spleen, and bones were collected. Blood was collected from all groups of mice via cardiac puncture and transferred to micro-blood collection tubes (BD, catalog #365967) for serum separation. The concentration of HDM-specific IgE in serum was determined using the Mouse Serum Anti-HDM IgE Antibody Detection Kit (Chondrex catalog #3037) according to the manufacturer's instructions. Briefly, 100 μL of serum sample diluted 1:10 or 1:60 and HDM-IgE standards diluted to a starting concentration of 50 ng / mL and further serially diluted 2-fold were added to the pre-coated plate provided with the kit and incubated overnight at 4°C. The plate was then washed three times with wash buffer and incubated with 100 μL of biotinylated HDM provided with the kit. The plate was then washed four times with wash buffer and incubated with 100 μL of streptavidin peroxidase (provided with the kit) at room temperature for 30 min. The plate was washed seven times, and 100 μL of TMB substrate solution (provided with the kit) was added to each sample. The plate was incubated in the dark for 25 min, followed by the addition of 50 μL of stop solution (2N sulfuric acid, provided with the kit). The absorbance was measured at 450 nm, and the HDM-IgE concentration was calculated based on the standard curve. Serum IgE values ​​are displayed as ng / mL. Statistical significance was determined using the Kronoswiss test and Dunn's post-hoc multiple comparison test in GraphPadPrism.

[0187] Spleens and femurs were also collected from mice. Spleens were homogenized using the back end of a 3 mL syringe on a 74-micron cell filter in 2 mL RPMI medium, and the single-cell suspension was transferred to 96-well plates. Bone marrow was extracted from the femurs by cutting both ends of each bone and placing each bone into a single well of a 96-well PCR plate (with an opening at the bottom of each well). The PCR plate was then placed on top of a 96-well 2 mL deep-well collection plate and centrifuged at 500 g for 4 min. The spleen sample was resuspended in 1 mL, and the bone marrow was resuspended in 0.5 mL of RBC lysis buffer and incubated at room temperature for 3 min. Then, 1–2 mL of PBS was added to inactivate the lysis buffer. Cells were centrifuged at 400 g for 4 min, the supernatant was decanted, and the pellet was resuspended in 1 mL of DPBS and filtered through a Millipore plate filter (100 μm) into a 2 mL deep-well plate. The cells were then centrifuged, and the spleen cells were resuspended in 1 mL, and the bone marrow was resuspended in 200 μL of PBS. One-tenth of the spleen cells and all bone marrow cells were then seeded into 96-well plates and stained with live / dead cell markers, followed by antibody staining with antibodies B220, CD138, IgM, IgG1, IgA, IgD, IgE (extracellular blockade), and "dump" (including TCRβ, CD200R3, Ly6G, CD49b, and CD11b). After staining, the cells were washed twice with MACS buffer, fixed for 15 minutes with BD Cytofix (catalog #554655) diluted 1:4 in PBS, resuspended in MACS buffer, and stored at 4°C. On the day of collection, the cells were washed, incubated for 10 minutes in BD Perm / wash buffer (catalog #554723), and stained with intracellular antibodies light chain κ, IgG1, and Intra IgE. Cells were then acquired in an LSR Tortessa instrument and analyzed using FlowJo software. Mature plasma cells were identified as live Dump- (Dump includes TCRb, CD200R3, Ly6G, CD49b, and CD11b) B220-light chain k+. The percentage reduction in plasma cells in each antibody-administered mouse was calculated using the following formula: 100 – (100 x plasma cell percentage / mean plasma cell percentage in the isotype group), where the plasma cell percentage is calculated relative to total live cells. The results are shown in Table 7 below.

[0188] result

[0189] In the chronic HDM model, anti-IL4RA treatment (group F) showed a trend toward decreasing serum HDM-specific IgE levels, with most samples exhibiting detectable HDM-specific IgE levels (see Table 7 and...). Figure 7Both anti-BCMA x anti-CD3 monotherapy (Group E) and anti-IL-4Rα combined with anti-BCMA x anti-CD3 therapy (Group G) eliminated serum HDM-specific IgE, with all mice showing levels below the lower limit of quantitation. These data suggest that anti-BCMA x anti-CD3 treatment is sufficient to significantly reduce serum HDM-specific IgE within 2 weeks after administration.

[0190] In the same experiment, anti-IL4RA treatment did not affect total bone marrow plasma cells ( Figure 8A ), and showed a trend of decreased IgE bone marrow plasma cells ( ), and exhibited a decreasing trend of IgE bone marrow plasma cells ( ). Figure 8B Compared with the untreated group and the allotype control group, anti-BCMA x anti-CD3 treatment alone resulted in a significant reduction in total and IgE bone marrow plasma cells at collection time (two weeks after REGN5459 administration) relative to the untreated group and the allotype control group. Figures 8A-8B Combination therapy with anti-IL-4Rα and anti-BCMA x anti-CD3 also significantly reduced total and IgE-specific bone marrow plasma cells to a level similar to that of anti-BCMA x anti-CD3 alone, indicating that the latter treatment was sufficient to deplete bone marrow plasma cells. Similar results were observed in the spleen; anti-IL-4Rα treatment had no effect on total or IgE-specific splenic plasma cells. Figure 8C-8D At collection time (two weeks after REGN5459 administration), anti-BCMA x anti-CD3 treatment alone resulted in a significant reduction in total splenic plasma cells compared to the untreated group and the isotype control group. Figure 8C-8D Combination therapy with anti-IL-4Rα and anti-BCMA x anti-CD3 also significantly reduced total splenic plasma cells to a similar degree as anti-BCMA x anti-CD3 alone, indicating that the latter treatment was sufficient to deplete splenic plasma cells. Treatment with anti-BCMA x anti-CD3 alone and the combination of anti-IL-4Rα and anti-BCMA x anti-CD3 both reduced IgE plasma cells. However, the reduction in IgE splenic plasma cells was statistically significant only compared to the group receiving HDM for 12 weeks without antibody treatment or to the isotype control group, and not statistically significant compared to the group receiving HDM for 15 weeks without antibody treatment.

[0191] Table 7: Effects of single anti-BMCA x anti-CD3 bispecific antibodies or their combination with anti-IL-4Rα antibodies on serum HDM-specific IgE levels

[0192]

[0193] Table 8: Effects of single anti-BMCA x anti-CD3 bispecific antibodies or their combination with anti-IL-4Rα antibodies on total and IgE levels in bone marrow and spleen plasma cells.

[0194]

[0195]

[0196] The scope of this invention is not limited to the specific embodiments described herein. In fact, various modifications of the invention will become apparent to those skilled in the art from the foregoing description and drawings, in addition to those embodiments described herein. Such modifications also fall within the scope of the appended claims.

Claims

1. Use of a therapeutically effective amount of an IL-4 / IL-13 pathway inhibitor and a plasma cell ablation agent in the preparation of a medicament for reducing or eliminating allergen-specific serum IgE in an individual with an allergic disease or condition, wherein the IL-4 / IL-13 pathway inhibitor is an anti-IL-4 receptor (IL-4R) antibody, wherein the anti-IL-4R antibody comprises (i) a heavy chain variable region (HCVR) consisting of the amino acid sequence of SEQ ID NO:1 and a light chain variable region (LCVR) consisting of the amino acid sequence of SEQ ID NO:2, or (ii) a heavy chain variable region (HCVR) consisting of the amino acid sequence of SEQ ID NO:43 and a light chain variable region (LCVR) consisting of the amino acid sequence of SEQ ID NO:44, and wherein the plasma cell ablation agent is an anti-BCMA / anti-CD3 bispecific antibody or an antigen-binding fragment thereof comprising (i) a first antigen-binding domain that specifically binds to BCMA and includes an HCVR consisting of the amino acid sequence of SEQ ID NO:12 and a light chain variable region consisting of the amino acid sequence of SEQ ID NO:

44. (ii) an LCVR consisting of the amino acid sequence of SEQ ID NO:20; and (ii) a second antigen-binding domain that specifically binds to CD3 and includes an HCVR consisting of the amino acid sequence of SEQ ID NO:28 or SEQ ID NO:36 and an LCVR consisting of the amino acid sequence of SEQ ID NO:

20.

2. Use of a therapeutically effective amount of an IL-4 / IL-13 pathway inhibitor and a plasma cell ablation agent in the preparation of a medicament for treating allergies or preventing allergic reactions to allergens or reducing their severity in individuals with allergic diseases or conditions, wherein the IL-4 / IL-13 pathway inhibitor is an anti-IL-4 receptor (IL-4R) antibody, wherein the anti-IL-4R antibody comprises (i) a heavy chain variable region (HCVR) consisting of the amino acid sequence of SEQ ID NO:1 and a light chain variable region (LCVR) consisting of the amino acid sequence of SEQ ID NO:2, or (ii) a heavy chain variable region (HCVR) consisting of the amino acid sequence of SEQ ID NO:43 and a light chain variable region (LCVR) consisting of the amino acid sequence of SEQ ID NO:44, and wherein the plasma cell ablation agent is an anti-BCMA / anti-CD3 bispecific antibody or an antigen-binding fragment thereof comprising (i) a first antigen-binding domain that specifically binds to BCMA and includes an HCVR consisting of the amino acid sequence of SEQ ID NO:12 and a light chain variable region consisting of the amino acid sequence of SEQ ID NO:

44. (ii) an LCVR consisting of the amino acid sequence of SEQ ID NO:20; and (ii) a second antigen-binding domain that specifically binds to CD3 and includes an HCVR consisting of the amino acid sequence of SEQ ID NO:28 or SEQ ID NO:36 and an LCVR consisting of the amino acid sequence of SEQ ID NO:

20.

3. The use according to any one of claims 1-2, wherein the allergic disease or condition is selected from allergic asthma, chronic urticaria, food allergy, pollen allergy, and allergies caused by non-food environmental allergens.

4. The use according to any one of claims 1-2, wherein the allergic disease or condition is hay fever.

5. The use according to any one of claims 1-2, wherein the individual is at risk of an allergic reaction to the allergen.

6. The use according to any one of claims 1-2, wherein the individual suffers from seasonal allergies.

7. The use according to any one of claims 1-2, wherein the individual suffers from severe allergies.

8. The use according to any one of claims 1-2, wherein the individual has an allergy caused by one or more allergens, said allergens being selected from milk, eggs, celery, sesame, wheat, soybeans, fish, fruits, shellfish, sugar, peanuts, dust, dust mites, pollen, insect venom, mold, animal dander, wool, latex, metals, household cleaners, detergents, cosmetics, fragrances, penicillin, sulfonamides, salicylates, therapeutic monoclonal antibodies, ragweed, and birch.

9. The use according to any one of claims 1-2, wherein the individual has an allergy caused by one or more allergens selected from dairy products, meat, legumes, tree nuts, animal fur, pharmaceuticals, and herbs.

10. The use according to any one of claims 1-2, wherein the allergen contained in or derived from food is selected from milk, eggs, celery, sesame, wheat, fruit, soybean, fish, shellfish, sugar, peanut and tree nuts.

11. The use according to any one of claims 1-2, wherein the allergen contained in or derived from food is selected from dairy products, meat and legumes.

12. The use according to any one of claims 1-2, wherein the allergen is a non-food environmental allergen selected from dust, dust mites, pollen, insect venom, mold, animal dander, wool, latex, metals, detergents, cosmetics, fragrances, penicillin, sulfonamides or salicylates, therapeutic monoclonal antibodies, ragweed and birch.

13. The use according to any one of claims 1-2, wherein the allergen is a non-food environmental allergen selected from animal fur, household cleaners, pharmaceuticals, and herbs.

14. Use of an IL-4 / IL-13 pathway inhibitor and a plasma cell ablation agent in the preparation of a medicament for increasing the efficacy and / or tolerability of an immunotherapy regimen for an individual with an allergic condition, wherein the medicament is administered to the individual prior to or concurrently with the immunotherapy regimen, wherein the IL-4 / IL-13 pathway inhibitor is an anti-IL-4 receptor (IL-4R) antibody, wherein the anti-IL-4R antibody comprises (i) a heavy chain variable region (HCVR) consisting of the amino acid sequence of SEQ ID NO:1 and a light chain variable region (LCVR) consisting of the amino acid sequence of SEQ ID NO:2, or (ii) a heavy chain variable region (HCVR) consisting of the amino acid sequence of SEQ ID NO:43 and a light chain variable region (LCVR) consisting of the amino acid sequence of SEQ ID NO:44, and wherein the plasma cell ablation agent is an anti-BCMA / anti-CD3 bispecific antibody or an antigen-binding fragment thereof comprising (i) a first antigen-binding domain that specifically binds to BCMA and includes an HCVR consisting of the amino acid sequence of SEQ ID NO:12 and a light chain variable region consisting of the amino acid sequence of SEQ ID NO:

44. (ii) an LCVR consisting of the amino acid sequence of SEQ ID NO:20; and (ii) a second antigen-binding domain that specifically binds to CD3 and includes an HCVR consisting of the amino acid sequence of SEQ ID NO:28 or SEQ ID NO:36 and an LCVR consisting of the amino acid sequence of SEQ ID NO:

20.

15. The use of claim 14, wherein the immunotherapy is an oral immunotherapy.

16. The use of claim 14, wherein the immunotherapy is a subcutaneous immunotherapy.

17. The use of any one of claims 14-16, wherein the individual suffers from a food allergy.

18. The use of claim 17, wherein the individual suffers from allergies to milk, eggs, celery, sesame, wheat, fruit, soybeans, fish, shellfish, sugar, peanuts, and tree nuts.

19. The use according to any one of claims 14-16, wherein the individual suffers from an allergy to dairy products, meat and legumes.

20. The use according to any one of claims 14-16, wherein the individual suffers from an allergy to a non-food allergen selected from dust, dust mites, pollen, insect venom, mold, animal dander, wool, latex, metals, detergents, cosmetics, fragrances, therapeutic monoclonal antibodies, ragweed, and birch.

21. Use according to any one of claims 14-16, wherein the individual suffers from an allergy to a non-food allergen selected from animal fur, household cleaners, pharmaceuticals, and herbs.

22. The use according to any one of claims 14-16, wherein the drug is administered prior to initiating an immunotherapy regimen.

23. The use according to any one of claims 14-16, wherein the drug is administered concurrently with an immunotherapy regimen.

24. Use according to any one of claims 1-2 and 14-16, wherein the anti-IL-4R antibody comprises: HCVR consisting of the amino acid sequence of SEQ ID NO:1 and LCVR consisting of the amino acid sequence of SEQ ID NO:

2.

25. The use of claim 24, wherein the anti-IL-4R antibody comprises a heavy chain and a light chain, wherein the heavy chain consists of the amino acid sequence of SEQ ID NO:

9.

26. The use of claim 24, wherein the anti-IL-4R antibody comprises a heavy chain and a light chain, wherein the light chain consists of the amino acid sequence of SEQ ID NO:

10.

27. The use of claim 24, wherein the anti-IL-4R antibody comprises a heavy chain and a light chain, wherein the heavy chain consists of the amino acid sequence of SEQ ID NO:9 and the light chain consists of the amino acid sequence of SEQ ID NO:

10.

28. Use according to any one of claims 1-2 and 14-16, wherein the IL-4 / IL-13 pathway inhibitor is dupilumab or its bioequivalence.

29. The use according to any one of claims 1-2 and 14-16, wherein the anti-BCMA / anti-CD3 bispecific antibody or its antigen-binding fragment comprises: The first antigen-binding domain includes an HCVR consisting of the amino acid sequence of SEQ ID NO:12 and an LCVR consisting of the amino acid sequence of SEQ ID NO:20; and the second antigen-binding domain includes an HCVR consisting of the amino acid sequence of SEQ ID NO:28 and an LCVR consisting of the amino acid sequence of SEQ ID NO:

20.

30. The use according to any one of claims 1-2 and 14-16, wherein the anti-BCMA / anti-CD3 bispecific antibody or its antigen-binding fragment comprises: The first antigen-binding domain includes an HCVR consisting of the amino acid sequence of SEQ ID NO:12 and an LCVR consisting of the amino acid sequence of SEQ ID NO:20; and the second antigen-binding domain includes an HCVR consisting of the amino acid sequence of SEQ ID NO:36 and an LCVR consisting of the amino acid sequence of SEQ ID NO:

20.

31. The use according to any one of claims 1-2 and 14-16, wherein at least one other therapeutic agent or therapy is administered to the individual.

32. The use of claim 31, wherein the other therapeutic agent is selected from IgE antagonists, antihistamines, anti-inflammatory drugs, corticosteroids, leukotriene antagonists, mast cell inhibitors, bronchodilators, decongestants, adrenaline, IL-1 antagonists, IL-5 antagonists, IL-31 antagonists, IL-33 antagonists, IL-25 antagonists, interferon-γ, TNF antagonists, and TSLP antagonists.

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