A method for treating or preventing allergic asthma by administering an IL-33 antagonist and / or an IL-4R antagonist.

Abstract

To provide a novel targeted therapy for the treatment and / or prevention of allergic asthma. [Solution] One particular method includes the step of administering a therapeutic composition comprising an interleukin-33 (IL-33) antagonist, for example, an anti-IL-33 antibody, to a subject in need. Another disclosed method includes the step of administering a therapeutic composition comprising an interleukin-4R (IL-4R) antagonist, for example, an anti-IL-4R antibody, to a subject in need. Yet another disclosed method includes the step of administering a first therapeutic composition comprising an interleukin-33 (IL-33) antagonist, for example, an anti-IL-33 antibody, and a second therapeutic composition comprising an interleukin-4 receptor (IL-4R) antagonist, for example, an anti-IL-4R antibody, to a subject in need.

Description

[Technical Field]

[0001] Related applications This application claims priority to U.S. Provisional Patent Application No. 62 / 952,996 filed December 23, 2019, and U.S. Provisional Patent Application No. 62 / 964,970 filed January 23, 2020. The entire disclosures of each of these applications are incorporated herein by reference.

[0002] The present invention relates to the treatment and / or prevention of allergic asthma and related conditions. More specifically, the present invention relates to the administration of an interleukin-33 (IL-33) antagonist for the treatment or prevention of allergic asthma in patients who require it. The present invention also relates to the administration of an interleukin-4 (IL-4R) antagonist for the treatment or prevention of allergic asthma in patients who require it. Finally, the present invention relates to the administration of an IL-33 (IL-33) antagonist and an interleukin-4 receptor (IL-4R) antagonist for the treatment or prevention of allergic asthma in patients who require it. [Background technology]

[0003] Asthma is a chronic inflammatory disease of the airways characterized by airway hyperresponsiveness, acute and chronic bronchoconstriction, airway edema, and mucus plugs. The inflammatory components of asthma are thought to involve many cell types, including mast cells, eosinophils, T lymphocytes, neutrophils, and epithelial cells, as well as their biological products. Patients with asthma often experience symptoms such as wheezing, shortness of breath, cough, and chest tightness. For most asthma patients, controller and bronchodilator regimens are used to provide long-term control. Inhaled corticosteroids (ICS) are considered the "golden rule" for controlling asthma symptoms, and inhaled β2-agonists are currently the most effective bronchodilators available.

[0004] Type 2-high asthma is the most common type among persistent asthma (Non-Patent Document 1). Type 2-high asthma includes overlapping phenotypes allergic asthma (characterized by increased expression of specific immunoglobulin E (IgE) to airborne allergens) and eosinophilic asthma (characterized by increased blood and / or airway / tissue eosinophilia) (Fahy, see above; Non-Patent Document 2; Non-Patent Document 3).

[0005] Allergic asthma is the most common type of asthma. Allergic sensitization is a powerful risk factor for the onset and severity of asthma in children and adults (Non-Patent Document 4). Current allergic asthma therapies that address the symptoms of the disease and ongoing inflammatory processes do not affect the underlying, dysregulated immune response and are thus very limited in controlling the progression of allergic asthma (Non-Patent Document 5).

[0006] There is a need in the art for new targeted therapies for the treatment and / or prevention of asthma, such as allergic asthma.

Prior Art Documents

Non-Patent Documents

[0007]

Non-Patent Document 1

Non-Patent Document 2

Non-Patent Document 3

Non-Patent Document 4

[0008] In one embodiment, a method for treating allergic asthma in a subject requiring it is provided, comprising the step of administering to the subject an antibody or antigen-binding fragment thereof, which specifically binds to interleukin-33 (IL-33) and comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs: 4, 6, and 8, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs: 12, 14, and 16. In one embodiment, an antibody or antigen-binding fragment thereof, which specifically binds to interleukin-33 (IL-33) and comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs: 4, 6, and 8, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs: 12, 14, and 16, is provided for use in treating allergic asthma in a subject requiring it.

[0009] In a particular exemplary embodiment, the antibody or its antigen-binding fragment comprises a heavy chain variable region (HCVR) containing the amino acid sequence of SEQ ID NO: 2 and a light chain variable region (LCVR) containing the amino acid sequence of SEQ ID NO: 10. According to a particular exemplary embodiment, the antibody or its antigen-binding fragment comprises REGN3500.

[0010] In certain exemplary embodiments, the antibody or its antigen-binding fragment is administered intravenously at a dose of 10 mg / kg. In certain exemplary embodiments, the antibody or its antigen-binding fragment is administered subcutaneously at doses of approximately 0.1 mg to approximately 600 mg, approximately 100 mg to approximately 400 mg, or approximately 300 mg. In certain exemplary embodiments, the antibody or its antigen-binding fragment is administered subcutaneously at an initial dose of approximately 600 mg or approximately 300 mg. In certain exemplary embodiments, the antibody or its antigen-binding fragment is administered subcutaneously at one or more second doses of approximately 300 mg.

[0011] In another embodiment, a method is provided for treating allergic asthma in a subject requiring the treatment, comprising the step of administering to the subject an antibody or antigen-binding fragment thereof that specifically binds to interleukin-4R (IL-4R) and comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs. 21, 22, and 23, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs. 24, 25, and 26. In one embodiment, an antibody or antigen-binding fragment thereof that specifically binds to interleukin-4R (IL-4R) and comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs. 21, 22, and 23, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs. 24, 25, and 26 is provided for use in treating allergic asthma in a subject requiring the treatment.

[0012] In certain exemplary embodiments, the antibody or its antigen-binding fragment comprises a heavy chain variable region (HCVR) containing the amino acid sequence of SEQ ID NO: 27 and a light chain variable region (LCVR) containing the amino acid sequence of SEQ ID NO: 28. In certain exemplary embodiments, the antibody or its antigen-binding fragment comprises dupilumab.

[0013] In certain exemplary embodiments, the antibody or its antigen-binding fragment is administered in doses of approximately 0.1 mg to approximately 600 mg, approximately 100 mg to approximately 400 mg, or approximately 300 mg. In certain exemplary embodiments, the antibody or its antigen-binding fragment is administered as an initial dose of approximately 600 mg. In certain exemplary embodiments, the antibody or its antigen-binding fragment is administered as one or more second doses of approximately 300 mg.

[0014] In a particular exemplary embodiment, the antibody or its antigen-binding fragment is administered once a week (q1w), every other week (q2w), every three weeks (q3w), or every four weeks (q4w). In a particular exemplary embodiment, the antibody or its antigen-binding fragment is administered every other week (q2w).

[0015] In certain exemplary embodiments, the antibody or its antigen-binding fragment is administered subcutaneously. In certain exemplary embodiments, the antibody or its antigen-binding fragment is administered subcutaneously using an auto-injector, needle and syringe, or pen-type delivery device.

[0016] In another embodiment, a method for treating allergic asthma in a subject requiring such treatment is provided, comprising the steps of administering to the subject an initial dose of approximately 600 mg of an antibody or antigen-binding fragment, and one or more subsequent doses of approximately 300 mg of the antibody or antigen-binding fragment, which specifically binds to interleukin-4R (IL-4R) and comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs. 21, 22, and 23, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs. 24, 25, and 26. In another embodiment, an initial dose of approximately 600 mg of an antibody or antigen-binding fragment, and one or more subsequent doses of approximately 300 mg of the antibody or antigen-binding fragment, are provided for use in treating allergic asthma in subjects requiring it, comprising three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs. 21, 22, and 23, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs. 24, 25, and 26.

[0017] In a particular exemplary embodiment, the antibody or its antigen-binding fragment includes a heavy chain variable region (HCVR) containing the amino acid sequence of SEQ ID NO: 27 and a light chain variable region (LCVR) containing the amino acid sequence of SEQ ID NO: 28.

[0018] In another embodiment, a method is provided for treating allergic asthma in a subject requiring such treatment, comprising the steps of administering to the subject a first antibody or antigen-binding fragment that specifically binds to interleukin-33 (IL-33) and comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs. 4, 6, and 8, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs. 12, 14, and 16, and a second antibody or antigen-binding fragment that specifically binds to the interleukin-4 receptor (IL-4R) and comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs. 21, 22, and 23, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs. 24, 25, and 26. In one embodiment, a first antibody or its antigen-binding fragment, which specifically binds to interleukin-33 (IL-33) and comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs. 4, 6, and 8, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs. 12, 14, and 16; and a second antibody or its antigen-binding fragment, which specifically binds to the interleukin-4 receptor (IL-4R) and comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs. 21, 22, and 23, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs. 24, 25, and 26, are provided for use in treating allergic asthma in subjects requiring it. It will be done.

[0019] In a particular exemplary embodiment, the first antibody or its antigen-binding fragment comprises a heavy chain variable region (HCVR) containing the amino acid sequence of SEQ ID NO: 2 and a light chain variable region (LCVR) containing the amino acid sequence of SEQ ID NO: 10. In a particular exemplary embodiment, the first antibody or its antigen-binding fragment comprises REGN3500.

[0020] In a particular exemplary embodiment, the second antibody or its antigen-binding fragment comprises a heavy chain variable region (HCVR) containing the amino acid sequence of SEQ ID NO: 27 and a light chain variable region (LCVR) containing the amino acid sequence of SEQ ID NO: 28. In a particular exemplary embodiment, the second antibody or its antigen-binding fragment comprises dupilumab.

[0021] In certain exemplary embodiments, the second antibody or its antigen-binding fragment is administered in doses of approximately 0.1 mg to approximately 600 mg, approximately 100 mg to approximately 400 mg, or approximately 300 mg. In certain exemplary embodiments, the second antibody or its antigen-binding fragment is administered as an initial dose of approximately 600 mg. In certain exemplary embodiments, the second antibody or its antigen-binding fragment is administered as one or more subsequent doses of approximately 300 mg of the antibody or its antigen-binding fragment.

[0022] In a particular exemplary embodiment, the second antibody or its antigen-binding fragment is administered once a week (q1w), every other week (q2w), every three weeks (q3w), or every four weeks (q4w). In a particular exemplary embodiment, the second antibody or its antigen-binding fragment is administered every other week (q2w).

[0023] In certain exemplary embodiments, the second antibody or its antigen-binding fragment is administered subcutaneously. In certain exemplary embodiments, the antibody or its antigen-binding fragment is administered subcutaneously using an auto-injector, needle and syringe, or pen-type delivery device.

[0024] In certain exemplary embodiments, the first antibody or its antigen-binding fragment is administered intravenously at a dose of 10 mg / kg. In certain exemplary embodiments, the first antibody or its antigen-binding fragment is administered subcutaneously at doses of approximately 0.1 mg to approximately 600 mg, approximately 100 mg to approximately 400 mg, or approximately 300 mg. In certain exemplary embodiments, the first antibody or its antigen-binding fragment is administered subcutaneously at an initial dose of approximately 600 mg or approximately 300 mg. In certain exemplary embodiments, the first antibody or its antigen-binding fragment is administered subcutaneously at one or more second doses of approximately 300 mg.

[0025] In another embodiment, a method is provided for treating allergic asthma in a subject requiring such treatment, comprising the steps of administering to a subject a first antibody or antigen-binding fragment, which specifically binds to interleukin-33 (IL-33) and comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs. 4, 6, and 8, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs. 12, 14, and 16, and is administered at a single dose of 10 mg / kg; and a second antibody or antigen-binding fragment, which specifically binds to the interleukin-4 receptor (IL-4R) and comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs. 21, 22, and 23, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs. 24, 25, and 26, and is administered at an initial dose of 600 mg / kg and one or more subsequent doses of approximately 300 mg. In one embodiment, interleukin-33 (IL-33) is specifically bound to three heavy chain complementary determination region (HCDR) sequences, including SEQ ID NOs. 4, 6, and 8, as well as SEQ ID NO. 12. A first antibody or antigen-binding fragment comprising three light chain complementary determinant region (LCDR) sequences including 14 and 16, administered at a single dose of 10 mg / kg, and a second antibody or antigen-binding fragment specifically binding to the interleukin-4 receptor (IL-4R) and comprising three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs. 21, 22, and 23, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs. 24, 25, and 26, administered at an initial dose of 600 mg / kg and one or more subsequent doses of approximately 300 mg, are provided for use in treating allergic asthma in subjects requiring it.

[0026] In certain exemplary embodiments, allergic asthma is mild allergic asthma. In certain exemplary embodiments, allergic asthma is mild persistent allergic asthma.

[0027] In certain exemplary embodiments, the subject is allergic to the dust mite allergen (HDM). In certain exemplary embodiments, the subject is a non-smoker. In certain exemplary embodiments, the subject is clinically stable and requires the use of an inhaled short-acting β2-agonist (SABA) as needed to control asthma symptoms.

[0028] In certain exemplary embodiments, loss of asthma control (LOAC) is reduced in the subjects. In certain exemplary embodiments, asthma symptoms selected from the group consisting of cough and wheezing, and the use of inhaled short-acting β2 agonists are reduced in the subjects.

[0029] In certain exemplary embodiments, one or more asthma-related parameters are improved in the subject. In certain exemplary embodiments, the asthma-related parameters are selected from the group consisting of forced expiratory volume in one second (FEV1), maximal expiratory flow rate (PEF), forced vital capacity (FVC), forced expiratory flow rate (FEF) of 25% to 75%, and a reduction in the frequency or dose of inhaled short-acting β2-agonist use in the subject. In certain exemplary embodiments, the FEV1 before bronchodilator administration in the subject is improved.

[0030] In certain exemplary embodiments, blood eosinophil levels decrease in the subjects.

[0031] In certain exemplary embodiments, one or both of the following scores are improved in the subjects: the Asthma Control Questionnaire Five-Question Version (ACQ-5) score and the Asthma Quality of Life Questionnaire with Standardized Activities (AQLQ) score.

[0032] In certain exemplary embodiments, the frequency or dosage of SABA use in the subject is reduced.

[0033] In certain exemplary embodiments, BAC-induced pneumonia is reduced in the subjects.

[0034] In certain exemplary embodiments, levels of type 2 cytokines are reduced in the subjects. In certain exemplary embodiments, type 2 cytokines are selected from the group consisting of IL-13 and IL-5. In certain exemplary embodiments, levels of type 2 cytokines are measured by determining the mRNA levels of one or more type 2 mediator genes, and the mRNA levels are reduced to at least about 50%, 60%, 70%, 80%, or 90%. In certain exemplary embodiments, levels of cytokines or chemokines selected from the group consisting of tumor necrosis factor-α (TNFα), TARC, lung and activating regulatory chemokines (PARC), CCL1, CCL26, FCER2, SIGLEC8, CCL17, and eotaxin-3 are reduced in the subjects.

[0035] In certain exemplary embodiments, the early allergen response (EAR) or late allergen response (LAR) is reduced in the subjects. In certain exemplary embodiments, FEV1 is improved by at least 20%, 30%, 40%, 50%, 60%, or 70% in the subjects. In certain exemplary embodiments, FeNO levels are reduced in the subjects. In certain exemplary embodiments, serum levels of sST2, IL-33, calcitonin, or matrix metalloproteinase-12 (MMP12) are reduced in the subjects. In certain exemplary embodiments, serum levels of CCL26, CCL17, or SIGLEC8 are reduced in the subjects. In certain exemplary embodiments, serum levels of ASAP1-IT1, AX747757, BC042385, PABPC1P2, AB209315, AX748268, TCEAL5, CCL13, CLC, CACNG8, GPR82, GATA1, PRSS33, FFAR3, LGALS12, ASB2, PTGDR2, IL-13, IL-5, PTGDS, or RD3 are reduced in the subjects.

[0036] In another embodiment, a method is provided for reducing cytokine or chemokine levels in a subject having allergic asthma, comprising the step of administering to the subject an antibody or antigen-binding fragment thereof that specifically binds to interleukin-33 (IL-33) and comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs: 4, 6, and 8, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs: 12, 14, and 16. In one embodiment, an antibody or antigen-binding fragment thereof that specifically binds to interleukin-33 (IL-33) and comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs: 4, 6, and 8, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs: 12, 14, and 16 is provided for use in reducing cytokine or chemokine levels in a subject having allergic asthma.

[0037] In certain exemplary embodiments, the cytokine is one or both of IL-13 and IL-5. In certain exemplary embodiments, the cytokine or chemokine is selected from the group consisting of TNFα, TARC, PARC, CCL1, CCL26, FCERV2, SIGLEC8, CCL17, and eotaxin-3.

[0038] In certain exemplary embodiments, serum levels of sST2, IL-33, calcitonin, or MMP12 are reduced in the subjects. In certain exemplary embodiments, serum levels of CCL26, CCL17, or SIGLEC8 are reduced in the subjects.

[0039] In certain exemplary embodiments, the anti-IL-33 antibody or its antigen-binding fragment comprises a heavy chain variable region (HCVR) containing the amino acid sequence of SEQ ID NO: 2 and a light chain variable region (LCVR) containing the amino acid sequence of SEQ ID NO: 10. In certain exemplary embodiments, the anti-IL-33 antibody or its antigen-binding fragment comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 18 and a light chain containing the amino acid sequence of SEQ ID NO: 20.

[0040] In a particular exemplary embodiment, the method further comprises administering to a subject an antibody or antigen-binding fragment thereof that specifically binds to IL-4R, wherein the antibody or antigen-binding fragment comprises three heavy chain complementary determinant region (HCDR) sequences, including SEQ ID NOs. 21, 22, and 23, and three light chain complementary determinant region (LCDR) sequences, including SEQ ID NOs. 24, 25, and 26.

[0041] In another embodiment, a method for reducing the expression of one or more allergic asthma signature genes in a subject having allergic asthma, wherein interlo A method is provided comprising the step of administering to a subject an antibody or antigen-binding fragment thereof that specifically binds to interleukin-33 (IL-33) and contains three heavy chain complementary determinant region (HCDR) sequences, including SEQ ID NOs: 4, 6, and 8, and three light chain complementary determinant region (LCDR) sequences, including SEQ ID NOs: 12, 14, and 16. In one embodiment, an antibody or antigen-binding fragment thereof that specifically binds to interleukin-33 (IL-33) and contains three heavy chain complementary determinant region (HCDR) sequences, including SEQ ID NOs: 4, 6, and 8, and three light chain complementary determinant region (LCDR) sequences, including SEQ ID NOs: 12, 14, and 16, is provided for reducing the expression of one or more allergic asthma signature genes in a subject having allergic asthma.

[0042] In certain exemplary embodiments, one or more allergic asthma signature genes include BC042385, AB209315, LOC100607117, BC035084, LOC145474, AX747853, TIMP1, NT5DC2, LOC541471, AREG, PTPN7, RUNDC3, XXYLT1, FAM159A, PTGDS, TESC, ITGB2-AS1, D0574721, and CLDN9. The group is selected from LOC100132052, AGAP7, NBEAL2, NTNG2, FLJ45445, KCNH3, POU51P3, OUG1, KIF21B, HSPA7, GAPT, BX6485Q2, PRR52, P1K3R6, LTC4S, CLEC11A, TRABD2A, DLGAP3, VDR, DKFZp686M11215, SIGLEC12, BC016361, BC052769, and RHOH. In a particular exemplary embodiment, one or more allergic asthma signature genes are selected from the group consisting of ASAP1-IT1, AX747757, BC042385, PABPC1P2, AB209315, AX748268, TCEAL5, CCL17, CCL13, CCL26, CLC, CACNG8, GPR82, GATA1, PRSS33, FFAR3, LGALS12, ASB2, PTGDR2, SIGLEC8, IL13, IL5, PTGDS, and RD3.

[0043] In certain exemplary embodiments, the anti-IL-33 antibody or its antigen-binding fragment comprises a heavy chain variable region (HCVR) containing the amino acid sequence of SEQ ID NO: 2 and a light chain variable region (LCVR) containing the amino acid sequence of SEQ ID NO: 10. In certain exemplary embodiments, the anti-IL-33 antibody or its antigen-binding fragment comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 18 and a light chain containing the amino acid sequence of SEQ ID NO: 20.

[0044] In a particular exemplary embodiment, the method further comprises administering to a subject an antibody or antigen-binding fragment thereof that specifically binds to IL-4R, wherein the antibody or antigen-binding fragment comprises three heavy chain complementary determinant region (HCDR) sequences, including SEQ ID NOs. 21, 22, and 23, and three light chain complementary determinant region (LCDR) sequences, including SEQ ID NOs. 24, 25, and 26.

[0045] In another embodiment, a method is provided for reducing the expression of any combination of type 2 inflammatory cytokines and type 2 chemokine signature genes in a subject having allergic asthma, comprising the step of administering to the subject an antibody or antigen-binding fragment thereof that specifically binds to interleukin-33 (IL-33) and comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs. 4, 6, and 8, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs. 12, 14, and 16. In one embodiment, an antibody or antigen-binding fragment thereof that specifically binds to interleukin-33 (IL-33) and comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs. 4, 6, and 8, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs. 12, 14, and 16 is provided for reducing the expression of any combination of type 2 inflammatory cytokines and type 2 chemokine signature genes in a subject having allergic asthma.

[0046] In a particular exemplary embodiment, the type 2 inflammatory cytokine and chemokine signature genes are selected from the group consisting of IL-5, CCL1, IL-13, GATA2, CCL26, FCER2, CACNG8, CLC, GATA1, LGALS12, SIGLEC8, GGT5, CCL17, and MMP10. In a particular exemplary embodiment, one or more type 2 inflammatory cytokine and chemokine signature genes are selected from the group consisting of IL-5, CCL1, IL-13, CCL26, FCER2, SIGLEC8, GGT5, and CCL17.

[0047] In certain exemplary embodiments, the anti-IL-33 antibody or its antigen-binding fragment comprises a heavy chain variable region (HCVR) containing the amino acid sequence of SEQ ID NO: 2 and a light chain variable region (LCVR) containing the amino acid sequence of SEQ ID NO: 10. In certain exemplary embodiments, the anti-IL-33 antibody or its antigen-binding fragment comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 18 and a light chain containing the amino acid sequence of SEQ ID NO: 20.

[0048] In a particular exemplary embodiment, the method further comprises administering to a subject an antibody or antigen-binding fragment thereof that specifically binds to IL-4R, wherein the antibody or antigen-binding fragment comprises three heavy chain complementary determinant region (HCDR) sequences, including SEQ ID NOs. 21, 22, and 23, and three light chain complementary determinant region (LCDR) sequences, including SEQ ID NOs. 24, 25, and 26.

[0049] In another embodiment, a method is provided for reducing the expression of one or more eosinophil signature genes in a subject having allergic asthma, comprising the step of administering to the subject an antibody or antigen-binding fragment thereof that specifically binds to interleukin-33 (IL-33) and comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs: 4, 6, and 8, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs: 12, 14, and 16. In one embodiment, an antibody or antigen-binding fragment thereof that specifically binds to interleukin-33 (IL-33) and comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs: 4, 6, and 8, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs: 12, 14, and 16 is provided for use in reducing the expression of one or more eosinophil signature genes in a subject having allergic asthma.

[0050] In a particular exemplary embodiment, one or more eosinophil signature genes are selected from the group consisting of IL1RL1, ADARB1, SIGLEC8, ASB2, VSTM1, SYNE1, CLC, PTPN7, and HDC.

[0051] In certain exemplary embodiments, the anti-IL-33 antibody or its antigen-binding fragment comprises a heavy chain variable region (HCVR) containing the amino acid sequence of SEQ ID NO: 2 and a light chain variable region (LCVR) containing the amino acid sequence of SEQ ID NO: 10. In certain exemplary embodiments, the anti-IL-33 antibody or its antigen-binding fragment comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 18 and a light chain containing the amino acid sequence of SEQ ID NO: 20.

[0052] In a particular exemplary embodiment, the method further comprises administering to a subject an antibody or antigen-binding fragment thereof that specifically binds to IL-4R, wherein the antibody or antigen-binding fragment comprises three heavy chain complementary determinant region (HCDR) sequences, including SEQ ID NOs. 21, 22, and 23, and three light chain complementary determinant region (LCDR) sequences, including SEQ ID NOs. 24, 25, and 26.

[0053] In another embodiment, in a subject having allergic asthma, one or more types of 2 A method for reducing the expression of type 2 inflammatory signature genes is provided, comprising the step of administering to a subject an antibody or antigen-binding fragment thereof that specifically binds to interleukin-33 (IL-33) and comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs. 4, 6, and 8, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs. 12, 14, and 16. In one embodiment, an antibody or antigen-binding fragment thereof that specifically binds to interleukin-33 (IL-33) and comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs. 4, 6, and 8, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs. 12, 14, and 16 is provided for use in a subject having allergic asthma to reduce the expression of one or more type 2 inflammatory signature genes.

[0054] In a particular exemplary embodiment, one or more type 2 inflammatory signature genes are selected from the group consisting of IL-4, IL-13, CCL26, CCL13, CCL17, CCL11, POSTN, IL-5, and IL-9.

[0055] In certain exemplary embodiments, the anti-IL-33 antibody or its antigen-binding fragment comprises a heavy chain variable region (HCVR) containing the amino acid sequence of SEQ ID NO: 2 and a light chain variable region (LCVR) containing the amino acid sequence of SEQ ID NO: 10. In certain exemplary embodiments, the anti-IL-33 antibody or its antigen-binding fragment comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 18 and a light chain containing the amino acid sequence of SEQ ID NO: 20.

[0056] In a particular exemplary embodiment, the method further comprises administering to a subject an antibody or antigen-binding fragment that specifically binds to IL-4R, wherein the antibody or antigen-binding fragment comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs. 21, 22, and 23, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs. 24, 25, and 26.

[0057] In another embodiment, a method is provided for reducing cytokine or chemokine levels in a subject having allergic asthma, comprising the step of administering to the subject an antibody or antigen-binding fragment thereof that specifically binds to the interleukin-4 receptor (IL-4R) and comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs. 21, 22, and 23, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs. 24, 25, and 26. In one embodiment, an antibody or antigen-binding fragment thereof that specifically binds to the interleukin-4 receptor (IL-4R) and comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs. 21, 22, and 23, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs. 24, 25, and 26 is provided for use in reducing cytokine or chemokine levels in a subject having allergic asthma.

[0058] In certain exemplary embodiments, the cytokines are one or both of IL-13 and IL-5.

[0059] In a particular exemplary embodiment, the cytokine or chemokine is selected from the group consisting of TNFα, TARC, PARC, CCL1, CCL26, FCERV2, SIGLEC8, CCL17, and eotaxin-3.

[0060] In certain exemplary embodiments, serum levels of sST2, IL-33, calcitonin, or MMP12 are reduced in the subjects. In certain exemplary embodiments, serum levels of CCL26, CCL17, or SIGLEC8 are reduced in the subjects.

[0061] In certain exemplary embodiments, the antibody or its antigen-binding fragment comprises a heavy chain variable region (HCVR) containing the amino acid sequence of SEQ ID NO: 27 and a light chain variable region (LCVR) containing the amino acid sequence of SEQ ID NO: 28. In certain exemplary embodiments, the antibody or its antigen-binding fragment comprises dupilumab.

[0062] In a particular exemplary embodiment, the method further comprises administering to a subject an antibody or antigen-binding fragment that specifically binds to IL-33, wherein the antibody or antigen-binding fragment comprises three heavy chain complementary determinant region (HCDR) sequences, including SEQ ID NOs: 4, 6, and 8, and three light chain complementary determinant region (LCDR) sequences, including SEQ ID NOs: 12, 14, and 16.

[0063] In another embodiment, a method is provided for reducing the expression of one or more allergic asthma signature genes in a subject having allergic asthma, comprising the step of administering to the subject an antibody or antigen-binding fragment thereof that specifically binds to the interleukin-4 receptor (IL-4R) and comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs. 21, 22, and 23, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs. 24, 25, and 26. In one embodiment, an antibody or antigen-binding fragment thereof that specifically binds to the interleukin-4 receptor (IL-4R) and comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs. 21, 22, and 23, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs. 24, 25, and 26 is provided for use in reducing the expression of one or more allergic asthma signature genes in a subject having allergic asthma.

[0064] In certain exemplary embodiments, one or more allergic asthma signature genes include BC042385, AB209315, LOC100607117, BC035084, LOC145474, AX747853, TIMP1, NT5DC2, LOC541471, AREG, PTPN7, RUNDC3, XXYLT1, FAM159A, PTGDS, TESC, ITGB2-AS1, D0574721, and CLDN9. The group is selected from LOC100132052, AGAP7, NBEAL2, NTNG2, FLJ45445, KCNH3, POU51P3, OUG1, KIF21B, HSPA7, GAPT, BX6485Q2, PRR52, P1K3R6, LTC4S, CLEC11A, TRABD2A, DLGAP3, VDR, DKFZp686M11215, SIGLEC12, BC016361, BC052769, and RHOH. In a particular exemplary embodiment, one or more allergic asthma signature genes are selected from the group consisting of ASAP1-IT1, AX747757, BC042385, PABPC1P2, AB209315, AX748268, TCEAL5, CCL17, CCL13, CCL26, CLC, CACNG8, GPR82, GATA1, PRSS33, FFAR3, LGALS12, ASB2, PTGDR2, SIGLEC8, IL13, IL5, PTGDS, and RD3.

[0065] In certain exemplary embodiments, the antibody or its antigen-binding fragment comprises a heavy chain variable region (HCVR) containing the amino acid sequence of SEQ ID NO: 27 and a light chain variable region (LCVR) containing the amino acid sequence of SEQ ID NO: 28. In certain exemplary embodiments, the antibody or its antigen-binding fragment comprises dupilumab.

[0066] In a particular exemplary embodiment, the method further comprises administering to a subject an antibody or antigen-binding fragment that specifically binds to IL-33, wherein the antibody or antigen-binding fragment comprises three heavy chain complementary determinant region (HCDR) sequences, including SEQ ID NOs: 4, 6, and 8, and three light chain complementary determinant region (LCDR) sequences, including SEQ ID NOs: 12, 14, and 16.

[0067] In another embodiment, a method is provided for reducing the expression of any combination of type 2 inflammatory cytokines and type 2 chemokine signature genes in a subject having allergic asthma, comprising the step of administering to the subject an antibody or antigen-binding fragment thereof that specifically binds to the interleukin-4 receptor (IL-4R) and comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs. 21, 22, and 23, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs. 24, 25, and 26. In one embodiment, an antibody or antigen-binding fragment thereof that specifically binds to the interleukin-4 receptor (IL-4R) and comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs. 21, 22, and 23, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs. 24, 25, and 26 is provided for use in reducing any combination of type 2 inflammatory cytokines and type 2 chemokine signature genes in a subject having allergic asthma.

[0068] In a particular exemplary embodiment, the type 2 inflammatory cytokine and chemokine signature genes are selected from the group consisting of IL-5, CCL1, IL-13, GATA2, CCL26, FCER2, CACNG8, CLC, GATA1, LGALS12, SIGLEC8, GGT5, CCL17, and MMP10. In a particular exemplary embodiment, one or more type 2 inflammatory cytokine and chemokine signature genes are selected from the group consisting of IL-5, CCL1, IL-13, CCL26, FCER2, SIGLEC8, GGT5, and CCL17.

[0069] In certain exemplary embodiments, the antibody or its antigen-binding fragment comprises a heavy chain variable region (HCVR) containing the amino acid sequence of SEQ ID NO: 27 and a light chain variable region (LCVR) containing the amino acid sequence of SEQ ID NO: 28. In certain exemplary embodiments, the antibody or its antigen-binding fragment comprises dupilumab.

[0070] In a particular exemplary embodiment, the method further comprises administering to a subject an antibody or antigen-binding fragment that specifically binds to IL-33, wherein the antibody or antigen-binding fragment comprises three heavy chain complementary determinant region (HCDR) sequences, including SEQ ID NOs: 4, 6, and 8, and three light chain complementary determinant region (LCDR) sequences, including SEQ ID NOs: 12, 14, and 16.

[0071] In another embodiment, a method is provided for reducing the expression of one or more eosinophil signature genes in a subject having allergic asthma, comprising the step of administering to the subject an antibody or antigen-binding fragment thereof that specifically binds to the interleukin-4 receptor (IL-4R) and comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs. 21, 22, and 23, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs. 24, 25, and 26. In one embodiment, an antibody or antigen-binding fragment thereof that specifically binds to the interleukin-4 receptor (IL-4R) and comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs. 21, 22, and 23, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs. 24, 25, and 26 is provided for use in reducing the expression of one or more eosinophil signature genes in a subject having allergic asthma.

[0072] In a particular exemplary embodiment, one or more eosinophil signature genes are selected from the group consisting of IL1RL1, ADARB1, SIGLEC8, ASB2, VSTM1, SYNE1, CLC, PTPN7, and HDC.

[0073] In a particular exemplary embodiment, the antibody or its antigen-binding fragment is SEQ ID NO: It includes a heavy chain variable region (HCVR) containing the amino acid sequence of 27 and a light chain variable region (LCVR) containing the amino acid sequence of SEQ ID NO: 28. In certain exemplary embodiments, the antibody or its antigen-binding fragment comprises dupilumab.

[0074] In a particular exemplary embodiment, the method further comprises administering to a subject an antibody or antigen-binding fragment that specifically binds to IL-33, wherein the antibody or antigen-binding fragment comprises three heavy chain complementary determinant region (HCDR) sequences, including SEQ ID NOs: 4, 6, and 8, and three light chain complementary determinant region (LCDR) sequences, including SEQ ID NOs: 12, 14, and 16.

[0075] In another embodiment, a method is provided for reducing the expression of one or more type 2 inflammatory signature genes in a subject having allergic asthma, comprising the step of administering to the subject an antibody or antigen-binding fragment thereof that specifically binds to the interleukin-4 receptor (IL-4R) and comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs. 21, 22, and 23, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs. 24, 25, and 26. In one embodiment, an antibody or antigen-binding fragment thereof that specifically binds to the interleukin-4 receptor (IL-4R) and comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs. 21, 22, and 23, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs. 24, 25, and 26 is provided for use in reducing the expression of one or more type 2 inflammatory signature genes in a subject having allergic asthma.

[0076] In a particular exemplary embodiment, one or more type 2 inflammatory signature genes are selected from the group consisting of IL-4, IL-13, CCL26, CCL13, CCL17, CCL11, POSTN, IL-5, and IL-9.

[0077] In certain exemplary embodiments, the antibody or its antigen-binding fragment comprises a heavy chain variable region (HCVR) containing the amino acid sequence of SEQ ID NO: 27 and a light chain variable region (LCVR) containing the amino acid sequence of SEQ ID NO: 28. In certain exemplary embodiments, the antibody or its antigen-binding fragment comprises dupilumab.

[0078] In a particular exemplary embodiment, the method further comprises administering to a subject an antibody or antigen-binding fragment that specifically binds to IL-33, wherein the antibody or antigen-binding fragment comprises three heavy chain complementary determinant region (HCDR) sequences, including SEQ ID NOs: 4, 6, and 8, and three light chain complementary determinant region (LCDR) sequences, including SEQ ID NOs: 12, 14, and 16.

[0079] The aforementioned and other features and advantages of the present invention will be better understood from the following detailed description of exemplary embodiments, which will be combined with the accompanying drawings. The files of the present invention include at least one drawing / photograph drawn in color. A copy of the present invention with the color drawing / photograph will be provided by the Office upon invoicing and payment of the required fees. [Brief explanation of the drawing]

[0080] [Figure 1]This figure shows a flowchart of an 8-week trial designed to evaluate the efficacy of treatment for bronchial allergen loading (BAC)-induced airway inflammation. The flowchart illustrates the following events: Sputum collection at baseline (immediately before BAC), after BAC (8 and 24 hours after BAC), at screening (before treatment), and at weeks 4 and 8 after the start of treatment. Changes in the sputum inflammation signature induced by BAC are evaluated by comparing the sputum signature at baseline and after BAC at screening (screening change), at week 4 (week 4 change), and at week 8 (week 8 change) after the start of treatment. The efficacy of treatment in the sputum inflammation signature is evaluated by determining the difference between the BAC-induced screening change and the BAC-induced week 4 change (change from screening to week 4), and the difference between the BAC-induced screening change and the BAC-induced week 8 change (change from screening to week 8). [Figure 2] This figure shows the charts and graphs from Part 1 of this study. Eligible patients (up to 32 in total) were randomized in a 1:1:1:1 ratio to receive either REGN3500, dupilumab, a combination of REGN3500 and dupilumab, or placebo. [Figure 3] This figure shows the charts and graphs from Part 2 of this study. Approximately six patients were administered fluticasone propionate by inhalation at a dose of 500 μg per day (2 puffs of 250 μg) twice daily for 4 days, starting on day 1 (a total of 8 doses). [Figure 4] This diagram illustrates the mechanism of action of IL-33 as an initiator and amplifier of innate and adaptive immunity. As shown in Figure 4, IL-33 is released after tissue damage. [Figure 5]This figure presents data showing that anti-IL-33 treatment reduces inflammation in a model of chronic dust mite disease (HDM) pneumonia. The figure demonstrates that anti-IL-33 treatment suppresses pro-inflammatory cytokines and chemokines. The data presents levels of pulmonary eosinophils and neutrophils in HDM models with and without anti-IL-33 treatment. A heatmap of the pulmonary cytokine gene panel presents levels of hIL-4, IL-5, IL-1b, TNFα, IFNg, GROa, and MCP-1. Alveolar SMA test data are similarly presented. [Figure 6] This diagram illustrates the mechanism of action of anti-IL-33 in reducing type 1 and type 2 inflammation. As demonstrated in this diagram, IL-33 promotes persistent and exacerbating pneumonia and remodeling. Anti-IL-33 reduces multiple components of chronic HDM-driven lung inflammation. [Figure 7] This figure shows a schematic diagram of the trial of Example 1 described herein. This diagram illustrates the bronchial allergen loading procedure in patients with mild asthma. Subjects are treated with placebo, placebo and REGN3500, dupilumab and placebo, or dupilumab and REGN3500. The effect of each treatment is determined by RNA sequencing of sputum cells after the patient inhales hypertonic saline and expectorates sputum. [Figure 8]This figure shows the expression of various signature genes associated with type 2 inflammation before allergen loading, 8 hours after allergen loading, and 24 hours after allergen loading. These results indicate that the top genes induced by bronchial allergen loading at the time of screening enhance type 2 inflammation, and that specific genes of interest include, but are not limited to, IL-4, IL-5, IL-13, IL-9, IL1RL1 (IL-33 receptor), Eot-3 (CCL26), TARC (CCL17), and FCER2. This list was obtained by screening for a mean relative change of more than 10 times and an FDR of less than 0.05. Allergen loading signals were reproducible, but their magnitude differed between groups. The identified supergenes (listed from top to bottom) included MMP10, WNT5A, CO1B, CD1A, CCL1, CCL17, PPP1R14A, IL-9, IL-5, IL-13, FCER2, CCL26, K3AA1755, GGT5, SIGLEC8, LGALS12, GATA1, CLC, CACNG8, BC015656, AKX05132, FFAR3, CACH1, IL1RL1, HPH4, CC5AML, GATA2, TAL1, HDC, NTRX1, and IL-4. [Figure 9] This figure shows the top type II inflammatory cytokine and chemokine signature genes induced by allergen loading and suppressed by REGN3500 at 8 or 24 hours (FC > 12 or p(adj) > 0.05). These results indicate that REGN3500 suppressed type II inflammatory cytokines and chemokines, including IL-5, IL-13, TARC, and eotaxin-3. Other genes of interest suppressed by REGN3500 and induced by bronchial allergen loading included CCL1, a ligand for CCR8 which attracts activated Th2 and Treg cells, CCL26, FCER2, SIGLEC8, and CCL17. [Figure 10]This figure shows the eosinophil gene signatures used to evaluate the therapeutic effect on sputum eosinophil levels. A set of 10 genes showed a high correlation with sputum eosinophil counts before and after allergen loading. This gene set includes ADARB1, ASB2, CLC, GLOD5, HDC, IL1RL1, PTPN7, SIGLEC8, SYNE1, and VSTM1. mRNA signatures were used to improve statistical performance in verifying the therapeutic effect size (fluticasone) against sputum % eosinophils. The genes were not exclusive to eosinophils, e.g., SIGLEC8 (expressed in eosinophils, basophils, and mast cells), HDC (expressed in mast cells), and VSTM1 (expressed in myeloid cells). [Figure 11] This figure shows the effect of REGN3500 on eosinophil signature genes in sputum, demonstrating suppression of eosinophil signature genes. Figure 11 also shows that REGN3500 had no effect on neutrophil signature genes. The genes presented in the data are ADARB1, ASB2, CLC, HDC, IL1RL1, PTPN7, SIGLEC8, SYNE1, and VSTM1. [Figure 12] This figure shows the effect of REGN3500 on type 2 inflammatory signature genes in sputum, demonstrating suppression of type 2 inflammatory signature genes. Figure 12 also shows that type 1 inflammatory signature genes were not induced by allergen loading. The data presented are for IL4, IL13, CCL26, CCL13, CCL17, CCL11, POSTN, IL5, and IL9. [Figure 13A] This figure presents data illustrating the mechanism by which increasing IL-33 levels promotes a self-persistent amplification loop that stimulates tissue deterioration. Figures 13A and 13B include data obtained in the HDM model. Increasing IL-33 levels promoted a self-persistent amplification loop that stimulates tissue deterioration. [Figure 13B]This figure presents data illustrating the mechanism by which increasing IL-33 levels promotes a self-persistent amplification loop that stimulates tissue deterioration. Figures 13A and 13B include data obtained in the HDM model. Increasing IL-33 levels promoted a self-persistent amplification loop that stimulates tissue deterioration. [Figure 14] This figure shows eosinophil gene signature scores across treatment arms. These arms include placebo, fluticasone, dupilumab, REGN3500, and combination therapy with dupilumab and REGN3500. Results are shown before and after bronchial allergen loading. These results indicate that both dupilumab and REGN3500 were able to reduce eosinophil gene signature scores after bronchial allergen loading. Combination therapy with dupilumab and REGN3500 was the most effective treatment for reducing eosinophil gene signature scores after bronchial allergen loading. [Figure 15] This figure shows the type 2 signature score across treatment arms. These arms include placebo, fluticasone, dupilumab, REGN3500, and combination therapy with dupilumab and REGN3500. Results are shown before and after bronchial allergen loading. These results indicate that the reduction in type 2 signature score is less pronounced in the REGN3500 treatment arm than in the fluticasone treatment arm. [Figure 16]This figure shows allergic asthma signature genes affected by REGN3500 (in 8 and / or 24 hours). Results are presented at screening and treatment, which were performed after bronchial allergen loading. The genes tested, from top to bottom, are BC042385, AB209315, LOC100607117, BC035084, LOC145474, AX747853, TIMP1, NT5DC2, LOC541471, AREG, PTPN7, RUNDC3, XXYLT1, FAM159A, PTGDS, TESC, ITGB2-AS1, D0574721, CLDN9, and LOC1001320. This includes 52, AGAP7, NBEAL2, NTNG2, FLJ45445, KCNH3, POU51P3, OUG1, KIF21B, HSPA7, GAPT, BX6485Q2, PRR52, P1K3R6, LTC4S, CLEC11A, TRABD2A, DLGAP3, VDR, DKFZp686M11215, SIGLEC12, BC016361, BC052769, and RHOH. [Figure 17] This figure shows the top allergic asthma signature genes induced by 24-hour bronchial allergen challenge and suppressed by REGN3500. Results are presented for screening and treatment, which were performed after bronchial allergen challenge. The genes shown, from top to bottom, include ASAP1-IT1, AX747757, BC042385, PABPC1P2, AB209315, AX748268, TCEAL5, CCL17, CCL13, CCL26, CLC, CACNG8, GPR82, GATA1, PRSS33, FFAR3, LGALS12, ASB2, PTGDR2, SIGLEC8, IL13, IL5, PTGDS, and RD3. [Modes for carrying out the invention]

[0081] Before describing the present invention, it should be understood that the present invention is not limited to the specific methods and experimental conditions described, as such methods and conditions may vary. It should also be understood that the scope of the present invention is limited only by the appended claims, and that the terms used herein are for the purpose of describing only specific embodiments and are not intended to limit them.

[0082] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art to which this invention pertains.

[0083] As used herein, the term “about” means, when used in reference to a specific enumerated number, that the value may vary by up to 1% from the enumerated value. For example, as used herein, the expression “about 100” includes 99 and 101, as well as all values ​​in between (e.g., 99.1, 99.2, 99.3, 99.4, etc.).

[0084] As used herein, the terms “to treat,” “to treat,” etc., mean to alleviate symptoms, to temporarily or permanently eliminate the causal relationship between symptoms and symptoms, or to prevent or delay the onset of symptoms of a specified disorder or condition.

[0085] Any methods and materials similar to or equivalent to those described herein may be used in the practice of the present invention, but typical methods and materials are described herein. All publications mentioned herein are incorporated herein in their entirety by reference.

[0086] Methods to reduce the incidence of allergic asthma exacerbations The present invention includes a method for reducing the incidence of allergic asthma exacerbations in subjects requiring it, comprising the step of administering a pharmaceutical composition comprising an interleukin-33 (IL-33) antagonist. Also provided is an interleukin-33 (IL-33) antagonist for use in reducing the incidence of allergic asthma exacerbations in subjects requiring it. The present invention also includes a method for reducing the incidence of allergic asthma exacerbations in subjects requiring it, comprising the step of administering a pharmaceutical composition comprising an interleukin-4 receptor (IL-4R) antagonist. Also provided is an interleukin-4 receptor (IL-4R) antagonist for use in reducing the incidence of allergic asthma exacerbations in subjects requiring it. The method characterized in the present invention is interleukin- The present invention further comprises administering a first therapeutic composition comprising an IL-33 (IL-33) antagonist and a second therapeutic composition comprising an interleukin-4 receptor (IL-4R) antagonist to a subject in need. Also provided are interleukin-33 (IL-33) antagonists and interleukin-4 receptor (IL-4R) antagonists for use in reducing the incidence of allergic asthma exacerbations in subjects in need. According to certain embodiments, the IL-33 antagonist is an antibody or its antigen-binding fragment that specifically binds to IL-33. Exemplary anti-IL-33 antibodies that can be used in the context of the methods or uses characterized in the present invention are described herein. According to certain embodiments, the IL-4R antagonist is an antibody or its antigen-binding fragment that specifically binds to IL-4R. Exemplary anti-IL-4R antibodies that can be used in the context of the methods or uses characterized in the present invention are described herein.

[0087] As used herein, the expression “asthma exacerbation” means an increase in the severity and / or frequency and / or duration of one or more symptoms or signs of asthma. “Asthma exacerbation” also includes any worsening of the respiratory health condition of the subject that requires and / or can be treated with therapeutic intervention for asthma (e.g., steroid therapy, inhaled corticosteroid therapy, hospitalization, etc.). There are two types of asthma exacerbation events: loss of asthma control (LOAC) events and severe exacerbation events.

[0088] In a particular embodiment, a loss of asthma control (LOAC) event is defined as one or more of the following: (a) a decrease of 30% or more from baseline in morning PEF for two consecutive days; (b) six or more additional palliative puffs of salbutamol / albuterol or levosalbutamol / levalbuterol in a 24-hour period (compared to baseline) for two consecutive days; (c) an increase in ICS of four times or more the last prescribed ICS dose (or ≥50% of the ICS dose prescribed in V2 if discontinuation of basic therapy is completed); (d) use of systemic (oral and / or parenteral) corticosteroid therapy; or (e) hospitalization or emergency room visit due to asthma.

[0089] In certain cases, an asthma exacerbation can be classified as a “severe asthma exacerbation event.” A severe asthma exacerbation event refers to an event requiring immediate intervention in the form of treatment with systemic or inhaled corticosteroids at a dose four times or more greater than the dose administered prior to the event. According to certain embodiments, a severe asthma exacerbation event is defined as an asthma exacerbation requiring systemic corticosteroid use for three days or more; or requiring hospitalization or emergency room visit due to asthma. Therefore, the general expression “asthma exacerbation” includes and encompasses more detailed subcategories of “severe asthma exacerbation.” Thus, it includes methods to reduce the incidence of severe asthma exacerbations in patients who require it.

[0090] A “reduced incidence” of asthma exacerbations means that subjects receiving a pharmaceutical composition containing an IL-4R antagonist experience fewer allergic asthma exacerbations after treatment than before treatment (i.e., at least one fewer exacerbation), or do not experience any allergic asthma exacerbations at least four weeks after the start of treatment with the pharmaceutical composition (e.g., 4, 6, 8, 12, 14 weeks or more). Alternatively, a “reduced incidence” of asthma exacerbations means that after administration of the pharmaceutical composition, the likelihood of a subject experiencing an asthma exacerbation is reduced by at least 10% (e.g., 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50% or more) compared to subjects not receiving the pharmaceutical composition.

[0091] The present invention includes a step of administering a pharmaceutical composition containing an IL-4R antagonist to a target, and one or more maintenance doses of inhaled corticosteroids (ICS) and / or This includes a method for reducing the incidence of allergic asthma exacerbations in subjects requiring it, comprising the step of administering to a subject one or more maintenance doses of a second controller, such as a long-acting β-agonist (LABA) or a leukotriene receptor antagonist (LTA). Also provided is a pharmaceutical composition for use comprising an interleukin-4 receptor (IL-4R) antagonist in combination with one or more maintenance doses of an inhaled corticosteroid (ICS) and / or a second controller, such as one or more maintenance doses of a long-acting β-agonist (LABA) or a leukotriene receptor antagonist (LTA), for reducing the incidence of allergic asthma exacerbations in subjects requiring it. The present invention includes a method for reducing the incidence of allergic asthma exacerbations in subjects requiring its use, comprising the steps of administering a pharmaceutical composition comprising an IL-33 antagonist to a subject and administering one or more maintenance doses of an inhaled corticosteroid (ICS) and / or a second controller, such as one or more maintenance doses of a long-acting β-agonist (LABA) or a leukotriene receptor antagonist (LTA), to the subject. Also provided is a pharmaceutical composition comprising an interleukin-33 receptor (IL-33) antagonist for use in combination with one or more maintenance doses of an inhaled corticosteroid (ICS) and / or a second controller, such as one or more maintenance doses of a long-acting β-agonist (LABA) or a leukotriene receptor antagonist (LTA), in order to reduce the incidence of allergic asthma exacerbations in subjects requiring its use.Suitable ICS include, but are not limited to, fluticasone (e.g., fluticasone propionate, e.g., Flovent®), budesonide, mometasone (e.g., mometasone furoate, e.g., Asmanex®), flunisolide (e.g., Aerobid®), dexamethasone acetate / phenobarbital / theophylline (e.g., Azmacort®), and beclomethasone dipropionate HFA (Qvar®). Suitable LABAs include, but are not limited to, salmeterol (e.g., Serevent®) and formoterol (e.g., Foradil®). Suitable LTAs include, but are not limited to, montelukast (e.g., Singulaire®) and zafirlukast (e.g., Accolate®).

[0092] The present invention includes a method for reducing the incidence of allergic asthma exacerbations in subjects requiring its use, comprising the steps of administering a pharmaceutical composition comprising one or both of an IL-4R antagonist and an IL-33 antagonist to a subject, and administering one or more palliative agents to a subject to eliminate or reduce one or more asthma-related symptoms. Also provided is a pharmaceutical composition for use comprising one or both of an IL-4R antagonist and an IL-33 antagonist, in combination with one or more maintenance doses of an inhaled corticosteroid (ICS) and / or a second controller, such as one or more maintenance doses of a long-acting β-agonist (LABA) or a leukotriene receptor antagonist (LTA), to reduce the incidence of allergic asthma exacerbations in subjects requiring its use. The present invention includes a method for reducing the incidence of allergic asthma exacerbations in subjects requiring it, comprising the steps of administering a pharmaceutical composition comprising one or both of an IL-4R antagonist and an IL-33 antagonist to a subject, and administering one or more palliative agents to a subject to eliminate or reduce one or more asthma-related symptoms. Also provided is a pharmaceutical composition for use, comprising one or both of an IL-4R antagonist and an IL-33 antagonist, in combination with one or more palliative agents to eliminate or reduce one or more asthma-related symptoms, thereby reducing the incidence of allergic asthma exacerbations in subjects requiring it. Suitable palliative agents include, but are not limited to, immediate These include active β2-adrenergic receptor agonists, such as albuterol (i.e., salbutamol, e.g., Proventil®, Ventolin®, Xopenex®, etc.), pirbuterol (e.g., Maxair®), and metaproterenol (e.g., Alupent®).

[0093] Methods to improve asthma-related parameters The present invention also includes a method for improving one or more asthma-related parameters in subjects requiring it, the method comprising the step of administering a pharmaceutical composition comprising an IL-33 antagonist to a subject. Also provided is a pharmaceutical composition comprising an IL-33 antagonist for use in improving one or more asthma-related parameters in subjects requiring it. The present invention further includes a method for improving one or more asthma-related parameters in subjects requiring it, the method comprising the step of administering a pharmaceutical composition comprising an IL-4R antagonist to a subject. Also provided is a pharmaceutical composition comprising an IL-4R antagonist for use in improving one or more asthma-related parameters in subjects requiring it. The present invention also includes a method for improving one or more asthma-related parameters in subjects requiring it, the method comprising the step of administering a first pharmaceutical composition comprising an IL-33 antagonist and a second pharmaceutical composition comprising an IL-4R antagonist to a subject. What is offered is a first pharmaceutical composition containing an IL-33 antagonist and a second pharmaceutical composition containing an IL-4R antagonist, for use in improving one or more asthma-related parameters in subjects requiring such improvement. (As previously stated) a reduction in the incidence of asthma exacerbations may correlate with an improvement in one or more asthma-related parameters; however, such a correlation is not necessarily observed in all cases.

[0094] Examples of "asthma-related parameters" include: (1) Percentage relative change from baseline (e.g., at week 12) in forced expiratory volume in one second (FEV1); (2) Percentage relative change from baseline (e.g., at week 12) as measured by forced expiratory rate in lung volume 25-75% (FEF25-75); (3) Annual rate of asthma control loss events during the treatment period; (4) Annual rate of severe exacerbation events during the treatment period; (5) Time to asthma control loss event during the treatment period; (6) Time to severe exacerbation event during the treatment period; (7) Time to asthma control loss event throughout the entire study period. (8) Time to an elephant; (9) Time to a severe exacerbation event throughout the entire study period; (10) i) Morning and evening asthma symptom scores, ii) ACQ-5 score, iii) AQLQ score, iv) Morning and evening PEF, v) Daily number of inhalations of salbutamol / albuterol or levosalmola / levalbuterol for symptom relief, vi) Change from baseline at week 12 in nocturnal awakenings; (11) i) 22 nasal sinus disease assessment items (SNOT-22), ii) Hospital Anxiety and Depression Score (HADS), iii) Change from baseline at weeks 12 and 24 in the EuroQual Questionnaire (EQ-5D-3L or EQ-5D-5L). "Improvement in asthma-related parameters" means an increase from baseline in one or more of the following: FEV1, AM PEF, or PM PEF, and / or a decrease from baseline in one or more of the following: daily albuterol / levalbuterol use, ACQ5 score, mean nocturnal wakefulness, or SNOT-22 score. As used herein, with respect to asthma-related parameters, the term "baseline" means the numerical value of asthma-related parameters for a patient before or at the time of administration of a pharmaceutical composition containing an IL-33 antagonist, the numerical value of asthma-related parameters for a patient before or at the time of administration of a pharmaceutical composition containing an IL-4R antagonist, or the numerical value of asthma-related parameters for a patient before or at the time of administration to a target of the first pharmaceutical composition containing an IL-33 antagonist and the second pharmaceutical composition containing an IL-4R antagonist.

[0095] To determine whether asthma-related parameters are “improving,” the parameters are quantified at baseline and at points after administration of the pharmaceutical compositions described herein. For example, asthma-related parameters can be measured on days 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, and 14 after initial treatment with the pharmaceutical composition, or at weeks 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or later. The difference between the parameter values ​​at a specific point in time after the start of treatment and the baseline parameter values ​​is used to determine whether there has been an "improvement" (e.g., an increase or decrease, depending on the specific parameter being measured) in the asthma-related parameters.

[0096] "Asthma-related parameters" also include changes (i.e., increases or decreases) in the expression of one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more) allergic asthma "signature genes" compared to baseline expression. As used herein, with respect to signature genes, the term "baseline" means the numerical value of the expression of one or more signature genes in a patient before or at the time of administration of a pharmaceutical composition containing an IL-33 antagonist, the numerical value of the expression of one or more signature genes in a patient before or at the time of administration of a pharmaceutical composition containing an IL-4R antagonist, or the numerical value of the expression of one or more signature genes in a patient before or at the time of administration to a subject of a first pharmaceutical composition containing an IL-33 antagonist and a second pharmaceutical composition containing an IL-4R antagonist. According to a particular exemplary embodiment, a patient is selected for treatment with a pharmaceutical composition comprising an IL-33 antagonist, a pharmaceutical composition comprising an IL-4R antagonist, or a first pharmaceutical composition comprising an IL-33 antagonist and a second pharmaceutical composition comprising an IL-4R antagonist, based on an increase or decrease in the expression of one or more signature genes.

[0097] In a particular exemplary embodiment, the expression of one or more signature genes is reduced compared to a baseline expression level, for example, the expression is reduced to a level where the baseline expression level is approximately 99%, approximately 98%, approximately 97%, approximately 96%, approximately 95%, approximately 94%, approximately 93%, approximately 92%, approximately 91%, approximately 90%, approximately 85%, approximately 80%, approximately 75%, approximately 70%, approximately 65%, approximately 60%, approximately 55%, approximately 50%, approximately 45%, approximately 40%, approximately 35%, approximately 30%, approximately 25%, approximately 20%, approximately 15%, approximately 10%, or approximately 5%, or any range between these numbers.

[0098] In a particular exemplary embodiment, the expression of one or more signature genes is increased compared to a baseline expression level, for example, the expression is reduced to a level where the baseline expression level is approximately 105%, 110%, 120%, 125%, 130%, 135%, 140%, 145%, 150%, 175%, 200%, 225%, 250%, 275%, 300%, 400%, or 500% or more, or any range between these numbers.

[0099] Suitable signature genes include allergic asthma signature genes, but are not limited to these; they also include type 2 inflammatory signature genes, cytokine signature genes, chemokine signature genes, eosinophil signature genes, and others.

[0100] Exemplary allergic asthma signature genes are shown in Figures 16 and 17, and This is not limited to just these, but also includes BC042385, AB209315, LOC100607117, BC035084, LOC145474, AX747853, TIMP1, NT5DC2, LOC541471, AREG, PTPN7, RUNDC3, XXYLT1, FAM159A, PTGDS, TESC, ITGB2-AS1, D0574721, CLDN9, LOC100132052, AGAP7, NBEAL2, NTNG2, FLJ45445, KCNH3, POU51P3, OUG1, KIF21B, HSPA7, GAPT, BX6485Q2, PRR52, P This includes 1K3R6, LTC4S, CLEC11A, TRABD2A, DLGAP3, VDR, DKFZp686M11215, SIGLEC12, BC016361, BC052769, RHOH, ASAP1-IT1, AX747757, BC042385, PABPC1P2, AB209315, AX748268, TCEAL5, CCL17, CCL13, CCL26, CLC, CACNG8, GPR82, GATA1, PRSS33, FFAR3, LGALS12, ASB2, PTGDR2, SIGLEC8, IL13, IL5, PTGDS, and RD3.

[0101] Exemplary type 2 inflammatory signature genes are shown in Figures 8 and 12, but are not limited to, IL-4, IL-5, IL-13, IL-9, IL1RL1 (IL-33 receptor), Eot-3 (CCL26), TARC (CCL17), FCER2, MMP10, WNT5A, CO1B, CD1A, CCL1, CCL17, PPP1R14A, IL-9, IL-5, IL-13, FCER2, CC This includes L26, K3AA1755, GGT5, SIGLEC8, LGALS12, GATA1, CLC, CACNG8, BC015656, AKX05132, FFAR3, CACH1, IL1RL1, HPH4, CC5AML, GATA2, TAL1, HDC, NTRX1, IL-4, IL4, IL13, CCL26, CCL13, CCL17, CCL11, POSTN, IL5, and IL9.

[0102] Exemplary cytokine signature genes and chemokine signature genes are shown in Figure 9, but are not limited to, IL-5, IL-13, TARC, eotaxin-3, CCL1, CCL26, FCER2, SIGLEC8, and CCL17.

[0103] Exemplary eosinophil signature genes, shown in Figure 11, include, but are not limited to, ADARB1, ASB2, CLC, HDC, IL1RL1, PTPN7, SIGLEC8, SYNE1, and VSTM1.

[0104] Signature genes, though not limited to these, can be detected in biological samples using any suitable means known in the art for detecting proteins, RNA (e.g., mRNA), and / or DNA, including, but not limited to, Northern blotting, Western blotting, Southern blotting, immunoprecipitation, in situ hybridization, PCR (e.g., RT-PCR), array technologies (e.g., Sequential Analysis of Gene Expression (SAGE), DNA microarrays, RNA sequencing, tiling arrays, etc.), and nuclease assays.

[0105] As used herein, “biological sample” includes, but is not limited to, cell cultures or extracts thereof; biopsy material obtained from mammals or extracts thereof; and blood, saliva, urine, feces, semen, tears, or other bodily fluids or extracts thereof. Exemplary biological samples include sputum and blood.

[0106] As used herein, the terms “to acquire” or “to obtain” mean to acquire ownership of a physical entity or value, such as a numerical value, by “directly acquiring” or “indirectly acquiring” a physical entity or value, such as an asthma-related parameter. "Direct acquisition" means performing a method (e.g., a synthesis or analytical method) to obtain a physical entity or value. "Indirect acquisition" means receiving a physical entity or value from another party or source (e.g., a third-party laboratory from which the physical entity or value is directly acquired). Direct acquisition of a physical entity includes performing a method that involves a physical change of a material existence, e.g., a physical change of starting materials. Exemplary changes include performing chemical reactions that involve creating a physical entity from two or more starting materials, shearing or fragmenting a substance, separating or purifying a substance, combining two or more separate entities into a mixture, or breaking or forming covalent or non-covalent bonds. Direct acquisition of a value includes performing a method that involves a physical change of a sample or another substance, e.g., performing an analytical method that involves a physical change of a substance, e.g., a sample, analyte, or reagent (sometimes referred to herein as "physical analysis").

[0107] Information obtained indirectly may be provided in the form of reports, supplied, for example, in written or electronic form, such as in online databases or applications ("App"). Reports or information may be provided, for example, by healthcare institutions, such as hospitals or clinics; or by healthcare providers, such as physicians or nurses.

[0108] Volume per second (FEV1) According to certain embodiments, administration or use of an IL-4R antagonist in a patient results in an increase in forced expiratory volume in one second (FEV1) from baseline. In some embodiments, administration or use of an IL-33 antagonist in a patient results in an increase in FEV1 from baseline. In other embodiments, administration or use of an IL-4R antagonist in combination with an IL-33 (IL-33) antagonist in a patient results in an increase in FEV1 from baseline. Methods for measuring FEV1 are known in the art. For example, FEV1 can be measured in a patient using a spirometer that conforms to the recommendations of the American Thoracic Society (ATS) / European Respiratory Society (ERS) 2005. The ATS / ERS Standardization of Spirometry can be used as a guideline. Spirometry is generally performed between 6 and 10 AM after at least 6 hours of albuterol retention. Pulmonary function tests are generally performed in a seated position, and the highest measurement is recorded for FEV1 (in liters).

[0109] According to certain embodiments, a method or use of treatment is provided that results in an increase of at least 0.05 L of FEV1 from baseline at 12 weeks after the initiation of treatment with a pharmaceutical composition comprising an anti-IL-33 antagonist, a pharmaceutical composition comprising an anti-IL-4R antagonist, or a pharmaceutical composition comprising an IL-33 antagonist and an IL-4R antagonist. For example, administration of an IL-33 antagonist, an IL-4R antagonist, or both an IL-33 antagonist and an IL-4R antagonist increases FEV1 from baseline to approximately 0.05L, 0.10L, 0.12L, 0.14L, 0.16L, 0.18L, 0.20L, 0.22L, 0.24L, 0.26L, 0.28L, 0.30L, 0.32L, 0.34L, 0.36L, 0.38L, 0.40L, 0.42L, 0.44L, 0.46L, 0.48L, 0.50L or more at week 12.

[0110] Bronchial allergen load According to several embodiments, administration or use of IL-33 antagonists in patients reduces bronchial allergen load (BAC)-induced pneumonia. According to several embodiments, administration or use of IL-4R antagonists in patients reduces BAC-induced pneumonia. According to several embodiments, administration or use of both IL-4R antagonists and IL-33 antagonists in patients reduces BAC-induced pneumonia. BAC is a model for testing asthma medications and has been used for over 30 years (D Iamant et al., Inhaled allergen bronchoprovocation tests. J Allergy Clin Immunol. 2013. Vol. 132: pp. 1045-1055 e1046; Fahy et al., Analysis of cellular and biochemical constituents of induced sputum after allergen challenge: a method for studying allergic airway inflammation. J Allergy Clin Immunol. 1994. Vol. 93: pp. 1031-1039; and Inman et al., Dose-dependent effects of inhaled mometasone furoate on Airway function and inflammation after allergen inhalation challenge. Am J Respir Crit Care Med. 2001. Vol. 164: pp. 569-574). BAC involves patient inhalation of an allergen that elicits a biphasic airway response, characterized by a rapid (30 minutes to 2 hours after allergen loading) and a slow (approximately 3-8 hours after allergen loading) decrease in FEV1. This model facilitates the assessment of the inflammatory response of allergies by measuring cellular content, cytokine production, and mRNA inflammatory signatures in bronchoalveolar lavage, bronchial biopsy, or induced sputum changes.

[0111] Sputum mRNA scale The induced sputum samples will be used in clinical trials for asthma to evaluate airway inflammation. In studies comparing sputum obtained from asthma patients with sputum obtained from normal controls, proteins and / or RNA were identified, including IL-33 and ST2 (Hamzaoui et al., Induced sputum levels of IL-33 and soluble ST2 in young asthmatic children. (J Asthma. 2013. Vol. 50: pp. 803-809) and Salter et al., IL-25 and IL-33 induce Type 2 inflammation in basophils from subjects with allergic asthma. Respir Res. 2016. Vol. 17: p. 5), eotaxin, and TARC (Heijink et al., Effect of ciclesonide treatment on allergen-induced changes in T cell regulation in asthma. Int Arch Allergy Immunol. 2008. Vol. 145: pp. 111-121) and Sekiya et al., Increased levels of a TH2-type CC chemokine). Thymus and activation-regulated chemokine (TARC)in serum and induced sputum of asthmatics. Allergy. 2002, Vol. 57: pp. 173-177), as well as IL-5 and IL-13 (Park et al., Interleukin-13 and interleukin-5 in induced sputum of eosinophilic bronchitis: comparison with asthma. Chest. 2005, Vol. 128: pp. 1921-1927, and Peters, MC, ZK Mekonnen, S. Yuan, NRB Hakta, PG Woodruff, and JV Fahy. Measures of gene expression in sputum cells can identify TH2-high and TH2-low subtypes of asthma.Elevated concentrations of sputum cytokines, such as IL-4, IL-5, and IL-13, were found (J Allergy Clin Immunol, 2014, Vol. 133: pp. 388-394). Sputum cytokines were elevated and associated with the presence and severity of asthma symptoms (Truyen et al., Evaluation of airway inflammation by quantitative Th1 / Th2 cytokine mRNA measurement in sputum of asthma patients. Thorax, 2006, 6). (Volume 1: pp. 202-208). In previous studies, BAC in patients with mild asthma caused a dramatic increase in the levels of type 2 cytokines, such as IL-13 and IL-5, in the lungs, up to approximately 10×. Treatment with inhaled corticosteroids significantly suppressed this BAC-mediated upregulation of mRNA levels of proteins and type 2 cytokines (Zuiker et al., Kinetics of TH2 biomarkers in sputum of asthmatics following inhaled allergen. Eur Clin Respir J. 2015. Vol. 2 and Zuiker et al., Sputum RNA signature in allergic asthmatics following allergen bronchoprovocation test. Eur Clin Respir J. 2016. Vol. 3: 31324).

[0112] According to some embodiments, administration or use of an IL-4R antagonist to a patient suppresses BAC-induced upregulation of mRNA levels of proteins and / or type 2 cytokines. According to some embodiments, administration or use of an IL-33 antagonist to a patient suppresses BAC-induced upregulation of mRNA levels of proteins and / or type 2 cytokines. According to some embodiments, administration or use of both an IL-4R antagonist and an IL-33 antagonist to a patient suppresses BAC-induced upregulation of mRNA levels of proteins and / or type 2 cytokines. According to several embodiments, administration or use of an IL-4R antagonist to a patient suppresses BAC-induced upregulation at the protein and / or mRNA level of any one of the following: CCL26, CCL17, SIGLEC8, IL-33, ST2, eotaxin, TARC, IL-4, IL-5, IL-13, ASAP1-IT1, AX747757, BC042385, PABPC1P2, AB209315, AX748268, TCEAL5, CCL13, CLC, CACNG8, GPR82, GATA1, PRSS33, FFAR3, LGALS12, ASB2, PTGDR2, PTGDS, or RD3. According to several embodiments, administration or use of an IL-33 antagonist to a patient suppresses BAC-induced upregulation at the protein and / or mRNA level of any one of the following: CCL26, CCL17, SIGLEC8, IL-33, ST2, eotaxin, TARC, IL-4, IL-5, IL-13, ASAP1-IT1, AX747757, BC042385, PABPC1P2, AB209315, AX748268, TCEAL5, CCL13, CLC, CACNG8, GPR82, GATA1, PRSS33, FFAR3, LGALS12, ASB2, PTGDR2, PTGDS, or RD3.According to several embodiments, administration or use of IL-4R antagonists and IL-33 antagonists to patients suppresses BAC-induced upregulation at the protein and / or mRNA level of any one of the following: CCL26, CCL17, SIGLEC8, IL-33, ST2, eotaxin, TARC, IL-4, IL-5, IL-13, ASAP1-IT1, AX747757, BC042385, PABPC1P2, AB209315, AX748268, TCEAL5, CCL13, CLC, CACNG8, GPR82, GATA1, PRSS33, FFAR3, LGALS12, ASB2, PTGDR2, PTGDS, or RD3.

[0113] Sputum cytokines and chemokines According to some embodiments, administration or use of an IL-4R antagonist to a patient suppresses BAC-induced increases in cytokines and chemokines associated with both the IL-33 and IL-4R pathways, including IL-13, IL-5, tumor necrosis factor-α (TNFα), TARC, pulmonary and activating modulated chemokines (PARC), CCL1, CCL26, FCER2, SIGLEC8, CCL17, and eotaxin-3. According to some embodiments, administration or use of an IL-33 antagonist to a patient suppresses BAC-induced increases in I The BAC-induced increase in cytokines and chemokines associated with both the IL-33 and IL-4R pathways, including L-13, IL-5, tumor necrosis factor-α (TNFα), TARC, lung and activating modulated chemokines (PARC), CCL1, CCL26, FCER2, SIGLEC8, CCL17, and eotaxin-3, is suppressed. According to several embodiments, the administration or use of IL-4R antagonists and IL-33 antagonists to patients suppresses the BAC-induced increase in cytokines and chemokines associated with both the IL-33 and IL-4R pathways, including IL-13, IL-5, tumor necrosis factor-α (TNFα), TARC, lung and activating modulated chemokines (PARC), CCL1, CCL26, FCER2, SIGLEC8, CCL17, and eotaxin-3. Previous studies have shown that cytokines and chemokines can be measured in sputum induced after BAC. Cytokines and chemokines associated with both the IL-33 and IL-4R pathways, including IL-13, IL-5, tumor necrosis factor-α (TNFα), TARC, pulmonary and activation-modulated chemokines (PARC), CCL1, CCL26, FCE2, SIGLEC8, CCL17, and eotaxin-3, are expected to be elevated after BAC.

[0114] Decrease in initial and delayed phase FEV1 after bronchial allergen exposure. Changes in lung function after BAC are a standard endpoint for most allergen loading tests evaluating the effects of inhaled corticosteroids. In sensitized patients, allergen inhalation elicits an acute phase response characterized by bronchoconstriction within 0–2 hours after exposure, also known as the early allergen response (EAR). This EAR is thought to primarily represent the release of pre-formed mast cell mediators and is usually unresponsive to steroids. The early allergen response is often followed by a late allergen response (LAR) which occurs approximately 3–8 hours after exposure. This LAR is seen in 50–60% of adult asthma patients. The LAR occurs concurrently with the initial influx of inflammatory cells and is generally responsive to steroids. According to some embodiments, administration or use of IL-4R antagonists in patients attenuates BAC-induced EAR or LAR, for example, as measured by FEV1. According to some embodiments, administration or use of an IL-33 antagonist to a patient attenuates BAC-induced EAR or LAR, for example, as measured by FEV1. According to some embodiments, administration or use of an IL-4R antagonist and an IL-33 antagonist to a patient attenuates BAC-induced EAR or LAR, for example, as measured by FEV1.

[0115] Exhaled Nitric Oxide Scale In BAC, sputum eosinophil counts have been shown to increase in asthma patients exhibiting a delayed-phase response. While a correlation between sputum eosinophil counts and FeNO has been reported, FeNO is not an eosinophil-specific marker and can be present in non-eosinophilic inflammation (Haldar et al., Mepolizumab and exacerbations of refractory eosinophilic asthma. N Engl J Med. 2009. Vol. 360: pp. 973-984). Furthermore, mRNA levels in bronchial tissue correlate with FeNO levels (Porsbjerg et al., IL-33 is related to Innate immune activation and sensitization to HDM in mild steroid-free asthma. Clin Exp Allergy. 2016. Vol. 46: pp. 564-574.

[0116] serum biomarkers Serum levels of sST2, IL-33, calcitonin, and matrix metalloproteinase-12 (MMP12) may increase after BAC. According to certain embodiments, administration or use of IL-4R antagonists to patients may increase sST2, IL-33, calcitonin, and matrix metalloproteinase-12, which are typically seen after BAC. To reduce the increase in serum levels of sST2, IL-33, calcitonin, and matrix metalloproteinase-12 (MMP12). According to certain embodiments, administration or use of an IL-33 antagonist to a patient reduces the increase in serum levels of sST2, IL-33, calcitonin, and matrix metalloproteinase-12 (MMP12) that is typically seen after BAC. According to certain embodiments, administration or use of an IL-4R antagonist and an IL-33 antagonist to a patient reduces the increase in serum levels of sST2, IL-33, calcitonin, and matrix metalloproteinase-12 (MMP12) that is typically seen after BAC.

[0117] FEF 25-75% According to certain embodiments, administration or use of an IL-33 antagonist, an IL-4R antagonist, or both an IL-33 antagonist and an IL-4R antagonist in a patient results in an increase from baseline FEF of 25–75%. Methods for measuring FEF are known in the art. For example, FEV1 can be measured in a patient using a spirometer that conforms to the recommendations of the American Thoracic Society (ATS) / European Respiratory Society (ERS) 2005. FEF 25–75 (between 25% and 75% of forced expiratory velocity) is the rate at which a person can empty the middle half of their airflow (i.e., forced vital capacity or FVC) during maximal exhalation (in liters per second). The parameter relates to the average flow from the point when 25 percent of FVC is exhaled to the point when 75 percent of FVC is exhaled. The subject's FEF 25–75% provides information regarding small airway function, such as the degree of small airway disease and / or inflammation. Changes in FEF 25–75 are an early indicator of obstructive pulmonary disease. In certain embodiments, improvement and / or increase in the FEF 25–75% parameter is an improvement of at least 10%, 25%, or 50% or more compared to baseline. In certain embodiments, the method of the present invention results in a normal FEF 25–75% value in the subject (e.g., a value ranging from an average of 50–60% to 130%).

[0118] Maximum expiratory flow rate in the morning and evening (AM PEF and PM PEF) According to certain embodiments, administration or use of an IL-33 antagonist, an IL-4R antagonist, or both an IL-33 antagonist and an IL-4R antagonist in a patient results in an increase from baseline in morning (AM) and / or evening (PM) maximal expiratory flow (AM PEF and / or PM PEF). Methods for measuring PEF are known in the art. For example, according to one method for measuring PEF, the patient is provided with an electronic PEF meter to record morning (AM) and evening (PM) PEF (as well as daily albuterol use, morning and evening asthma symptom scores, and the number of nighttime awakenings due to asthma symptoms requiring emergency medication). The patient is instructed on the use of the device, and a written instruction manual for the electronic PEF meter is provided to the patient. Furthermore, a healthcare professional can instruct the patient on how to record the appropriate variables on the electronic PEF meter. AM PEF is generally taken within 15 minutes after waking (between 6 a.m. and 10 a.m.) before administering any albuterol. PM PEF is generally performed in the evening (between 6 pm and 10 pm) before administering any albuterol. Subjects should attempt to withhold albuterol at least 6 hours before measuring their PEFs. Three PEF efforts are performed by the patient, and all three values ​​are recorded using an electronic PEF meter. Typically, the highest value is used for evaluation. Baseline AM PEF can be calculated as the mean AM measurement recorded 7 days prior to administration of the first dose of an IL-33 antagonist, an IL-4R antagonist, or a pharmaceutical composition containing both an IL-33 antagonist and an IL-4R antagonist, and baseline PM PEF can be calculated as the mean PM measurement recorded 7 days prior to administration of the first dose of an IL-33 antagonist, an IL-4R antagonist, or a pharmaceutical composition containing both an IL-33 antagonist and an IL-4R antagonist.

[0119] According to a particular exemplary embodiment, a method or use of treatment is provided that results in an increase in AM PEF and / or PM PEF from baseline at least 1.0 L / min at 12 weeks after the initiation of treatment with an anti-IL-33 antagonist, an IL-4R antagonist, or a pharmaceutical composition comprising an IL-33 antagonist and an IL-4R antagonist. For example, administration or use of an IL-33 antagonist, an IL-4R antagonist, or an IL-33 antagonist and an IL-4R antagonist in a patient requiring such treatment may result in an increase in PEF from baseline of approximately 0.5 L / min, 1.0 L / min, 1.5 L / min, 2.0 L / min, 2.5 L / min, 3.0 L / min, 3.5 L / min, 4.0 L / min, 4.5 L / min, 5.0 L / min, 5.5 L / min, 6.0 L / min, 6.5 L / min, 7.0 L / min, 7.5 L / min, 8.0 L / min, 8.5 L / min, 9.0 L / min, 9.5 L / min, 10.0 L / min, 10.5 L / min, 11.0 L / min, 12.0 L / min, 15 L / min, or 20 L / min or more at week 12.

[0120] Use of albuterol / levalbuterol According to certain embodiments, administration or use of an IL-33 antagonist, an IL-4R antagonist, or an IL-33 antagonist and an IL-4R antagonist to a patient results in a reduction from baseline in the daily use of albuterol or levalbuterol. The number of albuterol / levalbuterol inhalations can be recorded daily by the patient using a diary, a PEF meter, or other recording device. During treatment with the pharmaceutical compositions described herein, albuterol / levalbuterol may generally be used as needed for symptoms, not routinely or prophylactically. The baseline number of albuterol / levalbuterol inhalations / day can be calculated based on the mean seven days prior to administration of a first dose of the pharmaceutical composition containing an IL-33 antagonist, an IL-4R antagonist, or an IL-33 antagonist and an IL-4R antagonist.

[0121] In certain exemplary embodiments, a treatment method or use is provided that results in a reduction of albuterol / levalbuterol use from baseline by at least 0.25 puffs per day at 12 weeks after the initiation of treatment with an IL-33 antagonist, an IL-4R antagonist, or a pharmaceutical composition comprising an IL-33 antagonist and an IL-4R antagonist. For example, administration or use of an IL-33 antagonist, an IL-4R antagonist, or an IL-33 antagonist and an IL-4R antagonist in a patient requiring such treatment may result in a reduction in albuterol / levalbuterol use from baseline of approximately 0.25 puffs per day, 0.50 puffs per day, 0.75 puffs per day, 1.00 puffs per day, 1.25 puffs per day, 1.5 puffs per day, 1.75 puffs per day, 2.00 puffs per day, 2.25 puffs per day, 2.5 puffs per day, 2.75 puffs per day, or 3.00 puffs per day or more by week 12.

[0122] Use of OCS According to certain embodiments, the administration or use of an IL-33 antagonist, an IL-4R antagonist, or an IL-33 antagonist and an IL-4R antagonist to a patient may be used in conjunction with an OCS, such as oral prednisone. The number of OCS doses can be recorded daily by the patient using a diary, a PEF meter, or other recording device. During treatment with the pharmaceutical compositions described herein, occasional short-term use of prednisone can generally be used to control acute asthma episodes, such as episodes in which bronchodilators and other anti-inflammatory agents cannot control the symptoms. In other embodiments, prednisone is used concurrently with or as an alternative to ICS. Oral prednisone may be administered in doses of approximately 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, or 40 mg. OCS is administered once daily. Alternatively, it may be administered multiple times a day (for example, twice, three times, or four times a day), depending on the circumstances.

[0123] In certain exemplary embodiments, methods or uses are provided for reducing or eliminating subject dependence in the use of OCS. Reducing or eliminating steroid dependence is highly advantageous and desirable. In certain embodiments, a reduction of 50% or more (e.g., 50%, 60%, 70%, 80%, 90% or more) of the OCS dose is achieved over a certain period (e.g., at 240 weeks) after administration of IL-4R antibody therapy, IL-33 antibody therapy, or IL-33 antibody therapy in combination with IL-4R antibody therapy. In certain embodiments, OCS is substantially eliminated after the initial dose, 40 weeks, 45 weeks, 50 weeks, 52 weeks or more after the administration of the first dose. In other embodiments, the level of OCS use is reduced to less than 5 mg per day (e.g., less than 5 mg, 4 mg, 3 mg, 2 mg or less per day). In other embodiments, the dependence on the use of OCS is substantially eliminated 3 months, 6 months, 9 months, or 1 year after treatment with an IL-33 antagonist, an IL-4R antagonist, or both an IL-33 antagonist and an IL-4R antagonist.

[0124] 5-Item Asthma Control Questionnaire (ACQ) Score According to certain embodiments, administration or use of an IL-33 antagonist, an IL-4R antagonist, or both an IL-33 antagonist and an IL-4R antagonist in patients results in a reduction from baseline in the 5-item Asthma Control Questionnaire (ACQ5) score. The ACQ5 is an effective questionnaire for assessing asthma control.

[0125] According to a particular exemplary embodiment, a method or use of treatment is provided that results in a reduction of at least 0.10 points from baseline in the ACQ5 score at 12 weeks after the initiation of treatment with an IL-33 antagonist, an IL-4R antagonist, or a pharmaceutical composition comprising an IL-33 antagonist and an IL-4R antagonist. For example, administration or use of an IL-33 antagonist, an IL-4R antagonist, or an IL-33 antagonist and an IL-4R antagonist to a subject requiring it results in a reduction of approximately 0.10 points, 0.15 points, 0.20 points, 0.25 points, 0.30 points, 0.35 points, 0.40 points, 0.45 points, 0.50 points, 0.55 points, 0.60 points, 0.65 points, 0.70 points, 0.75 points, 0.80 points, 0.85 points or more from baseline in the ACQ score at 12 weeks.

[0126] Nocturnal awakening According to certain embodiments, administration or use of an IL-33 antagonist, an IL-4R antagonist, or both an IL-33 antagonist and an IL-4R antagonist to a patient results in a baseline reduction in the mean number of nocturnal awakenings.

[0127] In a particular embodiment, the method or use reduces the mean number of nocturnal awakenings per night from baseline to at least about 0.10 times by the 12th week after the start of treatment. For example, administration or use of an IL-33 antagonist, an IL-4R antagonist, or an IL-33 antagonist and an IL-4R antagonist to a patient requiring such treatment showed a decrease in the mean number of nocturnal awakenings per night from baseline at week 12, approximately 0.10 times, 0.15 times, 0.20 times, 0.25 times, 0.30 times, 0.35 times, 0.40 times, 0.45 times, 0.50 times, 0.55 times, 0.60 times, 0.65 times, 0.70 times, 0.75 times, 0.80 times, 0.85 times, 0.90 times, 0.95 times, 1.0 times, per night. This can result in a decrease of 2.0 times or more.

[0128] 22-item SNOT-22 score for evaluating nasal and paranasal sinus disease. According to certain embodiments, administration or use of IL-33 antagonists, IL-4R antagonists, or both IL-33 and IL-4R antagonists in patients results in a reduction from baseline of 22 nasal sinus disease assessment items (SNOT-22). SNOT-22 is an effective questionnaire for assessing the impact of chronic rhinosinusitis on quality of life (Hopkins et al., 2009, Clin. Otolaryngol. 34: pp. 447-454).

[0129] According to certain exemplary embodiments, a method or use of treatment is provided that results in a reduction of at least one point in the SNOT-22 score from baseline at 12 weeks after the initiation of treatment with an IL-33 antagonist, an IL-4R antagonist, or a pharmaceutical composition comprising an IL-33 antagonist and an IL-4R antagonist. For example, administration or use of an IL-33 antagonist, an IL-4R antagonist, or an IL-33 antagonist and an IL-4R antagonist to a subject requiring it may result in a reduction of approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 points or more in the SNOT-22 score from baseline at 12 weeks.

[0130] biomarkers In certain embodiments, administration or use of an IL-33 antagonist, an IL-4R antagonist, or both an IL-33 antagonist and an IL-4R antagonist in a patient improves lung function as measured by biomarkers. For example, biomarkers may be exhaled nitric oxide (FeNO), eotaxin-3, total IgE, periostin, or thymic and activating modulated chemokines (TARCs). In certain embodiments, the improvement in lung function is indicated by a decrease or increase at 4, 12, or 24 weeks post-treatment (if necessary).

[0131] Methods for treating asthma In some embodiments, the present invention provides a method for treating allergic asthma, including, for example, mild allergic asthma and mild persistent allergic asthma, in subjects where it is needed, the method comprising the step of administering to a subject an IL-33 antagonist, an IL-4R antagonist, or a pharmaceutical composition comprising an IL-33 antagonist and an IL-4R antagonist. Also provided is an anti-IL-33 antagonist, an IL-4R antagonist, or a pharmaceutical composition comprising an IL-33 antagonist and an IL-4R antagonist for treating allergic asthma, including, for example, mild allergic asthma and mild persistent allergic asthma, in subjects where it is needed. In certain embodiments, the method is useful for treating allergic asthma in subjects. In some embodiments, the method is useful for treating mild persistent allergic asthma in subjects.

[0132] As used herein, the term “asthma” can be used synonymously with “intermittent asthma” or “bronchial asthma.” “Asthma,” “bronchial asthma,” and “intermittent asthma” mean asthma that is characterized by one or any combination of the following: symptoms occurring two or several days per week; symptoms not interfering with daily activities; nocturnal symptoms occurring less than two days per month; or one or more pulmonary function tests (e.g., forced expiratory volume in one second (FEV1) greater than 80% and / or maximal expiratory flow (PEF)) being normal when the subject is not experiencing an asthma attack.

[0133] "Allergic asthma" refers to allergens, such as inhaled allergens (e.g., perennial allergens). This refers to asthma induced by airborne allergens (including seasonal airborne allergens), such as dust mites, pet dander, pollen, and fungi. In certain embodiments, the allergen is a dust mite (HDM) allergen (e.g., a perennial airborne allergen).

[0134] As used herein, "perennial air allergens" refers to airborne allergens that may be present in the environment throughout the year, such as dust mites, fungi, dandruff, etc. Perennial air allergens include, but are not limited to, Alternaria alternata, Aspergillus fumigatus, Aureobasidium pullulans, Candida albicans, Cladosporium herbarum, Dermatofagoides farinae, Dermatofagoides pteronyssinus, Mucor racemosus, Penicillium chrysogenum, Phoma betae, and Setomelanomma rostrata. This includes rostrata, Stemphylium herbarum, cat dander, dog dander, cow dander, chicken feathers, goose feathers, duck feathers, cockroaches (e.g., German cockroach, Eastern cockroach), mouse urine, peanut dust, and nut dust.

[0135] As used herein, “seasonal air allergens” means airborne allergens that are present in the environment seasonally, such as pollen and spores. Seasonal air allergens include, but are not limited to, tree pollen (e.g., birch, alder, cedar, hazel, hornbeam, horse chestnut, willow, poplar, linden, pine, maple, oak, olive, etc.), grass pollen (e.g., ryegrass, cattail, etc.), weed pollen (e.g., ragweed, plantain, nettle, mugwort, goosefoot, sorrel, etc.), and fungal spores that increase during certain seasons, temperatures, etc. (e.g., mold).

[0136] As used herein, the terms “persistent asthma” or “persistent bronchial asthma” mean asthma that is more severe than (bronchial) asthma / intermittent (bronchial) asthma. Subjects with persistent asthma or persistent bronchial asthma experience one or more of the following: symptoms for more than two days per week; symptoms that interfere with daily activities; nocturnal symptoms occurring for more than two days per month; or one or more pulmonary function tests that are not normal when the subject is not experiencing an asthma attack (e.g., forced expiratory volume in one second (FEV1) and / or maximum expiratory flow rate (PEF) less than 80%); the subject is dependent on asthma control medication on a daily basis; the subject has received systemic corticosteroids more than once since the last year in which a severe relapse of severe asthma occurred; or use of short-acting β-2 agonists for relief of asthma symptoms for more than two days per week.

[0137] Asthma / intermittent asthma, bronchial asthma / intermittent bronchial asthma, and persistent asthma / persistent bronchial asthma can be classified as "mild," "moderate," "severe," or "moderate to severe." "Mild intermittent asthma" or "mild intermittent bronchial asthma" is defined as having symptoms less than once per week and a forced expiratory volume in one second (FEV1) or maximum expiratory flow rate (PEF) of ≥80%. "Mild persistent asthma" or "mild persistent bronchial asthma" differs in that the frequency of symptoms is more than once per week but less than once per day, and the variability of FEV1 or PEF is <20% to 30%. "Moderate intermittent asthma" or "moderate intermittent bronchial asthma" is defined as having symptoms less than once per week and a forced expiratory volume in one second (FEV1) or maximum expiratory flow rate (PEF) of 60-80%. "Moderate persistent asthma" or "moderate persistent bronchial asthma" is defined as having daily symptoms, exacerbations that may affect activity and / or sleep, and once a week. "Asthma" is defined as having more than one episode of symptoms per week, daily use of an inhaled short-acting β-2 agonist, and a forced expiratory volume in one second (FEV1) or maximum expiratory flow rate (PEF) of 60-80%. "Severe intermittent asthma" or "severe intermittent bronchial asthma" is defined as having fewer than one episode of symptoms per week and a forced expiratory volume in one second (FEV1) or maximum expiratory flow rate (PEF) of 60%. "Severe persistent asthma" or "severe persistent bronchial asthma" is defined as having daily symptoms, frequent exacerbations that may affect activity and / or sleep, frequent nocturnal symptoms, limitation of physical activity, daily use of an inhaled short-acting β-2 agonist, and a forced expiratory volume in one second (FEV1) or maximum expiratory flow rate (PEF) of 60%. "Moderate to severe intermittent asthma" or "moderate to severe intermittent bronchial asthma" is defined as having symptoms between those of moderate intermittent asthma / moderate intermittent bronchial asthma and severe intermittent asthma / severe intermittent bronchial asthma. "Moderate to severe persistent asthma" or "moderate to severe persistent bronchial asthma" is defined as having symptoms between those of moderate persistent asthma / moderate persistent bronchial asthma and severe persistent asthma / severe persistent bronchial asthma.

[0138] As used herein, the term “poorly controlled asthma” refers to patients whose asthma is “not well controlled” or “very poorly controlled,” as defined in “Expert Panel Report 3: Guidelines for the Diagnosis and Management of Asthma,” National Heart, Blood and Lung Institute, NIH, August 28, 2007. “Poorly controlled asthma” is defined as having symptoms for more than two days per week, nocturnal awakenings one to three times per week, some limitations on normal activity, use of a short-acting β2-agonist for symptom control for more than two days per week, a predicted and / or best-ever FEV1 of 60–80%, an ATAQ score of 1–2, an ACQ score of 1.5 or higher, and an ACT score of 16–19. "Very poorly controlled asthma" is defined as having symptoms throughout the day, nocturnal awakenings four or more times per week, extreme limitation of normal activity, and use of short-acting β2-agonists several times a day for symptom control, with a predicted and / or personal best FEV1 of less than 60%, an ATAQ score of 3–4, an ACQ score of N / A, and an ACT score of 15 or less.

[0139] In some embodiments, the subject is the Global Initiative for Asthma Management (GINA). Asthma) 2009 guidelines, and one or more of the following criteria: i) Existing treatment with moderate to high doses of ICS / LABA (fluticasone dipropionate 250 μg twice daily or an equivalent daily dose of ICS) with a stable dose of ICS / LABA at least one month prior to the initial dose of an IL-4R antagonist, an IL-33 antagonist, or an initial dose of both an IL-33 antagonist and an IL-4R antagonist; ii) FEV1 of 40-80% of the predicted normal value prior to the initial dose of an IL-4R antagonist, an IL-33 antagonist, or an initial dose of both an IL-33 antagonist and an IL-4R antagonist; iii) ACQ-5 score prior to the initial dose of an IL-4R antagonist, an IL-33 antagonist, or an initial dose of both an IL-33 antagonist and an IL-4R antagonist. a) 1.5 or higher; iv) Reversibility of at least 12% and 200 mL of FEV1 after salbutamol / albuterol 200 μg to 400 μg (2 to 4 inhalations) before administration of the initial dose of IL-4R antagonist, IL-33 antagonist, or IL-33 antagonist and IL-4R antagonist; or v) If diagnosed by a physician based on having experienced one or more of the following events within the past year prior to administration of the initial dose of IL-4R antagonist, IL-33 antagonist, or IL-33 antagonist and IL-4R antagonist: (a) treatment with one or more systemic (oral or parenteral) steroid bursts for asthma exacerbation, or (b) hospitalization or emergency / urgent visit for asthma exacerbation, the subject is identified as having "moderate to severe poorly controlled" asthma. .

[0140] "Severe asthma" means asthma in which adequate control cannot be achieved, or will be lost, with high-dose treatment with inhaled corticosteroids and additional controllers (e.g., long-acting inhaled β2 agonists, montelukast, and / or theophylline) or with oral corticosteroid treatment (e.g., at least 6 months per year). In certain embodiments, severe asthma includes asthma treated with high-dose ICS and at least one additional controller (e.g., LABA, montelukast, or theophylline) or oral corticosteroids > 6 months / year, and at least one of the following occurs if treatment is reduced: ACT < 20 or ACQ > 1.5; at least two exacerbations in the past 12 months; at least one exacerbation in the past 12 months requiring hospital treatment or mechanical ventilation; or (if FEV1 / FVC is below the lower limit of normal) FEV1 < 80%.

[0141] "Steroid-dependent asthma" refers to asthma requiring one or more of the following treatments: frequent, short bursts of oral corticosteroid treatment in the past 12 months; regular use of high-dose inhaled corticosteroids in the past 12 months; regular use of injected long-acting corticosteroids; daily use of oral corticosteroids; every-other-day oral corticosteroids; or long-term use of oral corticosteroids within the past year.

[0142] "Oral corticosteroid-dependent asthma" refers to patients who have received ≥3 doses of 30-day oral corticosteroid (OCS) fills over a 12-month period and who have received a primary asthma diagnosis within 12 months of the first OCS fill. Patients with OCS-dependent asthma are receiving physician-prescribed LABA and high-dose ICS (total daily dose >500 μg fluticasone propionate dry powder equivalent) for at least 3 months (ICS and LABA may be part of a combination product or administered via separate inhalers); receiving additional maintenance asthma controller medications as part of standard medical practice, such as leukotriene receptor antagonists (LTRAs), theophylline, long-acting muscarinic antagonists (LAMAs), secondary ICS, and chromone; and being treated for asthma at doses between ≥7.5 and ≤30 mg (prednisone or prednisolone equivalent). You are receiving OCS for the following reasons; you are receiving an OCS dose administered every other day (or different doses every other day); you have evidence of asthma as described by a predicted normal pre-morning bronchodilator (BD) FEV1 < 80%; a reversible FEV1 ≥ 12% and ≥ 200 mL (15-30 minutes after 4 puffs of albuterol / salbutamol) after BD administration; or you have experienced one or any combination of the following: a history of at least one asthma exacerbation event within the last 12 months.

[0143] In one embodiment, a method for treating asthma is provided, comprising the steps of (a) selecting a patient whose blood eosinophil level is at least 300 cells per microliter; and (b) administering to the patient a pharmaceutical composition comprising an IL-33 antagonist, an IL-4R antagonist, or an IL-33 antagonist and an IL-4R antagonist. In one embodiment of the composition for use, the patient has a blood eosinophil level of at least 300 cells per microliter.

[0144] In another embodiment, a method for treating asthma is provided, comprising the steps of (a) selecting a patient whose blood eosinophil level is 150 to 299 cells per microliter; and (b) administering to the patient a pharmaceutical composition comprising an IL-33 antagonist, an IL-4R antagonist, or an IL-33 antagonist and an IL-4R antagonist. In one embodiment of the composition for use, the patient has a blood eosinophil level of at least 150 to 299 cells per microliter.

[0145] In another embodiment, a method for treating asthma is provided, comprising the steps of (a) selecting a patient whose blood eosinophil level is less than 150 cells per microliter; and (b) administering to the patient a pharmaceutical composition comprising an IL-33 antagonist, an IL-4R antagonist, or an IL-33 antagonist and an IL-4R antagonist. In one embodiment of the composition for use, the patient has a blood eosinophil level of less than 150 cells per microliter.

[0146] In one embodiment, a method for treating asthma is provided, comprising the steps of (a) selecting a patient exhibiting low levels of periostin; and (b) administering to the patient a pharmaceutical composition comprising an IL-33 antagonist, an IL-4R antagonist, or an IL-33 antagonist and an IL-4R antagonist. In one embodiment of the composition for use, the patient exhibits low levels of periostin.

[0147] In another embodiment, a method for treating asthma is provided, comprising the steps of (a) selecting a patient exhibiting high levels of periostin; and (b) administering to the patient a pharmaceutical composition comprising an IL-33 antagonist, an IL-4R antagonist, or an IL-33 antagonist and an IL-4R antagonist. In one embodiment of the composition for use, the patient exhibits high levels of periostin.

[0148] As used herein, “high levels of periostin” refers to blood periostin levels of approximately 60 ng / mL or higher, approximately 65 ng / mL or higher, approximately 70 ng / mL or higher, approximately 75 ng / mL or higher, or approximately 80 ng / mL or higher, approximately 85 ng / mL or higher, approximately 90 ng / mL or higher, approximately 95 ng / mL or higher, or approximately 100 ng / mL or higher. In detailed exemplary embodiments, high levels of periostin are approximately 75.0 ng / mL or higher or approximately 74.4 ng / mL or higher.

[0149] As used herein, “low levels of periostin” refers to blood periostin measurements of less than approximately 100 ng / mL, less than approximately 95 ng / mL, less than approximately 90 ng / mL, less than approximately 85 ng / mL, less than approximately 80 ng / mL, less than approximately 75 ng / mL, less than approximately 70 ng / mL, less than approximately 65 ng / mL, or less than approximately 60 ng / mL. In detailed exemplary embodiments, low levels of periostin are less than approximately 75.0 ng / mL or less than approximately 74.4 ng / mL.

[0150] In related embodiments, methods for treating asthma are provided, including add-on therapy to basic therapy. Also provided, in related embodiments, is an IL-33 antagonist for use in treating allergic asthma in patients, the IL-33 antagonist being used as add-on therapy to basic therapy. In certain embodiments, the IL-33 antagonist, IL-4R antagonist, or IL-33 antagonist and IL-4R antagonist are administered as add-on therapy to asthma patients receiving basic therapy for a period of time (e.g., one week, two weeks, three weeks, one month, two months, five months, twelve months, eighteen months, twenty-four months or longer) (also called the “stable phase”). In some embodiments, the basic therapy includes ICS and / or LABA.

[0151] In some embodiments, the present invention includes a method for reducing an asthmatic patient's dependence on ICS and / or LABA for the treatment of one or more exacerbations of allergic asthma, comprising the steps of: (a) selecting a patient having asthma that is not adequately controlled by basic asthma therapy including ICS, LABA, or a combination thereof; and administering to the patient a pharmaceutical composition comprising an IL-33 antagonist, an IL-4R antagonist, or an IL-33 antagonist and an IL-4R antagonist. In one embodiment of the composition for use, The patient has asthma that is not adequately controlled with basic asthma therapy, including ICS, LABA, or a combination thereof.

[0152] In some embodiments, the present invention encompasses methods for treating or alleviating asthma-related conditions or complications, such as chronic sinusitis, allergic rhinitis, allergic fungal sinusitis, allergic bronchopulmonary aspergillosis, unified airway disease, Churg-Strauss syndrome, vasculitis, chronic obstructive pulmonary disease (COPD), and exercise-induced bronchospasm. Also provided are, in subjects requiring it, an anti-IL-33 antagonist, an IL-4R antagonist, or a pharmaceutical composition comprising an IL-33 antagonist and an IL-4R antagonist for treating asthma-related conditions or complications, such as chronic sinusitis, allergic rhinitis, allergic fungal sinusitis, allergic bronchopulmonary aspergillosis, unified airway disease, Churg-Strauss syndrome, vasculitis, chronic obstructive pulmonary disease (COPD), and exercise-induced bronchospasm.

[0153] The present invention also includes methods for treating persistent asthma. Also provided are, in subjects requiring it, an anti-IL-33 antagonist, an IL-4R antagonist, or a pharmaceutical composition comprising an IL-33 antagonist and an IL-4R antagonist for treating persistent asthma. As used herein, the term “persistent asthma” means that a subject has symptoms at least once a week during the day and / or at night, and that these symptoms last from several hours to several days. In certain alternative embodiments, persistent asthma is “mild persistent” (e.g., severe symptoms that interfere with daily activities or sleep occur more than twice a week but less than daily, and / or lung function is normal or reversible with inhalation of a bronchodilator), “moderate persistent” (e.g., symptoms that interfere with sleep at least once a week and / or moderately impaired lung function occur daily), or “severe persistent” (e.g., symptoms persist despite correct use of approved medications and / or lung function is severely affected).

[0154] Interleukin-33 (IL-33) antagonists and interleukin-4 receptor (IL-4R) antagonists The methods disclosed herein may optionally include the step of administering a therapeutic composition comprising an IL-33 antagonist to a subject requiring it. As used herein, “IL-33 antagonist” is any agent that binds to or interacts with IL-33 and inhibits the normal biological signaling function of IL-33 when IL-33 is expressed in cells in vitro or in vivo.

[0155] The methods disclosed herein may optionally include the step of administering a therapeutic composition comprising an IL-4R antagonist to a subject requiring it. As used herein, “IL-4R antagonist” is any agent that binds to or interacts with IL-4R and inhibits the normal biological signaling function of IL-4R when IL-4R is expressed in cells in vitro or in vivo.

[0156] Examples of categories of IL-33 antagonists and IL-4R antagonists, not limited to these, include small molecule IL-33 antagonists, small molecule IL-4R antagonists, anti-IL-33 aptamers, anti-IL-4R aptamers, peptide-based IL-33 antagonists or peptide-based IL-4R antagonists (e.g., "peptibody" molecules), and antigen-binding fragments of antibodies or antibodies that specifically bind to human IL-33 or human IL-4R.

[0157] According to a particular embodiment, the IL-33 antagonist is described in other parts of this specification. The present invention includes an anti-IL-33 antibody or its antigen-binding fragment that can be used in the context of the methods characterized in the present invention. For example, in one embodiment, the IL-33 antagonist is an antibody or its antigen-binding fragment that specifically binds to IL-33 and comprises heavy chain and light chain (complementary determination region) CDR sequences from the heavy chain variable region (HCVR) and light chain variable region (LCVR) of SEQ ID NOs. 2 and 10, respectively. In another embodiment, the IL-33 antagonist is an antibody or its antigen-binding fragment that specifically binds to IL-33 and comprises the heavy chain and light chain CDR sequences of SEQ ID NOs. 4, 6 and 8 and SEQ ID NOs. 12, 14 and 16, respectively. In yet another embodiment, the IL-33 antagonist is an antibody or its antigen-binding fragment that specifically binds to IL-33 and comprises the HCVR / LCVR pairs of SEQ ID NOs. 2 and 10, respectively.

[0158] REGN3500 HCVR, DNA sequence: aggtgcagct ggtggagtct gggggaaact tggaacagcc tggggggtcc cttagactct cctgtacagc ctctggattc acctttagca gatctgccat gaactgggtc cgccgggctc cagggaaggg gctggagtgg gtctcaggaa ttagtggtag tggtggtcga acatactacg cagactccgt gaagggccgg ttcaccatct ccagagacaa ttccaagaat acgctatatc tgcaaatgaa cagcctgagc gccgaggaca cggccgcata ttactgtgcg aaagattcgt atactaccag ttggtacgga ggtatggacg tctggggcca cgggaccacg gtcaccgtct cctca (SEQ ID NO: 1).

[0159] REGN3500 HCVR, amino acid sequence: VQLVESGGNLEQPGGSLRLSCTASGFTFSRSAMNWVRRAPGKGLEWVSGISGSGGRTYYADSVKGRFTISRDNSKNTLYLQMNSLSAEDTAAYYCAKDSYTTSWYGGMDVWGHGTTVTVSS(Sequence ID 2).

[0160] REGN3500 HCDR1, DNA sequence: ggattcacctt tagcagatct gcc (sequence number 3).

[0161] REGN3500 HCDR1, amino acid sequence: GFTFSRSA (SEQ ID NO: 4).

[0162] REGN3500 HCDR2, DNA sequence: attagtggtag tggtggtcga aca (Sequence ID 5).

[0163] REGN3500 HCDR2, amino acid sequence: ISGSGGRT (Sequence ID 6).

[0164] REGN3500 HCDR3, DNA sequence: gcgaaagattc gtatactacc agttggtacg gaggtatgga cgtc (Sequence ID 7).

[0165] REGN3500 HCDR3, amino acid sequence: AKDSYTTSWYGGMDV (Sequence ID 8).

[0166] REGN3500 LCVR, DNA sequence: acatccagat gacccagtct ccatcttccg tgtctgcatc tgtaggagac agagtcacca tcacttgtcg ggcgagtcag ggtattttca gctggttagc ctggtatcag cagaaaccag gaaaagcccc taagctcctg atctatgctg cttccagttt acaaagtggg gtcccatcaa gattcagcgg cagtggatct gggacagatt tcactctcac catcagcagc ctgcagcctg aggattttgc aatttactat tgtcaacagg ctaacagtgt cccgatcacc ttcggccaag ggacacgact ggagattaaa cga (SEQ ID NO: 9).

[0167] REGN3500 LCVR, amino acid sequence: IQMTQSPSSVSASVGDRVTITCRASQGIFSWLAWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFAIYYCQQANSVPITFGQGTRLEIKR (Sequence ID 10).

[0168] REGN3500 LCDR1, DNA sequence: cagggtatttt cagctgg (Sequence ID 11).

[0169] REGN3500 LCDR1, amino acid sequence: QGIFSW (sequence number 12).

[0170] REGN3500 LCDR2, DNA sequence: gctgcttcc (sequence number 13).

[0171] REGN3500 LCDR2, amino acid sequence: AAS (Sequence ID 14).

[0172] REGN3500 LCDR3, DNA sequence: caacaggctaa cagtgtcccg atcacc (Sequence ID 15).

[0173] REGN3500 LCDR3, amino acid sequence: QQANSVPIT (sequence number 16).

[0174] In another embodiment, the IL-33 antagonist is a REGN3500 antibody, comprising the HC / LC pairs of SEQ ID NOs. 18 and 20, respectively.

[0175] REGN3500 heavy-chain DNA sequence: aggtgcagct ggtggagtct gggggaaact tggaacagcc tgggggtcc cttagactct cctgtacagc ctctggattc acctttagca gatctgccat gaactgggtc cgccgggctc cagggaaggg gctggagtgg gtctcaggaa ttagtggtag tggtggttcga acatactacg cagactccgt gaagggccgg ttcaccatct ccagagacaa ttccaagaat acgctatatc tgcaaatgaa cagcctgagc gccgaggaca cggccgcata ttactgtgcg aaagattcgt atactaccag ttggtacgga ggtatggacg tctggggcca cgggaccacg gtcaccgtct cctcagcctc caccaagggc ccatcggtct tccccctggc gccctgctcc aggagcacct ccgagagcac agccgc cctg ggctgcctgg tcaaggacta cttccccgaa ccggtgacgg tgtcgtggaa ctcaggcgcc ctgaccagcg gcgtgcacac cttcccggct gtcctacagt cctcaggact ctactccctc agcagcgtgg tgaccgtgcc ctccagcagc ttgggcacga agacctacac ctgcaacgta gatcacaagc ccagcaacac caaggtggac aagagagttg agtccaaata tggtccccca tgcccaccct gcccagcacc tgagttcctg gggggaccat cagtcttcct gttcccccca aaacccaagg acactctcat gatctcccgg acccctgagg tcacgtgcgt ggtggtggac gtgagccagg aagaccccga ggtccagttc aactggtacg tggatggcgt ggaggtgcat aatgccaaga caaagccgcg ggaggagcag ttcaacagca cgtaccgtgt ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaac ggcaaggagt acaagtgcaa ggtctccaac aaaggcctcc cgtcctccat cgagaaaacc atctccaaag ccaaagggca gccccgagag ccacaggtgt acaccctgcc cccatcccag gaggagatga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt ctaccccagc gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct cccgtgctgg actccgacgg ctccttcttc ctctacagca ggctcaccgt ggacaagagc aggtggcagg aggggaatgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac tacacacaga agtccctctc cctgtctctg ggtaaatga(SEQ ID NO: 17).

[0176] REGN3500 heavy chain amino acid sequence: VQLVESGGNLEQPGGSLRLSCTASGFTFSRSAMNWVRRAPGKGLEWVSGISGSGGRTYYADSVKGRFTISRDNSKNTLYLQMNSLSAEDTAAYYCAKDSYTTSWYGGMDVWGHG TTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCP PCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (Sequence ID 18).

[0177] REGN3500 light chain DNA sequence: acatccagat gacccagtct ccatcttccg tgtctgcatc tgtaggagac agagtcacca tcacttgtcg ggcgagtcag ggtattttca gctggttagc ctggtatcag cagaaaccag gaaaagcccc taagctcctg atctatgctg cttccagttt acaaagtggg gtcccatcaa gattcagcgg cagtggatct gggacagatt tcactctcac catcagcagc ctgcagcctg aggattttgc aatttactat tgtcaacagg ctaacagtgt cccgatcacc ttcggccaag ggacacgact ggagattaaa cgaactgtgg ctgcaccatc tgtcttcatc ttcccgccat ctgatgagca gttgaaatct ggaactgcct ctgttgtgtg cctgctgaat aacttctatc ccagagaggc caaagtacag tggaaggtgg ataacgccct ccaatcgggt aactcccagg agagtgtcac agagcaggac agcaaggaca gcacctacag ccctcagcagc accctgacgc tgagcaaagc agactacgag aaacacaaag tctacgcctg cgaagtcacc catcagggcc tgagctcgcc cgtcacaaag agcttcaaca ggggagagtg ttag (SEQ ID NO: 19).

[0178] REGN3500 light chain amino acid sequence: IQMTQSPSSVSASVGDRVTITCRASQGIFSWLAWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFAIYYCQQANSVPITFGQGTRLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (Sequence ID 20).

[0179] According to certain embodiments, the IL-4R antagonist comprises an anti-IL-4R antibody or its antigen-binding fragment that can be used in the context of the methods characterized in the present invention as described in other parts of this specification. For example, in one embodiment, the IL-4R antagonist is an antibody or its antigen-binding fragment that specifically binds to IL-4R and comprises heavy chain and light chain (complementary determination region) CDR sequences from the heavy chain variable region (HCVR) and light chain variable region (LCVR) of SEQ ID NOs. 27 and 28, respectively. In another embodiment, the IL-4R antagonist is an antibody or its antigen-binding fragment that specifically binds to IL-4R and comprises the heavy chain and light chain CDR sequences of SEQ ID NOs. 21, 22 and 23 and SEQ ID NOs. 24, 25 and 26, respectively. In yet another embodiment, the IL-4R antagonist is an antibody or its antigen-binding fragment that specifically binds to IL-4R and comprises the HCVR / LCVR pairs of SEQ ID NOs. 27 and 28, respectively.

[0180] Dupilumab HCDR1 amino acid sequence: GFTFRDYA (Sequence ID 21).

[0181] Dupilumab HCDR2 amino acid sequence: ISGSGGNT (Sequence ID 22).

[0182] Dupilumab HCDR3 amino acid sequence: AKDRLSITIRPRYYGL (Sequence ID 23).

[0183] Dupilumab LCDR1 amino acid sequence: QSLLYSIGYNY (Sequence ID 24).

[0184] Dupilumab LCDR2 amino acid sequence: LGS (Sequence ID 25).

[0185] Dupilumab LCDR3 amino acid sequence: MQALQTPYT(Sequence ID 26).

[0186] Dupilumab HCVR amino acid sequence: EVQLVESGGGLEQPGGSLRLSCAGSGFTFRDYAMTWVRQAPGKGLEWVSSISGSGGNTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDRLSITIRPRYYGLDVWGQGTTVTVS (Sequence ID 27).

[0187] Dupilumab LCVR amino acid sequence: DIVMTQSPLSLPVTPGEPASISCRSSQSLLYSIGYNYLDWYLQKSGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGFYYCMQALQTPYTFGQGTKLEIK (Sequence ID 28).

[0188] In another embodiment, the IL-4R antagonist is dupilumab. Dupilumab HC amino acid sequence: EVQLVESGGGLEQPGGSLRLSCAGSGFTFRDYAMTWVRQAPGKGLEWVSSISGSGGNTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDRLSITIRPRYYGLDVWGQGTTVTV SSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVF LFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG (Sequence ID 29) (Amino acids 1-124 = HCVR; Amino acids 125-451 = HC constants).

[0189] Dupilumab LC amino acid sequence: DIVMTQSPLSLPVTPGEPASISCRSSQSLLYSIGYNYLDWYLQKSGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGFYYCMQALQTPYTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 30) (amino acids 1 - 112 = LCVR; amino acids 112 - 219 = LC constant).

[0190] The term "human IL - 33" (hIL - 33) refers to a human cytokine receptor that specifically binds to interleukin - 33 (IL - 33). The term "human IL - 4R" (hIL - 4R) refers to a human cytokine receptor that specifically binds to interleukin - 4 (IL - 4), for example, IL - 4Rα.

[0191] The term "antibody" refers to an immunoglobulin molecule containing four polypeptide chains, two heavy (H) chains and two light (L) chains interconnected by disulfide bonds, as well as their multimers (e.g., IgM). Each heavy chain contains a heavy - chain variable region (abbreviated herein as HCVR or V H for short) and a heavy - chain constant region. The heavy - chain constant region contains three domains, C H 1, C H 2, and C H 3. Each light chain contains a light - chain variable region (abbreviated herein as LCVR or V L for short) and a light - chain constant region. The light - chain constant region contains one domain (C L 1). The V H and V L [[ID=2--7]]regions can be further subdivided into regions of hypervariability called complementarity - determining regions (CDRs) and can be dispersed in more conserved regions called framework regions (FRs). Each V H and V L has the following order: FR1 It consists of three CDRs and four FRs, arranged from the amino terminus to the carboxyl terminus as CDR1, FR2, CDR2, FR3, CDR3, FR4. In different embodiments, the FRs of the anti-IL-33 antibody, anti-IL-4R antibody, or their antigen-binding moieties may be identical to the human germline sequence, or they may be naturally or artificially modified. The amino acid consensus sequence can be defined based on side-by-side analysis of two or more CDRs.

[0192] The term “antibody” also includes the antigen-binding fragment of a complete antibody molecule. As used herein, the terms “antigen-binding portion” of an antibody, “antigen-binding fragment” of an antibody, etc., 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 may be derived from, for example, a complete antibody molecule using any suitable standard technique, e.g., protein digestion or DNA-coding antibody variability, and optionally recombinant genetic engineering techniques involving manipulation and expression of constant domains. 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. DNA can be sequenced and manipulated, for example, by using chemical or molecular biological techniques to sequence one or more variable and / or constant domains in appropriate configurations, or to introduce codons, create cysteine ​​residues, modify, add, or delete amino acids.

[0193] Examples of antigen-binding fragments, though not limited to these, 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 hypervariable regions of antibodies (e.g., isolated complementary determinant regions (CDRs), e.g., CDR3 peptides), or minimal recognition units consisting of amino acid residues mimicking bound FR3-CDR3-FR4 peptides. Other manipulated molecules, such as domain-specific antibodies, single-domain antibodies, domain deletion antibodies, chimeric antibodies, CDR-implanted antibodies, bispecific antibodies, triplicate antibodies, quadruplicate antibodies, minibodies, nanobodies (e.g., monovalent nanobodies, divalent nanobodies, etc.), small modular immunopharmaceuticals (SMIPs), and Shark variable IgNAR domains, also fall within the scope of the expression “antigen-binding fragment.”

[0194] Antibody antigen-binding fragments generally contain at least one variable domain. The variable domain can be of any size or amino acid composition and generally contains at least one CDR, which is a frame unit adjacent to or containing one or more framework sequences. L V related to the domain H In antigen-binding fragments having a domain, V H and V L Domains can be positioned relative to each other in any suitable arrangement. For example, a variable region can be a dimer, V H -V H , V H -V L or V L -V L It contains a dimer. Alternatively, the antigen-binding fragment of the antibody contains monomer V H or V L It can include a domain.

[0195] In certain embodiments, the antigen-binding fragment of an antibody may include at least one variable domain covalently bound to at least one constant domain. Exemplary arrangements of variable and constant domains that can be found within the antigen-binding fragments of antibodies described herein 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 H 3;(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;(xi i)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 LThis includes. In any configuration of variable and constant domains, including any of the exemplary configurations listed above, the variable and constant domains may be directly linked to each other or linked by a complete or partial hinge or linker region. The hinge region may consist of at least two amino acids (e.g., 5, 10, 15, 20, 40, 60 or more) that result in mobile or semi-mobile linkage between adjacent variable and / or constant domains in a single polypeptide molecule, and generally the hinge region may consist of 2 to 60 amino acids, generally between 5 to 50, or generally between 10 to 40 amino acids. Furthermore, the antigen-binding fragments of the antibodies described herein may consist of each other and / or one or more monomer V H Or V L In non-covalent associations with the domain (e.g., via disulfide bonds), the domain may contain any of the variable and constant domain configurations listed above, including homodimers or heterodimers (or other polymers).

[0196] Like complete antibody molecules, antigen-binding fragments may be monospecific or polyspecific (e.g., bispecific). A polyspecific antigen-binding fragment of an antibody generally contains at least two distinct variable domains, each capable of specifically binding to separate antigens or to different epitopes within the same antigen. Any polyspecific antibody format can be adapted for use in association with the antibody antigen-binding fragments described herein using routine techniques available in the art.

[0197] The constant region of an antibody is crucial in its ability to immobilize complement and mediate cell-dependent cytotoxicity. Therefore, antibody isotypes can be selected based on whether or not it is desirable for the antibody to mediate cytotoxicity.

[0198] The term "human antibody" includes antibodies having variable and constant regions derived from human germline immunoglobulin sequences. Nevertheless, the human antibodies characterized in the present invention may include, for example, CDRs, particularly CDR3, amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-directed mutagenesis in vitro or by somatic mutation in vivo). However, the term "human antibody" does not include antibodies in which a CDR sequence derived from the germline of another mammalian species, such as mouse, has been transplanted into a human framework sequence.

[0199] The term “recombinant human antibody” includes all human antibodies produced, expressed, created, or isolated by recombinant means, such as antibodies expressed using recombinant expression vectors transfected into host cells (as described below), antibodies isolated from recombinant, combinatorial human antibody libraries (as described below), antibodies isolated from animals transgenic for human immunoglobulin genes (e.g., mice) (see, e.g., Taylor et al. (1992) Nucl. Acids Res. Vol. 20: 6287-6295), or antibodies produced, expressed, created, or isolated by any other means involving splicing of human immunoglobulin gene sequences to other DNA sequences. Such recombinant human antibodies have variable and constant regions derived from human germline immunoglobulin sequences. However, in certain embodiments, such recombinant human antibodies are subjected to in vitro mutagenesis (or, if animals transgenic for human Ig sequences are used, in vivo somatic mutagenesis), and therefore the recombinant antibody is V H and V L The amino acid sequence of the region is human germline V H and V L When derived from or associated with a sequence, it is a sequence that cannot naturally exist in the human antibody germline repertoire in vivo.

[0200] Human antibodies can exist in two forms related to hinge heterogeneity. In one form, the immunoglobulin molecule contains a dimer consisting of four suitable chain constructs of approximately 150-160 kDa, linked by interchain heavy-chain disulfide bonds. In the second form, the dimer is not linked by interchain disulfide bonds, and a molecule of approximately 75-80 kDa is formed, consisting of covalently bonded light and heavy chains (half-antibodies). These forms are extremely difficult to separate even after affinity purification.

[0201] The frequency of the appearance of the second form in various untreated IgG isotypes is due, but not limited to, structural differences related to the hinge region isotype of the antibody. A single amino acid substitution in the hinge region of the human IgG4 hinge can significantly reduce the appearance of the second form (Angal et al. (1993) Molecular Immunology vol. 30: p. 105) to the level generally observed using the human IgG1 hinge. This invention relates to the hinge, C H 2, or C H This includes antibodies that have one or more mutations in three regions, which may be desirable, for example, in manufacturing, to improve the yield of the desired antibody form.

[0202] "Isolated antibody" means an antibody that has been identified, isolated, and / or recovered from at least one component of its natural environment. For example, an antibody that has been isolated or removed from at least one component of an organism, or an antibody that has been isolated or removed from a naturally occurring or naturally produced tissue or cell, is an "isolated antibody." Isolated antibodies also include antibodies in situ within recombinant cells. An isolated antibody is an antibody that has undergone at least one purification or isolation step. According to certain embodiments, an isolated antibody may be substantially free of other cellular material and / or chemicals.

[0203] The term "specifically binds" means that an antibody or its antigen-binding fragment forms a complex with an antigen that 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 and surface plasmon resonance. For example, an antibody that "specifically binds" to IL-33 or IL-4R as characterized in the present invention includes an antibody or a portion thereof that binds to IL-33 or IL-4R, respectively, and K D The levels are approximately less than 1000 nM, less than 500 nM, less than 300 nM, less than 200 nM, less than 100 nM, less than 90 nM, less than 80 nM, less than 70 nM, less than 60 nM, less than 50 nM, less than 40 nM, less than 30 nM, less than 20 nM, less than 10 nM, less than 5 nM, less than 4 nM, less than 3 nM, less than 2 nM, less than 1 nM, or less than 0.5 nM, as measured by surface plasmon resonance assays. However, isolated antibodies that specifically bind to human IL-33 or human IL-4R may exhibit cross-reactivity to other antigens, such as IL-33 or IL-4R molecules obtained from other (non-human) species.

[0204] Anti-IL-33 and anti-IL-4R antibodies useful for this method may contain one or more amino acid substitutions, insertions, and / or deletions (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 substitutions and / or 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 insertions and / or 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 deletions) in the framework of the heavy and light chain variable domains and / or the CDR region, compared to the corresponding germline sequence from which the antibody is derived. Such mutations can be readily identified by comparing the amino acid sequences disclosed herein with germline sequences available, for example, from public antibody sequence databases. The present invention includes methods involving the use of antibodies and their antigen-binding fragments derived from any of the amino acid sequences disclosed herein, and one or more frameworks. and / or one or more amino acids (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 for a tetrameric antibody, or 1, 2, 3, 4, 5, or 6 for the HCVR and LCVR of the antibody) within the CDR region are mutated to the corresponding residue in the germline sequence from which the antibody originates, or the corresponding residue in another human germline sequence, or to a conserved amino acid substitution of the corresponding germline residue (such sequence changes are collectively referred to herein as “germline mutations”). Those skilled in the art can readily produce a number of antibodies and antigen-binding fragments, including one or more individual germline mutations or combinations thereof, starting from the heavy and light chain variable region sequences disclosed herein. In certain embodiments, V H and / or V LAll of the framework and / or CDR residues within the domain are mutated back to residues found in the original germline sequence from which the antibody originates. In other embodiments, only certain residues, for example, only mutated residues found within the first eight amino acids of FR1 or within the last eight amino acids of FR4, or only mutated residues found within CDR1, CDR2, or CDR3, are mutated back to the original germline sequence. In other embodiments, one or more of the framework and / or CDR residues are mutated to corresponding residues in a different germline sequence (i.e., a germline sequence different from the germline sequence from which the antibody originally originates). Furthermore, the antibody may contain any combination of two or more germline mutations within the framework and / or CDR region, for example, certain individual residues are mutated to corresponding residues in a particular germline sequence, and certain other residues different from the original germline sequence are mutated to corresponding residues in a maintained or different germline sequence. After acquisition, antibodies and antigen-binding fragments containing one or more germline mutations can be readily tested for one or more desired properties, such as improved binding specificity, increased binding affinity, (in some cases) improved or enhanced biological properties of the antagonist or agonist, or decreased immunogenicity. The use of antibodies and antigen-binding fragments obtained in this general manner is encompassed within the present invention.

[0205] The present invention also includes methods involving the use of an anti-IL33 or anti-IL-4R antibody comprising any variant of the HCVR, LCVR, and / or CDR amino acid sequences disclosed herein, having one or more conservative substitutions. For example, the present invention includes the use of an anti-IL-4R antibody having an HCVR, LCVR, and / or CDR amino acid sequence, wherein any of the HCVR, LCVR, and / or CDR amino acid sequences disclosed herein is subjected to conservative amino acid substitutions such as 10 or fewer, 8 or fewer, 6 or fewer, or 4 or fewer.

[0206] The term "surface plasmon resonance" refers to an optical phenomenon that enables real-time interaction analysis by detecting changes in protein concentration within a biosensor matrix, for example, using the BIAcore® system (Biacore Life Sciences division of GE Healthcare, Piscataway, NJ).

[0207] The term “K D This refers to the equilibrium dissociation constant of a specific antibody-antigen interaction.

[0208] The term "epitope" refers to an antigenic determinant that interacts with a specific antigen-binding site in the variable region of an antibody molecule, known as a paratope. A single antigen may have more than one epitope. Therefore, different antibodies can bind to different regions on an antigen and have different biological effects. Epitopes can be structural or linear. Structural epitopes are produced by spatially parallel amino acids obtained from different segments of a linear polypeptide chain. Linear epitopes are produced by adjacent amino acids in a polypeptide chain. It is produced by acid residues. In certain circumstances, the epitope may contain sugar, phosphoryl, or sulfonyl groups in the antigen.

[0209] Manufacturing of human antibodies Methods for generating human antibodies in transgenic mice are known in the art. Any such known method can be used to produce human antibodies that specifically bind to human IL-33 or human IL-4R.

[0210] Using VELOCIMMUNE® technology (see, for example, U.S. Patent No. 6,596,541, Regeneron Pharmaceuticals) or any other known method for generating monoclonal antibodies, high-affinity chimeric antibodies to IL-33 or IL-4R having human variable regions and mouse constant regions are first isolated. VELOCIMMUNE® technology uses the generation of transgenic mice having a genome containing human heavy and light chain variable regions that are operably ligated to an endogenous mouse constant region locus, such that the mouse produces antibodies containing human variable and mouse constant regions in response to antigen stimulation. The DNA encoding the heavy and light chain variable regions of the antibody is isolated and operably ligated to the DNA encoding the human heavy and light chain constant regions. The DNA is then expressed in cells capable of expressing fully human antibodies.

[0211] Generally, VELOCIMMUNE® mice are exposed to the target antigen, and lymphoid cells (e.g., B cells) are recovered from the antibody-expressing mice. The lymphoid cells can be fused with myeloma cell lines to produce immortal hybridoma cell lines, which are then screened and selected to identify hybridoma cell lines that produce antibodies specific to the target antigen. DNA encoding the variable regions of the heavy and light chains can be isolated and ligated to the desired isotype constant regions of the heavy and light chains. Such antibody proteins can be produced in cells, e.g., CHO cells. Alternatively, DNA encoding antigen-specific chimeric antibodies or variable domains of the light and heavy chains can be isolated directly from antigen-specific lymphocytes.

[0212] First, a high-affinity chimeric antibody having a human variable region and a mouse constant region is isolated. The antibody is characterized and selected for desirable features, including affinity, selectivity, and epitopes, using standard procedures known to those skilled in the art. The mouse constant region is replaced with a desired human constant region to produce a fully human antibody characterized in the present invention, e.g., wild-type or modified IgG1 or IgG4. The selected constant region may vary depending on the specific use, and the high-affinity antigen-binding and target-specific features reside in the variable region.

[0213] Generally, antibodies that can be used in this method exhibit high affinity, as described above, when measured by binding to an antigen immobilized in a solid or dissolved phase. The mouse constant region is replaced with a desired human constant region to produce a fully human antibody characterized in the present invention. The selected constant region may vary depending on the specific use, and the high affinity antigen binding and target specificity characteristics reside in the variable region.

[0214] In one embodiment, a human antibody or antigen-binding fragment that specifically binds to IL-33 and can be used in the context of the method characterized in the present invention comprises three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) contained within a heavy chain variable region (HCVR) having the amino acid sequence of SEQ ID NO: 2. The antibody or antigen-binding fragment may also include three light chain CDRs (LCVR1, LCVR2, and LCVR3) contained within a light chain variable region (LCVR) having the amino acid sequence of SEQ ID NO: 10. In another embodiment, a human antibody that specifically binds to IL-4R and can be used in the context of the method characterized in the present invention. A heterobinding human antibody or its antigen-binding fragment contains three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) contained within a heavy chain variable region (HCVR) having the amino acid sequence of SEQ ID NO: 27. The antibody or antigen-binding fragment may also contain three light chain CDRs (LCVR1, LCVR2, and LCVR3) contained within a light chain variable region (LCVR) having the amino acid sequence of SEQ ID NO: 28.

[0215] Methods and techniques for identifying CDRs within HCVR and LCVR amino acid sequences are well known in the art and can be used to identify CDRs within the specified HCVR and / or LCVR amino acid sequences disclosed herein. Exemplary practices that can be used to identify CDR boundaries include, for example, the Kabat definition, the Chothia definition, and the AbM definition. In general terms, the Kabat definition is based on sequence diversity, the Chothia definition is based on the location of structural loop regions, and the AbM definition is a compromise between the Kabat and Chothia approaches. See, for example, Kabat, "Sequences of Proteins of Immunological Interest," National Institutes of Health, Bethesda, Md. (1991); Al-Lazikani et al., J.Mol.Biol. 273:927-948 (1997); and Martin et al., Proc.Natl.Acad.Sci.USA 86:9268-9272 (1989). Public databases are also available for identifying CDR sequences within antibodies.

[0216] In a particular embodiment, the antibody or its antigen-binding fragment comprises six CDRs (HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3) obtained from the heavy-chain and light-chain variable region amino acid sequence pairs (HCVR / LCVR) of SEQ ID NOs. 2 and 10.

[0217] In a particular embodiment, the antibody or its antigen-binding fragment comprises six CDRs (HCDR1 / HCDR2 / HCDR3 / LCDR1 / LCDR2 / LCDR3) having the amino acid sequences of SEQ ID NOs. 4 / 6 / 8 / 12 / 14 / 16.

[0218] In a particular embodiment, the antibody or its antigen-binding fragment comprises the HCVR / LCVR amino acid sequence pair of SEQ ID NOs: 2 and 10.

[0219] In one embodiment, the antibody is REGN3500 and contains the HCVR / LCVR amino acid sequence pairs of SEQ ID NOs: 2 and 10, and the heavy chain / light chain amino acid sequence pairs of SEQ ID NOs: 18 and 20.

[0220] In a particular embodiment, the antibody or its antigen-binding fragment comprises six CDRs (HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3) obtained from the heavy-chain and light-chain variable region amino acid sequence pairs (HCVR / LCVR) of SEQ ID NOs. 27 and 28.

[0221] In a particular embodiment, the antibody or its antigen-binding fragment comprises six CDRs (HCDR1 / HCDR2 / HCDR3 / LCDR1 / LCDR2 / LCDR3) having the amino acid sequences of SEQ ID NOs. 21 / 22 / 23 / 24 / 25 / 26.

[0222] In a particular embodiment, the antibody or its antigen-binding fragment comprises the HCVR / LCVR amino acid sequence pair of SEQ ID NOs. 27 and 28.

[0223] In one embodiment, the antibody is dupilumab, and the HCVR / It contains LCVR amino acid sequence pairs and heavy / light chain amino acid sequence pairs of SEQ ID NOs. 29 and 30.

[0224] Pharmaceutical composition The present invention includes a method comprising the step of administering an IL-33 antagonist, an IL-4R antagonist, or an IL-33 antagonist and an IL-4R antagonist to a patient, wherein the IL-33 antagonist, an IL-4R antagonist, or an IL-33 antagonist and an IL-4R antagonist are contained in a pharmaceutical composition. The present invention also includes an IL-33 antagonist, an IL-4R antagonist, or an IL-33 antagonist and an IL-4R antagonist for use, wherein the IL-33 antagonist, an IL-4R antagonist, or an IL-33 antagonist and an IL-4R antagonist are contained in a pharmaceutical composition. The pharmaceutical compositions characterized in the present invention are formulated with appropriate carriers, excipients, and other agents that provide appropriate mobility, delivery, resistance, etc. Numerous suitable formulations can be found in the prescription collection known to all pharmacists: Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, PA. These formulations include, for example, powders, pastes, ointments, gels, waxes, oils, lipids, lipid (cationic or anionic)-containing vesicles (e.g., LIPOFECTIN®), DNA conjugates, anhydrous absorbent pastes, oil-in-water and water-in-oil emulsions, carbowax emulsions (polyethylene glycol of various molecular weights), semi-solid gels, and semi-solid mixtures containing carbowax. See also Powell et al., "Compendium of excipients for parenteral formulations," PDA (1998), J. Pharm. Sci. Technol., Vol. 52: pp. 238–311.

[0225] The dosage of the antibody administered to a patient can vary depending on the patient's age and size, symptoms, condition, route of administration, etc. The dosage is generally calculated according to body weight or body surface area. Depending on the severity of the condition, the frequency and duration of treatment can be adjusted. An effective dosage and schedule for administering a pharmaceutical composition containing an anti-IL-33 antibody or an anti-IL-4R antibody can be determined based on experience. For example, the progression of a patient can be monitored by regular evaluations and appropriately adjusted dosages. Furthermore, interspecies scaling of dosages can be performed using methods well known in the art (e.g., Mordenti et al., 1991, Pharmaceut. Res. 8:1351).

[0226] Various delivery systems are known and can be used to administer the pharmaceutical compositions characterized in the present invention, for example, encapsulation in liposomes, microparticles, microcapsules, recombinant cells capable of expressing mutant viruses, receptor-mediated endocytosis (see, for example, Wu et al., 1987, J. Biol. Chem. 262:4429 - 4432). Methods of administration include, but are not limited to, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, intratracheal, epidural, and oral routes. The present composition can be administered by any convenient route, for example, by injection or bolus injection, by absorption through the epithelium or the back layer of the mucocutaneous (e.g., oral mucosa, rectal and intestinal mucosa, etc.), and can be administered together with other bioactive agents.

[0227] The pharmaceutical compositions characterized in the present invention can be delivered subcutaneously or intravenously using standard needles and syringes. Furthermore, with respect to subcutaneous delivery, pen-type delivery devices (e.g., pen-type autoinjectors) are readily applicable in delivering the pharmaceutical compositions characterized in the present invention. Such pen-type delivery devices can be reusable or disposable. Reusable pen-type delivery devices generally utilize replaceable cartridges containing the pharmaceutical composition. All of the pharmaceutical composition within the cartridge is administered, Once the cartridge is empty, the empty cartridge can be easily discarded and replaced with a new cartridge containing the pharmaceutical composition. The pen-type delivery device can then be reused. In a disposable pen-type delivery device, 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 empty of the pharmaceutical composition, the entire device is discarded.

[0228] Numerous reusable pen-type and auto-injector delivery devices have applications in the subcutaneous delivery of pharmaceutical compositions. Examples include, but are not limited to, AUTOPEN® (Owen Mumford, Inc., Woodstock, UK), DISETRONIC® pen (Disetronic Medical Systems, Bergdorf, Switzerland), HUMALOG MIX 75 / 25® pen, HUMALOG® pen, HUMALIN 70 / 30® pen (Eli Lilly and Co., Indianapolis, IN), NOVOPEN® I, II and III (Novo Nordisk, Copenhagen, Denmark), NOVOPEN JUNIOR® (Novo Nordisk, Copenhagen, Denmark), BD® pen (Becton Dickinson, Franklin Lakes, NJ), OPTIPEN®, OPTIPEN PRO®, and OPTIPEN®. This includes STARLET® and OPTICLIK® (Sanofi-Aventis, Frankfurt, Germany). Examples of disposable pen-type delivery devices that have applications in subcutaneous delivery of the pharmaceutical compositions characterized in the present invention include, but are not limited to, SOLOSTAR® pens (Sanofi-Aventis), FLEXPEN® (Novo Nordisk), and KWIKPEN® (Eli Lilly), SURECLICK® auto-injectors (Amgen, Thousand Oaks, CA), PENLET® (Haselmeier, Stuttgart, Germany), EPIPEN (Dey, LP), and HUMIRA® pens (Abbott Labs, Abbott Park, IL).Examples of high-volume delivery devices (e.g., high-volume injectors) include, but are not limited to, bolus injectors such as BD Libertas West SmartDose, Enable Injections, SteadyMed PatchPump, Sensile SenseTrial, YPsomed YpsoDose, and Bespak Lapas.

[0229] For direct administration into the sinuses, the pharmaceutical compositions characterized in this invention may be administered, for example, using a microcatheter (e.g., an endoscope and microcatheter), an aerozoizer, a powder dispenser, a nebulizer, or an inhaler. This method includes the administration of an IL-33 antagonist or an IL-4R antagonist in the form of an aerosolized formulation to a subject requiring such an antagonist. For example, an aerosolized antibody against IL-33 or IL-4R may be administered to a patient to treat asthma. The aerosolized antibody can be manufactured, for example, as described in U.S. Patent No. 8,178,098, which is incorporated herein by reference in its entirety.

[0230] In certain circumstances, pharmaceutical compositions can be delivered by a controlled-release system. In one embodiment, a pump can be used (see Langer, above; Sefton, 1987, CRC Crit.Ref. Biomed.Eng. Vol. 14: p. 201). In another embodiment, a polymer material can be used; see Medical Applications of Controlled Release, Langer and Wise (eds.), 1974, CRC Press, Boca Raton, Florida. In yet another embodiment, the controlled-release system is used to deliver the composition. It can be placed close to the target and therefore requires only a small fraction of the systemic dose (see, for example, Goodson, 1984, in Medical Applications of Controlled Release, Vol. 2, pp. 115-138, above). Other controlled-release systems are discussed in the review by Langer, 1990, Science Vol. 249: pp. 1527-1533.

[0231] Injectable preparations may include dosage forms for intravenous, subcutaneous, intradermal, and intramuscular injection, as well as intravenous drip infusion. These injectable preparations can be manufactured by known methods. For example, an injectable preparation may be manufactured by dissolving, suspending, or emulsifying, for example, an antibody or a salt thereof in a sterile aqueous or oily medium commonly used for injection. Examples of aqueous media for injection include physiological saline, isotonic solutions containing glucose, and other adjuvants, which can be used in combination with suitable solubilizers, such as alcohols (e.g., ethanol), polyhydric alcohols (e.g., propylene glycol, polyethylene glycol), and nonionic surfactants (e.g., polysorbate 80, HCO-50 (polyoxyethylene (50 mol) adduct of hydrogenated castor oil)). Examples of oily media include sesame oil and soybean oil, which may be used in combination with solubilizers, such as benzyl benzoate and benzyl alcohol. Therefore, the manufactured injections are generally filled into suitable ampoules.

[0232] Advantageously, the aforementioned pharmaceutical compositions for oral or parenteral use are manufactured into dosage forms in unit doses suitable for the dosage of the active ingredient. Such dosage forms in unit doses include, for example, tablets, pills, capsules, injections (ampoules), suppositories, and the like.

[0233] A model pharmaceutical composition containing an anti-IL-4R antibody that can be used in the present invention is disclosed, for example, in U.S. Patent Application Publication No. 2012 / 0097565.

[0234] Dosage The amount of IL-33 antagonist (e.g., anti-IL-33 antibody or its antigen-binding fragment) or IL-4R antagonist (e.g., anti-IL-4R antibody or its antigen-binding fragment) administered to a subject in accordance with the methods or uses characterized in the present invention is generally a therapeutically effective dose. As used herein, the term “therapeutically effective dose” means the amount of IL-33 antagonist or IL-4R antagonist that results in (a) a reduction in the incidence of allergic asthma exacerbations; (b) improvement in one or more allergic asthma-related parameters (as defined elsewhere herein); and / or (c) a detectable improvement in one or more symptoms or signs of an upper respiratory tract inflammatory condition. “Therapeuticly effective dose” also includes the amount of IL-33 antagonist or IL-4R antagonist that inhibits, prevents, reduces or delays the progression of allergic asthma in a subject.

[0235] In the case of anti-IL-33 antibodies or anti-IL-4R antibodies, the therapeutically effective dose is approximately 0.05 mg to 700 mg of anti-IL-33 antibody or anti-IL-4R antibody, for example, approximately 0.05 mg, approximately 0.1 mg, approximately 1.0 mg, approximately 1.5 mg, approximately 2.0 mg, approximately 3.0 mg, approximately 5.0 mg, approximately 7.0 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 15 0mg, about 160mg, about 170mg, about 180mg, about 190mg, about 200mg, about 210mg, about 220mg, about 230mg, about 240mg, about 250mg, about 260mg, about 270mg, about 280mg, about 290mg, about 300mg, about 310mg, about 320mg, about 330mg, about 340mg, about 350mg, about 360mg, about 370mg, about 380mg, about 390mg, about 400mg, about 410mg, about 420mg, about 430mg, about 440mg, about 450mg, about 460mg The dosage may be approximately 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, 600 mg, 610 mg, 620 mg, 630 mg, 640 mg, 650 mg, 660 mg, 670 mg, 680 mg, 690 mg, or 700 mg. In certain embodiments, 300 mg of anti-IL-4R antibody is administered.

[0236] The amount of IL-33 antagonist or IL-4R antagonist contained within an individual dose range can be expressed in terms of milligrams of antibody per kilogram of body weight (i.e., mg / kg). For example, an IL-4R antagonist may be administered to a patient in doses ranging from approximately 0.0001 to approximately 30 mg per kg of body weight. In the case of anti-IL-33 antibodies or anti-IL-4R antibodies, the therapeutically effective dose may be approximately 1 mg / kg, 2 mg / kg, 3 mg / kg, 4 mg / kg, 5 mg / kg, 6 mg / kg, 7 mg / kg, 8 mg / kg, 9 mg / kg, 10 mg / kg, 11 mg / kg, 12 mg / kg, 13 mg / kg, 14 mg / kg, 15 mg / kg, 16 mg / kg, 17 mg / kg, 18 mg / kg, 19 mg / kg, 20 mg / kg, 21 mg / kg, 22 mg / kg, 23 mg / kg, 24 mg / kg, 25 mg / kg, 26 mg / kg, 27 mg / kg, 28 mg / kg, 29 mg / kg, or 30 mg / kg of anti-IL-33 antibody or anti-IL-4R antibody. In a particular embodiment, an anti-IL-33 antibody at a dose of 10 mg / kg is administered.

[0237] In some embodiments, the dose of an IL-4R antagonist or IL-33 antagonist may vary depending on the eosinophil count. For example, the subjects may have a blood eosinophil count (high blood eosinophil count) of ≥300 cells / μL, or 300-499 cells / μL or ≥500 cells / μL (HEos); a blood eosinophil count of 200-299 cells / μL (moderate blood eosinophil count); or a blood eosinophil count of <200 cells / μL (low blood eosinophil count).

[0238] In some embodiments, the dose of an IL-4R antagonist or an IL-33 antagonist may vary depending on the periostin level. For example, the subjects may have high periostin levels (e.g., ≥75.0 ng / mL or 74.4 ng / mL) or low periostin levels (e.g., <75.0 ng / mL or <74.4 ng / mL).

[0239] In some embodiments, the method includes an initial dose of approximately 5 mg / kg to 15 mg / kg of IL-33 antagonist, for example, approximately 10 mg / kg of IL-33 antagonist. In certain embodiments, the method includes an initial dose of approximately 200 to 600 mg of IL-4R antagonist, for example, approximately 600 mg of IL-4R antagonist.

[0240] In certain embodiments, the method includes a single or more maintenance dose of approximately 200 to 300 mg of an IL-4R antagonist.

[0241] In certain embodiments, ICS and LABA are administered for the duration of administration of an IL-33 antagonist. In certain embodiments, ICS and LABA are administered for the duration of administration of an IL-4R antagonist.

[0242] In a particular embodiment, the initial dose comprises 600 mg of an anti-IL-4R antibody or its antigen-binding fragment, and one or more maintenance doses comprise 300 mg of the antibody or its antigen-binding fragment administered every other week.

[0243] In other embodiments, the initial dose comprises 600 mg of anti-IL-4R antibody or its antigen-binding fragment, and one or more maintenance doses are administered every four weeks. Contains 300 mg of the original conjugated fragment.

[0244] In other embodiments, the loading dose comprises 600 mg of an anti-IL-4R antibody or an antigen-binding fragment thereof, and the one or more maintenance doses comprise 300 mg of the antibody or an antigen-binding fragment thereof administered once a week.

[0245] In other embodiments, the loading dose comprises 600 mg of an anti-IL-4R antibody or an antigen-binding fragment thereof, and the one or more maintenance doses comprise 300 mg of the antibody or an antigen-binding fragment thereof administered every three weeks.

[0246] In one embodiment, the subject is 6 to <18 years old, and the IL-33 antibody or an antigen-binding fragment thereof or the IL-4R antibody or an antigen-binding fragment thereof is administered at 2 mg / kg or 4 mg / kg.

[0247] In one embodiment, the subject is 12 to <18 years old, and the IL-33 antibody or an antigen-binding fragment thereof or the IL-4R antibody or an antigen-binding fragment thereof is administered at 2 mg / kg or 4 mg / kg.

[0248] In one embodiment, the subject is 6 to <12 years old, and the IL-33 antibody or an antigen-binding fragment thereof or the IL-4R antibody or an antigen-binding fragment thereof is administered at 2 mg / kg or 4 mg / kg.

[0249] In one embodiment, the subject is 2 to <6 years old, and the IL-33 antibody or an antigen-binding fragment thereof or the IL-4R antibody or an antigen-binding fragment thereof is administered at 2 mg / kg or 4 mg / kg.

[0250] In yet other embodiments, the subject is <2 years old, and the IL-33 antibody or an antigen-binding fragment thereof or the IL-4R antibody or an antigen-binding fragment thereof is administered at 2 mg / kg or 4 mg / kg.

[0251] Combination therapy Certain embodiments of the methods characterized in the present invention include the step of administering to a target one or more additional therapeutic agents in combination with an IL-33 antagonist, one or more additional therapeutic agents in combination with an IL-4R antagonist, or one or more additional therapeutic agents in combination with an IL-33 antagonist and an IL-4R antagonist. Certain embodiments of the present invention include an IL-33 antagonist, an IL-4R antagonist, or an IL-33 antagonist and an IL-4R antagonist for use in combination with additional therapeutic agents. Certain embodiments of the present invention include a combination of an IL-33 antagonist, an IL-4R antagonist, or an IL-33 antagonist and an IL-4R antagonist with additional therapeutic agents for use. As used herein, the expression “in combination with” means that an additional therapeutic agent is administered before, after, or concurrently with a pharmaceutical composition comprising an IL-4R antagonist, an IL-33 antagonist, or an IL-33 antagonist and an IL-4R antagonist. In some embodiments, the term “in combination with” includes sequential or concurrent administration of an IL-33 antagonist, an IL-4R antagonist, or an IL-33 antagonist and an IL-4R antagonist, and the additional therapeutic agent. The present invention includes a method for treating asthma or a related condition or complication, or reducing at least one exacerbation, comprising the step of administering an IL-33 antagonist, an IL-4R antagonist, or an IL-33 antagonist and an IL-4R antagonist in combination with an additional therapeutic agent for additive or synergistic activity.

[0252] For example, when administered "before" an IL-33 antagonist, an IL-4R antagonist, or a pharmaceutical composition containing an IL-33 antagonist and an IL-4R antagonist, the additional therapeutic agent may be administered approximately 72 hours, 60 hours, 48 ​​hours, 36 hours, 24 hours, 12 hours, 10 hours, 8 hours, 6 hours, 4 hours, 2 hours, 1 hour, 30 minutes, 15 minutes, or 10 minutes before the administration of the IL-33 antagonist, an IL-4R antagonist, or a pharmaceutical composition containing an IL-33 antagonist and an IL-4R antagonist. When administered "after" an IL-33 antagonist, an IL-4R antagonist, or a pharmaceutical composition containing an IL-33 antagonist and an IL-4R antagonist, the additional therapeutic agent may be administered approximately 10 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, 10 hours, 12 hours, 24 hours, 36 hours, 48 ​​hours, 60 hours, or 72 hours after the administration of the IL-33 antagonist, an IL-4R antagonist, or a pharmaceutical composition containing an IL-33 antagonist and an IL-4R antagonist. "Contemporary" administration of an IL-33 antagonist, an IL-4R antagonist, or a pharmaceutical composition containing both an IL-33 antagonist and an IL-4R antagonist means that the additional therapeutic agent is administered to the subject in a separate dosage form within 5 minutes of (before, after, or simultaneously with) the administration of the IL-33 antagonist, an IL-4R antagonist, or a pharmaceutical composition containing both an IL-33 antagonist and an IL-4R antagonist, or that the additional therapeutic agent is administered to the subject as a combined single-dose formulation containing both the IL-33 antagonist and the IL-33 antagonist, an IL-4R antagonist, or both an IL-33 antagonist and an IL-4R antagonist.

[0253] Additional therapeutic agents include, for example, another IL-33 antagonist, another IL-4R antagonist, IL-1 antagonist (e.g., including the IL-1 antagonist described in U.S. Patent No. 6,927,044), IL-6 antagonist, IL-6R antagonist (e.g., including the anti-IL-6R antibody described in U.S. Patent No. 7,582,298), TNF antagonist, IL-8 antagonist, IL-9 antagonist, IL-17 antagonist, IL-5 antagonist, IgE antagonist, CD48 antagonist, leukotriene inhibitors, antifungal agents, NSAIDs, and short-acting β-antibodies. 2-agonists (e.g., bitolterol, fenoterol, isoprenaline, isoproterenol, levosalbutamol, levalbuterol, orciprenaline, metaproterenol, pirbuterol, procaterol, ritodrine, salbutamol, albuterol, or terbutaline), long-acting β2-agonists (e.g., salmeterol or formoterol), inhaled corticosteroids (e.g., fluticasone or budesonide), systemic corticosteroids (e.g., oral or intravenous), methylxanthines, nedocromyl sodium, cromolyn sodium, or combinations thereof. For example, in certain embodiments, a pharmaceutical composition comprising an IL-4R antagonist, an IL-33 antagonist, or an IL-33 antagonist and an IL-4R antagonist is administered in combination with a long-acting β2 agonist and an inhaled corticosteroid (e.g., fluticasone + salmeterol [e.g., Advair® (GlaxoSmithKline)]; or budesonide + formoterol [e.g., SYMBICORT® (Astra Zeneca)]).

[0254] Administration regimen According to certain embodiments, multiple doses of an IL-33 antagonist, an IL-4R antagonist, or an IL-33 antagonist and an IL-4R antagonist may be administered to a subject over a defined time course. Such a method or use involves administering multiple doses of an IL-33 antagonist, an IL-4R antagonist, or an IL-33 antagonist and an IL-4R antagonist sequentially to a subject. As used herein, “administer sequentially” means administering an IL-33 antagonist, an IL-4R The term "antagonist" or "IL-33 antagonist and IL-4R antagonist" means that each dose is administered to the subject at different times, for example, on different days separated by a predetermined interval (e.g., hours, days, weeks, or months). Methods or uses include administering to the patient sequentially a single initial dose of the IL-33 antagonist, IL-4R antagonist, or IL-33 antagonist and IL-4R antagonist, followed by one or more second doses of the IL-33 antagonist, IL-4R antagonist, or IL-33 antagonist and IL-4R antagonist, and optionally, a third dose of the IL-33 antagonist, IL-4R antagonist, or IL-33 antagonist and IL-4R antagonist.

[0255] The present invention includes a method or use comprising administering to a subject an IL-33 antagonist, an IL-4R antagonist, or a pharmaceutical composition comprising an IL-33 antagonist and an IL-4R antagonist at intervals of approximately four times per week, twice per week, once per week (q1w), every other week (once every two weeks or q2w), once every three weeks (every three weeks or q3w), once every four weeks (monthly or q4w), once every five weeks (q5w), once every six weeks (q6w), once every eight weeks (q8w), once every twelve weeks (q12w), or fewer doses until a therapeutic effect is achieved. In certain embodiments comprising the administration of a pharmaceutical composition comprising an anti-IL-33 antibody or an anti-IL-4R antibody, once-weekly doses of approximately 75 mg, 100 mg, 150 mg, 200 mg, or 300 mg may be used. In other embodiments involving the administration of a pharmaceutical composition containing an anti-IL-33 antibody or an anti-IL-4R antibody, doses of approximately 75 mg, 100 mg, 150 mg, 200 mg, or 300 mg can be administered every other week (once every two weeks). In other embodiments involving the administration of a pharmaceutical composition containing an anti-IL-33 antibody or an anti-IL-4R antibody, doses of approximately 75 mg, 100 mg, 150 mg, 200 mg, or 300 mg can be administered every three weeks. In other embodiments involving the administration of a pharmaceutical composition containing an anti-IL-33 antibody or an anti-IL-4R antibody, doses of approximately 75 mg, 100 mg, 150 mg, 200 mg, or 300 mg can be administered every four weeks (once a month). In other embodiments involving the administration of a pharmaceutical composition containing an anti-IL-33 antibody or an anti-IL-4R antibody, doses of approximately 75 mg, 100 mg, 150 mg, 200 mg, or 300 mg once every five weeks may be used. In other embodiments involving the administration of a pharmaceutical composition containing an anti-IL-33 antibody or an anti-IL-4R antibody, doses of approximately 75 mg, 100 mg, 150 mg, 200 mg, or 300 mg once every six weeks may be used. In other embodiments involving the administration of a pharmaceutical composition containing an anti-IL-33 antibody or an anti-IL-4R antibody, doses of approximately 75 mg, 100 mg, 150 mg, 200 mg, or 300 mg once every eight weeks may be used.In other embodiments, which involve the administration of a pharmaceutical composition containing an anti-IL-33 antibody or an anti-IL-4R antibody, doses of approximately 75 mg, 100 mg, 150 mg, 200 mg, or 300 mg once every 12 weeks may be used. In one embodiment, the route of administration is subcutaneous.

[0256] The term "week" refers to a period of (n × 7 days) ± 2 days, for example, (n × 7 days) ± 1 day, or (n × 7 days), where "n" is the number of weeks, for example, 1, 2, 3, 4, 5, 6, 8, 12 weeks or more.

[0257] The terms “initial dose,” “subsequent doses,” “second dose,” and “third dose” refer to the time series of IL-4R antagonist or IL-33 antagonist administrations. Therefore, the “initial dose” is the dose administered at the start of the treatment regimen (also referred to as the “baseline dose”); “subsequent doses” and “second dose” are doses administered after the initial dose; and “third dose” is the dose administered after the second dose. The initial dose, subsequent doses, second dose, and third dose may all comprise the same amount of IL-33 antagonist or IL-4R antagonist, but generally may differ from one another in terms of the number of administrations. However, in certain embodiments, the initial dose, subsequent doses, second dose The amounts of IL-33 antagonist or IL-4R antagonist contained in the dose and / or third dose vary from one another during treatment (e.g., adjusted by increasing or decreasing as needed). In certain embodiments, two or more doses (e.g., two, three, four, or five or more) are administered at the start of the treatment regimen as an “initial dose” or “loading dose,” followed by subsequent doses administered in fewer doses (e.g., “maintenance doses”). In one embodiment, the maintenance dose may be less than the loading dose or initial dose. For example, one or more loading doses of 600 mg of IL-4R antagonist may be followed by maintenance doses of approximately 75 mg to approximately 300 mg.

[0258] In one particular embodiment, the initial dose is approximately 400 mg to 600 mg of IL-4R antagonist. In another embodiment, the initial dose is 400 mg of IL-4R antagonist. In yet another embodiment, the initial dose is 600 mg of IL-4R antagonist.

[0259] In one particular embodiment, the subsequent dose is approximately 200 to 300 mg of IL-4R antagonist. In another embodiment, the subsequent dose is 200 mg of IL-4R antagonist. In yet another embodiment, the subsequent dose is 300 mg of IL-4R antagonist.

[0260] In one particular embodiment, the initial dose is approximately 5 mg / kg to 15 mg / kg of IL-33 antagonist. In another embodiment, the initial dose is 10 mg / kg of IL-33 antagonist.

[0261] In one particular embodiment, the subsequent dose is approximately 5 mg / kg to 15 mg / kg of IL-33 antagonist. In another embodiment, the subsequent dose is 5 mg / kg of IL-33 antagonist. In yet another embodiment, the subsequent dose is 10 mg / kg of IL-33 antagonist. In yet another embodiment, no subsequent dose is administered to the subject (for example, only the initial dose is administered to the subject).

[0262] In certain embodiments, the loading dose is twice the maintenance dose. In some embodiments, the initial dose is the same as the maintenance dose. In certain embodiments, the initial dose is a single dose.

[0263] In some embodiments, the initial dose comprises 300 mg of the antibody or its antigen-binding fragment, and one or more maintenance doses comprise 300 mg of the antibody or its antigen-binding fragment administered every other week.

[0264] In some embodiments, the initial dose comprises 300 mg of the antibody or its antigen-binding fragment, and one or more maintenance doses comprise 300 mg of the antibody or its antigen-binding fragment administered every four weeks.

[0265] In some embodiments, the subjects have mild allergic asthma, and the initial dose comprises 600 mg of the antibody or its antigen-binding fragment, with one or more maintenance doses comprising 300 mg of the antibody or its antigen-binding fragment administered every four weeks.

[0266] In some embodiments, the initial dose comprises 10 mg / kg of the antibody or its antigen-binding fragment, and one or more maintenance doses comprise 10 mg / kg of the antibody or its antigen-binding fragment administered every other week.

[0267] In some embodiments, the initial dose comprises 10 mg / kg of the antibody or its antigen-binding fragment, and one or more maintenance doses are administered every four weeks. Contains 10 mg / kg of conjugated fragment.

[0268] In some embodiments, the subjects have mild allergic asthma, and the initial dose contains 10 mg / kg of the antibody or its antigen-binding fragment.

[0269] In a model embodiment, each subsequent, second and / or third dose is administered 1 to 14 weeks (e.g., 1, 1 1 / 2, 2, 2 1 / 2, 3, 3 1 / 2, 4, 4 1 / 2, 5, 5 1 / 2, 6, 6 1 / 2, 7, 7 1 / 2, 8, 8 1 / 2, 9, 9 1 / 2, 10, 10 1 / 2, 11, 11 1 / 2, 12, 12 1 / 2, 13, 13 1 / 2, 14, 14 1 / 2 or more) weeks after the immediately preceding dose. The phrase “immediately preceding dose” means a dose of IL-33 antagonist or IL-4R antagonist administered to the patient in a series of multiple doses, in an order that does not interfere with the dose, before the administration of the next dose.

[0270] The method or use may include the step of administering a considerable number of second and / or third doses of an IL-33 antagonist or an IL-4R antagonist to a patient. For example, in one particular embodiment, only a single second dose is administered to the patient. In other embodiments, two or more (e.g., two, three, four, five, six, seven, eight or more) second doses are administered to the patient. For example, in one particular embodiment, only a single third dose is administered to the patient. In other embodiments, two or more (e.g., two, three, four, five, six, seven, eight or more) third doses are administered to the patient.

[0271] In embodiments including multiple second doses, each second dose may be administered in the same number of doses as the other second doses. For example, each second dose may be administered to the patient 1 to 2 weeks after the previous dose. Similarly, in embodiments including multiple third doses, each third dose may be administered in the same number of doses as the other third doses. For example, each third dose may be administered to the patient 2 to 4 weeks after the previous dose. Alternatively, the number of times the second and / or third doses are administered to the patient may vary during the treatment regimen. The number of doses may also be adjusted during treatment by the physician according to the individual patient's needs after clinical examinations.

[0272] The present invention includes a method comprising sequential administration to a patient of an IL-33 antagonist, an IL-4R antagonist, or an IL-33 antagonist and an IL-4R antagonist, along with additional therapeutic agents, for the treatment of asthma or a related condition. The present invention also comprises an IL-33 antagonist, an IL-4R antagonist, or an IL-33 antagonist and an IL-4R antagonist for use in a patient to treat asthma or a related condition, wherein the patient is treated by sequential administration of an IL-33 antagonist or an IL-33 antagonist and an IL-4R antagonist, along with additional therapeutic agents. In some embodiments, the method or use includes the step of administering one or more doses of an IL-33 antagonist or one or more doses of an IL-33 antagonist and an IL-4R antagonist, followed by one or more doses (e.g., two, three, four, five, six, seven, eight or more) of an additional therapeutic agent. For example, one or more doses of an IL-4R antagonist of about 75 mg to about 300 mg and / or one or more doses of an IL-33 antagonist of about 5 mg / kg to about 20 mg / kg may be administered, followed by one or more doses (e.g., two, three, four, five, six, seven, eight or more) of an additional therapeutic agent (e.g., as described in other parts of this specification, inhaled corticosteroids or β2-agonists or any other therapeutic agent) to treat, alleviate, reduce or relieve one or more symptoms of asthma. In some embodiments, an IL-33 antagonist and / or an IL-33 antagonist is administered in one or more doses (e.g., two, three, four, five, six, seven, eight or more) resulting in improvement of one or more asthma-related parameters, followed by a recurrence of at least one symptom of asthma. A second therapeutic agent is administered for prevention. Alternative embodiments involve the simultaneous administration of an IL-33 antagonist and / or an IL-4R antagonist, as well as an additional therapeutic agent. For example, one or more doses (e.g., two, three, four, five, six, seven, eight or more) of the IL-33 antagonist and / or IL-4R antagonist are administered, and the additional therapeutic agent is administered in separate doses at a similar or different number of times to the IL-33 antagonist and / or IL-4R antagonist. In some embodiments, the additional therapeutic agent is administered before, after, or concurrently with the IL-33 antagonist and / or IL-4R antagonist.

[0273] In certain embodiments, the IL-33 antagonist and / or IL-4R antagonist is administered every other week for 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48 weeks or longer. In other embodiments, the IL-33 antagonist and / or IL-4R antagonist is administered once every four weeks for 12, 16, 20, 24, 28, 32, 36, 40, 44, 48 weeks or longer. In detailed embodiments, the IL-33 antagonist and / or IL-4R antagonist is administered for at least 24 weeks.

[0274] The present invention includes a method for treating a subject with mild allergic asthma, comprising the step of administering a loading dose to a subject of an antibody or antigen-binding fragment thereof that specifically binds to IL-4R and / or an antibody or antigen-binding fragment thereof that specifically binds to IL-33. In certain embodiments, the method or use comprises the step of administering to a subject multiple maintenance doses of one or more antibodies or (their) antigen-binding fragments, the multiple maintenance doses being administered during the treatment period.

[0275] Treatment population For example, “subjects requiring it” may include, for example, subjects who, prior to treatment, exhibit (or exhibit) one or more asthma-related parameters, e.g., decreased FEV1 (e.g., less than 2.0 L), decreased FEF by 25-75%, decreased AM PEF (e.g., less than 400 L / min), decreased PM PEF (e.g., less than 400 L / min), an ACQ5 score of at least 2.5, at least one nocturnal awakening per night, and / or a SNOT-22 score of at least 20.

[0276] The method characterized in the present invention includes the step of administering a therapeutic composition comprising an IL-33 antagonist, an IL-4R antagonist, or both an IL-33 antagonist and an IL-4R antagonist to a subject in need. The phrase "subject in need" refers to a human or non-human animal exhibiting one or more symptoms or signs of asthma (e.g., allergic asthma), or a human or non-human animal diagnosed with asthma. In various embodiments, the method can be used to treat mild, moderate to severe, and severe allergic asthma in patients in need, including mild persistent allergic asthma.

[0277] In related embodiments, “patients who need it” may be subjects who have been prescribed or are currently receiving a SABA or ICS / LABA combination prior to being administered an IL-4R antagonist or an IL-33 antagonist and an IL-4R antagonist. Examples of SABAs include, but are not limited to, bitolterol, fenoterol, isoprenaline, isoproterenol, levosalbutamol, levalbuterol, orciprenaline, metaproterenol, pirbuterol, procaterol, ritodrine, salbutamol, albuterol, or terbutamol Tallinn is included.

[0278] Examples of ICS include, but are not limited to, mometasone furoate, budesonide, and fluticasone propionate. Examples of LABA include, but are not limited to, formoterol and salmeterol. Examples of ICS / LABA therapy include, but are not limited to, fluticasone / salmeterol combination therapy and budesonide / formoterol combination therapy. For example, the present invention includes a method comprising, where necessary, administering an IL-4R antagonist or both an IL-33 antagonist and an IL-4R antagonist to a patient who has received a course of SABA for two weeks or more immediately prior to the administration of an IL-4R antagonist and / or an IL-33 antagonist (such prior treatment is referred to herein as “basic treatment”). The present invention also includes a treatment method in which the basic treatment is continued in combination with the administration of an IL-4R antagonist and / or an IL-33 antagonist. In further embodiments, the amount of LABA is gradually reduced before or after the initiation of administration of an IL-4R antagonist and / or an IL-33 antagonist. In some embodiments, a method is provided for treating patients with mild persistent asthma for at least ≥12 months. In one embodiment, patients with mild persistent allergic asthma may be administered an IL-4R antagonist and / or an IL-33 antagonist according to this method.

[0279] In some embodiments, “patients in need of it” may be subjects with elevated levels of asthma-related biomarkers. Examples of asthma-related biomarkers include, but are not limited to, IgE, thymic and activating modulated chemokines (TARCs), eotaxin-3, CEA, YKL-40, and periostin. In some embodiments, “patients in need of it” may be subjects with blood eosinophil counts of ≥300 cells / μL, 150-299 cells / μL, or <150 cells / μL. In one embodiment, “patients in need of it” may be subjects with elevated levels of bronchial or airway inflammation, as measured by exhaled nitric oxide (FeNO).

[0280] In some embodiments, “patients who need it” are selected from groups consisting of ages 18 and over, ages 12 and over, ages 12–17 (12–<18), ages 6–11 (6–<12), and ages 2–5 (2–<6). In some embodiments, “patients who need it” are selected from groups consisting of adults, adolescents, and children. In some embodiments, “patients who need it” are selected from groups consisting of adults aged 18 and over, adolescents aged 12–17 (12–<18), children aged 6–11 (6–<12), and children aged 2–5 (2–<6). The subjects may be under 2 years old, for example, 12–23 months or 6–11 months.

[0281] Normal IgE levels in healthy subjects are less than approximately 100 kU / L (for example, as measured using the IMMUNOCAP® assay [Phadia, Inc. Portage, MI]). Therefore, the present invention includes a method comprising the steps of selecting a subject exhibiting elevated serum IgE levels exceeding approximately 100 kU / L, 150 kU / L, 500 kU / L, 1000 kU / L, 1500 kU / L, 2000 kU / L, 2500 kU / L, 3000 kU / L, 3500 kU / L, 4000 kU / L, 4500 kU / L, or 5000 kU / L, and administering to the subject a pharmaceutical composition containing a therapeutically effective amount of an IL-33 antagonist and / or an IL-4R antagonist.

[0282] In healthy subjects, TARC levels range from 106 ng / L to 431 ng / L, with an average of approximately 239 ng / L. (A model assessment for measuring TARC levels) The kit in question is a TARC quantitative ELISA kit provided by R&D Systems, Minneapolis, MN, under catalog number DDN00. Accordingly, the present invention includes a method comprising the steps of selecting a subject showing elevated TARC levels, where serum TARC levels exceed approximately 431 ng / L, approximately 500 ng / L, approximately 1000 ng / L, approximately 1500 ng / L, approximately 2000 ng / L, approximately 2500 ng / L, approximately 3000 ng / L, approximately 3500 ng / L, approximately 4000 ng / L, approximately 4500 ng / L, or approximately 5000 ng / L, and administering to the patient a pharmaceutical composition containing a therapeutically effective amount of an IL-33 antagonist and / or an IL-4R antagonist.

[0283] Eotaxin-3 belongs to a group of chemokines released by airway epithelial cells that are upregulated by Th2 cytokines IL-4 and IL-13 (Lilly et al., 1999, J. Allergy Clin. Immunol. 104: pp. 786-790). The present invention includes a method comprising the step of treating a patient with elevated eotaxin-3 levels, for example, greater than about 100 pg / ml, greater than about 150 pg / ml, greater than about 200 pg / ml, greater than about 300 pg / ml, or greater than about 350 pg / ml, by administering an IL-33 antagonist and / or an IL-4R antagonist. Serum eotaxin-3 levels can be measured, for example, by ELISA.

[0284] Exhaled nitric oxide (FeNO) is a biomarker of bronchial or airway inflammation. FeNO is produced by airway epithelial cells in response to inflammatory cytokines, including IL-4 and IL-13 (Alwing et al., 1993, Eur. Respir. J. 6: pp. 1368-1370). FeNO levels in healthy adults range from 2 to 30 parts per billion (ppb). A exemplary assay for measuring FeNO is Aerocrine. This is done using a NIOX device manufactured by AB Company, Solna, Sweden. The evaluation is performed before measuring vital capacity and after fasting for at least one hour. Included herein are methods comprising the step of administering an IL-33 antagonist and / or an IL-4R antagonist to a patient with elevated levels of exhaled NO (FeNO) exceeding, for example, approximately 30 ppb, approximately 31 ppb, approximately 32 ppb, approximately 33 ppb, approximately 34 ppb, or approximately 35 ppb.

[0285] Carcinoembryonic antigen (CEA) (also known as CEA cell adhesion molecule 5 [CEACAM5]) is a tumor marker found in association with non-neoplastic diseases of the lung (Marechal et al., 1988, Anticancer Res. Vol. 8: pp. 677-680). Serum CEA levels can be measured by ELISA. The present invention includes a method comprising administering an IL-33 antagonist and / or an IL-4R antagonist to a patient with elevated CEA levels, for example, greater than about 1.0 ng / ml, greater than about 1.5 ng / ml, greater than about 2.0 ng / ml, greater than about 2.5 ng / ml, greater than about 3.0 ng / ml, greater than about 4.0 ng / ml, or greater than about 5.0 ng / ml.

[0286] YKL-40 (derived from its N-terminal amino acids tyrosine (Y), lysine (K), and leucine (L), with a molecular weight of 40 kD) is a chitinase-like protein that has been found to be upregulated and associated with asthma exacerbations, IgE, and eosinophils (Tang et al., 2010, Eur.Respir.J. 35: pp. 757-760). Serum YKL-40 levels are measured, for example, by ELISA. This invention relates to YKL-40, for example, IL-33 antagonists and / or IL-4R antagonists in YKL-40 levels greater than approximately 40 ng / ml, greater than approximately 50 ng / ml, greater than approximately 100 ng / ml, greater than approximately 150 ng / ml, greater than approximately 200 ng / ml, or greater than approximately 250 ng / ml. The method includes the step of administering to a patient with an elevated level of -40.

[0287] Periostin is a secreted matrix cell protein associated with fibrosis, and its expression is upregulated by recombinant IL-4 and IL-13 in cultured bronchial epithelial cells and bronchial fibroblasts (Jia et al. (2012) J. Allergy Clin. Immunol. 130: 647). In human asthma patients, periostin expression levels correlate with reticular basement membrane thickness, an indicator of subepithelial fibrosis. (Same literature). Included herein is a method comprising the step of administering an IL-33 antagonist and / or an IL-4R antagonist to a patient with elevated periostin levels (e.g., ≥74.4 ng / mL).

[0288] Eosinophils and neutrophils in induced sputum are well-established direct markers of airway inflammation (Djukanovic et al., 2002, Eur. Respire. J. 37: 1S-2S). Sputum is induced by inhalation of hypertonic saline and processed for cell count according to methods known in the art, e.g., the European Respiratory Society guidelines.

[0289] In some embodiments, subjects are stratified into the following groups: blood eosinophil count (high blood eosinophil count) ≥ 300 cells / μL (HEos) or 300-499 cells / μL or ≥ 500 cells / μL, blood eosinophil count 200-299 cells / μL (moderate blood eosinophil count), or blood eosinophil count < 200 cells / μL (low blood eosinophil count), and are administered IL-33 antagonists and / or IL-4R antagonists in doses or drug regimens based on eosinophil levels.

[0290] In some embodiments, subjects have "eosinophilic phenotype" asthma, defined by a blood eosinophil count ≥ 150 cells / μL, a blood eosinophil count ≥ 300 cells / μL, or a blood eosinophil count ≥ 500 cells / μL, and are administered an IL-33 antagonist and / or an IL-4R antagonist.

[0291] In some embodiments, subjects have “periostin phenotype” asthma as defined by elevated blood periostin levels as defined herein and are administered an IL-33 antagonist and / or an IL-4R antagonist.

[0292] In some embodiments, “subjects requiring it” are clinically stable, non-smokers with mild, persistent allergic asthma who require only the use of inhaled short-acting β2 agonists (SABAs) when necessary to control asthma symptoms and are allergic to dust mite (HDM) allergens as determined by skin prick tests.

[0293] Methods for evaluating pharmacodynamic asthma-related parameters The present invention also includes a method for evaluating one or more pharmacodynamic asthma-related parameters in a subject requiring such evaluation, induced by the administration of an IL-33 antagonist, an IL-4R antagonist, or a pharmaceutical composition comprising an IL-33 antagonist and an IL-4R antagonist. A reduction in the incidence of allergic asthma exacerbations (as described above) or an improvement in one or more asthma-related parameters (as described above) may correlate with an improvement in one or more pharmacodynamic asthma-related parameters; however, such correlations are not necessarily observed in all cases.

[0294] Examples of "pharmacodynamic asthma-related parameters" include, for example, (a) biomarker expression levels; (b) serum protein and RNA analysis; and (c) induced eosinophils in sputum. This includes (d) neutrophil levels; (e) exhaled nitric oxide (FeNO); and (f) blood eosinophil count. "Improvement of pharmacodynamic asthma-related parameters" means, for example, a decrease from baseline in one or more biomarkers, such as periostin, TARC, eotaxin-3, or IgE; a decrease in sputum eosinophils or neutrophils; FeNO, periostin, or blood eosinophil count. As used herein, the term "baseline" with respect to pharmacodynamic asthma-related parameters means the numerical value of the pharmacodynamic asthma-related parameter for a patient before or at the time of administration of the pharmaceutical composition described herein.

[0295] To evaluate pharmacodynamic asthma-related parameters, the parameters are quantified at baseline and at points after administration of the pharmaceutical composition. For example, pharmacodynamic asthma-related parameters can be measured on days 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, and 14 after initial treatment with the pharmaceutical composition, or at weeks 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or later. The difference between the parameter values ​​at a specific point in time after the start of treatment and the parameter values ​​at baseline is used to determine whether there has been a change in pharmacodynamic asthma-related parameters, such as an "improvement" (e.g., an increase or decrease, depending on the specific parameter being measured).

[0296] In certain embodiments, administration of an IL-33 antagonist, an IL-4R antagonist, or both an IL-33 antagonist and an IL-4R antagonist to a patient results in changes in the expression of specific biomarkers, e.g., decreases or increases. Asthma-related biomarkers include, but are not limited to, (a) total IgE; (b) thymic and activating regulatory chemokines (TARCs); (c) YKL-40; (d) carcinoembryonic antigens in serum; (e) eotaxin-3 in plasma; and (f) periostin in serum. For example, administration of an IL-33 antagonist and / or an IL-4R antagonist to an asthma patient may result in one or more of the following: a decrease in TARC or eotaxin-3 levels, or a decrease in total serum IgE levels. This decrease can be detected at 1, 2, 3, 4, 5 weeks or more after administration of an IL-33 antagonist, an IL-4R antagonist, or both an IL-33 antagonist and an IL-4R antagonist. The expression of the biomarker can be assayed by methods known in the art. For example, protein levels can be measured by ELISA (enzyme-linked immunosorbent assay). RNA levels can be measured, for example, by reverse transcription (RT-PCR) linked to polymerase chain reaction.

[0297] As discussed above, biomarker expression can be assayed by detecting proteins or RNA in serum. Serum samples can also be used to monitor additional protein or RNA biomarkers associated with the response to treatment with IL-33 antagonists and / or IL-4R antagonists, IL-4 / IL-13 signaling, asthma, atopic dermatitis, or eosinophilic diseases (e.g., by measuring soluble IL-4Rα, IL-4, IL-13, periostin, etc.). In some embodiments, RNA samples are used to determine RNA levels (non-genetic analysis), e.g., the RNA levels of biomarkers, while in other embodiments, RNA samples are used for transcriptome sequencing (e.g., genetic analysis). [Examples]

[0298] The following examples are provided to give a complete disclosure and description of how the methods and compositions characterized in the present invention are prepared and used, and are not intended to limit the scope of what the inventors consider to be these inventions. (e.g., number of items used, quantity, temperature) While efforts are made to guarantee accuracy regarding (degrees, etc.), some experimental errors and deviations should be taken into consideration. Unless otherwise indicated, parts are parts by weight, molecular weight is the average molecular weight, temperature is in degrees Celsius, and pressure is atmospheric pressure or close to atmospheric pressure.

[0299] The exemplary IL-33 antagonist used in the following examples is a human anti-IL-33 antibody named SAR440340, also known as REGN3500 or, by its international generic name (INN), itepekimab. The exemplary IL-4R antagonist used in the following examples is a human anti-IL-4R antibody named dupilumab (i.e., DUPIXENT®). [Examples]

[0300] A randomized, placebo-controlled, parallel panel trial to evaluate the effects of REGN3500, dupilumab, and REGN3500 + dupilumab combinations on markers of inflammation after bronchial allergen loading in patients with allergic asthma. Overview and rationale for the test REGN3500 and dupilumab were fully human monoclonal antibodies (mAbs). Dupilumab was an anti-interleukin-4 receptor α subunit (IL-4Rα) mAb. REGN3500 targets IL-33, a pro-inflammatory cytokine that initiates and amplifies innate and adaptive inflammatory cascades (Cayrol et al., IL-33: an alarmin cytokine with crucial roles in innate immunity, inflammation and allergy. Curr Opin Immunol. 2014. Vol. 31: pp. 31-37).

[0301] This study aimed to evaluate the therapeutic effects of REGN3500, dupilumab, and REGN3500 + dupilumab combinations compared to placebo, as well as the effects of inhaled corticosteroids on the allergic inflammatory pathway induced by inhaled dust mite (HDM) bronchial allergen loading (BAC) in adult patients with mild asthma who are susceptible to HDM. The inhaled BAC model has been efficiently used in the development of asthma medications for 30 years (Diamant et al., Inhaled allergen bronchoprovocation tests. J Allergy Clin Immunol. 2013. Vol. 132: pp. 1045-1055 e1046; Fahy et al., Analysis of cellular and biochemical constituents of induced sputum after allergen challenge:a method for studying allergic airway inflammation.J Allergy Clin Immunol.1994.93:1031-1039;Inman et al., Dose-dependent effects of inhaled mometasone furoate on airway function and inflammation after allergen inhalation challenge.Am J Respir Crit Care Med. 2001. 164th year, Vol. 164: pp. 569-574).

[0302] Bronchial allergen overload involves the patient inhaling an allergen that elicits a biphasic airway response, characterized by a rapid (30 minutes to 2 hours after allergen overload) and slow (approximately 3 to 8 hours after allergen overload) decline in forced expiratory volume in one second (FEV1). This model facilitates the assessment of the inflammatory response of allergies by measuring cellular content, cytokine production, mRNA inflammatory signature in bronchoalveolar lavage, bronchial biopsy, or induced sputum changes (Zuiker et al., Sputum RNA signature in allergic asthmatics following allergen overload). Onchoprovocation test. Eur Clin Respir J. 2016. Vol. 3: 31324). As shown in Figure 7, this study utilized a BAC model to evaluate changes induced by treatment of allergic inflammation, as measured in induced sputum, with particular emphasis on changes in targeted and selected mRNA signatures.

[0303] A key feature of the study design was the requirement for pre-treatment BAC, which allowed for within-patient comparisons of the pre-treatment and post-treatment effects on BAC.

[0304] The objective of this study was to evaluate the effects of REGN3500, dupilumab, and REGN3500 + dupilumab combinations on molecular mechanisms involved in allergic airway inflammation in asthma, which are thought to contribute to the development of asthma. To achieve this goal, this study explored the expression of select inflammatory markers in the sputum of patients with mild allergic asthma induced by controlled BAC using HDM. By comparing the effects of treatment with REGN3500, dupilumab, or REGN3500 + dupilumab combinations on changes in the expression of inflammatory pathway molecules in sputum, data will be provided regarding whether the REGN3500 + dupilumab combination has an additional effect.

[0305] the goal The primary objective was to evaluate the effects of REGN3500, dupilumab, and REGN3500 + dupilumab combinations, compared to placebo, on changes in sputum inflammatory gene expression signatures induced after bronchial allergen loading (BAC) in adults with mild allergic asthma at 4 weeks post-treatment initiation, compared to screening.

[0306] The second objectives of this study were to evaluate the safety and tolerability of limited doses of REGN3500 and REGN3500 plus dupilumab combinations in adult patients with mild allergic asthma receiving BAC; to evaluate the pharmacokinetic (PK) profile of REGN3500 in adult patients with mild allergic asthma receiving BAC; to evaluate the immunogenicity of REGN3500 and dupilumab in adult patients with mild allergic asthma receiving continuous BAC; to evaluate the interception of REGN3500 by measuring serum total interleukin-33 (IL-33) levels in adult patients with mild allergic asthma receiving continuous BAC; and to evaluate the effect of fluticasone on changes in sputum inflammatory gene expression signatures induced after BAC in adults with mild allergic asthma, on day 4 after the start of treatment, compared to those at screening.

[0307] The exploratory objective of this study is to evaluate the effects of REGN3500, dupilumab, REGN3500 + dupilumab combination, and placebo on changes in sputum inflammatory gene expression signatures induced after BAC at 8 weeks post-treatment compared to screening; and on changes in sputum inflammatory cytokine proteins induced after BAC at 4 and 8 weeks post-treatment compared to screening. To evaluate the effect of sevofibrillator (SEB); to evaluate the potential effects of REGN3500, dupilumab, REGN3500 + dupilumab combination, and placebo on changes in serum markers of IL-33 pathway activation after BAC at weeks 4 and 8 after the start of treatment, compared to those at screening; to evaluate the effects of REGN3500 on pulmonary function parameters at weeks 4 and 8 after the start of treatment, compared to those at screening, including changes in FEV1 and exhaled nitric oxide (FeNO) before BAC, and changes in the area under the curve (AUC) of FEV1 0-2 hours and 3-8 hours after BAC. The objectives were to evaluate the effects of 0, dupilumab, the REGN3500 + dupilumab combination, and placebo; and to compare the effects of REGN3500, dupilumab, or the REGN3500 + dupilumab combination on changes in serum biomarkers such as sputum gene expression signature, sputum cytokines, IL-33 and IL-4R activity, and changes in FeNO and FEV1 AUC after BAC, at 4 and 8 weeks after initiation of treatment, compared to those at screening, with the effects of inhaled fluticasone propionate.

[0308] Test design This study evaluated the effects of REGN3500, dupilumab, and the REGN3500 + dupilumab combination on airway inflammation using an inhaled BAC model. The inhaled BAC model was a well-established and reproducible model of induced allergic bronchitis, enabling the evaluation of drug therapeutic effects at the functional, cellular, and molecular levels. Previous studies have demonstrated that this model can be used to assess the therapeutic response by measuring the mRNA of inflammatory genes in induced sputum (Zuiker et al., Sputum RNA signature in allergic asthmatics following allergen bronchoprovocation test. Eur Clin Respir J. 2016. Vol. 3: 31324). Several published reports have demonstrated that the inflammatory response after BAC leads to upregulation of type 2 cytokines in both bronchoalveolar fluid and sputum (Erin et al., Optimized dialysis and protease inhibition of sputum dithiothreitol supernatants. Am J Respir Crit Care Med. 2008. Vol. 177: pp. 132-141; Huang et al., IL-13 expression at the sites of allergen challenge in Patients with asthma. J Immunol. 1995. Vol. 155: pp. 2688-2694). The hypothesis of this study is that the acute upregulation of type 2 inflammation from the BAC model will allow for the evaluation of the effects of REGN3500, dupilumab, and the REGN3500 + dupilumab combination on this allergic airway inflammation.

[0309] This was a randomized, double-blind, two-part trial consisting of a placebo-controlled component (Part 1) and an open-label component (Part 2).

[0310] As shown in Figure 1, in Part 1 of this study, patients were randomized to receive either REGN3500 (intravenous (IV), single dose), dupilumab (subcutaneous (SC), two doses, 14 days apart), a combination of REGN3500 (IV, single dose) + dupilumab (SC, two doses, 14 days apart), or placebo. Enrolled patients underwent BAC during the screening period and at 4 and 8 weeks after administration of the first dose of the investigational drug(s). The effect of the investigational drug on BAC-induced pneumonia was evaluated by measuring sputum molecular signatures (mRNA and protein). Sputum molecular signatures were evaluated before BAC (baseline) and after BAC (post-BAC) at screening, and at 4 and 8 weeks after the start of treatment (Figure 1). The difference in BAC-induced sputum signature before and after BAC was evaluated at screening (screening change) and observed at weeks 4 and 8 post-treatment (week 4 change and week 8 change). Because both REGN3500 and dupilumab exhibit long half-lives, BAC changes were evaluated at both weeks 4 and 8 in the current study to monitor the durability of the investigational drug's potential impact on airway inflammation. The effect of investigational drug treatment on induced sputum gene signatures was assessed by the difference between the BAC-induced screening change and the BAC-induced week 4 change (change from screening to week 4), as well as the BAC-induced screening change and the BAC-induced week 8 change (change from screening to week 8). The evaluation was based on the difference between the changes in [the specified value].

[0311] The BAC model has been used in the development of effective and potent anti-inflammatory drugs for asthma, such as inhaled corticosteroids (Hansel et al., The allergen challenge. Clin Exp Allergy. 2002. Vol. 32: pp. 162-167; Inman et al., Dose-dependent effects of inhaled mometasone furoate on airway function). and inflammation after allergen inhalation challenge. Am J Respir Crit Care Med. 2001. Vol. 164: pp. 569-574, and Ravensberg et al., Validated safety predictions of airway responses to house dust mite in asthma. Clin Exp Allergy. 2007. Vol. 37: pp. 100-107. While the typical endpoint of these studies is the change in delayed-phase FEV1 (usually a decrease in FEV1 observed 4-8 hours after allergen exposure), this model has also demonstrated to be a good test for retesting the reproducibility of inflammatory markers measured in the airways after BAC (Fahy et al., Analysis of cellular and biochemical constituents of induced sputum after allergen challenge: a method for studying allergic). Airway inflammation. J Allergy Clin Immunol. 1994. Vol. 93: pp. 1031-1039. Inman et al., Dose-dependent effects of inhaled mometasone furoate on airway function and inflammation. after allergen inhalation challenge. Am J Respir Crit Care Med. 2001. Vol. 164: pp. 569-574, and Zuiker et al., Kinetics of TH2 biomarkers. in sputum of asthmatics following inhaled allergen. Eur Clin Respir J. 2015. Vol. 2).

[0312] To improve the ability to interpret results from a small number of patients, each patient underwent a BAC during the screening period. To be included in this study, each patient had to demonstrate both an early decline in FEV1 (0–30 minutes after BAC) and a late decline in FEV1 (3–8 hours after BAC). Previous studies have shown that patients with both early and late declines in FEV1 have higher levels of type 2 cytokines in the delayed phase compared to patients with no decline in FEV1 during this period. A second BAC in Part 1 of this study was performed approximately 4 weeks after administration of the first dose of the study treatment. A third BAC was proposed approximately 8 weeks after the start of treatment to provide an assessment of the durability of the effect. These data demonstrated the ability to create PK / pharmacodynamic (PD) models of the airway effects of REGN3500, dupilumab, and the REGN3500 + dupilumab combination.

[0313] The data obtained from six patients is expected to provide sufficient statistical power to detect the treatment effect. Due to potential technical difficulties in successfully completing all procedures, up to eight patients were enrolled in each treatment group. From the data obtained from six patients per group, the ability to detect a 2.5 to 5-fold increase in the size of the treatment effect on the expression of a specified mRNA cytokine is expected to be >80%.

[0314] As shown in Figure 3, in Part 2 of this study, patients received a screening BAC, followed by a short, open-label, high-dose inhaled fluticasone propionate. A second BAC was administered 4 days after the initiation of inhaled fluticasone propionate treatment. This part of the study examined the previously reported efficacy of corticosteroids against mRNA allergic inflammatory signatures. To confirm the result (Zuiker et al., SputumRNA signature in Allergic asthmatics following allergen bronchoprovocation test. Eur Clin Respir J. 2016. Vol. 3:31324) and served as a positive control to provide a control drug for evaluating the effect of investigational drugs on allergen-induced inflammation.

[0315] This was a Phase 1b trial consisting of two distinct parts. Part 1 lasted 42 weeks, excluding the screening period. Part 2 lasted 2 weeks, excluding the screening period. Parts 1 and 2 were conducted concurrently.

[0316] Part 1 of this study was a randomized, double-blind, placebo-controlled, double-dummy, parallel-group trial and measurement of delayed-phase inflammatory airway response to BAC using HDM in patients with mild persistent allergic asthma to evaluate the effects of REGN3500, dupilumab, REGN3500 + dupilumab combination, or placebo on pneumonia (as measured by sputum cytokine mRNA). Patients were randomized to one of the following treatment groups: REGN3500 (single IV dose); dupilumab (two SC doses, Q2W); REGN3500 (single IV dose) + dupilumab (two SC doses, Q2W); and placebo (single IV dose and two SC doses, Q2W).

[0317] The therapeutic effects on pneumonia and on the measurement of the delayed-phase inflammatory airway response to BAC by HDM were evaluated. Patients were followed from day 58 to the final study visit at day 293.

[0318] Part 2 involved open-label treatment with short-term inhaled fluticasone propionate. Patients received eight doses of inhaled fluticasone propionate over four days and were followed until the final study visit on day 15. The effect of fluticasone propionate on sputum cytokine mRNA and the delayed-phase airway response were measured and used as positive controls for comparison with measurements in Part 1.

[0319] Anti-drug antibody variables include status (positive or negative) and titer, as follows: total number of patients negative in ADA assays at all time points analyzed, total number of patients positive in ADA assays at all time points analyzed, total number of patients with pre-existing immunoreactivity, total number of patients with a treatment-boosted ADA response, and titer categories: low (titer < 1,000), medium (1,000 ≤ titer ≤ 10,000), and high (titer > 10,000).

[0320] screening All patients included in this study were clinically stable, non-smokers with mild, persistent allergic asthma with atopic dermatitis for HDM, as determined by skin prick testing, and who required only the use of inhaled short-acting β2 agonists when necessary to control asthma symptoms.

[0321] During the screening period (-28 to -1 day), pre-study procedures were performed on potential study patients. Inhaled HDM allergen loading was administered to patients who met all other enrollment criteria to provide a baseline assessment of airway allergic reactions and to determine eligibility. During the screening period (-28 to -1 day), patients were required to exhibit both initial and delayed-phase allergic reactions during the screening BAC.

[0322] Patients are tolerant of sputum induction and provide appropriate sputum samples during screening. It was necessary to provide a suitable sputum sample. The definition of a suitable sputum sample is provided in this study manual. Patients who were unable to produce a suitable sputum sample before BAC during the screening process could be rescreened for sputum induction with the approval of the investigator. Patients who were unable to produce sputum after BAC were ineligible to enroll in this study.

[0323] The allergen dose regimen for each individual patient at post-treatment BAC (days 29 and 57 after initiation of treatment in Part 1 and day 4 after initiation of treatment in Part 2) was calculated using the allergen dose required to demonstrate an appropriate EAR for that patient during the screening load. To be included in this study, patients had to demonstrate an appropriate LAR, defined as a decrease of 15% or more in FEV1 from pre-BAC (however, patients had to have a predicted FEV1 decrease of less than 25% or <1.4L). The procedure during the screening BAC was similar to the procedure performed in the treatment phases of this study.

[0324] Exam Part 1 As shown in Figure 2, eligible patients (up to 32 in total) were randomized in a 1:1:1:1 ratio to receive either REGN3500, dupilumab, a combination of REGN3500 and dupilumab, or placebo. Since the treatment groups in this study included the first clinical dose of the REGN3500 + dupilumab combination, Part 1 was conducted in two phases to address any potential safety issues. The first eight patients were randomized in a 1:1:1:1 ratio to receive the investigational drug. Day 1: Patients in the REGN3500 group received REGN3500 IV 10 mg / kg, followed by placebo SC in a dose balanced with dupilumab. Patients in the dupilumab group received placebo IV in a dose balanced with REGN3500, followed by a dupilumab SC loading dose of 600 mg (two 300 mg injections). Patients in the REGN3500 + dupilumab combination received REGN3500 10 mg / kg IV, followed by dupilumab SC 600 mg (two 300 mg injections). Patients in the placebo group received placebo IV in a dose balanced with REGN3500, followed by two placebo SC injections in a dose balanced with dupilumab.

[0325] On day 15 (week 2), patients randomized to either the dupilumab group or the REGN3500 + dupilumab combination group received 300 mg of dupilumab SC, while all other patients received a placebo SC at a dose equivalent to that of dupilumab.

[0326] These first eight patients completed the safety evaluation up to day 24 of the study (seventh visit). Regeneron Safety Monitoring Committee (RSOC) The Oversight Committee conducted an open-label safety review of the data. After the safety data was reviewed by the RSOC and further enrollment was approved, approximately 24 remaining patients were enrolled. These remaining patients were randomized and treated as described for the first 8 patients.

[0327] All enrolled patients returned to the clinic on day 24 ± 2 days and day 52 ± 2 days to provide baseline-induced sputum samples. If a suitable sputum sample could not be obtained, the patient may return to the clinic 72 hours later for a second attempt to generate baseline sputum. Sputum collection before BAC must be performed at least 72 hours prior to BAC. Based on a 72-hour follow-up of baseline-induced sputum samples, patients returned to the clinic on day 29 ± 2 days and day 57 ± 2 days, respectively, to have their FEV1 and FeNO measured before receiving BAC by HDM. Sputum induction was performed at both visits on day 29 ± 2 days (week 4) and day 57 ± 2 days (week 8), at 8 and 24 hours after inhaled BAC. Patients were monitored in the hospital at least 8 hours after BAC and by the investigator. Patients were permitted to leave the testing facility if deemed stable by their supervisor. The drug-free interval between BACs was at least 21 days.

[0328] Pulmonary function, assessed by vital capacity measurement, sputum mRNA, sputum cytokines, FeNO, and additional serum markers such as IL-33 and IL-4R activity were measured as detailed in the event schedule shown in Table 1 below. Patients were followed up at scheduled visits and after BAC until the final day of the study visit (day 293).

[0329] [Table 1-1] [Table 1-2]

[0330] Footnote Table 1 regarding the event schedule a. Informed consent can be obtained at separate visits before the screening period. b. Patients must sign a separate Informed Consent Form (ICF) before a sample is taken for DNA analysis. Patients who meet the inclusion / exclusion criteria are eligible to enroll in this study, regardless of whether they chose to participate in the genomics substudy. c. Administer REGN3500 (or a balanced dose of placebo) as a single intravenous dose on day 1. Administer dupilumab (or a balanced dose of placebo) as two intravenous injections on day 1 and one intravenous injection on day 15. The investigational drug should be administered intravenously before the investigational drug is administered via SC, with at least one hour between the IV administration and the SC administration. On days 1 and 15, observe the patient 8 hours prior to discharge after administration of the SC dose of dupilumab. The patient may eat or drink 2 hours after the completion of the infusion. d. All blood samples (for safety laboratory, PK, ADA, biomarkers, and future biomedical research) should be collected after at least eight hours of overnight fasting. e. Measurement of vital signs: e1. Vital signs should be collected before any blood draw. e2. On days 1 and 15 of the study, vital signs will be collected before administration of the investigational drug, immediately after (within 10 minutes) completion of IV infusion, before SC injection of the investigational drug, and 1, 2, 4, and 8 hours after completion of injection. f. Details of ECG measurement: f1. During a patient visit requiring blood sampling, an electrocardiogram will be performed, followed by blood collection. f2. On days 1 and 15, ECGs will be performed before IV infusion of the investigational drug, immediately after completion of IV infusion (within 10 minutes), and 4 hours after completion of SC injection. All blood samples for g.PK and ADA will be collected before the start of investigational drug administration. g1. On days 1 and 15, samples for the safety laboratory, ADA, and biomarkers will be collected (under fasting conditions) prior to the start of investigational drug administration. h. Clinical vital capacity measurements should be performed using a standard spirometer, in accordance with the standards for acceptable quality control set forth by the American Thoracic Society (Miller et al., Standardisation of spirometry. Eur Respir J. 2005. Vol. 26: pp. 319-338). i. FeNO should be administered after at least one hour of fasting. i1. FeNO measurement must be performed before vital capacity measurement and metacholine loading. i2. FeNO measurement must be performed before sputum induction. i3. On the loading days (3rd, 8th, and 12th visits), FeNO will be measured before BAC, and 8 and 23 hours after BAC. j. Days on which sputum can be collected are indicated in gray. j1. Sputum samples prior to allergen screening should be collected at least 48 hours after the methacholine loading administered at the first visit, and 72 hours before the BAC administered at the third visit. Sputum samples prior to other allergen screenings should be collected at least 72 hours before the BAC. If a sample prior to the BAC cannot be collected on the first attempt, a repeat attempt may be made at least 72 hours later. j2. Sputum should be collected on the BAC days (3rd, 8th, and 12th visits), 8 hours after the load, and again the following day, approximately 24 hours after the BAC (4th, 9th, and 13th visits). l. DNA samples should be collected on day 1. However, DNA may be collected at any visit during the course of the test. Whole blood for mRNA sample extraction should be collected only during the scheduled trial visit, prior to BAC, spirometry, or treatment of induced sputum on the scheduled day. Whole blood for RNA sample collection should be collected before drug administration on the investigational drug treatment day. n. Biomarker samples should be collected at specific times during the test, paying attention to the following: n1. Biomarker and total IL-33 samples on BAC days (3rd, 8th, and 12th visits) should be collected before BAC and 8 hours after BAC (after sputum sample collection 8 hours after BAC). n2. Biopsy samples taken the day after BAC (4th visit, 9th visit, and 13th visit) Omarker samples should be collected 24 hours after BAC, following the collection of sputum samples.

[0331] Exam Part 2 As shown in Figure 3, in Part 2 of this study, approximately six patients received fluticasone propionate inhaled at a dose of 500 μg per dose (2 puffs of 250 μg) twice daily for 4 days, starting on day 1 (a total of 8 doses). Patients were administered the drug at the clinic on day 1, and their inhalation technique was reviewed. Patients were sent home on days 2 and 3 to self-administer the drug. Induced sputum samples were collected on day 1 after the second dose of fluticasone. On day 4, patients were administered fluticasone propionate (in the clinic) before BAC treatment, and the patients self-administered the second dose the following day (in the clinic or at home) after sputum induction. After the 7th of the 8 doses of fluticasone propionate, patients underwent inhaled BAC using an HDM similar to that in Part 1 of the study. Induced sputum samples were collected on day 4, 8 hours and 24 hours after HDM loading. Pulmonary function, assessed by vital capacity measurement, sputum mRNA, sputum cytokines, FeNO, and additional serum markers of IL-33 and IL-4R activity were measured as described in Part 1. In cases where a patient was unable to return on day 4, the patient may continue administration of fluticasone propionate twice daily for up to two additional days. If the patient returned to the clinic, the procedure for day 4 was followed.

[0332] The patient maintained typical clinical activity throughout the entire period of approximately two weeks.

[0333] Part 2 of this study was conducted concurrently with Part 1. Patients who complete Part 2 can participate in Part 1 after a drug-free period of at least 21 days.

[0334] [Table 2-1] [Table 2-2]

[0335] Footnote Table 2 regarding the event schedule a. Informed consent can be obtained at separate visits before the screening period. b. Patients must sign a separate Informed Consent Form (ICF) before a sample is taken for DNA analysis. Patients who meet the inclusion / exclusion criteria are eligible to enroll in this study, regardless of whether they chose to participate in the genomics substudy. c. The patient begins self-administration of inhaled fluticasone propionate twice daily for four consecutive days, starting on day 1. The patient receives fluticasone propionate at the clinic on day 1 and at home on days 2 and 3. On day 4, the patient receives fluticasone propionate (in the clinic) before BAC treatment, and the patient self-administers a second dose the following day (in the clinic or at home) after inducing sputum. d. All blood samples (for safety testing, biomarkers, and future biomedical research) should be collected after at least eight hours of overnight fasting. e. Details of vital sign measurement: e1. Vital signs should be collected before any blood draw. e2. On day 1 of the trial, vital signs will be collected before administration of the investigational drug, immediately after the completion of fluticasone administration (within 10 minutes), and 1, 2, 4, 8, and 12 hours after the completion of fluticasone administration. f. Details of ECG measurement: f1. After performing an electrocardiogram, a blood sample is taken. f2. On day 1, an ECG should be performed before administration of fluticasone propionate and immediately after administration (within 10 minutes). g. Clinical vital capacity measurements shall be performed using a standard spirometer, in accordance with the standards for acceptable quality control of the American Thoracic Society (Miller et al., Standardisation of spirometry. Eur Respir J. 2005. Vol. 26: pp. 319-338). . h.FeNO should be administered after at least one hour of fasting. h1. FeNO measurement must be performed before vital capacity measurement and metacholine loading. h2.FeNO measurement must be performed before sputum induction. h3.FeNO is measured on the BAC day, before BAC and 8 and 23 hours after BAC. FeNO measurement on day 4.1 should be performed before administration. i. During the screening and treatment period, all pre-BAC sputum samples must be collected 72 hours prior to BAC. i1. During screening, pre-BAC induced sputum samples should be collected at least 72 hours prior to the BAC. If a pre-BAC sample cannot be collected on the first attempt at screening, a repeat may be performed at least 72 hours later. Sputum should be collected at the third visit, 8 hours after the load, and the following day, approximately 24 hours after the BAC (fourth visit). i2. Induced sputum samples (pre-BAC) are collected on day 1 after the second dose of fluticasone. Repeated pre-BAC sputum samples are not permitted in BAC treatment. i3. On day 4, patients are administered the first dose of fluticasone, followed by a BAC using HDM similar to that of Study Part 1. Induced sputum samples are collected on day 4, 8 hours and 24 hours after the BAC. The k.DNA sample should be collected on day 1, but it can be collected at any visit during the course of the test. l. Whole blood RNA samples should only be collected on the day of the visit, prior to BAC, spirometry, or treatment of induced sputum, as indicated in the table. Whole blood RNA samples should be collected before drug administration. m. Biomarker samples may be collected periodically during the test, as specified in the test chart, with the following precautions: Biomarkers and total IL-33 samples should be collected on day m1 of BAC before BAC and 8 hours after BAC; sputum samples were collected 8 hours after BAC. Biomarkers collected the day after m2.BAC should be collected 24 hours after BAC, after sputum sample collection.

[0336] Test population: The patient population in this study consisted of adult asthma patients allergic to HDM allergens. Numerous studies have demonstrated that BAC induces significant and reproducible upregulation of inflammatory gene signatures, which can be measured in induced sputum from patients with allergic asthma. Bronchial allergen loading (BAC) provides an opportunity to evaluate the potential effects of REGN3500 and dupilumab on pneumonia. When performed by skilled investigators in appropriate settings, BAC was safe and well-tolerated in the patient population proposed for this study. However, BAC was not performed in patients with severe or unstable asthma due to the risk of inducing severe, acute bronchoconstriction or anaphylaxis (Diamant et al., Inhaled allergen bronchoprovocation tests. J Allergy Clin Immunol. 2013. Vol. 132: pp. 1045-1055 e1046).

[0337] Demographic and baseline characteristics include standard demographics (e.g., age, sex, race, ethnicity, weight, height), medical and medication history for each patient, and disease characteristics, including biomarkers (total IL-33, sST2, calcitonin, and MMP12).

[0338] Enrollment included up to 38 patients from the two parts of this study in the UK (approximately 32 patients from Part 1 and approximately 6 patients from Part 2).

[0339] Approximately 38 non-smoking adult patients (male and female) aged 18-60 years with mild persistent allergic asthma were enrolled in this study. Patients included in this study should be clinically stable and require only short-acting β2-agonists as needed to control asthma symptoms. Patients included in this study should be allergic to HDM, as determined by skin prick testing.

[0340] Study cohort This study consisted of two parts. Patients in Part 1 of the study constituted Cohort 1, and patients in Part 2 constituted Cohort 2.

[0341] In Part 1, approximately 32 patients were randomized in a 1:1:1:1 ratio to receive either REGN3500 (single IV dose), dupilumab (two seroconcentrates, Q2W), a combination of REGN3500 (single IV dose) + dupilumab (two seroconcentrates, Q2W), or placebo (single IV dose and two seroconcentrates, Q2W), with approximately 8 patients per treatment group. These approximately 8 patients were then assigned to treatment groups to obtain data on 6 patients per treatment group within Cohort 1.

[0342] In Part 2, approximately six patients were treated openly with inhaled fluticasone propionate in Cohort 2.

[0343] Inclusion Criteria Patients must meet the following criteria to be eligible for enrollment in this study: 1. Male or female between 18 and 60 years of age (inclusive). 2. Pre-study body mass index (BMI) of 17–33 kg / m² at screening (BMI = weight [kg] / height [m]²). 3. Clinically stable, with a history of mild allergic asthma for at least 6 months, with typical symptoms including cough and wheezing, requiring treatment with short-acting β2 agonists only when necessary. Patients must be deemed healthy based on medical history, physical examination, vital signs, ECG, and laboratory safety testing performed at screening and / or before administration of the first dose of the investigational drug, apart from their asthma history. 4. Pre-bronchodilator forced expiratory volume in one second (FEV1) at screening is ≥70% of the predicted value. 5. Non-smoker or former smoker for at least 12 months. Patients must have a cumulative tobacco exposure of no more than 5 pack years, with each pack year = (number of cigarettes smoked per day × number of years of smoking) ÷ 20. 6. The patient must demonstrate atopic dermatitis for HDM, confirmed by a positive skin prick test at screening (a positive reaction should be defined as a wheal being 3 mm or larger than that of the negative control). 7. The patient must be a dual responder to inhaled BAC as manifested by positive allergen-induced early and late airway bronchoconstriction. EAR is defined as a decrease of at least 20% in FEV1 from the pre-load diluted baseline value for 30 minutes after inhaled BAC, and LAR is defined as a decrease of at least 215% in FEV1 from the diluted value at least three times, two of which must occur consecutively between 3 and 8 hours after administration of the final concentration of the allergen. 8. The patient must be tolerant of sputum induction and able to produce adequate sputum after BAC during the screening period (at 8 or 24 hours after BAC). 9. I understand the available alternative treatments and the risks associated with this study, and I agree to voluntarily participate by providing written informed consent to perform the study procedures. 10. I am willing to abide by the restrictions specified in the study protocol, including prohibited drugs and procedures. 11. At the time of screening, I have a positive response to the metacholine loading test (diluted value of metacholine PC20 ≤ 16 mg / mL) and This demonstrates a 220% reduction in FEV1 (compared to the previous example).

[0344] Exclusion criteria 1. A history of life-threatening asthma, defined as an asthma episode requiring intubation and / or associated with hypercapnia, respiratory arrest, and / or hypoxic episodes. 2. Hospitalization or emergency room admission for asthma within 12 months prior to screening. 3. An asthma exacerbation or respiratory infection within 4 weeks prior to screening or prior to the first dose of the investigational drug. 4. A history of other respiratory / lung diseases, e.g., a diagnosis of chronic obstructive pulmonary disease (COPD), cystic fibrosis, bronchiectasis, or α1-antitrypsin deficiency or restrictive lung disease, as defined by the Global Initiative for Chronic Obstructive Lung Disease (GOLD) guidelines (updated 2013). 5.5. After screening BAC, the patient has a FEV1 of less than 25% of their predicted values ​​and / or a decrease of FEV1 of <1.4L or a symptomatic decrease in FEV1 associated with dyspnea that is not resolved by bronchodilators within a reasonable time frame (approximately 30 minutes) after allergen exposure. 6. Has a history of severe allergies or anaphylactic reactions or significant intolerance to prescription or over-the-counter medications or foods. 7. Has received oral or systemic corticosteroids within the past 8 weeks or inhaled / nasal corticosteroids within the past 4 weeks and / or before randomization. 8. Within 4 weeks of screening or prior to the administration of the first dose of the investigational drug, the patient has used any other asthma medication in addition to a short-acting β-agonist (e.g., leukotriene receptor antagonist, muscarinic antagonist, terbutaline, theophylline β-blocker, digoxin, NSAID, MAO inhibitor, or tricyclic antidepressant). For approved medications or investigational biological agents (e.g., anti-IgE or anti-IL5), the patient has used them within 6 months of screening or within at least 5 half-lives, whichever is greater. 9. Prior to screening or prior to the administration of the first dose of the investigational drug, the patient has been treated with the investigational drug for a period of 8 weeks or longer (if known) or within 5 half-lives. 10. The patient has been treated with a live (attenuated) vaccine within 12 weeks prior to screening.11. Based on the MDRD formula, the estimated glomerular filtration rate (eGFR) at the time of screening is <60 mL / min / 1.73 m². 2 12. At the time of screening, HBsAg, HBcAb, or HCV is positive. 13. At the time of screening, there is a known history of human immunodeficiency virus (HIV) infection or HIV seropositivity. 14. At the time of screening, a blood test for tuberculosis (TB) is positive. 15. There is a history of tuberculosis or systemic fungal disease. 16. Currently or recently (within the past two months prior to screening), a bacterial infection, protozoal infection, viral infection, or parasitic infection has been diagnosed; there is a suspected parasitic infection or a high risk of parasitic infection. 17. There is a history of clinically significant neurological, endocrine, gastrointestinal, cardiovascular, hematological, hepatic, immunological, renal, or any other organ system disorder (apart from asthma or other mild allergic conditions, e.g., allergic rhinitis). Patients with a history of uncomplicated nephrolithiasis (kidney stones) may be enrolled in this study at the discretion of the investigator. 18. Have undergone major surgery in the past two months or are scheduled for surgery during this study or follow-up period. 19. History of cancer, excluding patients with basal cell carcinoma or cervical carcinoma in situ who have been adequately treated and patients with other malignancies that have been successfully treated in the >10 years prior to screening, provided that appropriate follow-up has not revealed evidence of recurrence during the screening period, as determined by the investigator and treating physician. 20. Positive results from a urinary drug test during screening or prior to randomization (e.g., amphetamine / methamphetamine, barbiturates, benzodiazepines, cannabinoids, cocaine, opiates, and cotinine), unless, in the opinion of the investigator, the positive test result could not be attributable to the patient's currently approved medications. 21. History of drug or alcohol abuse within one year prior to the screening visit. 22. Consumed alcohol within 48 hours prior to screening. 23. Unwillingness to comply with the alcohol consumption limits for this study. Alcohol use is limited to no more than two drinks per day (one drink is equivalent to 12 ounces of regular beer, 5 ounces of wine, or 1.5 ounces of 80-proof (40% alcohol) distilled spirits). Patients must refrain from drinking alcohol within 48 hours of their study visit, including the BAC. 24. Failure to comply with the caffeine intake restrictions of this study. Patients must refrain from caffeine for 8 hours prior to all study visits. Caffeine-free products are permitted. 25. Not of legal consent age, or mentally incapacitated or legally disqualified. 26. Any medical history that, in the opinion of the investigator, could confound the results of this study or where participation in this study could pose an additional risk to the patient. 27. A new or significant change in routine exercise within 4 weeks prior to the screening visit. Patients who do not intend to maintain a similar level of exercise during the study or who do not intend to refrain from significantly strenuous exercise during the study will be excluded. 28. Pregnant, planning to become pregnant, or breastfeeding. 29. A sexually active, childbearing woman who will not attempt to use highly effective contraception during this study, before the initiation of the first treatment, and at least four months after the last dose. Highly effective contraception includes the steady use of oral contraceptives (e.g., contraceptives containing estrogen / progesterone or high-dose progesterone) associated with ovulation suppression for at least two months prior to screening; intrauterine devices; intrauterine hormone-releasing systems; bilateral tubal ligation; vasectomy of the partner; and / or abstinence. Contraception is not required for male patients. 30. Known sensitivity to doxycycline and / or tetracycline or any of the components of the investigational formulation.

[0345] Experimental treatment Part 1 REGN3500 was supplied as a lyophilized powder. A placebo equivalent to REGN3500 (a placebo at a dose equal to REGN3500) was prepared using the same formulation as REGN3500, but without the addition of protein (i.e., the active substance, anti-IL-33 monoclonal antibody). Vials of REGN3500 or the placebo equivalent to REGN3500 were reconstituted with sterile water before injection. REGN3500 and the placebo equivalent to REGN3500 were administered intravenously on day 1 by the investigator or other qualified researcher.

[0346] Dupilumab was supplied into pre-filled syringes, each capable of delivering 2 mL of the 150 mg / mL (300 mg) solution of the investigational drug. A placebo equivalent to dupilumab (a placebo at a dose equal to dupilumab) was prepared in the same formulation as dupilumab, without the addition of protein. Dupilumab or a placebo at a dose equal to dupilumab was administered via SC by the investigator or other qualified researcher in two injections on day 1 (600 mg) and one injection on day 15 (300 mg). All SC injections were administered intraabdominally.

[0347] Patients will be randomly and equally assigned to the following treatment regimens: REGN3500: Day 1, REGN3500 IV 10 mg / kg + two injections of placebo SC at a dose balanced with dupilumab; Day 15, one injection of placebo SC at a dose balanced with dupilumab; Dupilumab: Day 1, two injections of placebo at a dose balanced with REGN3500 IV + dupilumab SC 300 mg (total loading dose 600 mg); Day 15, one injection of dupilumab SC 300 mg; REGN3500 + Dupilumab combination: Day 1, REGN3500 IV 10 mg / kg + two injections of dupilumab SC 300 mg (total loading dose of 600 mg); Day 15, one injection of dupilumab SC 300 mg; and placebo: Day 1, two injections of placebo IV at a dose balanced with REGN3500 + placebo SC at a dose balanced with dupilumab; Day 15, placebo S at a dose balanced with dupilumab. One of the single injections in Group C administered either REGN3500, dupilumab, a combination of REGN3500 and dupilumab, or placebo. All patients received an IV infusion first, followed by a SC injection. Patients were observed for at least one hour between the SC injection and the IV infusion.

[0348] Part 2 Fluticasone propionate (250 μg / puff) was administered by supplying it with a metered-dose inhaler, with a dose of 500 μg (2 puffs of 250 μg) inhaled twice daily from day 1 to day 4.

[0349] Treatment allocation In Part 1 of this study, randomization was performed in two separate phases. Eight patients were randomized in a 1:1:1:1 ratio to receive either REGN3500, dupilumab, REGN3500+dupilumab, or placebo, according to a central randomization scheme provided to designated study pharmacists (or qualified designated persons) via an automated voice response system (IVRS) / web automated response system (IWRS). After the first eight patients completed safety evaluations by day 24 of their study visits (7th visit), and the RSOC reviewed the safety data and approved further enrollment, approximately 24 additional patients could be randomized in a 1:1:1:1 ratio to receive either REGN3500, dupilumab, REGN3500+dupilumab, or placebo, similar to the first eight patients.

[0350] In Part 2 of this study, approximately six patients were enrolled and treated with fluticasone propionate in an open-label manner. Patients who complete Part 2 can participate in Part 1 after a drug-free period of at least 21 days.

[0351] In Part 1, trial patients, principal investigators, and site personnel remained blinded to all randomization assignments throughout the trial. Regeneron trial leaders, medical monitors, trial monitors, and any other Regeneron and contract research organization (CRO) personnel who regularly communicated with the research sites also remained blinded to all patient randomization assignments.

[0352] Selected individuals not involved in conducting the trial may have access to unblinded data as needed for safety reviews or other data reviews.

[0353] Blinded investigational drug kits coded using a drug identification number system were used. To maintain blinding, a list linking these codes, which were associated with product lot numbers, was not available to the individuals involved in conducting the trial.

[0354] The results of anti-drug antibodies and drug concentrations were not communicated to these facilities, and the sponsor's operations team did not allow the use of results relevant to patient identification until after the final database lock. The biochemical analysts, biochemical analysis team leaders, and clinical pharmacology leaders responsible for determining serum drug concentration levels, ADA, and biomarkers were not blinded to medication information.

[0355] Treatment procedures and accountability For Part 1 of this study, a drug identification number system was used to label blinded investigational drugs. A list associating drug identification numbers with product lot numbers was maintained by the group (or company) responsible for the packaging of the investigational drugs. To maintain blinding, these lists were not available to individuals involved in the conduct of the study. This automated system was also used for the use of IVRS (Intra-Voice Response System) in clinical trials, with particular emphasis on handling expiration dates (EMA / INS / GCP / 600788 / 2011, December 2013). The shelf life of the investigational drug was managed in accordance with the EMA Reflection Paper regarding its use. The drug label for dupilumab included the expiration date. The drug label for REGN3500 did not include the expiration date.

[0356] For Part 2, open-label investigational drugs were labeled with the product lot number and expiration date. Investigational drugs were stored at the facility at a temperature of 2°C to 8°C.

[0357] REGN3500, dupilumab, and a balanced dose of placebo for each investigational drug were shipped to the investigator or nominee at regular intervals or as needed during the study at a temperature of 2°C to 8°C. At specific points during the study (e.g., intermediate site monitoring visits), during site close-out visits, and after drug reconciliation and documentation by site monitors, all open-label and blinded investigational drugs were destroyed or returned to the sponsor or nominee.

[0358] All drug accountability records had to be kept up-to-date. Investigators had to be able to describe all open-label and blinded investigational drugs. These records should include the date, quantity, and study drug distributed to each patient, returned by each patient (where applicable), and disposed of at the site or returned to the sponsor or nominee. All accountability records had to be available for inspection by sponsor and regulatory inspectors; photocopies had to be provided to the sponsor at the end of the trial.

[0359] All medication adherence records had to be kept up-to-date and made available for inspection by the clinical trial sponsor and regulatory inspectors.

[0360] Concomitant medications and procedures Any treatment administered from the time of informed consent until the end of the treatment period was considered a concomitant medication. This included medications initiated before the study and those continued during the study. Any concomitant medication had to be reviewed and approved by the Regeneron Medical Monitor. Information on concomitant medications for each patient was recorded from screening to the end of the study.

[0361] The initiation of any new prescription medication treatment was prohibited from the time of screening until the final day of the trial visit unless agreed upon by the principal investigator or designated investigator and the medical monitor.

[0362] Furthermore, depending on the eligibility criteria, the following drugs and procedures are prohibited during this study: leukotriene receptor antagonists, muscarinic antagonists, terbutaline, theophylline, beta-blockers, digoxin, NSAIDs, MAO inhibitors, or tricyclic antidepressants, roflumilast, and cromoglycic acid; biological therapies (e.g., anti-IgE, anti-IL-5) or immunotherapies (subcutaneous immunotherapy (SCIT), sublingual immunotherapy (SLIT), or oral immunotherapy (OIT)); treatment with live (attenuated) vaccines; oral and systemic corticosteroids; treatment with inhaled or nasal inhaled corticosteroids; and any new day within four weeks prior to the visit. Initiating regular exercise or making a significant change to a previous daily exercise routine (patients must attempt to maintain a similar level of exercise during the study period and avoid significantly strenuous exercise during the study period); during this study, adult alcohol consumption is limited to no more than two drinks per day (one drink is equivalent to 12 ounces of regular beer, 5 ounces of wine, or 1.5 ounces of 80 proof (40% alcohol) spirits), and patients must refrain from drinking alcohol for 48 hours prior to screening and at the allergen test visit; Participants had to refrain from caffeine for 8 hours prior to each trial visit (caffeine-free products were permitted).

[0363] The following medications were approved: short-acting β-agonists; thyroid replacement therapy in patients receiving continuous treatment within the past six months prior to screening; vitamin and calcium supplements; over-the-counter antihistamines and decongestants; paracetamol (care should be taken to follow all guidance related to paracetamol administration and not to exceed the maximum approved daily dose); laxatives; antacids; and heat-sterilized vaccines.

[0364] Test Procedure For the sole purpose of determining eligibility for the trial or characterizing the baseline population: demographics, medical history, HIV, hepatitis and drug screening, HDM skin prick test, and metacholine loading, the following steps were performed.

[0365] Safety Procedures: Patient safety will be monitored by patient-reported adverse events (AEs) or by the investigator and by clinical laboratory tests (e.g., biochemistry, hematology, and urinalysis), vital signs, and standard 12-lead ECG auto-reading. Any clinically significant abnormalities (if any) will be monitored until they resolve or become clinically stable.

[0366] Vital signs: Vital signs, including temperature, blood pressure, pulse, and respiration, should be collected at a given time, at least 5 minutes after rest, and before administration, according to Tables 1 and 2.

[0367] Physical Examination: A thorough and complete physical examination, including height and weight, should be performed at some point in time according to Tables 1 and 2. Care should be taken to examine and evaluate any abnormalities that may be present, as indicated by the patient's medical history.

[0368] Electrocardiogram: An electrocardiogram should be performed during a visit requiring blood collection, before blood is drawn. A standard 12-lead ECG should be performed at the times specified in Tables 1 and 2. Record the heart rate, starting with the ventricular velocity, and record the PR, QRS, RR, and QT interval. Keep the ECG strip or report at its source. For ECG procedures, the 12-lead ECG should be systematically digitized and recorded after the patient has been in a supine position for at least 10 minutes. Place the electrodes in the same location for each ECG record throughout the study. Each ECG should consist of a 10-second recording of 12 leads simultaneously, and print out a single 12-lead ECG (25 mm / sec, 10 mm / mV) with the date, time, patient initials and number, research physician's signature, and assessments (HR, PR, QRS, RR, QT interval, and QTc), including at least three composites per lead. Reads should be used for immediate safety assessment. The medical opinion of the clinical trial physician and ECG values ​​will be recorded in the eCRF.

[0369] HDM Skin Prick Test: At the screening visit, a standard skin prick test using HDM allergens is performed to confirm inclusion criteria. Subsequently, a series of skin prick tests using diluted HDM solutions are performed to determine skin sensitivity. Skin sensitivity is used to determine the allergen dose regimen and to use it during the screening BAC.

[0370] Metacholine Challenge: At the screening visit, a metacholine challenge will be performed to confirm inclusion criteria and determine the allergen dose regimen used during the screening BAC. The metacholine challenge will be performed using a 2-minute resting ventilation protocol, following the ATS / ERS (1999) guidelines (Crapo RO et al., Guidelines for methacholine and exercise challenge testing-1999). official statement of the American Thor The acic Society will be conducted in accordance with the ATS Board of Directors, July 1999. Am J Respir Crit Care Med. January 2000; Vol. 161 (No. 1): pp. 309-329.

[0371] Clinical Testing Hematological, chemical, urinalysis, and pregnancy test samples were analyzed by a central laboratory. Detailed instructions for blood sample collection are provided in the laboratory manual offered to the testing facility. Samples for clinical testing are collected at the time of the visit, according to Table 1. The tests include the following:

[0372] blood chemistry Sodium, total protein, serum, total bilirubin Potassium, Creatinine, Total Cholesterol * Chloride, Blood urea nitrogen (BUN), Triglycerides Carbon dioxide, aspartate aminotransferase (AST), uric acid Calcium, alanine aminotransferase (ALT), creatine phosphokinase (CPK) Glucose (on an empty stomach), alkaline phosphatase, ionized calcium Albumin lactate dehydrogenase (LDH) * (Low-density lipoprotein [LDL] and high-density lipoprotein [HDL]) hematology Hemoglobin fractionation: Hematocrit neutrophils Red blood cells (RBCs) Lymphocytes White blood cells (WBCs) monocytes Red blood cell index basophil Platelet count, eosinophils Urine test Color Urine Sugar RBC Transparency of blood, heathlinism, and other casts. pH, bilirubin, bacteria Specific gravity Leukocyte esterase Epithelial cells Ketone nitrite crystals Protein WBC Yeast

[0373] Other clinical tests Patients were tested for follicle-stimulating hormone (FSH) levels (menopausal women only), and serum and urine pregnancy tests (women only) were performed at each time point shown in Tables 1 and 2. Samples were collected for evaluation of serum / plasma sST2, and total IL-33, calcitonin, MMP12, TARC, PARC, and eotaxin-3. Blood tests were performed for HIV, HBsAg, HBcAb, and HCV, as well as for TB. In addition, urine samples were collected for drug screening. Intact parathyroid hormone (iPTH) and 25-hydroxyvitamin D samples were collected in Part 1 (Table 1) but not in Part 2 of the study.

[0374] Pharmacokinetic and anti-drug antibody procedures Drug concentration samples were collected at the points shown in Table 1. Any unused samples may be used for exploratory biomarker studies.

[0375] Anti-drug antibody measurement and samples: Samples for ADA evaluation were collected at the points shown in Table 1. Results from any exploratory analyses were reported separately from the clinical trial reports. Unused samples collected for ADA analysis may be used for future biomedical research.

[0376] Pharmacodynamic procedures: Pharmacodynamic procedures include BAC, sputum induction, measurement of gene expression levels in sputum mRNA, clinical measurement of FEV1, FeNO, and measurement of circulating biomarkers.

[0377] Bronchial allergen load: Cockcroft et al., The links between allergen skin test sensitivity, airway responsiveness and airway Following the standard procedure using "response to allergen.Allergy. January 2005; Vol. 60 (No. 1): pp. 56-59," and to confirm the presence of early and late phase responses at the screening visits specified in Tables 1 and 2, BAC using HDM will be performed as detailed in this study manual. Post-treatment BAC will be performed using the dose regimen calculated from the screening BAC. EAR will be measured as the maximum decrease in FEV1 within the first 120 minutes (30 minutes if enrolled at screening). LAR will be measured as the decrease in FEV1 3-8 hours after allergen inhalation.

[0378] Sputum induction: Sputum induction should be performed at each time point according to Tables 1 and 2. Hypertonic saline (4.5% NaCl) should be sprayed through the mouth for 5 minutes in 4 cycles while the nose is closed with a clip, and inhaled. As a safety procedure, vital capacity should be measured for approximately 7 minutes, 5 minutes after each induction. Although some patients develop wheezing and dyspnea, sputum induction is generally well tolerated. Any airway constriction caused by sputum induction with hypertonic saline can be rapidly resolved with treatment with an inhaled short-acting β2 agonist (Wong et al., Safety of one method of sputum induction in asthmatic subjects. Am J Respir Crit Care Med. 1997. Vol. 156: pp. 299-303).

[0379] Measurement of gene expression levels in induced sputum: Induced sputum samples are collected at each time point according to Tables 1 and 2. Sputum is processed into RNA, and gene expression analysis is performed using Taqman assay, RNA-seq, or Nanostring. The genes tested are those thought to be related to the pathophysiology of asthma (type 1 and type 2 inflammation), countermeasures, and the mechanisms of action of REGN3500, dupilumab, and / or REGN3500 + dupilumab combination therapy. Molecular signatures for type 1 inflammation may include genes for IFNγ, CXCL9, CXCL10, CXCL11, IL-8, MPO, and neutrophil elastase. Molecular signatures for type 2 inflammation may include genes for IL-4, IL-5, IL-13, IL-9, CCL17, CCL26, CCL13, and CCL11. The list of genes being tested may be modified or expanded as additional potentially relevant or novel biomarkers may be discovered during this study.

[0380] Spirometry: Clinical spirometry was performed using a standard spirometer, in accordance with the American Thoracic Society's standards for acceptable quality control (Miller et al., Force. Standardisation of spirometry. Eur Respir J. 2005. Vol. 26: pp. 319-338). During the study (screening, treatment, and post-treatment period), patients were required to have their clinical spirometry (FEV1) measured at each scheduled study visit (Tables 1 and 2).

[0381] Exhaled nitric oxide: Measuring FeNO levels in asthma patients is used as a marker of airway inflammation. Exhaled nitric oxide is analyzed from exhaled condensate. Patients should be instructed to refrain from consuming nitrate-rich foods and drinks for at least 2 hours and to refrain from any food or beverages for at least 1 hour before FeNO measurement, and FeNO measurement should be performed before any vital capacity measurement. Throughout this study (screening, treatment, and post-treatment periods), patients are required to perform FeNO measurements as specified (Tables 1 and 2).

[0382] Circulating Biomarkers: Circulating biomarker samples will be collected at each time point according to Tables 1 and 2. Biomarker measurements will be performed in serum and plasma samples to determine the effects of biomarkers of inflammatory diseases on pathobiological or related physiological and pathogenic processes. The biomarkers tested are those thought to be related to the pathophysiology of the disease, its response, the mechanism of action of REGN3500 (and / or combination therapy), and its potential toxicity. Biomarkers tested in blood may include, but are not limited to, IL-33, soluble ST2, calcitonin, and MMP12.

[0383] Future Biomedical Research: Future biomedical research samples, as well as unused PK and ADA samples, will be stored until 15 years after the final date of the database lock. Unused samples can be used for future biomedical research, including research on inflammatory diseases. After 15 years, any remaining samples will be destroyed.

[0384] Genomics Substudy - Optional: Patients who consent to participate in a genomics substudy must sign a separate genomics substudy ICF before sample collection. Patients do not need to participate in a genomics substudy to enroll in the primary trial. DNA and whole blood RNA samples are included in the genomics substudy. Samples for DNA extraction should be collected on day 1 / baseline (pre-administration), but can be collected at any trial visit. Samples for whole blood RNA should be collected as specified in the trial chart. DNA samples for genomics substudies are double-encrypted as defined by the International Council of Harmonisation (ICH) guideline E15. Substudy samples may be stored for 15 years after the final date of the database lock and used for research purposes. The purpose of genomic analysis is to identify genomic associations with clinical or biomarker responses, other clinical outcome measures, and possible adverse events. Furthermore, associations between genomic variants and the prognosis or progression of other diseases can also be studied. These data may be used or combined with data collected from other studies to identify and validate genomic markers associated with the investigational drug or disease. Analysis may include sequencing of candidate genes and surrounding genomic regions or single nucleotide polymorphism (SNP) testing. Other methods, including whole exome sequencing, whole genome sequencing, DNA copy number variation (CNP) analysis, and transcriptome sequencing, may also be available. The list of methods may be expanded to include novel methodologies that may be developed during the course of the study or during sample storage.

[0385] Endpoint: Sputum mRNA scale Induced sputum samples have been used in clinical trials for asthma to assess airway inflammation. In studies comparing sputum obtained from asthma patients with sputum obtained from normal controls, the levels of IL-33 and ST2 were assessed by protein and / or RNA (Hamzaoui et al., Induced sputum levels of IL-33 and soluble ST2 in young asthmatic children. J Asthma. 2013. Vol. 50: pp. 803-809 and Salter et al., IL-25 a). nd IL-33 induce Type 2 inflammation in basophils from subjects with allergic asthma. Respir Res. 2016, Vol. 17: p. 5), eotaxin, TARC (Heijink et al., EE effect of ciclesonide treatment on allergen-induced changes in T cell Regulation in asthma. Int Arch Allergy Immunol. 2008. Vol. 145: pp. 111-121 and Sekiya et al., Increased levels of a TH2-type CC chemokine Elevated concentrations of thymus and activation-regulated chemokine (TARC)in serum and induced sputum of asthmatics (Allergy, 2002, Vol. 57: pp. 173-177), as well as IL-5 and IL-13 (Park et al., Interleukin-13 and interleukin-5 in induced sputum of eosinophilic bronchitis: comparison with asthma, Chest, 2005, Vol. 128: pp. 1921-1927, and Peters, MC, ZK Mekonnen, S. Yuan, NRB Hakta, PG Woodruff, and JV Fahy, Measures of gene expression in sputum cells can identify TH2-high and TH2-low subtypes of asthma, J Allergy Clin Immunol, 2014, Vol. 133: pp. 388-394). Sputum cytokines, such as IL-4, IL-5, and IL-13, are elevated and associated with the presence and severity of asthma symptoms (Truyen et al., Evaluation of airway inflammation by quantitative Th1 / Th2 cytokine mRNA measurement). in sputum of asthma patients.Thorax.2006.61:202-208).

[0386] Previous studies have shown that in patients with mild asthma, BAC acutely increased levels of type 2 cytokines, such as IL-13 and IL-5, in the lungs by approximately 10X. Treatment with inhaled corticosteroids significantly suppressed this BAC-mediated upregulation of mRNA levels of proteins and type 2 cytokines (Zuiker et al., Kinetics of TH2 biomarkers in sputum of asthmatics following inhaled allergen. Eur Clin Respir J. 2015. Vol. 2 and Zuiker et al., Sputum RNA signature in allergic asthmatics following allergen bronchoprovocation test. Eur Clin Respir J. 2016. Vol. 3:31324).

[0387] The endpoints in this study were designed to investigate the effects of REGN3500, dupilumab, and REGN3500 + dupilumab combination therapy on type 2 inflammatory gene expression. Furthermore, sputum mRNA measurements were analyzed to assess a broader gene expression profile, including genes involved in type 1 and type 2 inflammation and those reflecting changes in cellular content.

[0388] Sputum cytokines and chemokines Previous studies have shown that cytokines and chemokines can be measured in sputum induced after BAC. While the effect size of changes observed in previous studies suggests that measuring mRNA gene signatures may be superior to measuring protein signatures, this study uses cytokines as an exploratory endpoint. Samples were collected for evaluation of tokine and chemokine proteins. Cytokines and chemokines associated with the IL-33 and IL-4R pathways, including IL-13, IL-5, tumor necrosis factor-α (TNFα), TARC, pulmonary and activation-modulated chemokines (PARC), and eotaxin-3, were expected to be elevated after BAC. This increase in cytokines and chemokines was expected to be blunted by treatment with REGN3500 and / or dupilumab.

[0389] Decrease in FEV1 in the initial and delayed phases after bronchial allergen exposure. Changes in lung function after BAC were the standard endpoint for most allergen challenge tests evaluating the effects of inhaled corticosteroids. In sensitized patients, allergen inhalation induced an acute phase response, also known as the early allergen response (EAR), characterized by bronchoconstriction within 0–2 hours post-exposure. This EAR is thought to primarily represent the release of pre-formed mast cell mediators and was usually unresponsive to steroids. The early allergen response was often followed by a late allergen response (LAR), which occurred approximately 3–8 hours after exposure. This LAR is seen in 50–60% of adult asthma patients but was required for all patient enrollment in this study. The LAR occurred concurrently with the initial influx of inflammatory cells and generally responded to steroids.

[0390] When a crossover trial design was implemented with 12 patients, the reproducibility of these FEV1 endpoints was reported to be sufficient to demonstrate approximately 50% attenuation of EAR and / or LAR with >90% power. In this parallel design trial, the goal was to assess changes in sputum gene signatures that reflect changes in inflammatory stimuli that ultimately lead to changes in FEV1.

[0391] Exhaled Nitric Oxide Scale In BAC studies, sputum eosinophil counts have been shown to increase in asthma patients exhibiting a delayed-phase response. While an association between sputum eosinophil counts and FeNO has been reported, FeNO is not an eosinophil-specific marker and can be present in non-eosinophilic inflammation (Haldar et al., Mepolizumab and exacerbations of refractory eosinophilic asthma. N Engl J Med. 2009. Vol. 360: pp. 973-984). The strong correlation between baseline FeNO levels and REGN3500 and / or dupilumab response, as assessed by changes in mRNA levels of type 1 and type 2 genes, supports the usefulness of FeNO as a PD biomarker for responsive patients for future trials, as it is a relatively simple and inexpensive measure.

[0392] statistical analysis This study was driven by the suppression of sputum IL-13, IL-5, and ST2 mRNA in the case of aggressive treatment (rather than placebo) as the primary endpoint, rather than the change in LAR, which has traditionally been the primary endpoint of BAC trials (usually around 20 patients per treatment group). Six patients per treatment group demonstrated a power of >99% at a two-sided significance level of 0.05, detecting changes in IL-5 and IL-13 gene expression levels corresponding to the changes observed with inhaled corticosteroids (maximum observed effect size of 3.7 based on the inter-treatment difference compared to placebo). To ensure that sputum samples are generated after BAC and are appropriate for mRNA sizing, approximately eight patients per treatment group may be enrolled in Part 1 of this study.

[0393] Efficacy endpoints were analyzed using the maximum analysis population (FAS). For Part 1 of this study, the FAS included all randomized patients and assigned treatments. Based on the assumption of randomization.

[0394] Safety Analysis Population: The Safety Analysis Population (SAF) included all randomized patients who received at least one dose of the study drug. Patients were analyzed if treated. All safety analyses are presented together based on the SAF.

[0395] Pharmacokinetic analysis population: The PK analysis population included all treated patients who received any investigational drug and who had at least one post-administration, non-missing PK result after administration of the investigational drug.

[0396] Anti-drug antibody (ADA) analysis population: The ADA population included all treated patients who received any investigational drug and had at least one non-missing ADA result after the first dose of the investigational drug.

[0397] statistical methods For continuous variables, descriptive statistics included the following information: the number of patients reflected in the calculation (n), mean, median, standard deviation, Q1, Q3, minimum, and maximum. For categorical or rank data, frequency and percentage were shown for each category.

[0398] The breakdown of patients is as follows: total number of screened patients; patients who met the inclusion criteria for the target indication and signed the ICF; total number of randomized patients; patients who received a randomization number; total number of patients in each analysis population; total number of patients who discontinued the study and the reasons for discontinuation; list of patients who were treated but not randomized, patients who were randomized but not treated, and patients who, if randomized, were randomized but not treated; and list of patients who discontinued treatment prematurely due to the reasons for discontinuation.

[0399] Demographic and baseline characteristics are described descriptively for each treatment group and for all patients combined.

[0400] Primary analysis of efficacy: In this study, BAC using HDM was performed for each individual patient at screening and on day 29. The doubling of induced IL-13 mRNA from pre-BAC values ​​was obtained at 8 and 24 hours after loading (at screening and on day 29). The difference (8 and 24 hours) between the doubling of pre-BAC values ​​at screening and the doubling of pre-BAC values ​​at day 29 was calculated by computer for each patient. These doublings from baseline between screening and the day 29 visit were compared among the REGN3500 group, the dupilumab group, the REGN3500 + dupilumab combination group, and the placebo group. Furthermore, the effect of REGN3500 on a broader type 2 allergen signature was evaluated. Differences in doubling of mRNA expression levels were analyzed separately using the analysis of covariance model (ANCOVA). Descriptive statistics are also presented for treatment and doubling of mRNA levels at time and from baseline.

[0401] Secondary analysis of efficacy: For gene expression-based endpoints, the same analytical method as the primary analysis was used for continuous efficacy variables. The pharmacokinetic parameters of REGN3500 are summarized by arithmetic mean, standard deviation, coefficient of variation (CV(%)), minimum, Q1, median, Q3, maximum, and number of observations.

[0402] Safety Analysis: Treatment compliance / administration and all clinical safety variables were analyzed using SAF. The safety analysis was based on SAF. This included TEAE. This includes reports and other safety information (i.e., evaluation of clinical laboratory values, vital signs, and 12-lead ECG results). A summary of safety outcomes is presented for each treatment group. For safety variables, three observation periods were defined: the pre-treatment period was defined as the period from signing the ICF before the first dose of the investigational drug; the treatment period was defined as the period from the first dose of the investigational drug to the final dose plus 7 days; and the post-treatment period was defined as the period from the final dose plus 7 days after the final dose of the investigational drug. A treatment-induced adverse event (TEAE) was defined as one that was not present at baseline or one that worsened a pre-existing condition during the on-treatment period.

[0403] Vital signs: Vital signs (temperature, pulse, blood pressure, and respiratory rate) are presented together, showing the changes from baseline to the scheduled evaluation time using descriptive statistics.

[0404] Clinical Laboratory Tests: Clinical laboratory test results are presented summarized by baseline and by change from baseline to the planned evaluation time using descriptive statistics. Clinically significant abnormal laboratory values ​​(PCSV: potentially clinically significant) at any randomized time point are also shown. The number and percentage of patients with a significant value () are presented for each clinical test. Results for the target clinical test can be shown using a transition table based on baseline normal / abnormal values, as well as other tabular and graphical methods.

[0405] Treatment Exposure: The duration of exposure during this study is shown for each treatment group (integrated placebo) and calculated as (date of final investigational drug injection - date of first drug injection) + 7. The number (%) of patients exposed to the randomized and double-blind investigational drug is shown for each treatment group over a specific period. Furthermore, the duration of exposure during this study is summarized for each treatment group using the number of patients, mean, SD, minimum, Q1, median, Q3, and maximum.

[0406] Treatment compliance: Compliance with the investigational product as defined by the protocol was calculated as follows: Treatment compliance = (Number of investigational product injections during the exposure period) / (Number of planned investigational product injections during the exposure period) × 100%. Treatment compliance is presented within a specific range for each treatment group in Part 1 of this study.

[0407] Analysis of drug concentration data: Drug and total target concentrations for each sample collection period are summarized using descriptive statistics. The summary of drug and total target concentrations is shown by nominal time points (i.e., time points specified in the protocol). Plots of REGN3500 and total IL-33 concentrations are shown over time (linear and logarithmic scales). When the scale is linear, concentrations below the lower limit of quantification (LLOQ) were set to 0. In the logarithmic scaled figures, concentrations below LLOQ were interpolated as LLOQ / 2.

[0408] Analysis of anti-drug antibody data: The incidence of positive responses in ADA assays for REGN3500 and / or dupilumab was assessed by group as absolute occurrence (n) and percentage of patients (%). A list of all ADA titer levels is provided for patients who benefited from the ADA assay for REGN3500 and / or dupilumab. Plots of functional REGN3500 concentrations can be examined, and the potential impact of ADA on individual concentration-time profiles can be assessed. Assessment of the potential impact of ADA on safety and efficacy for REGN3500 and / or dupilumab can be explored.

[0409] Analysis of pharmacodynamic and biomarker data: Pharmacodynamic effects of REGN3500, dupilumab, and REGN3500+dupilumab on sputum gene expression (mRNA) outcomes, compared to the mean cytokine gene expression normalized to housekeeping gene expression. The log2-fold change (FCH) values ​​(SD, Q1, Q2, SEM, minimum, maximum, and number of observations) are summarized. Changes in gene expression were evaluated as changes after allergen inhalation induction and changes over time from pre-treatment baseline. The potential mitigating effects of REGN3500, dupilumab, or a combination of REGN3500 and dupilumab compared to placebo were investigated by comparing the increase in cytokine gene expression after allergen inhalation loading at weeks 4 and 8 to pre-treatment levels. The pharmacodynamic effects of REGN3500, dupilumab, and a combination of REGN3500 and dupilumab on FEV1 are summarized on day 29 as measured values ​​(SD, Q1, Q3, SEM, minimum, mean, median, maximum, and number of observations) and percentage changes from baseline (SD, Q1, Q3, SEM, minimum, mean, median, maximum, and number of observations) (clinical FEV1). The pharmacodynamic parameters (circulatory markers and FeNO) of REGN3500, dupilumab, and the combination of REGN3500 and dupilumab are summarized by measured values ​​(SD, Q1, Q3, SEM, minimum, mean, median, maximum, and number of observations), changes from baseline (SD, Q1, Q3, SEM, minimum, mean, median, maximum, and number of observations), and percentage changes from baseline (SD, Q1, Q3, SEM, minimum, mean, median, maximum, and number of observations). Correlation analyses were performed between baseline IL-33 concentrations and other baseline biomarkers (calcitonin, sST2, and MMP12, TARC, PARC, eotaxin-3, and FeNO). Scatter plots using Pearson or Spearman correlations are shown for each correlation analysis.

[0410] Additional Statistical Data Handling Guidelines: Definition of Baseline. Unless otherwise specified, the baseline assessment for all measurements is the most recent and valid measurement performed prior to administration of the investigational drug. For most variables, the procedure and assessment on Day 1 are considered baseline.

[0411] General rules for handling missing data: Unless otherwise specified below, missing samples or concentration values ​​will not be supplemented, but will remain missing in the calculation of the induced PK parameter. If the actual sampling period is missing but reasonable concentration values ​​have been measured, the scheduled protocol time may be used for the calculation of the induced PK parameter. Pre-administration missing values ​​due to SC administration will be set to 0 for PK calculation. If the start date of an AE or concomitant medication is incomplete or missing, it will be assumed to have occurred during or after ingestion of the study drug unless the incomplete date (e.g., month and year) clearly indicates that the event started before treatment. If a partial date indicates the same month or year as the date of ingestion of the study drug, the start date will be supplemented by the date of ingestion of the study drug; otherwise, the missing date or month will be supplemented by the first day or first month. Missing laboratory values, ECG data, vital sign data, or physical examination data will not be supplemented. Assessments received outside of the protocol's acceptable window will be indicated according to the CRF assessment recorded by the investigator. Ad-hoc assessments (laboratory values ​​or vital signs obtained during a clinical visit not included in the protocol, or while investigating or managing an adverse event) should be included in the list but not in the summary. If two or more clinical laboratory values ​​are available for a given visit, the first observation should be used in the summary, and all observations should be listed.

[0412] result Treatment with REGN3500 and dupilumab monotherapy significantly suppressed allergen-induced eosinophilic signatures and broader type 2 inflammatory signatures, including CCL26, CCL17, and SIGLEC8. Many of the genes suppressed by REGN3500 were also suppressed by dupilumab, despite their different dynamics. Several genes were suppressed in only one of two arms, suggesting that the two molecules overlap and modulate different pathways. The combination of the two molecules suppressed type 2 inflammation similar to that observed with monotherapy.

[0413] Figure 5 presents data showing that treatment with REGN3500 reduces inflammation in a model of chronic dust mite disease (HDM) pneumonia. Treatment with REGN3500 suppressed pro-inflammatory cytokines and chemokines. These results are based on data showing levels of pulmonary eosinophils and neutrophils in HDM models with and without anti-IL-33 treatment. The presented data include heatmaps of the pulmonary cytokine panel showing levels of hIL-4, IL-5, IL-1b, TNFα, IFNg, GROa, and MCP-1. Alveolar SMA testing was also performed.

[0414] Analysis of bronchial allergen loading molecular signatures in sputum is shown in Figure 8, which shows the expression of various signature genes associated with type 2 inflammation before allergen loading, 8 hours after allergen loading, and 24 hours after allergen loading. Top genes induced by bronchial allergen loading at the time of screening included IL-4, IL-5, IL-13, IL-9, IL1RL1 (IL-33 receptor), Eot-3 (CCL26), TARC (CCL17), and FCER2, which enhanced type 2 inflammation.

[0415] An analysis of the repression of various genes induced by allergen loading by REGN3500 is shown in Figure 9, which shows that various genes of type 2 inflammatory cytokines and chemokines induced by bronchial allergen loading, including IL-5, IL-13, Eot-3 (CCL26), and TARC (CCL17), were repressed by REGN3500. Other genes repressed by REGN3500 and induced by bronchial allergen loading included CCL1 (ligand for CCR8, which attracts activated Th2 and Treg cells), CCL26, FCER2, SIGLEC8, and CCL17.

[0416] Figure 10 shows the gene signatures used to evaluate the therapeutic effect on sputum eosinophil count. A set of 10 genes showed a high correlation with sputum eosinophil count before and after allergen loading. These genes included ADARB1, ASB2, CLC, GLOD5, HDC, IL1RL1, PTPN7, SIGLEC8, SYNE1, and VSTM1. These genes are not exclusive to eosinophils; for example, SIGLEC8 is expressed in eosinophils, basophils, and mast cells; HDC is expressed in mast cells; and VSTM1 is expressed in myeloid cells.

[0417] Figure 11 shows that REGN3500 treatment suppressed eosinophil signature genes in sputum. Data present for ADARB1, ASB2, CLC, HDC, IL1RL1, PTPN7, SIGLEC8, SYNE1, and VSTM1. The effects mediated by anti-IL-33 (REGN3500) treatment were not noteworthy regarding neutrophil signature genes.

[0418] Figure 12 shows that REGN3500 treatment suppressed type 2 inflammatory signature genes in sputum. The data shows IL-4, IL-13, CCL26, CCL13, CCL17, CCL11, POSTN, IL-5, and IL-9. Figure 12 also shows that type 1 inflammatory signature genes were not induced by allergen loading.

[0419] In summary, the data in Figures 8-12 showed that the decrease in serum eosinophils was due to a certain pharmacodynamic effect of anti-IL-33. No decrease mediated by anti-IL-33 in neutrophils was observed. Furthermore, no decrease mediated by anti-IL-33 in other circulating type II inflammatory mediators was observed.

[0420] Figure 14 shows that dupilumab and REGN3500 reduce eosinophils after bronchial allergen loading. This demonstrates that it was possible to reduce the gene signature score. Combination therapy with dupilumab and REGN3500 was the most effective treatment for reducing the eosinophil gene signature score after bronchial allergen loading. Figure 14 shows the eosinophil gene signature score across the treatment arms. These arms include placebo, fluticasone, dupilumab, REGN3500, and combination therapy with dupilumab and REGN3500. Results are shown before and after bronchial allergen loading.

[0421] Figure 15 shows that the reduction in type 2 signature scores was lower in the REGN3500 treatment arm than in the fluticasone treatment arm. Figure 15 shows type 2 signature scores across the treatment arms. These arms include placebo, fluticasone, dupilumab, REGN3500, and combination therapy with dupilumab and REGN3500. Results are shown before and after bronchial allergen loading.

[0422] Figure 16 presents data showing genes affected by various treatment arms 8 and 24 hours after bronchial allergen loading. Figure 16 shows genes affected by placebo, fluticasone, dupilumab, REGN3500, and combination therapy of dupilumab and REGN3500. Results are presented at screening and treatment, which occur after bronchial allergen loading. The genes tested, from top to bottom, are: BC042385, AB209315, LOC100607117, BC035084, LOC145474, AX747853, TIMP1, NT5DC2, LOC541471, AREG, PTPN7, RUNDC3, XXYLT1, FAM159A, PTGDS, TESC, ITGB2-AS1, D0574721, CLDN9, and LOC1001320. This includes 52, AGAP7, NBEAL2, NTNG2, FLJ45445, KCNH3, POU51P3, OUG1, KIF21B, HSPA7, GAPT, BX6485Q2, PRR52, P1K3R6, LTC4S, CLEC11A, TRABD2A, DLGAP3, VDR, DKFZp686M11215, SIGLEC12, BC016361, BC052769, and RHOH.

[0423] Figure 17 shows, from top to bottom, the top genes induced by bronchial allergen loading and suppressed by REGN3500 over 24 hours were ASAP1-IT1, AX747757, BC042385, PABPC1P2, AB209315, AX748268, TCEAL5, CCL17, CCL13, CCL26, CLC, CACNG8, GPR82, GATA1, PRSS33, FFAR3, LGALS12, ASB2, PTGDR2, SIGLEC8, IL13, IL5, PTGDS, and RD3. Figure 17 shows the genes induced by placebo, fluticasone, dupilumab, REGN3500, and combination therapy with dupilumab and REGN3500. Results are presented at screening and treatment, which occur after bronchial allergen loading.

Claims

1. A method for treating allergic asthma in a subject requiring such treatment, comprising the step of administering to the subject an antibody or antigen-binding fragment thereof, which specifically binds to interleukin-33 (IL-33) and comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs: 4, 6, and 8, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs: 12, 14, and 16.

2. The method according to claim 1, wherein the antibody or its antigen-binding fragment comprises a heavy chain variable region (HCVR) containing the amino acid sequence of SEQ ID NO: 2 and a light chain variable region (LCVR) containing the amino acid sequence of SEQ ID NO:

10.

3. The method according to claim 1 or 2, wherein the antibody or its antigen-binding fragment comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 18 and a light chain containing the amino acid sequence of SEQ ID NO:

20.

4. The method according to any one of claims 1 to 3, wherein the antibody or its antigen-binding fragment is administered intravenously at a dose of 10 mg / kg.

5. The method according to any one of claims 1 to 3, wherein the antibody or its antigen-binding fragment is administered subcutaneously in a dose of about 0.1 mg to about 600 mg, about 100 mg to about 400 mg, or about 300 mg.

6. The method according to any one of claims 1 to 3 or 5, wherein the antibody or its antigen-binding fragment is administered subcutaneously in an initial dose of about 600 mg or about 300 mg.

7. The method according to any one of claims 1 to 3, 5, or 6, wherein the antibody or its antigen-binding fragment is administered subcutaneously in one or more second doses of about 300 mg.

8. A method for treating allergic asthma in a subject requiring such treatment, comprising the step of administering to the subject an antibody or antigen-binding fragment thereof, which specifically binds to interleukin-4R (IL-4R) and comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs. 21, 22, and 23, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs. 24, 25, and 26.

9. The method according to claim 8, wherein the second antibody or its antigen-binding fragment comprises a heavy chain variable region (HCVR) containing the amino acid sequence of SEQ ID NO: 27 and a light chain variable region (LCVR) containing the amino acid sequence of SEQ ID NO:

28.

10. The method according to claim 8 or 9, wherein the antibody or its antigen-binding fragment comprises dupilumab.

11. The method according to any one of claims 8 to 10, wherein the antibody or its antigen-binding fragment is administered in a dose of about 0.1 mg to about 600 mg, about 100 mg to about 400 mg, or about 300 mg.

12. The method according to any one of claims 8 to 11, wherein the antibody or its antigen-binding fragment is administered in an initial dose of approximately 600 mg.

13. The method according to any one of claims 8 to 11, wherein the antibody or its antigen-binding fragment is administered in one or more second doses of about 300 mg.

14. The method according to any one of claims 1 to 13, wherein the antibody or its antigen-binding fragment is administered once a week (q1w), every other week (q2w), once every three weeks (q3w), or once every four weeks (q4w).

15. The method according to any one of claims 1 to 14, wherein the antibody or its antigen-binding fragment is administered every other week (q2w).

16. The method according to any one of claims 1 to 3 and 5 to 15, wherein the antibody or its antigen-binding fragment is administered subcutaneously.

17. The method according to any one of claims 1 to 3 and 5 to 16, wherein the antibody or its antigen-binding fragment is administered subcutaneously using an auto-injector, needle and syringe, or pen-type delivery device.

18. A method for treating allergic asthma in those who require it, An initial dose of approximately 600 mg of an antibody or its antigen-binding fragment that specifically binds to interleukin-4R (IL-4R) and contains three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs. 21, 22, and 23, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs. 24, 25, and 26; and One or more subsequent doses of the antibody or its antigen-binding fragment, approximately 300 mg The method comprising the step of administering to a target.

19. The method according to claim 18, wherein the antibody or its antigen-binding fragment comprises a heavy chain variable region (HCVR) containing the amino acid sequence of SEQ ID NO: 27 and a light chain variable region (LCVR) containing the amino acid sequence of SEQ ID NO:

28.

20. A method for treating allergic asthma in those who require it, A first antibody or its antigen-binding fragment that specifically binds to interleukin-33 (IL-33) and comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs: 4, 6, and 8, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs: 12, 14, and 16; and A second antibody or its antigen-binding fragment that specifically binds to the interleukin-4 receptor (IL-4R) and contains three heavy chain complementary determinant region (HCDR) sequences, including SEQ ID NOs. 21, 22, and 23, and three light chain complementary determinant region (LCDR) sequences, including SEQ ID NOs. 24, 25, and 26. The method comprising the step of administering to a target.

21. The method according to claim 20, wherein the first antibody or its antigen-binding fragment comprises a heavy chain variable region (HCVR) containing the amino acid sequence of SEQ ID NO: 2 and a light chain variable region (LCVR) containing the amino acid sequence of SEQ ID NO:

10.

22. The method according to claim 20 or 21, wherein the first antibody or its antigen-binding fragment comprises a heavy chain having the amino acid sequence of SEQ ID NO: 18 and a light chain having the amino acid sequence of SEQ ID NO:

20.

23. The method according to any one of claims 20 to 22, wherein the second antibody or antigen-binding fragment comprises a heavy chain variable region (HCVR) containing the amino acid sequence of SEQ ID NO: 27 and a light chain variable region (LCVR) containing the amino acid sequence of SEQ ID NO:

28.

24. The method according to any one of claims 20 to 23, wherein the second antibody or antigen-binding fragment comprises dupilumab.

25. The method according to any one of claims 20 to 24, wherein the second antibody or its antigen-binding fragment is administered in a dose of about 0.1 mg to about 600 mg, about 100 mg to about 400 mg, or about 300 mg.

26. The method according to any one of claims 20 to 25, wherein the second antibody or its antigen-binding fragment is administered in an initial dose of approximately 600 mg.

27. The method according to any one of claims 20 to 25, wherein the second antibody or its antigen-binding fragment is administered in one or more subsequent doses of about 300 mg of the antibody or its antigen-binding fragment.

28. The method according to any one of claims 20 to 27, wherein the second antibody or its antigen-binding fragment is administered once a week (q1w), every other week (q2w), once every three weeks (q3w), or once every four weeks (q4w).

29. The method according to any one of claims 20 to 28, wherein the second antibody or its antigen-binding fragment is administered every other week (q2w).

30. The method according to any one of claims 20 to 29, wherein the second antibody or its antigen-binding fragment is administered subcutaneously.

31. The method according to any one of claims 20 to 30, wherein the second antibody or its antigen-binding fragment is administered subcutaneously using an autoinjector, needle and syringe, or pen-type delivery device.

32. The method according to any one of claims 20 to 31, wherein the first antibody or its antigen-binding fragment is administered subcutaneously in a dose of about 0.1 mg to about 600 mg, about 100 mg to about 400 mg, or about 300 mg.

33. The method according to any one of claims 20 to 32, wherein the first antibody or its antigen-binding fragment is administered subcutaneously in an initial dose of about 600 mg or about 300 mg.

34. The method according to any one of claims 20 to 33, wherein the first antibody or its antigen-binding fragment is administered subcutaneously in one or more second doses of about 300 mg.

35. The method according to any one of claims 20 to 30, wherein the first antibody or its antigen-binding fragment is administered intravenously at a dose of 10 mg / kg.

36. A method for treating allergic asthma in those who require it, A first antibody or its antigen-binding fragment, which specifically binds to interleukin-33 (IL-33) and comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs. 4, 6, and 8, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs. 12, 14, and 16. A first antibody or its antigen-binding fragment administered at a single dose of 10 mg / kg; and It specifically binds to the interleukin-4 receptor (IL-4R), and sequence numbers 21 and 22 A second antibody or its antigen-binding fragment comprising three heavy chain complementary determinant region (HCDR) sequences including sequence numbers 23, and three light chain complementary determinant region (LCDR) sequences including sequence numbers 24, 25, and 26, The second antibody or its antigen-binding fragment is administered as an initial dose of 600 mg and subsequent doses of approximately 300 mg, once or more times. The method comprising the step of administering to a target.

37. The method according to any one of claims 1 to 36, wherein the allergic asthma is mild allergic asthma.

38. The method according to claim 37, wherein the allergic asthma is mild, persistent allergic asthma.

39. The method according to any one of claims 1 to 38 applies to individuals with an allergy to dust mite allergen (HDM).

40. The method according to any one of claims 1 to 39, wherein the subject is a non-smoker.

41. The method according to any one of claims 1 to 40, wherein the patient is clinically stable and requires the use of an inhaled short-acting β2-agonist (SABA) when necessary to control asthma symptoms.

42. The method according to any one of claims 1 to 41, wherein loss of asthma control (LOAC) is reduced in the subject.

43. The method according to any one of claims 1 to 42, wherein asthma symptoms selected from the group consisting of cough and wheezing, and the use of an inhaled short-acting β2 agonist are reduced in the subject.

44. The method according to any one of claims 1 to 43, wherein one or more asthma-related parameters are improved in the subject.

45. The method according to claim 44, wherein the asthma-related parameters are selected from the group consisting of forced expiratory volume in one second (FEV1), maximum expiratory flow rate (PEF), forced vital capacity (FVC), forced expiratory rate (FEF) of 25% to 75%, and a reduction in the frequency or dose of inhaled short-acting β2-agonist use in the subject.

46. The method according to claim 45, wherein FEV1 before administration of a bronchodilator is improved in the subject.

47. The method according to any one of claims 1 to 46, wherein the blood eosinophil level is reduced in the subject.

48. The method according to any one of claims 1 to 47, wherein one or both of the Asthma Control Questionnaire Five-Question Version (ACQ-5) score and the Asthma Quality of Life Questionnaire with Standardized Activities (AQLQ) score are improved in the subject.

49. The method according to any one of claims 1 to 48, wherein the frequency or dose of SABA use in the subject is reduced in the subject.

50. Bronchial allergen loading (BAC)-induced pneumonia is reduced in the subjects, claims 1 to The method described in any one of item 49.

51. The method according to any one of claims 1 to 50, wherein type 2 cytokine levels are reduced in the subject.

52. The method according to claim 51, wherein the type 2 cytokine is one or both of IL-13 and IL-5.

53. The method according to any one of claims 1 to 52, wherein cytokine or chemokine levels are reduced in the subject, and the cytokine or chemokine is selected from the group consisting of tumor necrosis factor-α (TNFα), thymic and activating modulated chemokines (TARC), lung and activating modulated chemokines (PARC), CCL1, CCL26, FCER2, SIGLEC8, CCL17, and eotaxin-3.

54. The method according to any one of claims 1 to 53, wherein the early allergen response (EAR) or late allergen response (LAR) is reduced in the subject.

55. The method according to any one of claims 1 to 54, wherein FEV1 is improved to at least 20%, 30%, 40%, 50%, 60%, or 70% in the subject.

56. The method according to any one of claims 1 to 55, wherein the FeNO level is reduced in the subject.

57. The method according to any one of claims 1 to 56, wherein serum levels of sST2, IL-33, calcitonin, or matrix metalloproteinase-12 (MMP12) are reduced in the subject.

58. The method according to any one of claims 1 to 57, wherein serum levels of CCL26, CCL17, or SIGLEC8 are reduced in the subject.

59. The method according to any one of claims 1 to 58, wherein serum levels of ASAP1-IT1, AX747757, BC042385, PABPC1P2, AB209315, AX748268, TCEAL5, CCL13, CLC, CACNG8, GPR82, GATA1, PRSS33, FFAR3, LGALS12, ASB2, PTGDR2, IL-13, IL-5, PTGDS, or RD3 are reduced in the subject.

60. A method for reducing cytokine or chemokine levels in a subject with allergic asthma, comprising the step of administering to the subject an antibody or antigen-binding fragment thereof, which specifically binds to interleukin-33 (IL-33) and comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs: 4, 6, and 8, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs: 12, 14, and 16.

61. The method according to claim 60, wherein the cytokine is one or both of IL-13 and IL-5.

62. The method according to claim 60 or 61, wherein the cytokine or chemokine is selected from the group consisting of TNFα, TARC, PARC, CCL1, CCL26, FCER2, SIGLEC8, CCL17, and eotaxin-3.

63. Serum levels of sST2, IL-33, calcitonin, or MMP12 were observed in the subjects. The method according to any one of claims 60 to 62, which reduces

64. The method according to any one of claims 60 to 63, wherein serum levels of CCL26, CCL17, or SIGLEC8 are reduced in the subject.

65. A method for reducing the expression of one or more allergic asthma signature genes in a subject having allergic asthma, comprising the step of administering to the subject an antibody or antigen-binding fragment thereof that specifically binds to interleukin-33 (IL-33) and comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs: 4, 6, and 8, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs: 12, 14, and 16.

66. One or more allergic asthma signature genes are BC042385, AB209315, LOC100607117, BC035084, LOC145474, AX747853, TIMP1, NT5DC2, LOC541471, AREG, PTPN7, RUNDC3, XXYLT1, FAM159A, PTGDS, TESC, ITGB2-AS1, D0574721, CLDN9, LOC100132052, The method according to claim 65, selected from the group consisting of AGAP7, NBEAL2, NTNG2, FLJ45445, KCNH3, POU51P3, OUG1, KIF21B, HSPA7, GAPT, BX6485Q2, PRR52, P1K3R6, LTC4S, CLEC11A, TRABD2A, DLGAP3, VDR, DKFZp686M11215, SIGLEC12, BC016361, BC052769, and RHOH.

67. The method according to claim 65 or 66, wherein one or more allergic asthma signature genes are selected from the group consisting of ASAP1-IT1, AX747757, BC042385, PABPC1P2, AB209315, AX748268, TCEAL5, CCL17, CCL13, CCL26, CLC, CACNG8, GPR82, GATA1, PRSS33, FFAR3, LGALS12, ASB2, PTGDR2, SIGLEC8, IL13, IL5, PTGDS, and RD3.

68. A method for reducing the expression of any combination of type 2 inflammatory cytokines and type 2 chemokine signature genes in a subject having allergic asthma, comprising the step of administering to the subject an antibody or antigen-binding fragment thereof that specifically binds to interleukin-33 (IL-33) and comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs: 4, 6, and 8, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs: 12, 14, and 16.

69. The method according to claim 68, wherein the type 2 inflammatory cytokine and chemokine signature gene is selected from the group consisting of IL-5, CCL1, IL-13, GATA2, CCL26, FCER2, CACNG8, CLC, GATA1, LGALS12, SIGLEC8, GGT5, CCL17, and MMP10.

70. The method according to claim 68 or 69, wherein one or more type 2 inflammatory cytokine and chemokine signature genes are selected from the group consisting of IL-5, CCL1, IL-13, CCL26, FCER2, SIGLEC8, GGT5, and CCL17.

71. A method for reducing the expression of one or more eosinophil signature genes in a subject with allergic asthma, comprising three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs: 4, 6, and 8, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs: 12, 14, and 16, which specifically bind to interleukin-33 (IL-33). The method comprising the step of administering an antibody or an antigen-binding fragment containing a sequence to a target.

72. The method according to claim 71, wherein one or more eosinophil signature genes are selected from the group consisting of IL1RL1, ADARB1, SIGLEC8, ASB2, VSTM1, SYNE1, CLC, PTPN7, and HDC.

73. A method for reducing the expression of one or more type 2 inflammatory signature genes in a subject having allergic asthma, comprising the step of administering to the subject an antibody or antigen-binding fragment thereof that specifically binds to interleukin-33 (IL-33) and comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs: 4, 6, and 8, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs: 12, 14, and 16.

74. The method according to claim 73, wherein one or more type 2 inflammatory signature genes are selected from the group consisting of IL-4, IL-13, CCL26, CCL13, CCL17, CCL11, POSTN, IL-5, and IL-9.

75. The method according to any one of claims 60 to 74, wherein the anti-IL-33 antibody or its antigen-binding fragment comprises a heavy chain variable region (HCVR) containing the amino acid sequence of SEQ ID NO: 2 and a light chain variable region (LCVR) containing the amino acid sequence of SEQ ID NO:

10.

76. The method according to any one of claims 60 to 75, wherein the anti-IL-33 antibody or its antigen-binding fragment comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 18 and a light chain containing the amino acid sequence of SEQ ID NO:

20.

77. A method for reducing cytokine or chemokine levels in a subject with allergic asthma, comprising the step of administering to the subject an antibody or antigen-binding fragment thereof, which specifically binds to the interleukin-4 receptor (IL-4R) and comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs. 21, 22, and 23, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs. 24, 25, and 26.

78. The method according to claim 77, wherein the cytokine is one or both of IL-13 and IL-5.

79. The method according to claim 77 or 78, wherein the cytokine or chemokine is selected from the group consisting of TNFα, TARC, PARC, CCL1, CCL26, FCER2, SIGLEC8, CCL17, and eotaxin-3.

80. The method according to any one of claims 77 to 79, wherein serum levels of sST2, IL-33, calcitonin, or MMP12 are reduced in the subject.

81. The method according to any one of claims 77 to 80, wherein serum levels of CCL26, CCL17, or SIGLEC8 are reduced in the subject.

82. A method for reducing the expression of one or more allergic asthma signature genes in a subject with allergic asthma, comprising three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs. 21, 22, and 23, and three light chain complementary determinant regions including SEQ ID NOs. 24, 25, and 26, which specifically bind to the interleukin-4 receptor (IL-4R). The method comprising the step of administering an antibody or an antigen-binding fragment containing a target-determining region (LCDR) sequence to a target.

83. One or more allergic asthma signature genes are BC042385, AB209315, LOC100607117, BC035084, LOC145474, AX747853, TIMP1, NT5DC2, LOC541471, AREG, PTPN7, RUNDC3, XXYLT1, FAM159A, PTGDS, TESC, ITGB2-AS1, D0574721, CLDN9, LOC100132052, The method according to claim 82, selected from the group consisting of AGAP7, NBEAL2, NTNG2, FLJ45445, KCNH3, POU51P3, OUG1, KIF21B, HSPA7, GAPT, BX6485Q2, PRR52, P1K3R6, LTC4S, CLEC11A, TRABD2A, DLGAP3, VDR, DKFZp686M11215, SIGLEC12, BC016361, BC052769, and RHOH.

84. The method according to claim 82 or 83, wherein one or more allergic asthma signature genes are selected from the group consisting of ASAP1-IT1, AX747757, BC042385, PABPC1P2, AB209315, AX748268, TCEAL5, CCL17, CCL13, CCL26, CLC, CACNG8, GPR82, GATA1, PRSS33, FFAR3, LGALS12, ASB2, PTGDR2, SIGLEC8, IL13, IL5, PTGDS, and RD3.

85. A method for reducing the expression of any combination of type 2 inflammatory cytokines and type 2 chemokine signature genes in a subject having allergic asthma, comprising the step of administering to the subject an antibody or antigen-binding fragment thereof that specifically binds to the interleukin-4 receptor (IL-4R) and comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs. 21, 22, and 23, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs. 24, 25, and 26.

86. The method according to claim 85, wherein the type 2 inflammatory cytokine and chemokine signature gene is selected from the group consisting of IL-5, CCL1, IL-13, GATA2, CCL26, FCER2, CACNG8, CLC, GATA1, LGALS12, SIGLEC8, GGT5, CCL17, and MMP10.

87. The method according to claim 85 or 86, wherein one or more type 2 inflammatory cytokines and chemokine signature genes are selected from the group consisting of IL-5, CCL1, IL-13, CCL26, FCER2, SIGLEC8, GGT5, and CCL17.

88. A method for reducing the expression of one or more eosinophil signature genes in a subject having allergic asthma, comprising the step of administering to the subject an antibody or antigen-binding fragment thereof that specifically binds to the interleukin-4 receptor (IL-4R) and comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs. 21, 22, and 23, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs. 24, 25, and 26.

89. The method according to claim 88, wherein one or more eosinophil signature genes are selected from the group consisting of IL1RL1, ADARB1, SIGLEC8, ASB2, VSTM1, SYNE1, CLC, PTPN7, and HDC.

90. A method for reducing the expression of one or more type 2 inflammatory signature genes in subjects with allergic asthma, wherein the method relates to the interleukin-4 receptor (IL-4 The method comprising the step of administering to a subject an antibody or its antigen-binding fragment that specifically binds to R) and comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs. 21, 22, and 23, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs. 24, 25, and 26.

91. The method according to claim 90, wherein one or more type 2 inflammatory signature genes are selected from the group consisting of IL-4, IL-13, CCL26, CCL13, CCL17, CCL11, POSTN, IL-5, and IL-9.

92. The method according to any one of claims 77 to 91, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region (HCVR) containing the amino acid sequence of SEQ ID NO: 27 and a light chain variable region (LCVR) containing the amino acid sequence of SEQ ID NO:

28.

93. The method according to any one of claims 77 to 92, wherein the antibody or its antigen-binding fragment comprises dupilumab.

94. The method according to any one of claims 60 to 76, further comprising the step of administering to a subject an antibody or antigen-binding fragment thereof that specifically binds to IL-4R, wherein the antibody or antigen-binding fragment comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs. 21, 22, and 23, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs. 24, 25, and 26.

95. The method according to any one of claims 77 to 93, further comprising the step of administering to a subject an antibody or antigen-binding fragment thereof that specifically binds to IL-33, wherein the antibody or antigen-binding fragment comprises three heavy chain complementary determinant region (HCDR) sequences including SEQ ID NOs: 4, 6, and 8, and three light chain complementary determinant region (LCDR) sequences including SEQ ID NOs: 12, 14, and 16.

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