Methods for treating or preventing asthma by administering an IL-4R antagonist

Abstract

Methods for treating or preventing asthma and related conditions are provided. A method for reducing the incidence of one or more asthma exacerbations in a subject in need thereof comprises administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising an interleukin-4 receptor (IL-4R) antagonist, wherein the asthma exacerbation is selected from the group consisting of: (a) a 30% or greater decrease from baseline in morning peak expiratory flow (PEF) for two consecutive days; (b) six or more additional exacerbation puffs of albuterol or levalbuterol (compared to baseline) in a 24-hour period for two consecutive days; and (c) (i) systemic (oral and / or parenteral) steroid treatment, or (ii) an increase in inhaled corticosteroid to at least four times the last dose received before discontinuation, or (iii) an asthma exacerbation requiring hospitalization.

Description

[Technical Field]

[0001] Related Applications This application claims the benefit of priority to U.S. Provisional Application No. 61 / 691,625, filed August 21, 2012; U.S. Provisional Application No. 61 / 758,097, filed January 29, 2013; U.S. Provisional Application No. 61 / 761,279, filed February 6, 2013; U.S. Provisional Application No. 61 / 783,796, filed March 14, 2013; U.S. Provisional Application No. 61 / 805,797, filed March 27, 2013; and French Application No. 1356994, filed July 16, 2013. The contents of each of the foregoing applications are incorporated herein by reference in their entirety.

[0002] FIELD OF THE INVENTION The present invention relates to the treatment and / or prevention of asthma and related conditions. More particularly, the present invention relates to the administration of interleukin-4 receptor (IL-4R) antagonists to treat or prevent asthma in patients in need thereof. [Background technology]

[0003] background Asthma is a chronic inflammatory disease of the airways characterized by airway hyperresponsiveness, acute and chronic bronchoconstriction, airway edema, and mucus plugging. The inflammatory component of asthma is 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 most frequently present with wheezing, shortness of breath, cough, and chest tightness. For most patients, a regimen of controller and bronchodilator therapy provides adequate long-term control. Inhaled corticosteroids (ICS) are considered the "gold standard" for managing asthma symptoms, and inhaled beta2-agonists are the most effective bronchodilators currently available. Studies have shown that combination therapy of ICS with an inhaled long-acting beta2-agonist (LABA) provides better asthma control than high-dose ICS alone. As a result, combination therapy has become the recommended treatment for subjects not managed with low-dose ICS alone.

[0004] Nevertheless, it is estimated that 5% to 10% of the asthma population will have symptomatic disease despite maximum recommended treatment with a combination of anti-inflammatory and bronchodilator medications. Furthermore, this severe asthma population accounts for up to 50% of total healthcare costs due to hospitalizations, emergency service use, and emergency department visits. Because many of these patients respond poorly to ICS due to a number of cellular and molecular mechanisms, there is an unmet need for new therapies in this severe asthma population. Furthermore, the long-term adverse effects of systemic and inhaled corticosteroids on bone metabolism, adrenal function, and child growth lead to attempts to utilize minimal corticosteroids. While the majority of asthma patients are reasonably well controlled with current treatments, patients with severe corticosteroid-resistant asthma have few therapeutic treatment options that can adequately manage their disease. The consequences of nonresponse or lack of adherence to treatment are loss of asthma control and, ultimately, asthma exacerbations.

[0005] One of the reasons for the poor response to pharmacotherapy in some patients with severe asthma may be the heterogeneity of the disease. There is growing interest in understanding these unique phenotypes, as targeted therapies are more likely to be successful in patients with similar underlying pathological features. Recent therapeutic approaches in asthma have focused on attempting to manage T helper cell-2 responses. Upregulation of interleukin-4 (IL-4) and interleukin-13 (IL-13) has been identified as important inflammatory components of asthma disease progression. It has been shown. Summary of the Invention [Problem to be solved by the invention]

[0006] Thus, there is a need in the art for new targeted therapies for the treatment and / or prevention of asthma. [Means for solving the problem]

[0007] Summary of the Invention According to one aspect of the present invention, there is provided a method for reducing the incidence of asthma exacerbations in a subject in need thereof. In a related aspect, there is provided a method for improving one or more asthma-related parameters in a subject in need thereof. In yet another aspect of the present invention, there is provided a method for treating asthma, e.g., moderate to severe eosinophilic asthma, in a subject in need thereof.

[0008] Methods featured in the invention involve administering to a subject a therapeutically effective amount of a pharmaceutical composition comprising an interleukin-4 receptor (IL-4R) antagonist. According to certain embodiments, the IL-4R antagonist is an antibody or antigen-binding fragment thereof that specifically binds to IL-4R. Exemplary anti-IL-4R antibodies that may be used in the context of the methods of the invention are described elsewhere herein, including in Example 1. For example, in one embodiment, the IL-4R antagonist is an antibody or antigen-binding fragment thereof that specifically binds to IL-4R and comprises heavy and light chain (complementarity-determining region) CDR sequences from the heavy chain variable region (HCVR) and light chain variable region (LCVR) of SEQ ID NOs: 162 and 164, respectively.

[0009] In one embodiment, a method is provided for reducing the incidence of one or more asthma exacerbations in a subject in need thereof by administering an antibody or antigen-binding fragment thereof that specifically binds to IL-4R. An asthma exacerbation can be one or more of the following: (a) a 30% or greater decrease from baseline in morning peak expiratory flow (PEF) for two consecutive days; (b) six or more additional reliever puffs of albuterol or levalbuterol (compared to baseline) in a 24-hour period for two consecutive days; and (c) an asthma exacerbation requiring (i) systemic (oral and / or parenteral) steroid treatment, or (ii) an increase in inhaled corticosteroid to at least four times the last dose received before discontinuation, or hospitalization.

[0010] In various embodiments, a method for improving one or more asthma-related parameters comprises administering a therapeutically effective amount of an IL-4R antagonist to a subject in need thereof, wherein the improvement in the asthma-related parameter is defined as one of the following: an increase from baseline in FEV1; an increase from baseline in AM PEF; an increase from baseline in PM PEF; a decrease from baseline in albuterol / levalbuterol use; a decrease from baseline in nighttime awakenings; and / or a decrease from baseline in SNOT-22 score. Examples of asthma-related parameters include: (a) forced expiratory volume in 1 second (FEV1); (b) peak expiratory flow (PEF), including morning PEF (AM PEF) and evening PEF (PM PEF); (c) use of inhaled bronchodilators, such as albuterol or levalbuterol; (d) five-item Asthma Control Questionnaire (ACQ5) score; (d) nighttime awakenings; and (e) 22-item Sino-Nasal Outcome Test (SNOT-22) score. In one embodiment, improvement in asthma-related parameters is a decrease in FEV1 of at least 0.10 L from baseline. In one embodiment, the improvement in asthma-related parameter is an increase in AM PEF of at least 10.0 L / min from baseline. In one embodiment, the improvement in asthma-related parameter is an increase in PM PEF of at least 1.0 L / min from baseline. In one embodiment, the improvement in asthma-related parameter is a reduction in albuterol / levalbuterol use of at least 1 puff per day from baseline. In one embodiment, the improvement in asthma-related parameter is a reduction in ACQ5 score of at least 0.5 points from baseline. In one embodiment, the improvement in asthma-related parameter is a reduction in nighttime awakenings of at least 0.2 times per night from baseline. In one embodiment, the improvement in asthma-related parameter is a reduction in SNOT-22 score of at least 5 points from baseline.

[0011] The present invention also provides a method for reducing the incidence of asthma exacerbations or improving one or more asthma-related parameters in a subject in need thereof, comprising administering to the subject in need thereof a single initial dose of a pharmaceutical composition comprising an IL-4R antagonist (e.g., an anti-IL-4R antibody or an antigen-binding fragment thereof), followed sequentially by one or more secondary doses of the pharmaceutical composition comprising the IL-4R antagonist. The pharmaceutical composition comprising the IL-4R antagonist can be administered to the subject in need thereof subcutaneously, intranasally, or intravenously.

[0012] According to certain embodiments, the present invention provides a method for reducing the incidence of asthma exacerbations or improving one or more asthma-related parameters in a subject in need thereof, comprising administering to the subject about 75 to about 300 mg of a pharmaceutical composition comprising an antibody or antigen-binding fragment thereof that specifically binds to IL-4R. According to this aspect, the pharmaceutical composition can be administered to the subject at a dosing frequency of, for example, once a week.

[0013] The present invention further encompasses methods of treating asthma (e.g., eosinophilic asthma, moderate to severe eosinophilic asthma, etc.) by selecting a subject exhibiting one or more symptoms or signs of asthma and administering to the patient a pharmaceutical composition comprising an IL-4R antagonist (e.g., an anti-IL-4R antibody or an antigen-binding fragment thereof), wherein the subject exhibits one or more of the following symptoms or signs of asthma: (1) the subject is receiving fluticasone / salmeterol combination therapy (250 / 50 μg twice daily (BID) or 500 / 50 μg BID) or budesonide / formoterol combination therapy (160 / 9 μg BID or 320 / 9 μg BID), and (2) the subject is receiving fluticasone / salmeterol combination therapy (250 / 50 μg twice daily (BID) or 500 / 50 μg BID) or budesonide / formoterol combination therapy (160 / 9 μg BID or 320 / 9 μg BID). (2) the subject has a blood eosinophil count greater than or equal to 300 cells / μL; (3) the subject has a sputum eosinophil count greater than or equal to 3%; (4) the subject has elevated levels of IgE, thymus and activation regulation chemokine (TARC), eotaxin-3, carcinoembryonic antigen (CEA), YKL-40, or periostin; (5) the subject has elevated levels of fractional exhaled nitric oxide (FeNO); and / or (6) the subject has a 5-item Asthma Control Questionnaire (ACQ5) score greater than or equal to 1.0.

[0014] Featured embodiments of the present invention relate to the above-described methods of treatment, further comprising administering a second therapeutic agent in combination with the IL-4R antagonist. The second therapeutic agent may be administered to a subject in need thereof before, after, or simultaneously with the IL-4R antagonist. Exemplary second therapeutic agents include, but are not limited to, one or more of the following in combination: an IL-1 inhibitor, an IL-5 inhibitor, an IL-8 inhibitor, an IgE inhibitor, a tumor necrosis factor (TNF) inhibitor, a corticosteroid, a long-acting base pair inhibitor, or an IL-4R antagonist. Ta2-agonist and leukotriene inhibitor.

[0015] In another aspect, the present invention provides a method for reducing or eliminating an asthma patient's dependence on background asthma therapy, the method comprising: selecting a patient with moderate to severe asthma that is uncontrolled or partially controlled with background asthma therapy; administering a prescribed dose of an IL-4R antagonist to the patient while maintaining the patient's background asthma therapy; and gradually reducing the dosage of one or more components of the background therapy over a subsequent treatment period while continuing administration of the IL-4R antagonist. In certain embodiments, the background therapy comprises an inhaled corticosteroid (ICS), a long-acting beta-agonist (LABA), or a combination of an ICS and a LABA. In some embodiments, the background therapy is gradually reduced or discontinued over a period of 2 to 8 weeks. In some embodiments, one component of the background therapy is eliminated after the initial treatment period. In one embodiment, the background therapy is gradually reduced over the subsequent treatment period.

[0016] In yet another aspect, the present invention provides a method for identifying and treating patients with moderate to severe asthma by selecting patients with elevated levels of biomarkers such as thymus and activation-regulated chemokine (TARC), IgE, eotaxin-3, periostin, carcinoembryonic antigen (CEA), or YKL-40, or with increased levels of fractional exhaled nitric oxide (FeNO); and administering a therapeutically effective amount of an IL-4R antagonist to the patient.

[0017] In another aspect, the invention features a method for monitoring the effectiveness of a treatment of moderate to severe asthma in a subject, e.g., by (a) determining the expression level of one or both of TARC or eotaxin-3, or the total serum level of IgE, in a biological sample obtained from the subject before treatment with an IL-4R antagonist; (b) determining the expression level of a biomarker in a biological sample obtained from the subject after treatment with an IL-4R antagonist; (c) comparing the expression level determined in step (a) with the level in step (b), and (d) concluding that the treatment is effective if the level determined in step (b) is lower than the level determined in step (a), or concluding that the treatment is not effective if the level determined in step (b) is the same as or higher than the level determined in step (a).

[0018] In one embodiment, the biomarker is FeNO, and treatment with the IL-4R antagonist is determined to be effective if FeNO levels decrease after administration of the antagonist.

[0019] The expression level of the biomarker can be determined, for example, one week, two weeks, three weeks, four weeks, five weeks, or more after administration of the IL-4R antagonist, and compared to the expression level before administration of the antagonist. The dose or dosing regimen of the IL-4R antagonist (e.g., an anti-IL4R antibody) can be adjusted after the determination. For example, if biomarker expression does not decrease within one week, two weeks, three weeks, four weeks, five weeks, or more after administration of the antagonist, treatment with the antagonist can be stopped, or the dose of the antagonist can be increased. If biomarker expression decreases after administration of the antagonist, the dosage of the antagonist can be maintained or reduced, for example, to identify a minimum effective dose. In some embodiments, treatment is maintained at a minimum effective dose.

[0020] In another aspect, the present invention provides a method for treating IL-4R deficiency, for example, after administration of an IL-4R antagonist to a subject. The present invention features a method for monitoring a subject's response to treatment with an IL-4R antagonist by obtaining information regarding the expression levels of one or both of biomarkers, e.g., TARC or eotaxin-3, or the total serum level of IgE, in a biological sample from the subject, wherein the subject has moderate to severe asthma; and providing instructions that treatment should be continued if the expression levels of the biomarkers are decreased compared to the levels before treatment with the IL-4R antagonist. In one embodiment, the biomarker is FeNO, and if it is determined that the FeNO level is decreased after administration of the antibody, instructions are provided to continue treatment with the IL-4R antagonist.

[0021] The present invention also includes the IL-4R antagonists disclosed herein for use in the manufacture of a medicament for the treatment and / or prevention of asthma (e.g., eosinophilic asthma, moderate to severe eosinophilic asthma, etc.), or for treating any of the other indications or conditions disclosed herein.

[0022] The present invention also includes the IL-4R antagonists disclosed herein for use in the treatment and / or prevention of asthma (e.g., eosinophilic asthma, moderate to severe eosinophilic asthma, etc.), or for the treatment and / or prevention of other indications or conditions disclosed herein.

[0023] The present invention includes pharmaceutical compositions comprising anti-IL4R antibody antagonists or antigen-binding fragments thereof for use in the treatment and / or prevention of asthma and related conditions.

[0024] The present invention also includes pharmaceutical compositions comprising an anti-IL4R antibody antagonist or an antigen-binding fragment thereof for use in reducing the incidence of one or more asthma exacerbations in a subject in need thereof.

[0025] Additionally, the present invention includes pharmaceutical compositions comprising an anti-IL4R antibody antagonist or an antigen-binding fragment thereof for use in improving one or more asthma-related parameters in a subject in need thereof.

[0026] The present invention includes pharmaceutical compositions comprising an anti-IL4R antibody antagonist or an antigen-binding fragment thereof for use in the treatment of asthma and related conditions in patients with elevated levels of a biomarker selected from the group consisting of thymus and activation-regulated chemokine (TARC), IgE, eotaxin-3, periostin, carcinoembryonic antigen (CEA), YKL-40, and exhaled nitric oxide (FeNO).

[0027] The present invention further includes a pharmaceutical composition comprising an anti-IL4R antibody antagonist or an antigen-binding fragment thereof for use in treating asthma or moderate to severe eosinophilic asthma in a subject in need thereof, wherein the treatment comprises testing the patient for the presence of a blood eosinophil level of at least 300 cells per microliter and / or a sputum eosinophil level of at least 3%, and initiating / continuing administration of the pharmaceutical composition if said blood eosinophil level and / or sputum eosinophil level are found.

[0028] Other embodiments of the present invention will become apparent from consideration of the following detailed description. [Brief explanation of the drawings]

[0029] [Figure 1] Figure 1 is a graph showing a Kaplan-Meier plot of time to asthma exacerbation in patients treated with placebo (open circles) compared with patients treated with anti-IL-4R antibody mAb1 (asterisks). The effect of treatment with anti-IL-4R antibody mAb1 persists over a period of time (including after 8 weeks) during which patients are at higher risk of developing an exacerbation due to steroid withdrawal. The vertical dashed line indicates LABA withdrawal. [Figure 2]Figure 2 is a graph showing the mean change from baseline in forced expiratory volume in 1 second (FEV1) in patients treated with placebo (open triangles) compared to patients treated with anti-IL-4R antibody mAb1 (filled circles). The vertical dashed line indicates LABA withdrawal. [Figure 3] FIG. 3 is a graph showing the mean change from baseline in morning peak expiratory flow (AM PEF) in liters per minute in patients treated with placebo (open triangles) compared with patients treated with anti-IL-4R antibody mAb1 (filled circles). [Figure 4] FIG. 4 is a graph showing the mean change from baseline in nighttime peak expiratory flow rate (PM PEF) in liters per minute in patients treated with placebo (open triangles) compared to patients treated with anti-IL-4R antibody mAb1 (filled circles). [Figure 5] FIG. 5 is a graph showing the mean change from baseline in albuterol use in inhalations per day in patients treated with placebo (open triangles) compared to patients treated with anti-IL-4R antibody mAb1 (filled circles). [Figure 6] Figure 6 is a graph showing the mean change from baseline in 5-item Asthma Control Questionnaire (ACQ5) scores in patients treated with placebo (open triangles) compared to patients treated with anti-IL-4R antibody mAb 1 (filled circles). The vertical dashed line indicates LABA withdrawal. [Figure 7] 7 is a graph showing the mean change from baseline in the number of nocturnal awakenings per night in patients treated with placebo (open triangles) compared to patients treated with anti-IL-4R antibody mAb 1 (filled circles). The vertical dashed line indicates LABA withdrawal. [Figure 8] 8 is a graph showing the mean percentage change from baseline in TARC by visits at weeks 0, 1, 4, 8, and 12 for the placebo-treated mITT population (filled circles) compared with patients treated with anti-IL-4R antibody mAb1 (filled squares). The vertical dashed line indicates LABA withdrawal. [Figure 9]9 is a graph showing the mean percentage change from baseline in eotaxin-3 by visits at weeks 0, 1, 4, 8, and 12 for the placebo-treated mITT population (filled circles) compared with patients treated with the anti-IL-4R antibody mAb1 (filled squares). The vertical dashed line indicates LABA withdrawal. [Figure 10] 10 is a graph showing the mean percentage change from baseline in total IgE by visits at weeks 0, 1, 4, 8, and 12 in the placebo-treated mITT population (filled circles) compared to patients treated with anti-IL-4R antibody mAb1 (filled squares). The vertical dashed line indicates LABA withdrawal. [Figure 11] FIG. 11 is a graph showing the mean percentage change from baseline in periostin by visits at weeks 0, 1, 4, 8, and 12 in mITT patients treated with placebo (filled circles) compared with patients treated with anti-IL-4R antibody mAb1 (filled squares). [Figure 12] FIG. 12 is a graph showing the mean percentage change from baseline in carcinoembryonic antigen (CEA) by visits at weeks 0, 1, 4, 8, and 12 in the placebo-treated mITT population (filled circles) compared with patients treated with anti-IL-4R antibody mAb1 (filled squares). [Figure 13] Figure 13 is a graph showing the mean percentage change from baseline in YKL-40 by visits at weeks 0, 1, 4, 8, and 12 in the placebo-treated mITT population (filled circles) compared with patients treated with the anti-IL-4R antibody mAb1 (filled squares). [Figure 14] FIG. 14 is a graph showing the mean percentage change from baseline in blood eosinophils by visits at weeks 0, 1, 2, 4, 6, 8, and 12 in the placebo-treated mITT population (filled circles) compared to patients treated with anti-IL-4R antibody mAb1 (filled squares). [Figure 15]15 is a graph showing the mean percentage change from baseline in exhaled nitric oxide (NO) levels by visits at weeks 0, 4, 8, and 12 in the placebo-treated mITT population (filled circles) compared to patients treated with anti-IL-4R antibody mAb1 (filled squares). The vertical dashed line indicates LABA withdrawal. [Figure 16] FIG. 16 is a scatter plot of the change from baseline in FEV1(L) of the ratio of exhaled nitric oxide (FeNO) (PPB) to baseline at week 12 in the placebo-treated mITT population (open circles and full line) compared with patients treated with the anti-IL-4R antibody mAb1 (plus signs and dotted line). [Figure 17] FIG. 17 is a scatter plot of AM-PEF (L / min) change from baseline in FeNO (PPB) at week 12 relative to baseline in the placebo-treated mITT population (open circles and solid line) compared with patients treated with anti-IL-4R antibody mAb1 (plus signs and dotted line). [Figure 18] FIG. 18 is a scatter plot of change from baseline in PM-PEF (L / min) at week 12 relative to baseline FeNO (PPB) in the placebo-treated mITT population (open circles and solid line) compared with patients treated with anti-IL-4R antibody mAb1 (plus signs and dotted line). [Figure 19] Figure 19 is a scatter plot of change in FEV1 from baseline at week 12 (L) versus blood eosinophil count (GIGA / L) in the placebo-treated mITT population (open circles and solid line) compared with patients treated with anti-IL-4R antibody mAb1 (plus signs and dotted line). [Figure 20] Figure 20 is a scatter plot of the change from baseline in ACQ at week 12 versus blood eosinophil count (GIGA / L) in the placebo-treated mITT population (open circles and solid line) compared with patients treated with the anti-IL-4R antibody mAb1 (plus signs and dotted line). [Figure 21]FIG. 21 is a scatter plot of the change from baseline in daily albuterol / levalbuterol use at week 12 in relation to blood eosinophil count (GIGA / L) in the placebo-treated mITT population (open circles and solid line) compared with patients treated with anti-IL-4R antibody mAb1 (plus signs and dotted line). [Figure 22] Figure 22 is a scatter plot of the change from baseline ACQ at week 12 relative to baseline periostin in the placebo-treated mITT population (open circles and solid line) compared with patients treated with anti-IL-4R antibody mAb1 (plus signs and dotted line). [Figure 23] Figure 23 is a scatter plot of the change from baseline in ACQ for YKL-40 at week 12 in the placebo-treated mITT population (open circles and solid line) compared with patients treated with the anti-IL-4R antibody mAb1 (plus signs and dotted line). [Figure 24] FIG. 24 is a schematic diagram of timing and dosing regimens for treatment of asthma patients. [Figure 25] FIG. 25 is a schematic diagram describing the patient demographics of a randomized, placebo-controlled, double-blind, parallel-group study of either 300 mg mAb1 or placebo administered subcutaneously once weekly for 12 weeks in patients with persistent moderate to severe eosinophilic asthma that was partially controlled / uncontrolled with inhaled corticosteroid (ICS) and long-acting beta-2 agonist (LABA) treatment. [Figure 26] 26A and 26B are scatter plots of morning (A) and evening (B) asthma symptoms measured over 12 weeks following administration of placebo (open triangles) or mAb1 (filled circles). [Figure 27] 27 is a graph showing serum IgE levels in humanized IL-4 / IL-4R mice (IL-4hu / hu IL-4Rαhu / hu) after house dust mite (HDM) exposure and treatment with either anti-IL-4R antibody or IL-13Ra2-Fc decoy receptor molecule, or mock treatment. Measurements were performed on samples taken on day 40 (24 hours before the first dose of treatment) and at the end of the experiment on day 85. [Figure 28] Figure 28 is a graph showing serum IgE levels in wild-type (Balb / c) mice after house dust mite (HDM) exposure and treatment with either an isotype control, an anti-IL-4R antibody, or an IL-13Ra2-Fc decoy receptor molecule, or sham treatment. [Figure 29] FIG. 29 is a graph showing the collagen content (expressed in μg / lobe) of lungs of humanized IL-4 / IL-4R mice after HDM exposure and the indicated treatments. [Figure 30] FIG. 30 is a graph showing lung collagen content (expressed in μg / lobe) of wild-type mice following HDM exposure and the indicated treatments. [Figure 31] FIG. 31A is a graph showing the levels of eosinophils and neutrophils in humanized IL-4 / IL-4R mice after HDM exposure and the indicated treatments, and FIG. 31B is a graph showing the levels of resident dendritic cells and inflammatory dendritic cells in humanized IL-4 / IL-4R mice after HDM exposure and the indicated treatments. DETAILED DESCRIPTION OF THE INVENTION

[0030] Detailed Description Before describing the present invention, it is to be understood that this invention is not limited to the particular methods and experimental conditions described, as such methods and conditions may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope of the present invention will be limited only by the appended claims.

[0031] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0032] As used herein, the term "about" when used in reference to a particular stated numerical value means that the numerical value may vary by no more than 1% from the stated value. For example, as used herein, the expression "about 100" includes 99 and 101 and all values ​​therebetween (e.g., 99.1, 99.2, 99.3, 99.4, etc.).

[0033] As used herein, the terms "treat," "treating," and the like mean to alleviate the symptoms, eliminate the cause of the symptoms, either temporarily or permanently, or prevent or delay the appearance of the symptoms of the named disorder or condition.

[0034] Although any methods and materials similar or equivalent to those described herein can be used in the practice of the present invention, the preferred methods and materials are now described. All publications mentioned herein are incorporated by reference in their entirety.

[0035] Methods for reducing the incidence of asthma exacerbations The present invention includes a method for reducing the incidence of asthma exacerbations in a subject in need thereof, the method comprising administering to the subject a pharmaceutical composition comprising an interleukin-4 receptor (IL-4R) antagonist. As used herein, the phrase "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 deterioration in a subject's respiratory health that requires and / or is treatable by asthma intervention (e.g., steroid treatment, inhaled corticosteroid treatment, hospitalization, etc.). According to certain embodiments of the present invention, an asthma exacerbation is defined as one or more of the following: (a) a 30% or greater decrease from baseline in morning peak expiratory flow ("AM PEF", as defined elsewhere herein) for two consecutive days; (b) six or more additional exacerbation medication puffs of albuterol or levalbuterol (compared to baseline) in a 24-hour period for two consecutive days; and (c) (i) systemic (oral and / or parenteral) steroid treatment, or (ii) an increase in inhaled corticosteroids to at least four times baseline levels. or (iii) a worsening of asthma requiring hospitalization (e.g., as determined by a physician or other medical practitioner).

[0036] In certain instances, an asthma exacerbation can be classified as a "severe asthma exacerbation." A "severe asthma exacerbation" means an event that requires immediate intervention in the form of treatment with either systemic corticosteroids or inhaled corticosteroids at four or more times the dose taken prior to the event. Thus, the general expression "asthma exacerbation" includes and encompasses the more specific subcategory of "severe asthma exacerbation." Accordingly, the present invention includes methods for reducing the incidence of severe asthma exacerbations in patients in need thereof.

[0037] A "reduced incidence" of asthma exacerbations means that a subject administered a pharmaceutical composition of the invention experiences fewer asthma exacerbations (i.e., at least one less exacerbation) after treatment than before treatment, or does not experience an asthma exacerbation for at least four weeks (e.g., 4, 6, 8, 12, 14 or more weeks) after initiation of treatment with a pharmaceutical composition of the invention. Alternatively, a "reduced incidence" of asthma exacerbations means that the likelihood that a subject will experience an asthma exacerbation after administration of a composition of the invention is reduced by at least 10% (e.g., 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50% or more) compared to a subject not administered a pharmaceutical composition of the invention.

[0038] Methods for improving asthma-related parameters The present invention also includes a method for improving one or more asthma-related parameters in a subject in need thereof, comprising administering to the subject a pharmaceutical composition comprising an interleukin-4 receptor (IL-4R) antagonist. For purposes of the present invention, a reduction in the incidence of asthma exacerbations (as described above) may correlate with an improvement in one or more asthma-related parameters; however, such a correlation may not be observed in all cases.

[0039] Examples of "asthma-related parameters" include: (a) forced expiratory volume in 1 second (FEV1); (b) peak expiratory flow (PEF), including morning PEF (AM PEF) and evening PEF (PM PEF); (c) use of inhaled bronchodilators, such as albuterol or levalbuterol; (d) 5-item Asthma Control Questionnaire (ACQ5) score; (d) nighttime awakenings; and (e) 22-item Sinus Nose Evaluation Test (SNOT-22) score. "Improvement of asthma-related parameters" refers to an increase from baseline in one or more of FEV1, AM PEF, or PM PEF, and / or a decrease from baseline in one or more daily albuterol / levalbuterol use, ACQ5 score, mean nighttime awakenings, or SNOT-22 score. As used herein, the term "baseline" with respect to an asthma-related parameter refers to the value of the asthma-related parameter for a patient before or at the time of administration of a pharmaceutical composition of the invention.

[0040] To determine whether an asthma-related parameter is "improved," the parameter is quantified at baseline and at a time point after administration of the pharmaceutical composition of the present invention. For example, the asthma-related parameter can be measured 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, or 14 days after initial treatment with the pharmaceutical composition of the present invention, or 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, or more. The difference between the value of the parameter at a particular time point after the start of treatment and the value of the parameter at baseline is used to establish whether there has been an "improvement" (e.g., an increase or decrease, as the case may be, depending on the specific parameter being measured) of the asthma-related parameter.

[0041] As used herein, the terms "acquire" or "acquiring" refer to possession of a physical entity or to obtaining a value, such as a numerical value, by "directly obtaining" or "indirectly obtaining" the physical entity or value, such as an asthma-related parameter. "Directly obtaining" means performing a process (e.g., performing a synthetic or analytical method) to obtain the physical entity or value. "Indirectly obtaining" refers to receiving a physical entity or value from another party or source (e.g., a third-party laboratory that directly obtained the physical entity or value). Directly obtaining a physical entity includes performing a process that involves a physical change in a physical substance, e.g., a starting material. Exemplary changes include creating a physical entity from two or more starting materials, shearing or fragmenting a material, separating or purifying a material, combining two or more separate entities into a mixture, and performing a chemical reaction that involves breaking or forming a covalent or non-covalent bond. Obtaining a value directly includes performing a process that involves a physical change in a sample or another substance, for example, performing an analytical process (sometimes referred to herein as a "physical analysis") that involves a physical change in a substance, such as a sample, analyte, or reagent.

[0042] Indirectly obtained information may be provided in the form of a report obtained, for example, in paper or electronic form, such as from an online database or application ("App"). This report or information may be provided, for example, by a health care organization, such as a hospital or clinic; or by a health care provider, such as a doctor or nurse.

[0043] Forced expiratory volume in 1 second (FEV1)According to certain embodiments of the present invention, administration of an IL-4R antagonist to a patient results in an increase from baseline in forced expiratory volume in 1 second (FEV1). Methods for measuring FEV1 are known in the art. For example, a spirometer conforming to the 2005 American Thoracic Society (ATS) / European Respiratory Society (ERS) recommendations can be used to measure FEV1 in a patient. The ATS / ERS Standardization of Spirometry can be used as a guideline. Spirometry is typically performed between 6 and 10 AM after at least 6 hours of albuterol withdrawal. Pulmonary function testing is typically performed in the seated position, and the highest reading is recorded as FEV1 (liters).

[0044] The present invention includes methods of treatment that result in an increase in FEV1 from baseline of at least 0.05 L at 12 weeks after initiation of treatment with a pharmaceutical composition comprising an anti-IL-4R antagonist. For example, according to the present invention, administration of an IL-4R antagonist to a subject in need thereof results in an increase in FEV1 from baseline of about 0.05 L, 0.10 L, 0.12 L, 0.14 L, 0.16 L, 0.18 L, 0.20 L, 0.22 L, 0.24 L, 0.26 L, 0.28 L, 0.30 L, 0.32 L, 0.34 L, 0.36 L, 0.38 L, 0.40 L, 0.42 L, 0.44 L, 0.46 L, 0.48 L, 0.50 L, or more at 12 weeks.

[0045] Morning and evening peak expiratory flow (AM PEF and PM PEF)According to certain embodiments of the present invention, administration of an IL-4R antagonist to a patient results in an increase from baseline in morning (AM) and / or evening (PM) peak 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, a patient is provided with an electronic PEF meter to read 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 rescue medication). The patient is instructed on the use of the device, and written instructions for use of the electronic PEF meter are provided to the patient. Additionally, a medical professional may instruct the patient on how to record the relevant variables on the electronic PEF meter. The subject may be instructed to take an AM PEF (Between 6 AM and 10 AM) within 15 minutes of waking up, typically before taking any albuterol. A PM PEF (Between 6 PM and 10 PM) is typically taken in the evening, typically before taking any albuterol. Subjects should attempt to withhold albuterol for at least 6 hours before measuring their PEF. Three PEF attempts are performed by the patient, and all three values ​​are recorded by an electronic PEF meter. Typically, the highest value is used for evaluation. A baseline AM PEF can be calculated as the average AM measurement recorded during the 7 days prior to administration of the first dose of a pharmaceutical composition containing an IL-4R antagonist, and a baseline PM PEF can be calculated as the average PM measurement recorded during the 7 days prior to administration of the first dose of a pharmaceutical composition containing an IL-4R antagonist.

[0046] The present invention includes methods of treatment that result in an increase in AM PEF and / or PM PEF from baseline of at least 1.0 L / min 12 weeks after initiation of treatment with a pharmaceutical composition comprising an anti-IL-4R antagonist. For example, according to the present invention, administration of an IL-4R antagonist to a subject in need thereof results in an increase in PEF from baseline at 12 weeks of about 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, 20 L / min, or more.

[0047] Albuterol / Levalbuterol Uses According to certain embodiments of the present invention, administration of an IL-4R antagonist to a patient results in a reduction from baseline in daily albuterol or levalbuterol use. The number of albuterol / levalbuterol inhalations can be recorded daily by the patient using a diary, PEF meter, or other recording device. During treatment with the pharmaceutical composition of the present invention, albuterol / levalbuterol use can be non-regular or non-prophylactic, typically as needed for symptoms. The baseline number of albuterol / levalbuterol inhalations / day can be calculated based on an average over the seven days prior to administration of the first dose of a pharmaceutical composition comprising an IL-4R antagonist.

[0048] The present invention encompasses methods of treatment that result in a reduction in albuterol / levalbuterol use from baseline of at least 0.25 puffs per day at 12 weeks after initiation of treatment with a pharmaceutical composition comprising an anti-IL-4R antagonist. For example, according to the present invention, administration of an IL-4R antagonist to a subject in need thereof results in a reduction in albuterol / levalbuterol use from baseline of about 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, 3.00 puffs per day, or more at 12 weeks.

[0049] Five-item Asthma Control Questionnaire (ACQ) score According to certain embodiments of the invention, administration of an IL-4R antagonist to a patient results in a reduction from baseline in the 5-item Asthma Control Questionnaire (ACQ5) score. The ACQ5 is a validated questionnaire for assessing asthma control.

[0050] The present invention encompasses methods of treatment that result in a reduction in ACQ5 score from baseline of at least 0.10 points at 12 weeks after initiation of treatment with a pharmaceutical composition comprising an anti-IL-4R antagonist. For example, according to the present invention, administration of an IL-4R antagonist to a subject in need thereof results ..., 0.15 points, 0.20 points, 0.25 points, 0.30 points, 0.35 points, 0.40 points, 0.45 points, or 0.50 points at 12 weeks. , 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 reduction in ACQ score from baseline.

[0051] Nighttime awakenings According to certain embodiments of the present invention, administration of an IL-4R antagonist to a patient results in a reduction from baseline in the mean number of nighttime awakenings.

[0052] The present invention includes methods of treatment that result in a reduction in the mean number of nighttime awakenings from baseline of at least about 0.10 per night at 12 weeks after initiation of treatment with a pharmaceutical composition comprising an anti-IL-4R antagonist. For example, according to the present invention, administration of an IL-4R antagonist to a subject in need thereof results in a reduction in the mean number of nighttime awakenings from baseline of approximately 0.10 per night, 0.15 per night, 0.20 per night, 0.25 per night, 0.30 per night, 0.35 per night, 0.40 per night, 0.45 per night, 0.50 per night, 0.55 per night, 0.60 per night, 0.65 per night, 0.70 per night, 0.75 per night, 0.80 per night, 0.85 per night, 0.90 per night, 0.95 per night, 1.0 per night, 2.0 per night, or more at 12 weeks.

[0053] 22-item Sinus Nose Evaluation Test (SNOT-22) score According to certain embodiments of the present invention, administration of an IL-4R antagonist to a patient results in a reduction from baseline in the 22-item Sinus Nose Evaluation Test (SNOT-22), a validated questionnaire for assessing the impact of chronic sinusitis on quality of life (Hopkins et al. 2009, Clin. Otolaryngol. 34:447-454).

[0054] The present invention includes methods of treatment that result in a decrease in SNOT-22 score from baseline of at least 1 point at week 12 after initiation of treatment with a pharmaceutical composition comprising an anti-IL-4R antagonist. For example, according to the present invention, administration of an IL-4R antagonist to a subject in need thereof results in a decrease in SNOT-22 score from baseline of about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or more points at week 12.

[0055] Methods for Treating Asthma According to certain embodiments, the present invention provides a method for treating asthma (including, for example, eosinophilic asthma) in a subject in need thereof, wherein the method comprises administering to the subject a pharmaceutical composition comprising an interleukin-4 receptor (IL-4R) antagonist. In certain embodiments, the method of the present invention is useful for treating moderate to severe eosinophilic asthma (e.g., persistent moderate to severe eosinophilic asthma) in a subject.

[0056] According to the present invention, a subject is identified as having moderate to severe eosinophilic asthma if the subject exhibits a blood eosinophil level of at least 300 cells per microliter and / or a sputum eosinophil level of at least 3%. Any method known and available in the art for measuring blood and / or sputum eosinophil levels will, in the context of the present invention, identify a subject as having moderate to severe eosinophilic asthma and therefore as a suitable subject for the treatment methods of the present invention.

[0057] According to a related aspect of the invention, there is provided a method for treating asthma comprising: (a) selecting a patient exhibiting a blood eosinophil level of at least 300 cells per microliter and / or a sputum eosinophil level of at least 3%; and (b) administering to the patient a pharmaceutical composition comprising an IL-4R antagonist.

[0058] In another aspect, a method is provided for reducing or eliminating an asthma patient's dependence on inhaled corticosteroids (ICS) and / or long-acting beta-agonists (LABAs) during the treatment of moderate to severe asthma. In certain embodiments, the method comprises: selecting a patient with moderate to severe asthma that is uncontrolled or partially controlled with background asthma therapy; administering a prescribed dose of an IL-4R antagonist, preferably an anti-IL-4R antibody, to the patient during the initial treatment period, while maintaining the patient's background asthma therapy during the initial treatment period; and gradually reducing the dosage of one or more components of the background therapy over a subsequent treatment period while continuing to administer the IL-4R antagonist. "Background therapy" refers to standard or conventional therapeutic agents known in the art used to treat asthma. In certain embodiments, the background therapy includes ICS, LABA, or a combination of both. In some embodiments, the dosage of ICS and / or LABA is eliminated or completely suspended during the initial treatment period. For example, a LABA such as salmeterol or formoterol may be administered during an initial treatment period and then stopped or withdrawn entirely during a subsequent treatment period.

[0059] An example of a treatment regimen for a patient with moderate to severe asthma is shown in Figure 24, in which an IL-4R antagonist is administered to a patient with moderate to severe asthma. During the initial treatment period (also called the "stable period"), a LABA and an ICS are administered to the patient as background therapy. During the subsequent treatment period (also called the "drug-off period"), the administration of the LABA is stopped, i.e., the LABA is discontinued or eliminated. The ICS is gradually reduced over the subsequent treatment period until it is eliminated.

[0060] In a related aspect, methods are provided for treating asthma, including add-on therapy to a background therapy while the systemic background therapy is withdrawn. In certain embodiments, an IL-4R antagonist is administered as add-on therapy to an asthma patient who has been on background therapy for a certain period of time (e.g., 1 week, 2 weeks, 3 weeks, 1 month, 2 months, 5 months, 12 months, 18 months, 24 months, or more) (also referred to as a "stable period"). In some embodiments, the background therapy includes an ICS and / or a LABA. The stable period is followed by a background therapy withdrawal period, in which one or more components of the background therapy are withdrawn, reduced, or eliminated, while the add-on therapy continues. In some embodiments, the background therapy can be reduced by about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, or more during the withdrawal period. The drug holiday can last 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks or more. In a preferred embodiment, background therapy can be reduced by about 5% during the drug holiday, and the drug holiday can last 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks or more. In a preferred embodiment, background therapy can be reduced by about 10% during the drug holiday, and the drug holiday can last 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks or more. In a preferred embodiment, background therapy can be reduced by about 20% during the drug holiday, and the drug holiday can last 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks or more. In a preferred embodiment, background therapy can be reduced by about 30% during the drug holiday, and the drug holiday can last 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks or more.In a preferred embodiment, background therapy can be reduced by about 40% during the drug holiday, and the drug holiday can last 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks or more. In a preferred embodiment, background therapy can be reduced by about 50% or more during the drug holiday, and the drug holiday can last 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks or more. obtain.

[0061] In some other embodiments, the invention encompasses methods of treating or alleviating conditions or complications associated with asthma, such as chronic sinusitis, allergic rhinitis, allergic fungal rhinosinusitis, allergic bronchopulmonary aspergillosis, unified airway disease, Churg-Strauss syndrome, vasculitis, chronic obstructive pulmonary disease (COPD), and exercise-induced bronchospasm.

[0062] The present invention also includes a method for treating persistent asthma. As used herein, the term "persistent asthma" means having symptoms at least once a week during the day and / or at night, and the symptoms last for several hours to several days. In certain alternative embodiments, persistent asthma is "mildly persistent" (e.g., symptoms are severe enough to interfere with daily activities or sleep, occurring more than twice a week but less than daily, and / or lung function is normal or reversible with inhaled bronchodilators), "moderately persistent" (e.g., sleep is disturbed at least weekly, and / or lung function is moderately abnormal, but symptoms occur daily), or "severely persistent" (e.g., symptoms persist despite the correct use of approved medications and / or lung function is severely affected).

[0063] Interleukin-4 receptor antagonist The methods of the present invention involve administering to a subject in need thereof a therapeutic composition comprising an interleukin-4 receptor (IL-4R) antagonist. As used herein, an "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 the IL-4R is expressed on a cell in vitro or in vivo. Non-limiting examples of categories of IL-4R antagonists include small molecule IL-4R antagonists, anti-IL-4R aptamers, peptide-based IL-4R antagonists (e.g., "peptibody" molecules), and antibodies or antigen-binding fragments of antibodies that specifically bind to human IL-4R.

[0064] The term "human IL4R" (hIL-4R) refers to a human cytokine receptor that specifically binds interleukin-4 (IL-4), such as IL-4Rα (SEQ ID NO: 274).

[0065] The term "antibody" refers to an immunoglobulin molecule, as well as multimers thereof (e.g., IgM), which comprise four polypeptide chains, two heavy (H) chains and two light (L) chains, interconnected by disulfide bonds. Each heavy chain contains a heavy chain variable region (herein referred to as HCVR or V H Abbreviated as The heavy chain constant region is made up of three domains: C H 1. C H 2, and C H Each light chain comprises a light chain variable region (referred to herein as LCVR or V L The light chain constant region comprises one domain (C L 1) V H and V L region can be further subdivided into regions of hypervariability called complementarity-determining regions (CDRs) that are interspersed with more conserved regions called framework regions (FRs). H and V Lis composed of three CDRs and four FRs arranged in the following order from amino terminus to carboxy terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. In different embodiments, the FRs of an anti-IL-4R antibody (or antigen-binding fragment thereof) may be identical to human germline sequences or may be naturally or artificially modified. An amino acid consensus sequence may be defined based on side-by-side analysis of two or more CDRs.

[0066] The term "antibody" also includes antigen-binding fragments of whole antibody molecules. As used herein, the terms "antigen-binding portion" of an antibody, "antigen-binding fragment" of an antibody, etc. refer to an antibody that is specific for an antigen. Antigen-binding fragments include any naturally occurring, enzymatically obtainable, synthetic, or genetically engineered polypeptide or glycoprotein that selectively binds to form a complex. Antigen-binding fragments of antibodies can be derived, for example, from intact antibody molecules using any suitable standard techniques, such as proteolytic digestion or recombinant genetic engineering techniques, including the manipulation and expression of DNA encoding the variable and optionally constant domains of the antibody. Such DNA is known and / or readily available, for example, from commercial sources, DNA libraries (including, for example, phage-antibody libraries), or can be synthesized. The DNA can be sequenced and manipulated, for example, chemically or by using molecular biology techniques, to arrange one or more variable and / or constant domains in the appropriate configuration, or to introduce codons, create cysteine ​​residues, modify, add, or delete amino acids, etc.

[0067] Non-limiting examples of antigen-binding fragments include: (i) Fab fragments; (ii) F(ab')2 fragments; (iii) Fd fragments; (iv) Fv fragments; (v) single-chain Fv (scFv) molecules; (vi) dAb fragments; and (vii) minimal recognition units consisting of amino acid residues that mimic the hypervariable regions of an antibody (e.g., isolated complementarity-determining regions (CDRs), e.g., CDR3 peptides), or constrained FR3-CDR3-FR4 peptides). Other engineered molecules, such as domain-specific antibodies, single domain antibodies, domain-deleted antibodies, chimeric antibodies, CDR-grafted antibodies, diabodies, triabodies, tetrabodies, minibodies, nanobodies (e.g., monovalent nanobodies, bivalent nanobodies, etc.), small modular immunopharmaceuticals (SMIPs), and shark variable IgNAR domains are also encompassed within the expression "antigen-binding fragment".

[0068] Antigen-binding fragments of antibodies typically contain at least one variable domain, which may be of any size or amino acid composition and generally contains at least one CDR adjacent to or in-frame with one or more framework sequences. L V associated with the domain H In an antigen-binding fragment having a domain, V H and V L The domains can be positioned relative to each other in any suitable configuration. For example, the variable region can be a dimer, with the V H -V H , V H -V L or V L -V L Alternatively, the antigen-binding fragment of an antibody may comprise a monomeric V H or V L It may include a domain.

[0069] In certain embodiments, an antigen-binding fragment of an antibody may comprise at least one variable domain covalently linked to at least one constant domain. Non-limiting exemplary configurations of variable and constant domains that may be found in an antigen-binding fragment of an antibody of the invention 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;(xii) V L -C H 1-C H 2-C H 3;(xiii)V L -C H 2-C H 3; and (xiv) V L -C LIn 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 may be linked by a full-length or partial hinge or linker region. The hinge region may consist of at least two (e.g., 5, 10, 15, 20, 40, 60 or more) amino acids that create a flexible or semi-flexible linkage between adjacent variable and / or constant domains in a single polypeptide molecule; preferably, the hinge region may consist of between 2 and 60 amino acids, preferably 5 to 50, or preferably 10 to 40 amino acids. Furthermore, antigen-binding fragments of antibodies of the invention may be linked to each other and / or to one or more monomeric V H Or V L The variable domains may comprise homodimers or heterodimers (or other multimers) of any of the above-listed variable and constant domain configurations non-covalently associated (e.g., by disulfide bonds) with the domains.

[0070] Like intact antibody molecules, antigen-binding fragments can be monospecific or multispecific (e.g., bispecific). Multispecific antigen-binding fragments of antibodies typically contain at least two different variable domains, where each variable domain can specifically bind to a separate antigen or a different epitope on the same antigen. Any multispecific antibody format can be adapted for use in the context of the antigen-binding fragments of antibodies of the present invention using routine techniques available in the art.

[0071] The constant region of an antibody is important in the ability of the antibody to fix complement and mediate cell-dependent cytotoxicity. Thus, the antibody isotype can be selected based on whether it is desirable for the antibody to mediate cytotoxicity.

[0072] The term "human antibody" includes antibodies having variable and constant regions derived from human germline immunoglobulin sequences. Nevertheless, human antibodies featured in the invention may include mutations, e.g., in the CDRs, and particularly CDR3, at amino acid residues not encoded by human germline immunoglobulin sequences (e.g., introduced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo). However, the term "human antibody" does not include antibodies in which CDR sequences derived from the germline of another mammalian species, such as a mouse, have been grafted onto human framework sequences.

[0073] The term "recombinant human antibody" refers to any human antibody that is produced, expressed, made, or isolated by recombinant means, such as antibodies expressed using a recombinant expression vector transfected into a host cell (described further below), antibodies isolated from a recombinant combinatorial human antibody library (described further below), antibodies isolated from an animal (e.g., a mouse) transgenic for human immunoglobulin genes (e.g., Taylor & Francis Group, 1999). (See, e.g., J. J. et al. (1992) Nucl. Acids Res. 20:6287-6295), or any other means involving splicing of human immunoglobulin gene sequences into 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 have been subjected to in vitro mutagenesis (or, when animals transgenic for human Ig sequences are used, in vivo somatic mutagenesis), thus altering the V and constant regions of the recombinant antibody. H and V L The amino acid sequence of the region is human germline V H and V L Derived from sequences and human germline V H and V L Although related in sequence, they may not naturally occur within the human antibody germline repertoire in vivo.

[0074] Human antibodies can exist in two forms related to hinge heterogeneity. In one form, the immunoglobulin molecule contains a stable four-chain construct of approximately 150-160 kDa, in which dimers are held together by interchain heavy chain disulfide bonds. In the second form, the dimers are not linked via interchain disulfide bonds, and a molecule of approximately 75-80 kDa is formed, composed of covalently coupled light and heavy chains (half antibodies). These forms have been very difficult to separate, even after affinity purification.

[0075] The frequency of occurrence of the second form in various intact IgG isotypes is due to, but not limited to, structural differences associated with antibody hinge region isotypes. A single amino acid substitution in the hinge region of a human IgG4 hinge can significantly reduce the occurrence of the second form to levels typically observed using a human IgG1 hinge (Angal et al. al. (1993) Molecular Immunology 30:105). The present invention provides a hinge, C H 2 or C H Antibodies with one or more mutations in three regions are included, which may be desirable in manufacturing, for example, to improve yield of the desired antibody form.

[0076] An "isolated antibody" means an antibody that has been identified and separated and / or recovered from at least one component of its natural environment. For example, an antibody that has been separated or removed from at least one component of an organism, or from a tissue or cell in which it naturally occurs or is naturally produced, is an "isolated antibody" for purposes of the present invention. An isolated antibody also includes an antibody in situ within a recombinant cell. An isolated antibody is an antibody that has been subjected to 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.

[0077] The term "specifically binds," or the like, means that an antibody or antigen-binding fragment thereof 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, surface plasmon resonance, and the like. For example, an antibody that "specifically binds" to IL-4R as used in the context of the present invention is one that binds to IL-4R or a portion thereof with a K of less than about 1000 nM, less than about 500 nM, less than about 300 nM, less than about 200 nM, less than about 100 nM, less than about 90 nM, less than about 80 nM, less than about 70 nM, less than about 60 nM, less than about 50 nM, less than about 40 nM, less than about 30 nM, less than about 20 nM, less than about 10 nM, less than about 5 nM, less than about 4 nM, less than about 3 nM, less than about 2 nM, less than about 1 nM, or less than about 0.5 nM, as measured in a surface plasmon resonance assay. D However, it does not contain antibodies that bind to human IL-4. An isolated antibody that specifically binds IL-4R may have cross-reactivity to other antigens, such as IL-4R molecules from other (non-human) species.

[0078] Anti-IL-4R antibodies useful for the methods of the present invention 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 and / or CDR regions of the heavy and light chain variable domains compared to the corresponding germline sequences from which the antibody is derived. Such mutations can be readily ascertained by comparing the amino acid sequences disclosed herein to germline sequences available, for example, from public antibody sequence databases. The present invention includes methods involving the use of antibodies and antigen-binding fragments thereof derived from any of the amino acid sequences disclosed herein, wherein 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 one or more framework and / or CDR regions (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acids) are mutated to the corresponding residue in the germline sequence from which the antibody was derived, or to the corresponding residue in another human germline sequence, or to a conservative amino acid substitution of the corresponding germline residue (such sequence changes are collectively referred to herein as "germline mutations"). Starting with the heavy and light chain variable region sequences disclosed herein, one of skill in the art can readily produce numerous antibodies and antigen-binding fragments containing one or more individual germline mutations or combinations thereof. In certain embodiments, V H and / or V LAll framework and / or CDR residues within a domain are backmutated to residues found in the original germline sequence from which the antibody was derived. In other embodiments, only specific residues are backmutated to the original germline sequence, e.g., only mutated residues found within the first eight amino acids of FR1 or the last eight residues of FR4, or only mutated residues found in CDR1, CDR2, or CDR3. In other embodiments, one or more of the framework and / or CDR residues are backmutated to a different germline sequence (i.e., different from the germline sequence from which the antibody was originally derived). In addition, antibodies of the present invention may contain any combination of two or more germline mutations within the framework and / or CDR regions, e.g., where certain individual residues are mutated to the corresponding residue in a particular germline sequence, while certain other residues that differ from the original germline sequence are either maintained or mutated to the corresponding residue in a different germline sequence. Once obtained, 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, improved or enhanced antagonist or agonist biological properties (as the case may be), reduced immunogenicity, etc. Uses of antibodies and antigen-binding fragments obtained in this general manner are encompassed within the present invention.

[0079] The present invention also includes methods that involve the use of anti-IL-4R antibodies that contain variants of any 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 anti-IL-4R antibodies that have HCVR, LCVR, and / or CDR amino acid sequences that contain, for example, 10 or fewer, 8 or fewer, 6 or fewer, 4 or fewer, etc., conservative amino acid substitutions compared to any of the HCVR, LCVR, and / or CDR amino acid sequences disclosed herein.

[0080] The term "surface plasmon resonance" is used in, for example, BIAcore TMThis refers to an optical phenomenon that allows for the analysis of real-time interactions by detecting changes in protein concentration within a biosensor matrix using a system (Biacore Life Sciences division of GE Healthcare, Piscataway, NJ).

[0081] The term “K D " refers to the equilibrium dissociation constant of a particular antibody-antigen interaction.

[0082] 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 the paratope. A single antigen can have more than one epitope. Thus, different antibodies may bind to different regions of an antigen and have different biological effects. Epitopes can be either conformational or linear. Conformational epitopes are formed by spatially adjacent amino acids from different segments of a linear polypeptide chain. Linear epitopes are formed by adjacent amino acid residues in a polypeptide chain. In certain circumstances, epitopes can include saccharide, phosphoryl, or sulfonyl moieties on the antigen.

[0083] Human antibody production Methods for generating human antibodies in transgenic mice are known in the art. Any such method can be used in the context of the present invention to generate human antibodies that specifically bind to human IL-4R.

[0084] Velocimmune TM Using techniques (see, e.g., U.S. Pat. No. 6,596,541, Regeneron Pharmaceuticals) or any other known method for generating monoclonal antibodies, a high affinity chimeric antibody against IL-4R having a human variable region and a mouse constant region is first isolated. (R) The technique involves inducing mice to produce antibodies containing human variable regions and mouse constant regions in response to antigen stimulation. The present invention involves the generation of transgenic mice having genomes containing human heavy and light chain variable regions operably linked to endogenous mouse constant region gene loci, such that the DNA encoding the heavy and light chain variable regions of the antibody is isolated and operably linked to DNA encoding human heavy and light chain constant regions, and the DNA is then expressed in cells capable of expressing fully human antibodies.

[0085] In general, VELOCIMMUNE (R) Mice are exposed to the antigen of interest and develop antibodies. Lymphatic cells (e.g., B cells) that express the antibody are harvested from the mouse. The lymphatic cells may be fused with a myeloma cell line to produce an immortal hybridoma cell line, and such hybridoma cell lines are screened and selected to identify hybridoma cell lines that produce antibodies specific to the antigen of interest. DNA encoding the heavy and light chain variable regions may be isolated and linked to the desired isotype constant regions of the heavy and light chains. Such antibody proteins may be produced in cells such as CHO cells. Alternatively, DNA encoding the antigen-specific chimeric antibody or the light and heavy chain variable domains may be isolated directly from the antigen-specific lymphocytes.

[0086] 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 desired characteristics, including affinity, selectivity, epitope, etc., using standard procedures known to those skilled in the art. The mouse constant region is replaced with a desired human constant region to generate the fully human antibody featured in the present invention, e.g., wild-type or modified IgG1 or IgG4. While the constant region selected can vary depending on the specific application, the characteristics of high-affinity antigen binding and target specificity reside in the variable region.

[0087] Generally, antibodies that can be used in the methods of the present invention have high affinity, as described above, when measured by binding to antigen either immobilized on a solid phase or in solution phase. The mouse constant regions are replaced with human constant regions as desired to generate the fully human antibodies featured in the present invention. While the constant region selected can vary depending on the particular application, the characteristics of high affinity antigen binding and target specificity reside in the variable regions.

[0088] Specific examples of antibodies or antigen-binding fragments of antibodies that specifically bind to IL-4R that can be used in the context of the methods of the present invention include any antibody or antigen-binding fragment that comprises three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) contained within a heavy chain variable region (HCVR) having an amino acid sequence selected from the group consisting of SEQ ID NOs: 2, 18, 22, 26, 42, 46, 50, 66, 70, 74, 90, 94, 98, 114, 118, 122, 138, 142, 146, 162, 166, 170, 186, 190, 194, 210, 214, 218, 234, 238, 242, 258, and 262. The antibody or antigen-binding fragment may comprise three light chain CDRs (LCVR1, LCVR2, LCVR3) contained within a light chain variable region (LCVR) having an amino acid sequence selected from the group consisting of SEQ ID NOs: 10, 20, 24, 34, 44, 48, 58, 68, 72, 82, 92, 96, 106, 116, 120, 130, 140, 144, 154, 164, 168, 178, 188, 192, 202, 212, 216, 226, 236, 240, 250, 260, and 264. Methods and techniques for identifying CDRs within HCVR and LCVR amino acid sequences are well known in the art and may be used to identify CDRs within particular HCVR and / or LCVR amino acid sequences disclosed herein. Exemplary conventions used to identify CDR boundaries include, for example, the Kabat definition, the Chothia definition, and the AbM definition. Roughly speaking, the Kabat definition is based on sequence variability, 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, e.g., 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.

[0089] In certain embodiments of the invention, the antibody or antigen-binding fragment thereof is selected from the group consisting of SEQ ID NOs: 2 / 10, 18 / 20, 22 / 24, 26 / 34, 42 / 44, 46 / 48, 50 / 58, 66 / 68, 70 / 72, 74 / 82, 90 / 92, 94 / 96, 98 / 106, 114 / 116, 118 / 120, 122 / 130, 138 / 140, 142 / 144, 146 / 154, 162 / 164, 166 / 168, 170 / 172, 174 / 176, 178 / 179, 180 / 181, 182 / 183, 184 / 185, 186 / 187, 188 / 189, 189 / 190, 191 / 192, 192 / 193, 193 / 194, 194 / 195, 195 / 196, 196 / 197, 197 / 198, 198 / 200, 200 / 201, 201 / 202, 201 / 203, 201 / 204, 201 / 205, 201 / 206, 201 / 207, 201 / 208, 202 / 203, 202 / 204, 202 / 205, 203 / 204, 204 / 205, 205 / 206, 206 / 207, 208 / 209, 210 / 211, 211 / 212, 212 / It comprises six CDRs (HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3) from heavy chain and light chain variable region amino acid sequence pairs (HCVR / LCVR) selected from the group consisting of 178, 186 / 188, 190 / 192, 194 / 202, 210 / 212, 214 / 216, 218 / 226, 234 / 236, 238 / 240, 242 / 250, 258 / 260, and 262 / 264.

[0090] In certain embodiments of the invention, the antibody or antigen-binding fragment thereof is selected from the group consisting of SEQ ID NOs: 4 / 6 / 8 / 12 / 14 / 16; 28 / 30 / 32 / 36 / 38 / 40; 52 / 54 / 56 / 60 / 62 / 64; 76 / 78 / 80 / 84 / 86 / 88; 100 / 102 / 104 / 108 / 110 / 112; 124 / 126 / 128 / 132 / 134 / 136; 148 / 150 / 152 / 156 / 1 It contains six CDRs (HCDR1 / HCDR2 / HCDR3 / LCDR1 / LCDR2 / LCDR3) having amino acid sequences selected from the group consisting of: 58 / 160; 172 / 174 / 176 / 180 / 182 / 184; 196 / 198 / 200 / 204 / 206 / 208; 220 / 222 / 224 / 228 / 230 / 232; and 244 / 246 / 248 / 252 / 254 / 256.

[0091] In certain embodiments of the invention, the antibody or antigen-binding fragment thereof is selected from the group consisting of SEQ ID NOs: 2 / 10, 18 / 20, 22 / 24, 26 / 34, 42 / 44, 46 / 48, 50 / 58, 66 / 68, 70 / 72, 74 / 82, 90 / 92, 94 / 96, 98 / 106, 114 / 116, 118 / 120, 122 / 130, 138 / 140, 142 / 144 4, 146 / 154, 162 / 164, 166 / 168, 170 / 178, 186 / 188, 190 / 192, 194 / 202, 210 / 212, 214 / 216, 218 / 226, 234 / 236, 238 / 240, 242 / 250, 258 / 260, and 262 / 264.

[0092] Pharmaceutical Composition The present invention includes methods comprising administering an IL-4R antagonist to a patient, wherein the IL-4R antagonist is contained within a pharmaceutical composition. The pharmaceutical compositions featured in the present invention are formulated with suitable carriers, additives, and other agents that provide for appropriate transport, delivery, tolerance, etc. Many suitable formulations can be found in formularies known to all pharmacists: Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, PA. These formulations include, for example, powders, pastes, ointments, jellies, waxes, oils, lipids, lipid (cationic or anionic)-containing vesicles (e.g., LIPOFECTIN). TM ), DNA conjugates, anhydrous absorbent pastes, oil-in-water and water-in-oil emulsions, emulsions carbowax (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 52:238-311.

[0093] The dose of the antibody administered to a patient according to the method of the present invention may vary depending on the age and size of the patient, symptoms, condition, route of administration, etc. The preferred dose is typically calculated according to body weight or body surface area. Depending on the severity of the condition, the frequency and duration of treatment may be adjusted. Effective dosages and administration schedules for pharmaceutical compositions containing anti-IL-4R antibodies can be empirically determined; for example, a patient's progress can be monitored by periodic evaluation, and the dosage can be adjusted accordingly. 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).

[0094] Various delivery systems are known and can be used to administer the pharmaceutical compositions featured in the present invention. For example, liposome encapsulation, microparticles, microcapsules, recombinant cells capable of expressing mutant viruses, and receptor-mediated endocytosis (see, e.g., Wu et al., 1987, J. Biol. Chem. 262:4429-4432). Administration methods include, but are not limited to, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, intratracheal, epidural, and oral routes. The compositions can be administered by any conventional route, such as by infusion or bolus injection, by absorption through epithelial or mucocutaneous linings (e.g., oral mucosa, rectal and intestinal mucosa, etc.), and can be administered together with other biologically active agents.

[0095] The pharmaceutical compositions of the present invention can be delivered subcutaneously or intravenously using a standard needle and syringe. Furthermore, for subcutaneous delivery, pen delivery devices are readily useful for delivering the pharmaceutical compositions of the present invention. Such pen delivery devices can be reusable or disposable. Reusable pen delivery devices generally utilize a replaceable cartridge containing the pharmaceutical composition. Once the pharmaceutical composition in the cartridge has been administered and the cartridge is empty, the empty cartridge can be easily discarded and replaced with a new cartridge containing the pharmaceutical composition. The pen delivery device can then be reused. Disposable pen delivery devices do not have a replaceable cartridge. Rather, the disposable pen device comes pre-filled with the pharmaceutical composition held in a reservoir within the device. Once the reservoir is emptied of the pharmaceutical composition, the entire device is discarded.

[0096] A number of reusable pen-type autoinjector delivery devices have utility in the subcutaneous delivery of the pharmaceutical compositions of the present invention. Examples include, but are not limited to, the AUTOPEN TM (Owen Mumford, Inc., Woodstock, UK), DISETRONIC TM Pen (Disetronic Medical Systems, Bergdorf, Switzerland), HUMALOG MIX 75 / 25 TM Pen, HUMALOG TM Pen, HUMALIN 70 / 30 TM Pen (Eli Lilly and Co., Indianapolis, IN), NOVOPEN TM I, II and III (Novo Nordisk, Copenhagen, Denmark), NOVOPEN JUNIOR TM (Novo Nordisk, Copenhagen, Denmark), BD TM Penn (Becton Dickinson, Franklin Lakes, NJ), OPTIPEN TM , OPTIPEN PRO TM , OPTIPEN STARLET TM, and OPTICLIK TM (sanofi-aventis, Frankfurt, Germany). Examples of disposable pen delivery devices that have utility in the subcutaneous delivery of pharmaceutical compositions of the present invention include, but are not limited to, SOLOSTAR TM Pen (sanofi-aventis), FLEXPEN TM (Novo Nordisk), and KWIKPEN TM (Eli Lilly), SURECLICK TM Autoinjector (Amgen, Thousand Oaks, CA), PENLET TM (Haselmeier, Stuttgart, Germany), EPIPEN (Dey, LP), and HUMIRA TM Penn (Abbott Labs, Abbott Park IL) is one example.

[0097] For direct administration to the paranasal sinuses, the pharmaceutical compositions of the present invention can be administered, for example, via a microcatheter (e.g., an endoscope and a microcatheter), an aerosolizer, or the like. The IL-4R antagonist may be administered using a powder dispenser, a powder dispenser, a nebulizer, or an inhaler. The method comprises administering the IL-4R antagonist in an aerosolized formulation to a subject in need thereof. For example, an aerosolized antibody against IL-4R may be administered to treat asthma in a patient. Aerosolized antibodies may be produced, for example, as described in U.S. Pat. No. 8,178,098, which is incorporated herein in its entirety.

[0098] In certain circumstances, pharmaceutical compositions can be delivered in controlled release systems. In one embodiment, a pump can be used (see Langer, supra; Sefton, 1987, CRC Crit. Ref. Biomed. Eng. 14:201). In another embodiment, a polymeric material can be used; see, e.g., Medical Applications of Controlled Release, Langer and Wise (eds.), 1974, CRC Press, Boca Raton, Florida. In yet another embodiment, a controlled release system can be placed in close proximity to the target of the composition, thus requiring only a fraction of the systemic dose (see, e.g., Goodson, 1984, in Medical Applications of Controlled Release, supra, vol. 2, pp. 115-138). Other controlled release systems are described in Langer, 1990, Science 249:1527-1533.

[0099] Injectable preparations may include dosage forms for intravenous, subcutaneous, intradermal, and intramuscular injections, infusions, and the like. These injectable preparations can be prepared by known methods. For example, injectable preparations can be prepared by dissolving, suspending, or emulsifying the above-mentioned antibody or a salt thereof in a sterile aqueous or oily medium conventionally used for injections. Aqueous media for injection include, for example, physiological saline, isotonic solutions containing glucose and other adjuvants, and the like, which may be used in combination with appropriate solubilizers such as alcohols (e.g., ethanol), polyalcohols (e.g., propylene glycol, polyethylene glycol), nonionic surfactants [e.g., polysorbate 80, HCO-50 (polyoxyethylene (50 mol) adduct of hydrogenated castor oil)], and the like. Oily media include, for example, sesame oil, soybean oil, and the like, which may be used in combination with solubilizers such as benzyl benzoate, benzyl alcohol, and the like. Therefore, injectable preparations are preferably filled into appropriate ampoules.

[0100] Advantageously, the above-mentioned pharmaceutical compositions for oral or parenteral use are prepared into dosage forms with unit doses suitable for the dose of the active ingredient, such as tablets, pills, capsules, injections (ampoules), suppositories, etc.

[0101] Exemplary pharmaceutical compositions comprising anti-IL-4R antibodies that can be used in the context of the present invention are disclosed, for example, in US Patent Application Publication No. 2012 / 0097565.

[0102] Dosage The amount of IL-4R antagonist (e.g., anti-IL-4R antibody) administered to a subject according to the methods of the present invention is generally a therapeutically effective amount. As used herein, the phrase "therapeutically effective amount" refers to an amount of IL-4R antagonist that results in one or more of: (a) a reduction in the incidence of asthma exacerbations; (b) an improvement in one or more asthma-related parameters (as defined elsewhere herein); and / or (c) a detectable improvement in one or more symptoms or signs of an upper airway inflammatory condition. A "therapeutically effective amount" also includes an amount of IL-4R antagonist that inhibits, prevents, reduces, or delays the progression of asthma in a subject.

[0103] In the case of an anti-IL-4R antibody, the therapeutically effective amount is about 0.05 mg to about 600 mg of the anti-IL-4R antibody, for example, about 0.05 mg, about 0.1 mg, about 1.0 mg, about 1.5 mg, about 2.0 mg, about 3.0 mg, about 5.0 mg, about 7.0 mg, about 10 mg, about 20 mg, about 30 mg, about 40 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 110 mg, about 120 mg, about 130 mg, about 140 mg, about 150 mg, about 160 mg, about 170 mg, about 180 mg, about 190 mg, about 200 mg, about 210 mg, about 220 mg, about 230 mg, about 240 mg The dose may be about 250 mg, about 260 mg, about 270 mg, about 280 mg, about 290 mg, about 300 mg, about 310 mg, about 320 mg, about 330 mg, about 340 mg, about 350 mg, about 360 mg, about 370 mg, about 380 mg, about 390 mg, about 400 mg, about 410 mg, about 420 mg, about 430 mg, about 440 mg, about 450 mg, about 460 mg, about 470 mg, about 480 mg, about 490 mg, about 500 mg, about 510 mg, about 520 mg, about 530 mg, about 540 mg, about 550 mg, about 560 mg, about 570 mg, about 580 mg, about 590 mg, or about 600 mg. In certain embodiments, 300 mg of anti-IL-4R antibody is administered.

[0104] The amount of IL-4R antagonist contained within an individual dose can be expressed in milligrams of antibody per kg of patient body weight (i.e., mg / kg). For example, the IL-4R antagonist can be administered to a patient at a dose of about 0.0001 to about 10 mg / kg of patient body weight.

[0105] Combination therapy According to certain embodiments, the methods of the present invention comprise administering to a subject one or more additional therapeutic agents in combination with an IL-4R antagonist. As used herein, the phrase "in combination" means that the additional therapeutic agent is administered before, after, or simultaneously with a pharmaceutical composition comprising an IL-4R antagonist. In some embodiments, the term "in combination" includes sequential or simultaneous administration of an IL-4R antagonist and a second therapeutic agent. The present invention includes methods for treating asthma or related conditions or complications, or reducing at least one exacerbation, comprising administering an IL-4R antagonist in combination with a second therapeutic agent for additive or synergistic activity.

[0106] For example, when administered "before" a pharmaceutical composition comprising an IL-4R antagonist, an additional therapeutic agent may be administered about 72 hours, about 60 hours, about 48 hours, about 36 hours, about 24 hours, about 12 hours, about 10 hours, about 8 hours, about 6 hours, about 4 hours, about 2 hours, about 1 hour, about 30 minutes, about 15 minutes, or about 10 minutes before administration of the pharmaceutical composition comprising an IL-4R antagonist. When administered "after" a pharmaceutical composition comprising an IL-4R antagonist, an additional therapeutic agent may be administered 10 minutes, about 15 minutes, about 30 minutes, about 1 hour, about 2 hours, about 4 hours, about 6 hours, about 8 hours, about 10 hours, about 12 hours, about 24 hours, about 36 hours, about 48 hours, about 60 hours, or about 72 hours after administration of the pharmaceutical composition comprising an IL-4R antagonist. Administration "concurrently with" a pharmaceutical composition comprising an IL-4R antagonist means that the additional therapeutic agent is administered to the subject in a separate dosage form within less than 5 minutes of (before, after, or simultaneously with) administration of the pharmaceutical composition comprising the IL-4R antagonist, or is administered to the subject as a single combined dosage formulation comprising both the additional therapeutic agent and the IL-4R antagonist.

[0107] The additional therapeutic agent may be, for example, another IL-4R antagonist, an IL-1 antagonist (including, for example, the IL-1 antagonists described in U.S. Pat. No. 6,927,044), an IL-6 antagonist, an IL-6R antagonist (including, for example, the anti-IL-6R antibodies described in U.S. Pat. No. 7,582,298), a TNF antagonist, an IL-8 antagonist, an IL-9 antagonist, an IL-17 antagonist, an IL-5 antagonist, an IgE antagonist, a CD48 antagonist, a leukotriene antagonist, or the like. antifungals, NSAIDs, long-acting beta-2 agonists (e.g., salmetin, The IL-4R antagonist may be a long-acting beta-2 agonist or a combination thereof. For example, in certain embodiments, the IL-4R antagonist may be a long-acting beta-2 agonist or a combination thereof. combinations containing steroids (e.g., fluticasone plus salmeterol [e.g., Advair (R) (GlaxoSmithKline)]; or budesonide + formoterol [e.g. For example, Symbicort (R) (Astra Zeneca)]) do.

[0108] Dosage regimen According to certain embodiments of the present invention, multiple doses of an IL-4R antagonist may be administered to a subject over a defined time course. Such methods include sequentially administering multiple doses of an IL-4R antagonist to a subject. As used herein, "sequentially administering" means that each dose of an IL-4R antagonist is administered to a subject at different times, e.g., on different days, separated by a predetermined interval (e.g., hours, days, weeks, or months). The present invention includes methods comprising sequentially administering to a patient a single initial dose of an IL-4R antagonist, followed by one or more secondary doses of the IL-4R antagonist, and optionally followed by one or more tertiary doses of the IL-4R antagonist.

[0109] The present invention includes methods comprising administering a pharmaceutical composition comprising an IL-4R antagonist to a subject at a dosing frequency of approximately 4 times per week, 2 times per week, once per week, once per 2 weeks, once per 3 weeks, once per 4 weeks, once per 5 weeks, once per 6 weeks, once per 8 weeks, once per 12 weeks, or less, so long as a therapeutic response is achieved. In certain embodiments involving the administration of a pharmaceutical composition comprising an anti-IL-4R antibody, a once-weekly dose in an amount of about 75 mg, 150 mg, or 300 mg may be used. In other embodiments involving the administration of a pharmaceutical composition comprising an anti-IL-4R antibody, a once-every-2-week dose in an amount of about 75 mg, 150 mg, or 300 mg may be used. In other embodiments involving the administration of a pharmaceutical composition comprising an anti-IL-4R antibody, a once-every-3-week dose in an amount of about 75 mg, 150 mg, or 300 mg may be used. In other embodiments involving the administration of a pharmaceutical composition comprising an anti-IL-4R antibody, a four-weekly dose of about 75 mg, 150 mg, or 300 mg may be used. In other embodiments involving the administration of a pharmaceutical composition comprising an anti-IL-4R antibody, a five-weekly dose of about 75 mg, 150 mg, or 300 mg may be used. In other embodiments involving the administration of a pharmaceutical composition comprising an anti-IL-4R antibody, a six-weekly dose of about 75 mg, 150 mg, or 300 mg may be used. In other embodiments involving the administration of a pharmaceutical composition comprising an anti-IL-4R antibody, a eight-weekly dose of about 75 mg, 150 mg, or 300 mg may be used. In other embodiments involving the administration of a pharmaceutical composition comprising an anti-IL-4R antibody, a 12-weekly dose of about 75 mg, 150 mg, or 300 mg may be used. The preferred route of administration is subcutaneous.

[0110] The term "week" or "weeks" refers to a period of (n x 7 days) ± 2 days, preferably (n x 7 days) ± 1 day, more preferably (n x 7 days), where "n" designates the number of weeks, e.g., 1, 2, 3, 4, 5, 6, 8, 12 or more.

[0111] The terms "initial dose," "secondary dose," and "tertiary dose" refer to the time sequence of administration of an IL-4R antagonist. Thus, an "initial dose" is a dose administered at the beginning of a treatment regimen (also called a "baseline dose"); a "secondary dose" is a dose administered after the initial dose. and a "tertiary dose" is a dose administered after the secondary dose. The initial, secondary, and tertiary doses may all contain the same amount of IL-4R antagonist, but generally may differ from one another in terms of frequency of administration. However, in certain embodiments, the amount of IL-4R antagonist contained in the initial, secondary, and / or tertiary doses varies from one another over the course of treatment (e.g., adjusted upward or downward as appropriate). In certain embodiments, two or more (e.g., 2, 3, 4, or 5) doses are administered as "loading doses" at the beginning of a treatment regimen, followed by subsequent doses (e.g., maintenance doses) administered at a lower frequency. In one embodiment, the maintenance dose may be lower than the loading dose. For example, one or more loading doses of 600 mg of IL-4R antagonist may be administered, followed by a maintenance dose of about 75 mg to about 300 mg.

[0112] In one exemplary embodiment of the invention, the secondary and / or tertiary doses are each 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 weeks or more later). The phrase "immediately preceding dose" refers to a dose of an IL-4R antagonist administered to a patient prior to administration of the next dose in the sequence, without any intervening doses, in a series of multiple doses.

[0113] These methods can include administering any number of secondary and / or tertiary doses of an IL-4R antagonist to the patient. For example, in certain embodiments, only a single secondary dose is administered to the patient. In other embodiments, two or more (e.g., 2, 3, 4, 5, 6, 7, 8, or more) secondary doses are administered to the patient. Similarly, in certain embodiments, only a single tertiary dose is administered to the patient. In other embodiments, two or more (e.g., 2, 3, 4, 5, 6, 7, 8, or more) tertiary doses are administered to the patient.

[0114] In embodiments involving multiple secondary doses, each secondary dose may be administered at the same frequency as the other secondary doses. For example, each secondary dose may be administered to the patient 1-2 weeks after the immediately preceding dose. Similarly, in embodiments involving multiple tertiary doses, each tertiary dose may be administered at the same frequency as the other tertiary doses. For example, each tertiary dose may be administered to the patient 2-4 weeks after the immediately preceding dose. Alternatively, the frequency with which the secondary and / or tertiary doses are administered to the patient may vary during the course of treatment. The frequency of administration may also be adjusted during the course of treatment by a physician depending on the needs of the individual patient after clinical testing.

[0115] The present invention includes methods for treating asthma or a related condition, comprising sequential administration of an IL-4R antagonist and a second therapeutic agent to a patient. In some embodiments, the present invention comprises administering one or more doses of an IL-4R antagonist, followed by one or more doses (e.g., 2, 3, 4, 5, 6, 7, 8, or more) of a second therapeutic agent. For example, to treat, alleviate, reduce, or ameliorate one or more symptoms of asthma, one or more doses of about 75 mg to about 300 mg of an IL-4R antagonist may be administered, followed by one or more doses (e.g., 2, 3, 4, 5, 6, 7, 8, or more) of a second therapeutic agent (e.g., an inhaled corticosteroid or beta2-agonist or any other therapeutic agent described elsewhere herein). In some embodiments, the IL-4R antagonist is administered in one or more doses (e.g., 2, 3, 4, 5, 6, 7, 8, or more) that result in improvement of one or more asthma-related parameters, followed by administration of a second therapeutic agent to prevent recurrence of at least one asthma symptom. An alternative embodiment relates to simultaneous administration of the IL-4R antagonist and the second therapeutic agent. For example, one or more doses (e.g., 2, 3, 4, 5, 6, 7, 8, or more) of the IL-4R antagonist are administered, and the second therapeutic agent is administered in a dose-dependent manner. In some embodiments, the second therapeutic agent is administered before, after, or simultaneously with the IL-4R antagonist.

[0116] Treatment population The methods of the present invention include administering a therapeutic composition comprising an IL-4R antagonist to a subject in need thereof. The phrase "subject in need thereof" refers to a human or non-human animal that exhibits one or more symptoms or signs of asthma (e.g., eosinophilic asthma, including moderate to severe eosinophilic asthma) or has been diagnosed with asthma. For example, a "subject in need thereof" can include a subject that exhibits (or has exhibited) one or more asthma-related parameters, such as, for example, a poor FEV1 (e.g., less than 2.0 L), a poor AM PEF (e.g., less than 400 L / min), a poor PM PEF (e.g., less than 400 L / min), an ACQ5 score of at least 2.5, at least one nighttime awakening per night, and / or a SNOT-22 score of at least 20, prior to treatment. In various embodiments, these methods can be used to treat mild, moderate-to-severe, and severe asthma in patients in need thereof.

[0117] In a related embodiment, the "subject in need thereof" is administered an inhaled corticosteroid (ICS) / long-acting beta2-adrenergic receptor antagonist (IL-4R antagonist) prior to receiving the IL-4R antagonist. The subject may have been prescribed or is currently taking a combination of IL-4R antagonists (LABAs). Examples of ICS / LABA treatments include fluticasone / salmeterol combination treatment and budesonide / formotorol combination treatment. For example, the present invention includes methods comprising administering an IL-4R antagonist to a patient who has been receiving a regular course of ICS / LABA for two or more weeks immediately prior to administration of the IL-4R antagonist (such prior treatment is referred to herein as "background treatment"). The present invention includes treatment methods in which the background treatment is discontinued at the time of or immediately prior to (e.g., one day to two weeks before) the first administration of the IL-4R antagonist. Alternatively, the background treatment may be continued in combination with administration of the IL-4R antagonist. In yet other embodiments, the amount of the ICS component, the amount of the LABA component, or both, is gradually reduced before or after the initiation of IL-4R antagonist administration. In some embodiments, the present invention includes a method for treating a patient with persistent asthma for at least 12 months or more. In one embodiment, a patient with persistent asthma may be resistant to treatment with a therapeutic agent such as a corticosteroid and may be administered an IL-4R antagonist according to the present invention.

[0118] In some embodiments, a "subject in need thereof" may be a subject with elevated levels of an asthma-associated biomarker. Examples of asthma-associated biomarkers include, but are not limited to, IgE, thymus and activation-regulated chemokine (TARC), eotaxin-3, CEA, YKL-40, and periostin. In some embodiments, a "subject in need thereof" may be a subject with a blood eosinophil count of 300 / μL or greater or a sputum eosinophil level of 3% or greater. In one embodiment, a "subject in need thereof" may be a subject with elevated levels of bronchial or airway inflammation as measured by fraction of exhaled nitric oxide (FeNO).

[0119] For purposes of the present invention, a normal IgE level in a healthy subject is less than about 100 kU / L (e.g., ImmunoCAP (R) Assay [Phadia, Inc. Portage, Thus, the present invention relates to elevated serum IgE levels (greater than about 100 kU / L, greater than about 150 kU / L, greater than about 500 kU / L, greater than about 1000 kU / L, greater than about 1500 kU / L, greater than about 2000 kU / L, The methods include selecting a subject exhibiting a serum IgE level greater than about 2500 kU / L, greater than about 3000 kU / L, greater than about 3500 kU / L, greater than about 4000 kU / L, greater than about 4500 kU / L, or greater than about 5000 kU / L), and administering to the subject a pharmaceutical composition comprising a therapeutically effective amount of an IL-4R antagonist.

[0120] TARC levels in healthy subjects range from 106 ng / L to 431 ng / L, with an average of about 239 ng / L. (An exemplary assay system for measuring TARC levels is the TARC quantitative ELISA kit provided by R&D Systems, Minneapolis, MN, under catalog number DDN00.) Accordingly, the present invention includes methods comprising selecting a subject exhibiting elevated TARC levels, where the serum TARC level is greater than about 431 ng / L, greater than about 500 ng / L, greater than about 1000 ng / L, greater than about 1500 ng / L, greater than about 2000 ng / L, greater than about 2500 ng / L, greater than about 3000 ng / L, greater than about 3500 ng / L, greater than about 4000 ng / L, greater than about 4500 ng / L, or greater than about 5000 ng / L, and administering to the subject a pharmaceutical composition comprising a therapeutically effective amount of an IL-4R antagonist.

[0121] Eotaxin-3 belongs to a group of chemokines released by airway epithelial cells and is upregulated by the Th2 cytokines IL-4 and IL-13 (Lilly et al. 1999, J. Allergy Clin. Immunol. 104:786-790). The present invention includes methods for treating patients with elevated eotaxin-3 levels, such as 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, comprising administering an IL-4R antagonist. Serum eotaxin-3 levels are measured, for example, by ELISA.

[0122] Periostin is an extracellular matrix protein involved in Th2-mediated inflammatory processes. Periostin levels are found to be upregulated in patients with asthma (Jia et al 2012 J Allergy Clin Immunol.130:647-654.e10.doi:10.1016 / j.jaci.2012.06.025.Epub 2012 Aug 1). The present invention includes methods for treating patients with elevated levels of periostin, comprising administering an IL-4R antagonist.

[0123] Fractional exhaled NO (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. 6:1368-1370). FeNO levels are 2-30 parts per billion (ppb) in healthy adults. An exemplary assay for measuring FeNO is by using a NIOX instrument by Aerocrine AB, Solna, Sweden. Assessment can be performed prior to spirometry and after at least one hour of fasting. The present invention includes methods comprising administering an IL-4R antagonist to a patient with elevated levels of exhaled NO (FeNO), such as greater than about 30 ppb, greater than about 31 ppb, greater than about 32 ppb, greater than about 33 ppb, greater than about 34 ppb, or greater than about 35 ppb.

[0124] Carcinoembryonic antigen (CEA) is a tumor marker that correlates with non-neoplastic lung diseases (Marechal et al. 1988, Anticancer Res. 8:677-680). Serum CEA levels can be measured by ELISA. The present invention provides CEA levels 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, and the like. The present invention also includes methods comprising administering an IL-4R antagonist to a patient having high, or elevated, levels of CEA, such as greater than about 5.0 ng / ml.

[0125] YKL-40 (derived from its N-terminal amino acids tyrosine (Y), lysine (K), and leucine (L) and its molecular weight of 40 kD) is a chitinase-like protein that is upregulated and correlates with asthma exacerbations, IgE, and eosinophils (Tang et al. 2010 Eur. Respir. J. 35:757-760). Serum YKL-40 levels are measured, for example, by ELISA. The present invention includes methods comprising administering an IL-4R antagonist to a patient with elevated YKL-40 levels, such as greater than about 40 ng / ml, greater than about 50 ng / ml, greater than about 100 ng / ml, greater than about 150 ng / ml, greater than about 200 ng / ml, or greater than about 250 ng / ml.

[0126] Induced sputum eosinophils and neutrophils 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 counting according to methods known in the art, e.g., European Respiratory Society guidelines. The present invention includes methods comprising administering an IL-4R antagonist to a patient with elevated levels of sputum eosinophils, such as greater than about 2.5% or greater than about 3%.

[0127] Methods for assessing pharmacodynamic asthma-related parameters The present invention also includes methods for assessing one or more pharmacodynamic asthma-related parameters resulting from administration of a pharmaceutical composition comprising an interleukin-4 receptor (IL-4R) antagonist in a subject in need thereof. A reduction in the incidence of 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 a correlation may not be observed in all cases.

[0128] Examples of "pharmacodynamic asthma-related parameters" include, for example: (a) biomarker expression levels; (b) serum protein and RNA analysis; (c) induced sputum eosinophil and neutrophil levels; (d) exhaled nitric oxide (FeNO); and (e) blood eosinophil count. An "improvement in a pharmacodynamic asthma-related parameter" refers to, for example, a decrease from baseline in one or more biomarkers such as TARC, eotaxin-3, or IgE, a decrease in sputum eosinophils or neutrophils, FeNO, or blood eosinophil count. The term "baseline," as used herein with respect to a pharmacodynamic asthma-related parameter, refers to the value of the pharmacodynamic asthma-related parameter for a patient before or at the time of administration of a pharmaceutical composition featured in the present invention.

[0129] To evaluate pharmacodynamic asthma-related parameters, parameters are quantified at baseline and at the time point after administration of the pharmaceutical compositions of the present invention.For example, pharmacodynamic asthma-related parameters can be measured 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 14 days after initial treatment with the pharmaceutical compositions of the present invention, or 3 weeks after, 4 weeks after, 5 weeks after, 6 weeks after, 7 weeks after, 8 weeks after, 9 weeks after, 10 weeks after, 11 weeks after, 12 weeks after, 14 weeks after, 3 weeks after, 4 weeks after, 5 weeks after, 6 weeks after, 7 weeks after, 8 weeks after, 9 weeks after, 10 weeks after, 11 weeks after, 12 weeks after, 13 weeks after, 14 weeks after, 15 weeks after, 16 weeks after, 17 weeks after, 18 weeks after, 19 weeks after, 20 weeks after, 21 weeks after, 22 weeks after, 23 weeks after, 24 weeks after or more after. The difference between the parameter value at a particular time point after the start of treatment and the parameter value at baseline is used to establish whether there has been an "improvement"-like change (e.g., an increase or decrease, as the case may be, depending on the particular parameter being measured) in the pharmacodynamic asthma-related parameter.

[0130] In certain embodiments, administration of an IL-4R antagonist to a patient causes changes, such as a decrease or increase, in the expression of certain biomarkers. Asthma-related biomarkers include: (a) total IgE; (b) thymus and activation-regulated chemokine (TARC); (c) YKL-40; and (d) carcinoembryonic antigen (CEA, also known as CEA cell adhesion molecule 5 [CEACAM5]) in serum, and (e) eotaxin-3 in plasma. For example, administration of an IL-4R antagonist to an asthma patient can cause one or more of a decrease in TARC or eotaxin-3 levels, or a decrease in total serum IgE levels. The decrease can be detected 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, or more after administration of the IL-4R antagonist. Biomarker expression can be assayed using methods known in the art. For example, protein levels can be measured by ELISA (enzyme-linked immunosorbent assay), or RNA levels can be measured by reverse transcription coupled with polymerase chain reaction (RT-PCR).

[0131] As discussed above, biomarker expression can be assayed by detection of protein or RNA in serum. Serum samples can also be used to monitor additional protein or RNA biomarkers associated with response to treatment with an IL-4R antagonist, IL-4 / IL-13 signaling, asthma, atopic, or eosinophilic disease (e.g., by measuring soluble IL-4Rα, IL-4, IL-13, periostin). In some embodiments, RNA samples are used to determine RNA levels, e.g., biomarker RNA levels (non-genetic analysis); and in other embodiments, RNA samples are used for transcriptome sequencing (e.g., genetic analysis). [Example]

[0132] The following examples are presented so as to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the methods and compositions featured in this invention, and are not intended to limit the scope of what the inventors regard as their invention. Efforts have been made to ensure accuracy with respect to numbers used (e.g., amounts, temperatures, etc.), but some experimental error and deviation should be accounted for. Unless otherwise indicated, parts are parts by weight, molecular weight is average molecular weight, temperature is in degrees Celsius, and pressure is at or near atmospheric.

[0133] Example 1. Generation of human antibodies against human IL-4R Human anti-hIL-4R antibodies were produced as described in U.S. Patent No. 7,608,693. Table 1 shows the heavy and light chain variable region amino acid sequence pairs and sequence identifiers for the CDR amino acid sequences of selected anti-IL-4R antibodies, as well as their corresponding antibody designations.

[0134] [Table 1]

[0135] [Table 2]

[0136] The exemplary IL-4R antagonist used in the following examples is the human anti-IL-4R antibody designated H1H098-b in Table 1 (also referred to herein as "mAb1").

[0137] Example 2: Clinical Trial of Subcutaneously Administered Anti-IL-4R Antibody (mAb1) in Patients with Persistent Moderate to Severe Eosinophilic Asthma, Including Asthma Patients with Chronic Hypertrophic Eosinophilic Sinusitis A. Research Objectives and Overview A randomized, placebo-controlled, double-blind, parallel-group study was conducted in patients with persistent moderate-to-severe eosinophilic asthma partially or uncontrolled with inhaled corticosteroids (ICS) and long-acting beta-agonists (LABAs). Patients received either 300 mg mAb1 or placebo subcutaneously once weekly for 12 weeks. The primary objective of the study was to examine the effect of mAb1 administered subcutaneously once weekly for 12 weeks compared with placebo on reducing the incidence of asthma exacerbations in patients with persistent moderate-to-severe eosinophilic asthma. Secondary objectives of the study were to evaluate the safety and tolerability of mAb1 administered subcutaneously once weekly for 12 weeks in patients with persistent moderate-to-severe eosinophilic asthma and to evaluate mAb1 serum concentrations after 12 weeks of subcutaneous dosing in patients with persistent moderate-to-severe eosinophilic asthma.

[0138] Prior to screening, patients had to have been on a stable dose of one of the following doses and regimens of ICS / LABA combination therapy (also called "background therapy") for at least one month: Fluticasone / salmeterol combination therapy - Advair (R) Diskus-Dry Powder Inhaler (DPI): 250 / 5 0ug twice daily (BID) or 500 / 50ug BID; or - Advair(R) HFA - Metered dose inhaler (MDI): 230 / 42ug BID or 460 / 42ug BID; or Budesonide / formoterol combination therapy (Symbicort (R) 160 / 9u g BID or 320 / 9ug BID); or Mometasone / formoterol combination therapy (Dulera (R) 200 / 10ug B ID or 400 / 10ug BID).

[0139] Patients continuing budesonide / formoterol or mometasone / formoterol were switched to equivalent doses of fluticasone / salmeterol at the time of randomization (day 1), and patients continuing fluticasone / salmeterol remained on background therapy.

[0140] Patients who met the inclusion and exclusion criteria (see below) were randomized to one of the following treatments: mAb1 300 mg administered subcutaneously once weekly for 12 weeks; or placebo administered subcutaneously once weekly for 12 weeks.

[0141] The study included a 2-week screening period, a 12-week treatment period including a 4-week background treatment stabilization period and an 8-week background treatment washout period after randomization, followed by an 8-week post-treatment follow-up period.

[0142] Algorithm for withdrawal of background therapy (ICS / LABA): Patients remained on twice-daily (BID) fluticasone / salmeterol background therapy for 4 weeks after initiating add-on therapy or treatment with 300 mg mAb1 (or placebo). Four weeks after randomization, patients were switched from BID fluticasone / salmeterol combination therapy to equivalent ICS doses of fluticasone monotherapy (250 μg or 500 μg Flovent). (R) Diskus-BIDDPI formulation; or 220ug if Flovent 440ug BID (R) (including HFA-MDI formulations) The LABA component (i.e., salmeterol) was discontinued. If, at subsequent visits, the patient did not meet any criteria for an asthma exacerbation (as defined below), starting at week 6, the fluticasone dose was reduced by approximately 50%. If no asthma exacerbations occurred, ICS withdrawal proceeded according to the following schedule:

[0143] [Table 3]

[0144] Upon completion of 12 weeks of treatment with study medication (or after early discontinuation), patients were set back on their original dose of fluticasone / salmeterol, budesonide / formoterol, or mometasone / formoterol (dose at study entry) and albuterol or levalbuterol as needed to manage symptoms for an additional 8 weeks of off-study medication before the final safety assessment.

[0145] Adult patients were included in the study based on the following criteria: (1) a physician diagnosis of persistent asthma for at least 12 months according to the Global Initiative for Asthma (GINA) 2009 guidelines, whose airway inflammation may be eosinophilic; and (2) whose asthma was partially controlled or not controlled with inhaled corticosteroid / long-acting beta-agonist combination therapy according to the following criteria: (i) fluticasone / salvage for at least 1 month prior to screening. (ii) a stable dose of either meterol combination therapy (DPI formulation: 250 / 50 μg BID or 500 / 50 μg BID or MDI: 230 / 42 μg BID or 460 / 42 μg BID), or budesonide / formoterol combination therapy (160 / 9 μg BID or 320 / 9 μg BID), or mometasone / formoterol combination therapy (200 / 10 μg BID or 400 / 10 μg BID) during the screening period; (iii) a Juniper Asthma Control Questionnaire (5-question version, ACQ) score of 1.5 or greater and 3.0 or less at screening; (iv) a FEV1 during the screening period (maximum of 3 attempts) and at randomization prior to the first dose (maximum of 3 attempts). 50% or greater predicted normal; (v) treatment with one or more systemic (oral and / or parenteral) steroid bursts for asthma exacerbation, or hospitalization or emergency department visit for asthma exacerbation, within 2 years prior to screening; and (vi) a documented history of a reversible FEV1 of at least 12% and 200 mL (maximum of 3 attempts) after albuterol 200 μg to 400 μg (2 to 4 inhalations) during the baseline-screening period, or a documented history of a positive methacholine challenge (PD20 methacholine 8 mg or less) within 12 months prior to screening. (R) , SYMBIC ORT (R) or DULERA (R) Patients who were partially controlled or not controlled on moderate-to-high dose combination therapy with EGFR-1000 or EGFR-10000, and who had blood eosinophils equal to or greater than 300 cells per microliter or sputum eosinophils equal to or greater than 3% during the screening phase were included in the study.

[0146] Patients who met all inclusion criteria were screened for the following exclusion criteria: (1) patients under 18 years of age or over 65 years of age; (2) clinically relevant abnormal laboratory values ​​suggesting an unknown disease and requiring further evaluation; (3) chronic obstructive pulmonary disease (COPD) and / or other lung diseases impairing pulmonary function tests; (4) patients requiring beta-adrenergic receptor blockers for any reason; (5) current smokers or those who had quit within 6 months prior to screening; (6) a history of smoking more than 10 packs per year; and (7) hospitalization due to an asthma exacerbation within 2 months prior to screening. or emergency treatment visit; (8) planned initiation of allergen immunotherapy within the study period; (9) exposure to another study antibody within the prescreening period of less than 5 half-lives of the antibody but more than 30 days, or at least 6 months if the half-life of the antibody is not known; (10) previous enrollment in the current study; (11) the patient was an employee of the investigator, his / her family, or the testing laboratory; (12) known or suspected non-compliance, alcohol, or drug abuse; (13) inability to follow study procedures (e.g., due to language problems or psychological disorders) (14) reversal of sleep pattern (e.g., night workers); (15) treatment with drugs known to prolong the QTc interval; (16) concomitant severe illness that contraindicates the use of ICS (e.g., active or inactive pulmonary tuberculosis) or LABA (e.g., diabetes, cardiovascular disease, hypertension, hyperthyroidism, thyrotoxicosis, etc.); (17) use of injectable glucocorticoids or oral systemic glucocorticoids for more than three courses within two months prior to screening or within six months prior to screening; (18) use of a nonsteroidal controller alone or (19) patients receiving prohibited concomitant medications (listed below); (20) known allergy to doxycycline or related compounds; (21) pregnancy or intention to become pregnant during the course of the study, breastfeeding, or not intending to use an effective method of contraception; and (22) recent history of parasitic infection or travel to a parasite-endemic area within 6 months prior to screening.

[0147] Patients were maintained on a stable dose of background asthma medication for the first 4 weeks of the study, after which The dose of background therapy was gradually reduced. First, the long-acting beta-agonist component of the background therapy was discontinued at week 4, and then the inhaled corticosteroid dose was reduced by half every 2 weeks until week 12. Patients continued on study treatment until the end of the study or until discontinuation due to an asthma exacerbation or any other reason.

[0148] B. Test Procedures Study Medications: Sterile mAb1 150 mg / mL solution for subcutaneous (SC) injection was provided in 5 mL glass vials. Each vial contained a withdrawable volume of 2 mL. A 300 mg dose was administered subcutaneously at the test site once weekly in the morning for 12 weeks. Placebo: Sterile placebo for SC injection was provided in identically matched 5 mL glass vials. Each vial contained a withdrawable volume of 2 mL. Placebo was administered subcutaneously at the test site once weekly in the morning for 12 weeks.

[0149] The following concomitant medications were not allowed during the study period: fluticasone / salmeterol combination therapy administered per the protocol or any other inhaled steroid other than fluticasone (or budesonide / formoterol or mometasone / formoterol during the screening period); systemic or ocular steroids; LABAs other than the salmeterol component of fluticasone / salmeterol combination therapy administered per the protocol; any other ICS / LABA combinations other than those indicated above; any inhaled anticholinergics (e.g., ipratropium bromide or tiotropium); methylxanthines (theophylline, aminophylline); cromones; anti-IgE therapy; lipoxygenase inhibitors; and leukotriene receptor antagonists or leukotriene synthesis inhibitors.

[0150] C. Treatment Efficacy The primary endpoint of this study was the occurrence of asthma exacerbations, defined as any of the following: (1) a 30% or greater decrease from baseline in morning peak expiratory flow (PEF) for 2 consecutive days; or (2) 6 or more additional puffs of albuterol or levalbuterol medication in a 24-hour period (compared to baseline) for 2 consecutive days; or (3) an investigator-determined worsening of asthma requiring (a) systemic (oral and / or parenteral) steroid treatment, or (b) a 4-fold or greater increase in the last dose of ICS received before discontinuation from the study, or (c) hospitalization.

[0151] Secondary endpoints of the study included mean changes from baseline in the following parameters: (1) forced expiratory volume in 1 second (FEV1) in liters measured at each visit; (2) morning and evening peak expiratory flow rates in liters / minute (AM PEF and PM PEF) measured daily; (3) daily albuterol / levalbuteral use in inhalations / day; (4) 5-item Asthma Control Questionnaire (ACQ5) score at each visit; and (5) nocturnal awakenings (number per night) measured daily, and (6) 22-item Sinus Nose Test (SNOT-22), assessed at baseline and at the treatment endpoint (week 12) to assess upper respiratory tract symptoms. Secondary endpoints also included the proportion of patients with a combined asthma event, defined as a 30% or greater decrease from baseline in morning PEF for two consecutive days, along with six or more additional puffs of albuterol or levalbuterol in a 24-hour period (compared to baseline) for two consecutive days. PEF, ACQ5, asthma symptom score, nighttime awakenings, and acute asthma treatment use were obtained via electronic diaries. Mean daily nighttime awakenings, ranging from 0 to 10, were averaged from the previous 7 days. Morning and evening asthma symptom scores consisted of unvalidated patient-reported outcomes assessed on a 5-point Likert-type scale, with higher scores indicating worse outcomes (Table 2). Patients recorded a global symptom score twice daily before measuring PEF. Data are presented as the average over the 7 days prior to the specified time point (e.g., Figure 26). (See 26A and 26B).

[0152] [Table 4]

[0153] D. Adverse Event Monitoring Safety was assessed throughout the study by monitoring adverse events and serious adverse events.

[0154] An Adverse Event (AE) is any untoward medical occurrence in a subject administered a medicinal product or in a clinical investigational subject. Thus, an AE can be any untoward and unintended sign (including abnormal laboratory findings), symptom, or disease that is temporally related to the use of the medicinal (investigational) product, whether or not it is considered related to that medicinal product. AEs also include: any worsening of a pre-existing condition (i.e., a clinically significant change in either frequency and / or intensity) that is temporally related to the use of the study drug; an abnormal laboratory finding that is considered clinically significant by the investigator; and any adverse medical occurrence.

[0155] A Serious Adverse Event (SAE) is any untoward medical occurrence that, at any dose, results in death; is life-threatening; requires hospitalization or an extension of an existing hospitalization; results in persistent or significant disability / incapacity; is a congenital abnormality / birth defect; or is a serious medical event.

[0156] E. Statistical methods For the primary analysis of the proportion of patients experiencing an asthma exacerbation, a logistic regression model was used to compare the SAR group with placebo. The model included terms for treatment and stratification factors (prior ICS / LABA combination treatment dose). The primary analysis was based on the modified intent-to-treat (mITT) population, which included all randomized patients who received at least one dose of the investigational medication (IMP). A stratified chi-square test was also used to support the primary analysis.

[0157] For secondary efficacy endpoints, except for SNOT-22, change from baseline was analyzed using a mixed-effects model with repeated measures (MMRM) approach. The model included change from baseline value through week 12 as the response variable and factors for treatment (determined effect), stratification factors, visit, treatment-by-visit interaction, baseline value, and baseline-by-visit interaction. Statistical estimates for treatment comparisons for change from baseline at week 12 were derived from the mixed-effects model. Change from baseline in SNOT-22 was analyzed using analysis of covariance (ANCOVA), with end-of-treatment measurements used to impute missing data. Pharmacodynamic effects were assessed using the MMRM model in a post-hoc manner. Because there was only one primary endpoint and analysis, no adjustment for multiplicity was made. Safety variables, including AEs, laboratory parameters, vital signs, ECGs, clinical laboratory findings, and physical examinations, were summarized using descriptive statistics.

[0158] Demographic and clinical characteristics were summarized using descriptive characteristics. Plots of secondary and pharmacodynamic variables are shown as mean change from baseline over time with standard error. Comparisons of treatment effects from MMRM analyses are based on least-squares mean change from baseline at week 12 (95% confidence interval [CI]).

[0159] F. Results The results observed for all 104 randomized patients (screened from 491) who completed or discontinued the study treatment period are summarized below. All randomized patients were exposed to study treatment and included in the mITT population. Baseline characteristics were similar between groups. Demographic and clinical characteristics were also similar between the two groups (Table 3). Patients were treated with either 300 mg subcutaneous mAb1 once weekly or placebo, as shown above. The study treatment period was completed by 86.5% and 67.3% of mAb1 and placebo patients, respectively (Figure 25). The most common reason for discontinuation was lack of efficacy, which was more frequent with placebo (21.2%) than with mAb1 (1.9%).

[0160] [Table 5]

[0161] (i) Primary efficacy endpoint The incidence of asthma exacerbations in the placebo and mAb1 treatment groups is shown in Table 4.

[0162] [Table 6]

[0163] There were a total of 26 asthma exacerbations during the treatment period, and no patients were hospitalized due to an asthma exacerbation. In the placebo group, 23 patients (44.2%) experienced an asthma exacerbation, while only 3 patients (5.8%) in the mAb1-treated group experienced an asthma exacerbation. The odds ratio was 0.077 (p<0.0001), and the relative risk reduction was approximately 87%.

[0164] Of the 26 asthma exacerbations experienced during the study, 9 were considered severe, as evidenced by the need for immediate intervention in the form of treatment with either systemic corticosteroids or inhaled corticosteroids at four or more times the dose taken before the event. The incidence of severe asthma exacerbations is summarized in Table 5.

[0165] [Table 7]

[0166] As shown in Table 5, eight severe asthma exacerbations occurred in the placebo group and only one severe asthma exacerbation occurred in the mAb1 treatment group. The remaining 15 asthma exacerbations in the placebo group and two in the mAb1 group met the protocol definition of an exacerbation based on decreased morning PEF and / or increased albuterol / levalbuterol use. As shown in Table 6, within the active treatment group, sustained improvements relative to baseline were observed over the course of the study for all parameters despite steroid withdrawal.

[0167] [Table 8]

[0168] With mAb1, the time to exacerbation was longer and the risk of exacerbation was reduced compared with placebo (Figure 1). Kaplan-Meier plot analysis of the time to asthma exacerbation revealed that the effect of treatment with mAb1 persisted for a long period, including after 8 weeks, when patients were at higher risk of experiencing an exacerbation due to steroid withdrawal (Figure 1).

[0169] Only one patient in the placebo group had a complex asthma event, defined as a 30% or greater decrease from baseline in morning PEF for 2 consecutive days, along with 6 or more additional puffs of albuterol or levalbuterol in a 24-hour period (compared to baseline) for 2 consecutive days.

[0170] (ii) Other efficacy endpoints Pulmonary function parameters (FEV1, AM PEF, and PM PEF), asthma symptom-based endpoints (ACQ score, nighttime awakenings), and albuterol use were assessed for each patient at each visit. The observed results (weekly change from baseline) for these parameters are shown in Figures 2-7, respectively. Additionally, SNOT-22 scores were assessed at baseline and end of treatment. Mean values ​​at baseline and 12 weeks (LOCF) for all parameters, along with the mean differences between treatment groups (ANOVA model for SNOT-22), are summarized in Table 7. In Table 7, the column labeled "Difference vs. Placebo" indicates the placebo-corrected value from baseline, which takes into account the observed change in the value of the parameter compared to the change observed for that parameter in the placebo-treated group.

[0171] [Table 9]

[0172] Treatment with mAb1 produced a significant change from baseline in FEV1 at week 1 that was maintained through week 12 despite LABA and ICS withdrawal (Figure 2), with a slight decrease in FEV1 at week 5 coincident with LABA withdrawal. Similar improvements were observed in morning PEF, but less in evening PEF (Figures 3 and 4). The least squares (LS) mean change from baseline in FEV1 through week 12 was -0.22 L for placebo and 0.05 L for the mAb1 group (p=0.0009).

[0173] ACQ5 scores improved in both treatment groups at week 1 (Figure 6). However, ACQ5 improved further with mAb1 between weeks 1 and 4, whereas the placebo effect stabilized and maintained the difference through week 12.

[0174] Morning symptom scores increased with placebo from baseline to week 12. With mAb1, there was an initial decrease that remained below baseline through week 12 (Figure 26A). A similar pattern (greater variation) was observed in evening asthma scores (Figure 26B).

[0175] Nighttime awakenings remained stable in the placebo group until week 6, then increased from week 6 to week 12. In contrast, nighttime awakenings decreased in the mAb1 group by week 1 and remained improved relative to baseline until week 12 (Figure 7).

[0176] Changes in albuterol / levalbuterol use (Figure 5) were similar to other secondary endpoints: with placebo, there was an initial decrease followed by a return to baseline. With mAb1, the initial decrease was maintained over time.

[0177] There was a non-significant difference between SNOT-22 values ​​at baseline, with the mean placebo score being 26.24 and the mean mAb1 score being 39.02. At week 12, the LS mean change was a slight increase of 0.23 points for the placebo group and a mean decrease (improvement) of 8.26 points for the mAb1 group. This corresponds to a magnitude of improvement of 8.49 points for the mAb1 group (p=0.0027).

[0178] [Table 10]

[0179] [Table 11]

[0180] For all secondary endpoints, except for nighttime PEF and nocturnal awakenings, the 12-week measurements favored mAb1 treatment and were significant (Tables 7 and 8). Significant improvements with mAb1 were also observed for the three SNOT-22 items related to upper respiratory tract disease (Table 9).

[0181] (iii) Safety mAb1 was generally safe and well tolerated. Treatment-emergent adverse events (TEAEs) were reported similarly by 40 (76.9%) placebo-treated patients and 42 (80.8%) mAb1-treated patients (Table 10). TEAEs were nonspecific, generally mild to moderate in intensity, and most resolved by the end of the study. Increased reporting of the following TEAEs was seen with mAb1 compared to placebo: injection site reactions were reported by 15 (28.8%) mAb1 patients and 5 (9.6%) placebo patients; nasopharyngitis was reported by 7 (13.5%) mAb1 patients and 2 (3.8%) placebo patients; headache was reported by 6 (11.5%) mAb1 patients and 3 (5.8%) placebo patients, and nausea was reported by 4 (7.7%) mAb1 patients and 1 (1.9%) placebo patient.

[0182] [Table 12]

[0183] No deaths were reported during the study period. Four treatment-emergent serious adverse events (SAEs) were reported: one mAb1 patient experienced bipolar disorder, and three placebo patients experienced SAEs of asthma with pneumonia, gunshot wound with left pneumothorax, and right ankle fracture. None of these SAEs were considered related to IMP, and all but the new ankle fracture resolved by the end of the study. There were no deaths.

[0184] A total of six patients discontinued the study due to TEAEs: three patients in the mAb1 group (bipolar disorder, wheezing asthma, and angioedema) and three patients in the placebo group (upper respiratory tract infection, psoriasis, and asthma). The TEAE of angioedema occurred in a 42-year-old African-American woman after the ninth dose of study treatment as a pruritic, generalized rash observed at and distant from the injection site. This persisted for 1 week and resolved after discontinuation of study treatment and treatment with prednisone and diphenhydramine. It was considered treatment-related. This AE followed a milder rash at the injection site after the first and sixth doses of study treatment.

[0185] Among the most common AEs occurring in three or more patients in either treatment group (Table 10), injection site reactions, nasopharyngitis, nausea, and headache occurred more frequently with mAb1 than with placebo. Clinically significant changes in vital signs, physical examination, laboratory tests, or ECG findings in either group are reported.

[0186] G. Conclusion Significant improvements were observed in lung function and other asthma control parameters. Early and sustained efficacy was observed despite background treatment interruption. An approximately 87% relative reduction (p<0.0001) in the primary endpoint of the incidence of asthma exacerbations in patients with persistent moderate-to-severe asthma and eosinophilia was observed after 12 weeks of treatment with mAb1 300 mg once weekly (5.8%) compared with placebo (44.2%). As shown in Table 7, clinically meaningful and statistically significant (unadjusted for multiplicity) improvements with treatment compared with placebo were observed in lung function parameters (FEV1, PEF AM), Asthma Symptom Score (ACQ), and albuterol use. Beneficial trends were observed for PEF PM (p=0.0567) and nocturnal awakenings (p=0.0518). Statistically significant (unadjusted for multiplicity) improvements were also observed in SNOT-22 scores. Within the active treatment group, sustained improvements over baseline were observed during the course of the study for all parameters despite withdrawal of LABA and ICS. mAb1 was generally safe and well tolerated.

[0187] Example 3: Biomarker studies Biomarker analysis was performed on samples collected from subjects participating in a clinical trial of mAb1 (see Example 2 above). Specifically, serum / plasma biomarkers associated with TH2 inflammation, such as thymus and activation chemokine (TARC; CCL17), immunoglobulin E (IgE), eotaxin-3, periostin, carcinoembryonic antigen (CEA), YKL-40, and blood eosinophils, were measured in patient samples at baseline and at various time points after the initiation of study treatment. Baseline levels of these biomarkers were evaluated for their potential predictive value for treatment response. Additionally, exhaled NO (FeNO) and induced sputum eosinophils and neutrophils were measured as biomarkers of bronchial inflammation. Exhaled nitric oxide assessment was performed using a NIOX instrument (Aerocrine AB, Solna, Sweden) before spirometry and after at least 1 hour of fasting. Biomarkers were analyzed using a mixture model, and least squares means derived from the model are reported below.

[0188] Asthma subjects (N=104) received either mAb1 (300 mg) or placebo subcutaneously on days 1, 8, 15, 22, 29, 36, 43, 50, 57, 64, 71, and 78 (i.e., 12 weekly doses) of the study (see Example 2 herein). Samples for biomarker analysis were collected from antibody- and placebo-treated subjects at weeks 0, 1, 4, 8, and 12. Antigen-specific IgE was measured using Phadiatop®. (R) Test was detected using

[0189] TARC, eotaxin-3, and IgE remained unchanged in response to placebo (Figures 8, 9, and 10). In contrast, rapid decreases in TARC (mean % change +0.3% vs. -22.7%; p=0.0003) (Figure 8) and eotaxin-3 (mean % change 12.69% vs. -39.62%; p<0.0001) (Figure 9) were observed within 1 week in patients treated with mAb1 and persisted through 12 weeks: TARC: +7.6% vs. -26.0% (p=0.0005); eotaxin-3: +5.13% vs. -45.67% (p<0.0001).

[0190] TARC levels responded within 1 week after exposure to mAb1 administered subcutaneously at 300 mg. TARC levels remained constant at approximately 50% of baseline levels in mAb1-treated subjects despite ICS withdrawal. This data demonstrates that TARC expression is more directly related to IL-4R signaling than FEV1 changes (which declined in parallel with ICS withdrawal [after 4 weeks]) and that IL-4R blockade induces a shift toward TH1 characteristics, as observed, for example, with IFN-gamma administration. This is particularly relevant in patients requiring long-term treatment and who are prone to TH1-type immunity. In patients at risk for infectious disease, it may be possible to titrate mAb1 dose using TARC (and e.g., CXCL10).

[0191] Total serum IgE also decreased after mAb1 treatment. The total serum IgE response was more heterogeneous and delayed compared with the TARC response. The mean (SD) baseline IgE levels were 694.68 IU / L (1837.82) for the placebo group (n = 52) and 657.66 (1482.25) for the mAb1 group (n = 52), while the median values ​​were 169.95 for the placebo group and 206.15 for the mAb1 group. Despite this heterogeneity, a trend toward decreased IgE was observed in mAb1-exposed patients compared with placebo, but this trend only began at week 4. Serum IgE was significantly reduced in the mAb1 group compared to placebo (mean % change, +13.5% vs. 10.1%; p=0.0325), beginning at week 4 and continuing to decrease through week 12 (mean % change, -36.8% for REGN668 / SAR231893 vs. -5.5% for placebo; p<0.0001) (Figure 10).

[0192] Changes from baseline and placebo at week 12 for FeNO, TARC, eotaxin-3, and IgE all favored mAb1 (all P<0.001) (Table 11). No differences from baseline or between treatments were observed for YKL-40 or CEA.

[0193] [Table 13]

[0194] There was a transient decrease in periostin levels, followed by an increase upon LABA / ICS withdrawal (Figure 11). Administration of mAb1 delayed the increase but did not prevent the increase above baseline. No consistent treatment effect was observed for CEA (Figure 12) and YKL-40 (Figure 13). Blood eosinophil counts remained unchanged through week 6, then increased at weeks 8 and 12 (Figure 14). Peripheral blood eosinophil counts remained unchanged with placebo throughout treatment. The difference between treatments was not significant, and only in a few patients treated with mAb1 was a borderline increase driven by a larger blood eosinophil rise. Little or no increase was observed in the majority of patients (Table 12).

[0195] [Table 14]

[0196] Only three mAb1 patients experienced asthma exacerbations during the study, so no conclusions could be drawn regarding the association between baseline biomarker levels and asthma exacerbations.

[0197] mAb1 treatment was also associated with a significant decrease from baseline in FeNO at week 4, and despite ICS withdrawal, FeNO remained below baseline through week 12 (mean % change at week 12: 35.0 for placebo vs. -28.7 for mAb1; p<0.0001) (Figure 15). In contrast, placebo FeNO values ​​remained stable through week 8 and then increased consistent with ICS withdrawal.

[0198] Improvement in forced expiratory volume in 1 second (FEV1) was associated with a reduction in FeNO (r=-0.40) at 12 weeks. 8, p=0.009) (Figure 16). Similarly, improvements in AM-PEF and PM-PEF correlated with FeNO reduction (Figures 17 and 18). Other correlations with FeNO were not significant. See Table 13.

[0199] [Table 15]

[0200] Scatterplot analysis of baseline eosinophils vs. change from baseline in FEV1 at Week 12 did not appear to indicate an association between baseline eosinophils and treatment effect, as measured by change from baseline in FEV1 at Week 12 in the study population (baseline eosinophils ≥ 0.3 Giga / L) (Figure 19). Baseline eosinophils correlated with decreased ACQ (Figure 20) and decreased albuterol / levalbuterol use (Figure 21). Baseline periostin and YKL-40 correlated with decreased ACQ (Figures 22 and 23).

[0201] The change from baseline in FEV1 at week 12 worsened with ICS withdrawal (starting at week 4). A similar analysis showed no association between baseline TARC or IgE and change from baseline in FEV1 at week 12 in the study population (baseline eosinophils ≥ 0.3 Giga / L).

[0202] summary These results indicate that mAb1 significantly reduced serum biomarkers related to Th2 inflammation (TARC, eotaxin-3, and IgE) and bronchial inflammation (FeNO) in adult asthmatics. The correlation between FeNO reduction and FEV1 improvement was consistent with IL-4 / Suggest a relationship between IL-13-mediated anti-inflammatory activity and improved lung function in moderate to severe uncontrolled asthma.

[0203] The present invention is not to be limited in scope by the specific embodiments described herein. Indeed, various modifications in addition to those described herein will become apparent to those skilled in the art from the foregoing description and accompanying figures. Such modifications are intended to be within the scope of the appended claims.

[0204] Example 4: Blockade of the IL-4 / IL-13 signaling pathway inhibits IgE production and airway remodeling in a mouse model of house dust mite-induced eosinophilic asthma introduction House dust mite allergen (HDM) has been shown to induce Th2 immune responses, including the influx of Th2 cells into the lungs and IL-4-induced transendothelial migration of eosinophils into the lungs. Eosinophils are the main effector cells in allergic reactions, and the release of granule components (including IL-4) from eosinophils contributes to inflammation. In asthmatics, Th2-driven production of IL-4 promotes eosinophil migration from the blood to the lungs via eotaxin, a potent eosinophil chemotactic factor (Mochizuki et al., J. Immunol., 1998, 160(1):60-68). Furthermore, when localized at the site of inflammation, eosinophils produce and secrete IL-4, thereby contributing to ongoing Th2-driven inflammation (Bjerke et al., Respir. Med., 1996, 90(5):271-277). In patients with allergic asthma, HDM challenge increased serum IgE and Th2 cytokine levels up to 5 weeks after allergen exposure (van de Pol et al., Allergy, 2012, 67(1):67-73).

[0205] In this example, we evaluated the pharmacodynamic effects of anti-IL-4R antibodies on markers of airway inflammation in mice using an HDM-induced model of chronic asthma. Furthermore, because collagen deposition correlates with the degree of airway remodeling, we evaluated the effects of anti-IL-4R antibodies on airway collagen deposition in this model.

[0206] Materials and Methods Two different anti-IL-4Rα antibodies were used in the experiments of this example: "mAb1," a fully human monoclonal antibody specific for human IL-4Rα (i.e., the anti-IL-4R antibody used in other examples presented herein); and "anti-mIL-4Rα," a murine monoclonal antibody specific for the murine IL-4Rα protein. mAb1 does not cross-react with murine IL-4Rα; therefore, mAb1 was used in combination with a humanized mouse IL-4Rα (IL-4Rα) engineered so that both the human IL-4 and IL-4Rα ectodomains were replaced with the corresponding murine sequences in the mouse. hu / hu IL-4Rα hu / hu ) was evaluated. The mouse anti-mouse IL-4Rα antibody, "anti-mIL-4Rα," was tested in wild-type (Balb / c) mice. It acts as a decoy receptor, blocking IL-13 signaling by IL-13 cytokine sequestration. A murine IL-13Rα2-mFc fusion protein was also tested in these experiments.

[0207] For the HDM-induced asthma model, mice were sensitized with intranasal application of HDM (50 μg in 20 μL of PBS per mouse) daily for 10 days, followed by rest (a 2-week recovery period). Allergen challenge was administered by intranasal application of HDM (50 μg in 20 μL of PBS per mouse) three times a week for 8 weeks. For each administration of HDM, mice were lightly anesthetized with isoflurane during either the sensitization or exposure period.

[0208] Mice were allowed to acclimate in the experimental facility for a minimum of 5 days before the start of experimental procedures. During the entire duration of the experiment, animals remained housed in the experimental facility under standard conditions with a 12-hour day / night cycle and free access to food and water. The number of mice per cage was limited to a maximum of 5 mice.

[0209] A total of 48 humanized mice (IL-4) were engineered to replace the human ectodomains of the human IL-4 ligand and IL-4Rα with their corresponding mouse sequences. hu / hu IL-4Rαhu / hu ) was used in two experiments. hu / hu IL-4Rα hu / hu Mice were of mixed background C57Bl / 6NTac (75%) / 129S6SvEvTac (25%). Additionally, 20 wild-type littermate mice of the same mixed background were used in one of three experiments. In each experiment, mice were sensitized with HDM (or PBS in the control group) daily for 10 days, followed by rest from days 11 to 29. From day 30, animals were exposed to HDM three times a week for 8 weeks until day 81, and then euthanized on day 85 for analysis. Mice were divided into six experimental groups as follows: (1) Non-sensitized, untreated PBS was applied intranasally during the sensitization and challenge periods. Mice were not treated with antibodies (IL-4 hu / hu IL-4Rα hu / hu Mice, n=9; wild type Litter n=5); (2) HDM sensitized, untreated :HDM was applied intranasally during the sensitization and challenge periods. Mice were not treated with antibodies (IL-4 hu / hu IL-4Rα hu / hu Mice, n = 7; Live littermates, n=5); (3) HDM sensitization, treatment with anti-mIL-4Rα HDM was applied intranasally during the sensitization and challenge periods. Mice were intraperitoneally (ip) injected with anti-mIL-4Rα at a dose of 50 mg / kg twice a week from week 7 to week 12 for a total of 12 doses over a 6-week period (wild-type littermates, n = 5); (4) HDM sensitization, treatment with anti-human mAb1 HDM was applied intranasally during the sensitization and challenge periods. Mice were injected intraperitoneally with mAb1 at a dose of 50 mg / kg twice a week from week 7 to week 12 for a total of 12 doses over a 6-week period (IL-4 hu / hu IL-4Rα hu / hu Ma Us, n=12); (5) HDM sensitization, treated with mouse IL-13Rα2-mFc fusion protein HDM was applied intranasally during the sensitization and challenge periods. Mice were intraperitoneally injected with IL-13Rα2-mFc at a dose of 25 mg / kg twice a week for a 6-week period from week 7 to week 12 for a total of 12 doses (IL-4hu / hu IL-4Rα hu / hu Mice, n = 7; wild-type littermates, n = 5 ); (6) HDM sensitization, treated with isotype control antibody HDM was applied intranasally during the sensitization and challenge periods. Mice were injected intraperitoneally with an isotype control Ab (IL-4) at a dose of 50 mg / kg twice a week for a total of 12 doses over a 6-week period from week 7 to week 12. hu / hu IL-4Rα hu / hu mice, n = 7).

[0210] Mice were euthanized on day 85, blood was collected for serum immunoglobulin level assays, and lungs (one lobe) were analyzed for i) bronchoalveolar lavage (BAL) fluid, ii) digested single-cell suspension samples for flow cytometry analysis, iii) fixed formalin specimens for staining and histological analysis, or iv) Sir to quantify collagen content per lung lobe. col TM One was used to generate samples for analysis using a collagen assay.

[0211] BAL lavage fluid was obtained from euthanized animals by first exposing the trachea and introducing a 23G lavage tube through a small incision in the tracheal wall. Sterile PBS (1 mL) was then instilled into the lungs, and BAL lavage fluid was withdrawn through the lavage tube using a syringe. 100 μL was loaded onto a Cytospin, which was spun at 500 rpm for 5 minutes to extract the cells onto a microscope slide. The slide was dried and stained with H&E to visualize the eosinophils.

[0212] Serum IgE levels were quantified using a commercially available ELISA kit. Briefly, serially diluted serum samples were incubated with an anti-IgE capture antibody in a 96-well plate, and IgE was detected with a biotinylated anti-mouse IgE secondary antibody. HRP-labeled purified mouse IgE was used as a standard.

[0213] HDM-specific IgG1 serum levels were quantified by ELISA. Briefly, HDM-coated plates were incubated with serially diluted sera, followed by incubation with anti-mouse IgG1 HRP-conjugated antibody. Relative levels of IgG1 serum levels are expressed as titer units (OD450 multiplied by the dilution factor required to achieve OD450 ≤ 0.5). Collected lung lobes were flash-frozen in liquid nitrogen and stored at -80°C until the extraction step. To extract collagen, lungs were homogenized in ice-cold NaCl / NaHCO3 solution. The cells were washed with Acid-Salt Wash Reagent to remove unbound Sircol dye and then mixed with Alkaline Reagent. 200 μL of each sample was transferred to a 96-well plate, and the OD at 555 nm was measured. A collagen standard was used for final quantification of collagen content in each sample.

[0214] Lungs were collected from euthanized mice and kept on ice in complete DMEM medium until digested with a mixture of collagenase and DNAse in HBSS buffer for 20 min at 37°C. Collagenase activity was quenched by adding 0.5 M EDTA, samples were centrifuged, and red blood cells were lysed using ACK buffer. The resulting cell suspension for each sample was divided into three separate pools and stained with antibody mixture 1 (anti-CD11c-APC Ab, anti-SiglecF-PE Ab, anti-F4 / 80-FITC Ab, anti-CD45-PerCp-Cy5.5 Ab), mixture 2 (anti-CD11c-APC Ab, anti-CD11b-PerCp-Cy5.5 Ab, anti-CD103-FITC Ab, anti-MHCII-PE Ab), or mixture 3 (anti-CD19-PE Ab, anti-Ly6G-APC Ab, anti-CD3-FITC, anti-CD11b-PerCp-Cy5.5 Ab) for 25 minutes at 4°C. The stained cells were fixed with Cytofix / Cytoperm solution for 30 minutes at 4°C and stored in PBS until flow cytometry analysis using a FACSCanto (BD Biosciences).

[0215] From a HDM-induced chronic model of eosinophilic asthma (EA), left lung lobes were harvested from four mice per group for microarray analysis of gene expression using GeneChip® technology. Gene expression levels in mice sensitized and challenged with HDM and then treated with isotype control Ab were compared to those in mice sensitized and challenged with mock (PBS) and not treated with antibody. The threshold for gene expression change was set at >1.5-fold. Mice sensitized and challenged with HDM were then compared. The set of genes identified as differentially expressed in the anti-IL-4Rα treated group was further analyzed in comparison with the isotype control treated group. The threshold for a change in gene expression in the IL-4Rα-Ab treated group compared with the isotype control treated group was set at >2-fold.

[0216] result HDM sensitization and challenge resulted in increased levels of IgE and HDM-specific IgG1. The IgE increase was completely blocked by both anti-IL-4Rα Abs but not by IL-13Rα2-Fc treatment (Figures 27A and 27B); HDM-specific IgG1 levels were unaffected by either treatment (data not shown).

[0217] HDM sensitization and challenge also resulted in increased collagen content in the lungs of mice. Collagen content in the lungs of mice treated with both IL-4Rα Ab and IL-13Rα2-Fc protein was reduced to the level observed in sham-sensitized and challenged mice (Figures 28A and 28B).

[0218] Furthermore, mAb1 treatment prevented the influx of eosinophils, neutrophils, and inflammatory dendritic cells into the lungs (FIG. 29, panels A and B).

[0219] HDM-induced IL-4 treated with isotype control antibody hu / hu IL-4Rα hu / hu Microarray analysis of mRNA isolated from lung tissue of mice shows that sham-sensitized and sham-sensitized Compared to exposed mice, 1468 genes were differentially expressed (826 up-regulated and 642 down-regulated genes). hu / hu IL-4Rα hu / hu Treatment of mice with mAb1 resulted in expression changes in only 521 genes (sham sensitization). The mAb1-treated group (HDM-sensitized / exposed mice) effectively blocked approximately 65% ​​of the genes affected by HDM sensitization / exposure (>1.5-fold change, p<0.05). Particularly interesting was the finding that mAb1 mediated down-regulation of gene expression for several members of the IL-1 cytokine family, specifically IL1α (2.9-fold), IL-33 (2.6-fold), and IL-18 binding protein (1.5-fold). IL-1β gene expression was not increased in the HDM-induced isotype control-treated group (compared to sham-sensitized mice) but was reduced (1.5-fold) in the mAb1-treated group. Gene expression of the Th1 inflammatory cytokines IL-12β and IFN-γ was also down-regulated by mAb1 compared to the isotype control-treated group. In particular, eight genes encoding chemokine ligands involved in cell homing and trafficking were downregulated in the mAb1-treated group when compared with the isotype control-treated group: Ccl11 (approximately 9-fold decrease), Ccl8 and Cxcl2 (both approximately 5-fold decrease), Cxcl1, Ccl7, Ccl6 (all approximately 3-fold decrease), Ccl2 and Ccl9 (approximately 2-fold decrease).

[0220] conclusion This example demonstrates that blocking IL-4 signaling through type I and type II receptors with anti-IL-4Rα antibodies suppresses inflammatory and fibrotic changes in the lungs of HDM-exposed mice, as well as HDM-driven genetic changes.

[0221] Other embodiments are within the claims.

Claims

1. 1. A pharmaceutical composition comprising an antibody or antigen-binding fragment thereof for use in reducing the incidence of one or more asthma exacerbations in a subject suffering from severe persistent asthma, comprising: The antibody or antigen-binding fragment thereof specifically binds to the interleukin-4 receptor (IL-4R) and comprises the heavy chain complementarity determining region (HCDR) sequences of SEQ ID NOs: 148, 150, and 152 and the light chain complementarity determining region (LCDR) sequences of SEQ ID NOs: 156, 158, and 160; and the pharmaceutical composition is an add-on maintenance treatment administered in combination with a background treatment; and The pharmaceutical composition, wherein the subject has an eosinophil phenotype comprising a blood eosinophil level of at least 300 cells / microliter and / or a sputum eosinophil level of at least 3%.

2. The asthma exacerbation may be: (a) a 30% or greater decrease from baseline in morning peak expiratory flow (PEF) for two consecutive days; (b) 6 or more additional puffs of albuterol or levalbuterol in a 24-hour period (compared to baseline) on 2 consecutive days; and (c) Asthma exacerbation requiring: (i) systemic (oral and / or parenteral) steroid treatment, or (ii) an increase in inhaled corticosteroids of at least four-fold above the last dose received before discontinuation, or (iii) hospitalization, 2. A pharmaceutical composition comprising an antibody or an antigen-binding fragment thereof for use according to claim 1, selected from the group consisting of:

3. The pharmaceutical composition comprising an antibody or antigen-binding fragment thereof for use according to claim 1, wherein the pharmaceutical composition comprises 75 mg to 600 mg of the antibody or antigen-binding fragment thereof.

4. 2. The pharmaceutical composition comprising an antibody or antigen-binding fragment thereof for use according to claim 1, wherein the pharmaceutical composition is administered to a subject at a dosing frequency of once every two weeks.

5. 2. The pharmaceutical composition comprising an antibody or antigen-binding fragment thereof for use according to claim 1, wherein the background treatment is selected from the group consisting of TNF inhibitors, IL-1 inhibitors, IL-5 inhibitors, IL-8 inhibitors, IgE inhibitors, leukotriene inhibitors, corticosteroids, methylxanthines, NSAIDs, nedocromil sodium, cromolyn sodium, long-acting beta 2 agonists and antifungals, or combinations thereof.

6. The pharmaceutical composition comprising an antibody or antigen-binding fragment thereof for use according to claim 1, wherein the antibody or antigen-binding fragment thereof comprises the heavy chain variable region (HCVR) / light chain variable region (LCVR) sequence pair of SEQ ID NO: 162 / 164.

7. 1. A pharmaceutical composition comprising an antibody or antigen-binding fragment thereof for use in improving one or more asthma-related parameters in a subject suffering from severe persistent asthma, comprising: The antibody or antigen-binding fragment thereof specifically binds to the interleukin-4 receptor (IL-4R) and comprises the heavy chain complementarity determining region (HCDR) sequences of SEQ ID NOs: 148, 150, and 152, and the light chain complementarity determining region (LCDR) sequences of SEQ ID NOs: 156, 158, and 160, and said pharmaceutical composition being an add-on maintenance treatment administered in combination with a background treatment; The improvement in the asthma-related parameter is: (a) an increase from baseline in forced expiratory volume in 1 second (FEV1) of at least 0.10 L; (b) an increase from baseline in morning peak expiratory flow (AM PEF) of at least 10.0 L / min; (c) an increase from baseline in nighttime peak expiratory flow (PM PEF) of at least 1.0 L / min; (d) a reduction from baseline in daily albuterol or levalbuterol use of at least one inhalation / day; (e) a reduction from baseline of at least 0.5 points in the 5-item Asthma Control Questionnaire (ACQ5) score; (f) a reduction from baseline in daily measured nocturnal awakenings (number per night) of at least 0.2 awakenings per night; and (g) a reduction of at least 5 points from baseline in the Sinus Nose and Nasal Tract Assessment Test (SNOT-22) score; and The pharmaceutical composition, wherein the subject has an eosinophil phenotype comprising a blood eosinophil level of at least 300 cells / microliter and / or a sputum eosinophil level of at least 3%.

8. The pharmaceutical composition comprising an antibody or antigen-binding fragment thereof for use according to claim 7, wherein the pharmaceutical composition comprises 75 mg to 600 mg of the antibody or antigen-binding fragment thereof.

9. One or more asthma exacerbations in subjects with severe persistent asthma 1. A pharmaceutical composition comprising an antibody or antigen-binding fragment thereof for use in reducing the incidence, or in treating or reducing the incidence of asthma exacerbations, or in improving one or more asthma-related parameters in a subject suffering from severe persistent asthma, comprising: The antibody administered to the subject comprises a single initial dose of the pharmaceutical composition followed by one or more secondary doses: the subject has an eosinophil phenotype comprising a blood eosinophil level of at least 300 cells / microliter and / or a sputum eosinophil level of at least 3%, and the antibody or antigen-binding fragment thereof is prescribed as an add-on maintenance treatment administered in combination with background therapy; The pharmaceutical composition, wherein the antibody or antigen-binding fragment thereof specifically binds to the interleukin-4 receptor (IL-4R) and comprises the heavy chain complementarity determining region (HCDR) sequences of SEQ ID NOs: 148, 150 and 152, and the light chain complementarity determining region (LCDR) sequences of SEQ ID NOs: 156, 158 and 160.

10. 10. The pharmaceutical composition comprising an antibody or antigen-binding fragment thereof for use according to claim 9, wherein the initial dose and the secondary dose of the pharmaceutical composition each comprise 75 mg to 600 mg of the antibody or antigen-binding fragment thereof.

11. The pharmaceutical composition comprising an antibody or antigen-binding fragment thereof for use according to claim 10, wherein each of said secondary doses is administered 1 to 8 weeks after the immediately preceding dose.

12. 1. A pharmaceutical composition comprising an antibody or antigen-binding fragment thereof for use in treating severe persistent asthma in a subject, comprising: The subject suffers from severe persistent asthma and exhibits one or both of a blood eosinophil level of at least 300 cells / microliter and a sputum eosinophil level of at least 3%; The pharmaceutical composition, wherein the antibody or antigen-binding fragment thereof specifically binds to the interleukin-4 receptor (IL-4R) and comprises the heavy chain complementarity determining region (HCDR) sequences of SEQ ID NOs: 148, 150 and 152, and the light chain complementarity determining region (LCDR) sequences of SEQ ID NOs: 156, 158 and 160.

13. 1. A pharmaceutical composition comprising an antibody or antigen-binding fragment thereof for use in reducing or eliminating the dependency of a patient with severe persistent asthma on background therapy comprising inhaled corticosteroids (ICS) and / or long acting beta agonists (LABAs) to treat one or more asthma exacerbations, The subject has severe persistent asthma that is partially controlled or uncontrolled on background asthma therapy, including ICS, LABA, or a combination thereof, and an eosinophil phenotype that includes a blood eosinophil level of at least 300 cells / microliter, and / or a sputum eosinophil level of at least 3%, administering a prescribed dose of the antibody or antigen-binding fragment thereof, formulated to be administered at a prescribed frequency during the initial treatment while maintaining the patient's background asthma therapy during the initial treatment; continuing to administer an antibody or antigen-binding fragment thereof at the prescribed frequency and dosage used during said initial treatment, while gradually reducing or eliminating ICS and / or LABA administered to said subject over a subsequent treatment period; and The pharmaceutical composition, wherein the antibody or antigen-binding fragment thereof specifically binds to the interleukin-4 receptor (IL-4R) and comprises the heavy chain complementarity determining region (HCDR) sequences of SEQ ID NOs: 148, 150 and 152, and the light chain complementarity determining region (LCDR) sequences of SEQ ID NOs: 156, 158 and 160.

14. 14. The pharmaceutical composition comprising an antibody or antigen-binding fragment thereof for use according to claim 13, wherein the ICS is fluticasone, budesonide, or mometasone.

15. 1. A pharmaceutical composition comprising an antibody or antigen-binding fragment thereof for use in treating severe persistent asthma in a patient, comprising: the subject has elevated levels of elevated biomarkers selected from the group consisting of thymus and activation-regulated chemokine (TARC), IgE, eotaxin-3, periostin, carcinoembryonic antigen (CEA), YKL-40, and exhaled nitric oxide (FeNO), the subject having an eosinophil phenotype comprising a blood eosinophil level of at least 300 cells / microliter and / or a sputum eosinophil level of at least 3%; and the antibody or antigen-binding fragment thereof is formulated as an add-on maintenance treatment administered in combination with a background therapy; The pharmaceutical composition, wherein the antibody or antigen-binding fragment thereof specifically binds to the interleukin-4 receptor (IL-4R) and comprises the heavy chain complementarity determining region (HCDR) sequences of SEQ ID NOs: 148, 150 and 152, and the light chain complementarity determining region (LCDR) sequences of SEQ ID NOs: 156, 158 and 160.

16. 1. A pharmaceutical composition comprising an antibody or antigen-binding fragment thereof for use in treating severe persistent asthma in a subject whose asthma is not adequately controlled by moderate to high doses of inhaled corticosteroids and a second controller medication, comprising: The antibody or antigen-binding fragment thereof is administered to a subject in a single initial dose followed by one or more secondary doses; the antibody or antigen-binding fragment thereof specifically binds to the interleukin-4 receptor (IL-4R) and comprises the heavy chain complementarity determining region (HCDR) sequences of SEQ ID NOs: 148, 150 and 152, and the light chain complementarity determining region (LCDR) sequences of SEQ ID NOs: 156, 158 and 160; The antibody or antigen-binding fragment thereof is prescribed as an add-on maintenance treatment to medium to high doses of inhaled corticosteroids (ICS) and a second controller medication; and The pharmaceutical composition, wherein the subject has an eosinophil phenotype comprising a blood eosinophil level of at least 300 cells / microliter and / or a sputum eosinophil level of at least 3%.

17. 17. The pharmaceutical composition comprising an antibody or antigen-binding fragment thereof for use according to claim 16, wherein a systemic corticosteroid is administered to the subject before, after, or simultaneously with the pharmaceutical composition.

18. 17. The pharmaceutical composition comprising an antibody or antigen-binding fragment thereof for use according to claim 16, wherein the initial dose and the secondary dose of the pharmaceutical composition each comprise the same amount of the antibody or antigen-binding fragment thereof.

19. The pharmaceutical composition comprising an antibody or antigen-binding fragment thereof for use according to claim 16, wherein the initial dose and the secondary dose of the pharmaceutical composition are administered once every two weeks.

20. The pharmaceutical composition comprising an antibody or antigen-binding fragment thereof for use according to claim 16, wherein the initial dose and the secondary dose of the pharmaceutical composition are administered once every two weeks.

21. The pharmaceutical composition comprising an antibody or an antigen-binding fragment thereof for use according to claim 1, wherein the pharmaceutical composition is administered systemically, subcutaneously, intravenously or intranasally to a subject.

22. The pharmaceutical composition comprising an antibody or antigen-binding fragment thereof for use according to claim 7, wherein the pharmaceutical composition is administered systemically, subcutaneously, intravenously or intranasally to a subject.

23. The pharmaceutical composition comprising an antibody or antigen-binding fragment thereof for use according to claim 9, wherein the pharmaceutical composition is administered systemically, subcutaneously, intravenously or intranasally to a subject.

24. The background therapy may include TNF inhibitors, IL-1 inhibitors, IL-5 inhibitors, IL-8 inhibitors, IgE inhibitors, leukotriene inhibitors, corticosteroids, methylxanthines, NSAIDs, nedocromil sodium, cromolyn sodium, long-acting beta 2 10. A pharmaceutical composition comprising an antibody or an antigen-binding fragment thereof for use according to claim 9, selected from the group consisting of agonists and antifungal agents, or combinations thereof.

25. 10. The pharmaceutical composition comprising an antibody or antigen-binding fragment thereof for use according to claim 9, wherein at least eight secondary doses of the antibody or antigen-binding fragment thereof are administered to the subject, each of the secondary doses being administered two weeks after the immediately preceding dose.

26. 14. The pharmaceutical composition comprising an antibody or antigen-binding fragment thereof for use according to claim 13, wherein the LABA is salmeterol or formoterol.

27. 14. The pharmaceutical composition comprising an antibody or antigen-binding fragment thereof for use according to claim 13, wherein the ICS / LABA combination is fluticasone / salmeterol, budesonide / formoterol, or budesonide / formoterol.

28. 14. The pharmaceutical composition comprising an antibody or antigen-binding fragment thereof for use according to claim 13, wherein background therapy includes LABA and ICS, or LABA only, and the patient eliminates doses of LABA at the end of the subsequent treatment period.

29. 14. The pharmaceutical composition comprising an antibody or antigen-binding fragment thereof for use according to claim 13, wherein the dosage of LABA and / or ICS is gradually reduced or eliminated over a period of 2 to 8 weeks.

30. 1. A pharmaceutical composition comprising an antibody or antigen-binding fragment thereof for use in reducing the incidence of one or more asthma exacerbations in a subject suffering from severe persistent asthma, comprising: The antibody or antigen-binding fragment thereof specifically binds to the interleukin 4 receptor (IL-4R) and comprises a heavy chain variable region (HCVR) / light chain variable region (LCVR) having the sequence pair SEQ ID NO: 162 / 164; The antibody or antigen-binding fragment thereof is formulated as an add-on maintenance treatment administered in combination with a background therapy; and The pharmaceutical composition, wherein the subject has an eosinophil phenotype comprising a blood eosinophil level of at least 300 cells / microliter and / or a sputum eosinophil level of at least 3%.

31. 22. The pharmaceutical composition comprising an antibody or antigen-binding fragment thereof for use according to claim 21, wherein the pharmaceutical composition is administered subcutaneously by needle and syringe, a pen delivery device, or an auto-injector.

32. 23. The pharmaceutical composition comprising an antibody or antigen-binding fragment thereof for use according to claim 22, wherein the pharmaceutical composition is administered subcutaneously by needle and syringe, a pen delivery device, or an auto-injector.

33. 24. The pharmaceutical composition comprising an antibody or antigen-binding fragment thereof for use according to claim 23, wherein the pharmaceutical composition is administered subcutaneously by needle and syringe, a pen delivery device, or an auto-injector.

34. The antibody or antigen-binding fragment thereof comprises the heavy chain variable region (HCVR) / light chain variable region (LCVR) sequence pair of SEQ ID NO: 162 / 164; A pharmaceutical composition comprising an antibody or an antigen-binding fragment thereof for use as claimed in claim 1.

35. The pharmaceutical composition comprises 75 mg to 600 mg of the antibody or antigen-binding fragment thereof. A pharmaceutical composition comprising an antibody or an antigen-binding fragment thereof for use according to any one of claims 12 to 34.

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