Biopharmaceutical compositions and methods for pediatric patients

Abstract

The present disclosure relates to compositions and related methods for treating interleukin 5 (IL-5) mediated diseases in a pediatric subject.

Description

[0001] This application is a divisional application of the invention patent application with the application date of May 8, 2018, application number 201880051204.4, and invention name “Biopharmaceutical compositions and methods for pediatric patients”. Technical Field

[0002] The present disclosure relates to compositions and related methods for treating interleukin 5 (IL-5) mediated diseases in pediatric patients. Background Art

[0003] IL-5 is a secretory protein. IL-5 plays a role in many different diseases, such as asthma, mild asthma, moderate asthma, severe asthma, mild eosinophilic asthma, moderate eosinophilic asthma, severe eosinophilic asthma, uncontrolled eosinophilic asthma, eosinophilic asthma, subeosinophilic asthma, chronic obstructive pulmonary disease, eosinophilic granulomatosis with polyangiitis, hypereosinophilic syndrome, nasal polyps, bullous pemphigoid, eosinophilic esophagitis, atopic dermatitis, moderate atopic dermatitis, and severe atopic dermatitis. These serious diseases affect hundreds of millions of people worldwide, including pediatric patients.

[0004] This means that there is a need for compositions suitable for treating IL-5 mediated diseases in pediatric patients.Such compositions and related methods are provided by the present disclosure. Summary of the Invention

[0005] One aspect of the present disclosure is a method of treating a disease in a pediatric subject comprising the steps of: a) identifying a pediatric subject weighing less than 40 kg having a disease selected from the group consisting of asthma, mild asthma, moderate asthma, severe asthma, mild eosinophilic asthma, moderate eosinophilic asthma, severe eosinophilic asthma, uncontrolled eosinophilic asthma, eosinophilic asthma, subeosinophilic asthma, chronic obstructive pulmonary disease, eosinophilic granulomatosis with polyangiitis, hypereosinophilic syndrome, nasal polyposis, bullous pemphigoid, eosinophilic esophagitis, atopic dermatitis, moderate atopic dermatitis, and severe atopic dermatitis; and b) subcutaneously administering to the pediatric subject a therapeutically effective amount of an antibody comprising the heavy chain amino acid sequence set forth in SEQ ID NO: 1 and the light chain amino acid sequence set forth in SEQ ID NO: 2, wherein the therapeutically effective amount of the antibody provides about 10.1960 ± 10.1960 % of the total dose. A maximum plasma concentration (Cmax) of the antibody of 0.3345 μg / mL, and an area under the curve [0-infinity] value of approximately 454.39 ± 15.8876 μg*day / mL; thereby treating the disease in pediatric subjects.

[0006] Another aspect of the present disclosure is a method of treating a disease in a pediatric subject comprising the steps of: a) identifying a pediatric subject weighing greater than or equal to 40 kg who has a disease selected from the group consisting of asthma, mild asthma, moderate asthma, severe asthma, mild eosinophilic asthma, moderate eosinophilic asthma, severe eosinophilic asthma, uncontrolled eosinophilic asthma, eosinophilic asthma, subeosinophilic asthma, chronic obstructive pulmonary disease, eosinophilic granulomatosis with polyangiitis, hypereosinophilic syndrome, nasal polyposis, bullous pemphigoid, eosinophilic esophagitis, atopic dermatitis, moderate atopic dermatitis, and severe atopic dermatitis; and b) subcutaneously administering to the pediatric subject a therapeutically effective amount of an antibody comprising the heavy chain amino acid sequence set forth in SEQ ID NO: 1 and the light chain amino acid sequence set forth in SEQ ID NO: 2, wherein the therapeutically effective amount of the antibody provides approximately 16.3412 ± 0.6364 A maximum plasma concentration (Cmax) of the antibody of about 675.20 ± 35.8980 μg / mL, and an area under the curve [0-infinity] value of about 675.20 ± 35.8980 μg*day / mL; thereby treating the disease in pediatric subjects.

[0007] Another aspect of the present disclosure is a method of reducing the absolute blood eosinophil count in a pediatric subject comprising the steps of: a) identifying a pediatric subject weighing less than 40 kg who has a condition selected from the group consisting of asthma, mild asthma, moderate asthma, severe asthma, mild eosinophilic asthma, moderate eosinophilic asthma, severe eosinophilic asthma, uncontrolled eosinophilic asthma, eosinophilic asthma, subeosinophilic asthma, chronic obstructive pulmonary disease, eosinophilic granulomatosis with polyangiitis, hypereosinophilic syndrome, nasal polyposis, bullous pemphigoid, eosinophilic esophagitis, and atopic dermatitis; and b) subcutaneously administering to the pediatric subject a therapeutically effective amount of an antibody comprising the heavy chain amino acid sequence set forth in SEQ ID NO: 1 and the light chain amino acid sequence set forth in SEQ ID NO: 2, wherein the therapeutically effective amount of the antibody provides approximately 10.1960 ± 0.3345 The invention provides a maximum plasma concentration (Cmax) of the antibody of about 1.5 μg / mL and an area under the curve [0-infinity] value of about 454.39 ± 15.8876 μg*day / mL; thereby reducing the absolute blood eosinophil count in pediatric subjects.

[0008] Another aspect of the present disclosure is a method of reducing the absolute blood eosinophil count in a pediatric subject comprising the steps of: a) identifying a pediatric subject weighing greater than or equal to 40 kg who has a condition selected from the group consisting of asthma, mild asthma, moderate asthma, severe asthma, mild eosinophilic asthma, moderate eosinophilic asthma, severe eosinophilic asthma, uncontrolled eosinophilic asthma, eosinophilic asthma, subeosinophilic asthma, chronic obstructive pulmonary disease, eosinophilic granulomatosis with polyangiitis, hypereosinophilic syndrome, nasal polyposis, bullous pemphigoid, eosinophilic esophagitis, and atopic dermatitis; and b) subcutaneously administering to the pediatric subject a therapeutically effective amount of an antibody comprising the heavy chain amino acid sequence set forth in SEQ ID NO: 1 and the light chain amino acid sequence set forth in SEQ ID NO: 2, wherein the therapeutically effective amount of the antibody provides about 16.3412 ± 16.3412 eosinophils. A maximum plasma concentration (Cmax) of the antibody of 0.6364 μg / mL, and an area under the curve [0-infinity] value of approximately 675.20 ± 35.8980 μg*day / mL; thereby reducing absolute blood eosinophil counts in pediatric subjects.

[0009] Another aspect of the disclosure is a composition for treating a disease selected from the group consisting of asthma, mild asthma, moderate asthma, severe asthma, mild eosinophilic asthma, moderate eosinophilic asthma, severe eosinophilic asthma, uncontrolled eosinophilic asthma, eosinophilic asthma, subeosinophilic asthma, chronic obstructive pulmonary disease, eosinophilic granulomatosis with polyangiitis, hypereosinophilic syndrome, nasal polyposis, bullous pemphigoid, eosinophilic esophagitis, atopic dermatitis, moderate atopic dermatitis, and severe atopic dermatitis in a pediatric subject weighing less than 40 kg, wherein the composition comprises an antibody comprising the heavy chain amino acid sequence set forth in SEQ ID NO: 1 and the light chain amino acid sequence set forth in SEQ ID NO: 2, and a therapeutically effective amount of the antibody administered subcutaneously to the pediatric subject provides approximately 10.1960 ± 0.3345 The maximum plasma concentration (Cmax) of the antibody was approximately 454.39 ± 15.8876 μg*day / mL, and the area under the curve [0-infinity] value was approximately 454.39 ± 15.8876 μg*day / mL.

[0010] Another aspect of the disclosure is a composition for treating a disease selected from the group consisting of asthma, mild asthma, moderate asthma, severe asthma, mild eosinophilic asthma, moderate eosinophilic asthma, severe eosinophilic asthma, uncontrolled eosinophilic asthma, eosinophilic asthma, subeosinophilic asthma, chronic obstructive pulmonary disease, eosinophilic granulomatosis with polyangiitis, hypereosinophilic syndrome, nasal polyposis, bullous pemphigoid, eosinophilic esophagitis, atopic dermatitis, moderate atopic dermatitis, and severe atopic dermatitis in a pediatric subject weighing greater than or equal to 40 kg, wherein the composition comprises an antibody comprising the heavy chain amino acid sequence set forth in SEQ ID NO: 1 and the light chain amino acid sequence set forth in SEQ ID NO: 2, and a therapeutically effective amount of the antibody administered subcutaneously to the pediatric subject provides approximately 16.3412 ± 0.6364 The maximum plasma concentration (Cmax) of the antibody was 1.5 μg / mL, and the area under the curve [0-infinity] value was approximately 675.20±35.8980 μg*day / mL.

[0011] Another aspect of the present disclosure is a composition for reducing the absolute blood eosinophil count in a pediatric subject weighing less than 40 kg, wherein the pediatric subject has a disease selected from the group consisting of asthma, mild asthma, moderate asthma, severe asthma, mild eosinophilic asthma, moderate eosinophilic asthma, severe eosinophilic asthma, uncontrolled eosinophilic asthma, eosinophilic asthma, subeosinophilic asthma, chronic obstructive pulmonary disease, eosinophilic granulomatosis with polyangiitis, hypereosinophilic syndrome, nasal polyposis, bullous pemphigoid, eosinophilic esophagitis, atopic dermatitis, moderate atopic dermatitis, and severe atopic dermatitis, and the composition comprises an antibody comprising the heavy chain amino acid sequence set forth in SEQ ID NO: 1 and the light chain amino acid sequence set forth in SEQ ID NO: 2, and a therapeutically effective amount of the antibody administered subcutaneously to the pediatric subject provides approximately 10.1960 ± 0.3345 The maximum plasma concentration (Cmax) of the antibody was approximately 454.39 ± 15.8876 μg*day / mL, and the area under the curve [0-infinity] value was approximately 454.39 ± 15.8876 μg*day / mL.

[0012] Another aspect of the present disclosure is a composition for reducing the absolute blood eosinophil count in a pediatric subject weighing greater than or equal to 40 kg, wherein the pediatric subject has a disease selected from the group consisting of asthma, mild asthma, moderate asthma, severe asthma, mild eosinophilic asthma, moderate eosinophilic asthma, severe eosinophilic asthma, uncontrolled eosinophilic asthma, eosinophilic asthma, subeosinophilic asthma, chronic obstructive pulmonary disease, eosinophilic granulomatosis with polyangiitis, hypereosinophilic syndrome, nasal polyposis, bullous pemphigoid, eosinophilic esophagitis, atopic dermatitis, moderate atopic dermatitis, and severe atopic dermatitis, and the composition comprises an antibody comprising the heavy chain amino acid sequence set forth in SEQ ID NO: 1 and the light chain amino acid sequence set forth in SEQ ID NO: 2, and a therapeutically effective amount of the antibody administered subcutaneously to the pediatric subject provides approximately 16.3412 ± 0.6364 The maximum plasma concentration (Cmax) of the antibody was 1.5 μg / mL, and the area under the curve [0-infinity] value was approximately 675.20±35.8980 μg*day / mL. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 :box Figure 1-1 Diagnostic flowcharts for clinical practice – a preliminary introduction.

[0014] Figure 2 :box Figure 3 -5. A step-by-step approach to controlling symptoms and minimizing future risks.

[0015] Figure 3 : Ratio of blood eosinophil count to baseline (geometric mean 95% confidence interval). DETAILED DESCRIPTION

[0016] The present disclosure provides compositions and methods for treating interleukin 5 (IL-5)-mediated diseases in pediatric patients, and related subject matter.

[0017] As used herein, the term "asthma" refers to an inflammatory disease of the airways characterized by reversible airway obstruction and bronchospasm. Common symptoms include wheezing, coughing, chest tightness, and shortness of breath. Asthma is a heterogeneous disease generally characterized by chronic airway inflammation. It is defined by a history of respiratory symptoms such as wheezing, shortness of breath, chest tightness, and coughing that vary in intensity over time, along with variable expiratory airflow limitation.

[0018] In the methods of the present disclosure, asthma diagnosis in a subject can be performed according to the guidelines provided by the Global Initiative for Asthma (GINA), Global Strategy for Asthma Management and Prevention (2016 Update) document. Those of ordinary skill in the art will be familiar with the GINA clinical practice diagnostic flowchart shown below ( Figure 1 ), as well as diagnostic criteria for asthma in adults, adolescents, and children aged 6-11 years (Table 1), as well as other aspects of the guidelines (e.g., regarding pregnant women, etc.). See also Tables 2 and 3.

[0019] In the methods of the present disclosure, "asthma" can be "mild asthma," "moderate asthma," or "severe asthma." In the methods of the present disclosure, asthma severity can be assessed according to the GINA guidelines. In particular, asthma severity can be assessed retrospectively based on the level of treatment required to control symptoms and acute exacerbations. For example, an assessment can be made once a patient has been on controller therapy for several months, and, if appropriate, a step-down of treatment has been attempted to find the patient's lowest effective treatment level. Asthma severity is not a static characteristic and can change over months or years.

[0020] When patients have been taking conventional controller therapy for several months, asthma severity can be assessed: • “Mild asthma” is treated with Step 1 or Step 2 (see Figure 2 ;box Figure 3 -5), that is, asthma that is well controlled with either a single, as-needed reliever medication or with a low-intensity controller such as a low-dose ICS, a leukotriene receptor antagonist, or a chromone. • "Moderate asthma" is treated with Step 3 (see Figure 2 ;box Figure 3 -5), such as well-controlled asthma with low-dose ICS / LABA. • “Severe asthma” is defined as requiring Step 4 or Step 5 treatment (see Figure 2 ;box Figure 3-5), such as high-dose ICS / LABA, to prevent them from becoming 'uncontrolled' asthma, or maintaining 'uncontrolled' asthma despite such treatment. Although many patients with uncontrolled asthma may be difficult to treat due to inadequate or inappropriate treatment, or long-term compliance problems or comorbidities (such as chronic rhinosinusitis or obesity), the European Respiratory Society / American Thoracic Society Task Force on Severe Asthma believes that the definition of "severe asthma" should be retained for patients with refractory asthma and those in whom the response to treatment of comorbidities is incomplete. Table 4 can also be used as a reference in the asthma severity assessment process.

[0021] In the methods of the present disclosure, "asthma" can be "mild eosinophilic asthma," "moderate eosinophilic asthma," or "severe eosinophilic asthma."

[0022] "Mild eosinophilic asthma" is mild asthma with an eosinophilic phenotype. For example, a subject with mild eosinophilic asthma may have mild asthma and blood eosinophils greater than or equal to 150 eosinophils / μL of blood in the past 12 months, greater than or equal to 200 eosinophils / μL of blood in the past 12 months, greater than or equal to 300 eosinophils / μL of blood in the past 12 months, or greater than or equal to 350 eosinophils / μL of blood in the past 12 months.

[0023] "Moderate eosinophilic asthma" is moderate asthma with an eosinophilic phenotype. For example, a subject with moderate eosinophilic asthma may have moderate asthma and blood eosinophils greater than or equal to 150 eosinophils / μL of blood in the past 12 months, greater than or equal to 200 eosinophils / μL of blood in the past 12 months, greater than or equal to 300 eosinophils / μL of blood in the past 12 months, or greater than or equal to 350 eosinophils / μL of blood in the past 12 months.

[0024] "Severe eosinophilic asthma" is severe asthma with an eosinophilic phenotype. For example, a subject with severe eosinophilic asthma may have severe asthma and blood eosinophils greater than or equal to 150 eosinophils / μL of blood in the past 12 months, greater than or equal to 200 eosinophils / μL of blood in the past 12 months, greater than or equal to 300 eosinophils / μL of blood in the past 12 months (preferred), or greater than or equal to 350 eosinophils / μL of blood in the past 12 months.

[0025] A subject with severe eosinophilic asthma may also meet one or more of the criteria described in Table 5.

[0026] Importantly, subjects with severe eosinophilic asthma according to these criteria may have less than 150 eosinophils / μL of blood at the start of treatment.

[0027] Mepolizumab is a monoclonal antibody comprising the heavy chain amino acid sequence set forth in SEQ ID NO: 1 and the light chain amino acid sequence set forth in SEQ ID NO: 2. Mepolizumab and antigen binding proteins, particularly antibody molecules, comprising the heavy chain CDRs and light chain CDRs of mepolizumab (e.g., SEQ ID NOs: 5-10), or the heavy chain variable region and light chain variable region of mepolizumab (e.g., SEQ ID NO: 3 and SEQ ID NO: 4), can be used to treat patients according to the methods of the present disclosure. Mepolizumab specifically binds to human IL-5 (SEQ ID NO: 11) and antagonizes the activity of IL-5R receptors (including SEQ ID NOs: 12 and 13). NUCALA™ is an example of an FDA-approved pharmaceutical composition comprising mepolizumab.

[0028] Mepolizumab or a composition of the present disclosure comprising mepolizumab may be indicated for add-on maintenance treatment of severe eosinophilic asthma in pediatric patients as identified by blood eosinophils greater than or equal to 300 cells / μL in the past 12 months, and / or blood eosinophils greater than or equal to 150 cells / μL at the start of treatment, and / or blood eosinophils less than 150 cells / μL at the start of treatment. Alternatively, mepolizumab or a composition of the present disclosure comprising mepolizumab may be indicated for add-on maintenance treatment of severe eosinophilic asthma in pediatric patients as identified by blood eosinophils greater than or equal to 300 cells / μL in the past 12 months, and / or blood eosinophils greater than or equal to 150 cells / μL at the start of treatment. Mepolizumab or a composition comprising mepolizumab of the present disclosure may be indicated for additional maintenance treatment of severe eosinophilic asthma in pediatric patients, as identified by blood eosinophils greater than or equal to 300 cells / μL in the past 12 months, and / or blood eosinophils less than 150 cells / μL at the start of treatment. Such pediatric patients may be between 11 and 6 years old (inclusive). Treatment with mepolizumab or a composition comprising mepolizumab of the present disclosure may reduce acute exacerbations of asthma in pediatric patients (e.g., patients with a history of acute exacerbations). When treatment with mepolizumab or a composition comprising mepolizumab of the present disclosure is indicated, the methods of the present disclosure may be used (i.e., such treatment with mepolizumab may be combined with the methods of the present disclosure). Treatment with mepolizumab or a composition comprising mepolizumab of the present disclosure may: a) Produces a reduction in the frequency of acute exacerbations. Compared to placebo, treatment with mepolizumab or a composition comprising mepolizumab of the present disclosure can reduce the rate of: 1) clinically significant acute exacerbations, 2) acute exacerbations requiring hospitalization or ED visits, and 3) acute exacerbations requiring hospitalization. This benefit could potentially lead to a reduction in morbidity and fatal events due to asthma. b) Producing a reduction in daily OCS dose: Treatment with mepolizumab or a composition comprising mepolizumab of the present disclosure can allow subjects to reduce their daily dose of concomitant corticosteroids without experiencing loss of asthma control. Compared to subjects treated with placebo, subjects treated with mepolizumab or a composition comprising mepolizumab of the present disclosure can achieve a median percentage reduction from baseline in daily oral corticosteroid (OCS) dose. In addition, subjects treated with mepolizumab or a composition comprising mepolizumab of the present disclosure, or an antigen-binding composition of the present disclosure, can achieve a reduction in OCS dose compared to subjects treated with placebo. c) Producing improvements in lung function: Clinically relevant changes in pre- and post-bronchodilator FEV1 can be demonstrated by treatment with mepolizumab or a composition of the disclosure comprising mepolizumab compared to placebo. Any improvement in lung function is of particular clinical importance in this subject population, as most are on maximal asthma therapy including high-dose ICS (inhaled corticosteroids) and / or OCS plus controller medication. d) produces an improvement in asthma control: A statistically significant and clinically relevant improvement can be observed in the ACQ-5 or ACQ-7 with mepolizumab or a composition of the disclosure comprising mepolizumab compared to placebo, indicating that subjects can achieve asthma control by adding mepolizumab or a composition of the disclosure comprising mepolizumab to their existing asthma treatment. e) Producing improvements in quality of life: Demonstrating statistically significant and clinically relevant changes in SGRQ scores with mepolizumab or a composition of the disclosure comprising mepolizumab compared to placebo. Subjects can experience significant improvements in asthma symptoms and the ability to perform daily activities. f) Efficacy and durability of pharmacodynamic effects: Sustained reductions in asthma exacerbations and blood eosinophils, as well as improvements in lung function, asthma control, and quality of life, are observed over a period of treatment duration (e.g., 4, 8, 9, 12, 16, and 20 weeks), without the development of tolerance. and g) Producing a Reduction in Blood Eosinophils. Treatment with a composition comprising mepolizumab, or a composition of the disclosure comprising mepolizumab, can result in a rapid reduction in blood eosinophils in a subject.

[0029] In the methods of the present disclosure, "asthma" can be "severe asthma." In the methods of the present disclosure, "asthma" can also be "mild asthma," "moderate asthma," "severe asthma," "mild eosinophilic asthma," "moderate eosinophilic asthma," or "severe eosinophilic asthma," as discussed above. Treatment with a composition comprising mepolizumab or a composition comprising mepolizumab of the present disclosure can be used to treat these conditions according to the methods of the present disclosure.

[0030] In the methods of the present disclosure, “asthma” can be “uncontrolled eosinophilic asthma.” A subject with uncontrolled eosinophilic asthma meets the criteria described in Table 6.

[0031] Treatment with a composition comprising mepolizumab, or a composition of the disclosure comprising mepolizumab, can be used to treat uncontrolled eosinophilic asthma according to the methods of the disclosure.

[0032] In the methods of the present disclosure, "asthma" can be "eosinophilic asthma." A subject with eosinophilic asthma meets the criteria described in Table 7.

[0033] In the methods of the present disclosure, “asthma” may be “subeosinophilic asthma.” A subject with subeosinophilic asthma meets the criteria described in Table 8.

[0034] Treatment with a composition comprising mepolizumab, or a composition of the disclosure comprising mepolizumab, can be used to treat eosinophilic asthma according to the methods of the disclosure, and can also be used to treat subeosinophilic asthma.

[0035] As used herein, the term "bullous pemphigoid" (BP) refers to an acute or chronic autoimmune skin disease involving blisters, more appropriately referred to as bullous formations, in the space between the skin layers of the epidermis and dermis. BP is the most common autoimmune blistering skin disease. It characteristically affects the elderly (>70 years old), with an annual incidence of 5 to 35 per million. The incidence of BP is increasing rapidly with an average annual rate of 17%. BP often begins with extremely itchy skin lesions, similar to eczema or hives, before the appearance of vesicles and blisters. In 10%-30% of patients, BP also involves the oral mucosa. Disease severity can be determined using the Autoimmune Bullous Skin Condition Intensity Score (ABSIS), which assesses the affected area and disease activity. The disease is caused by an autoimmune response to structural components of the junction-adhesion complex, resulting in damage to the dermal-epidermal junction accompanied by subepidermal blister formation. Specifically, autoreactive B and T cell responses to the hemidesmosome antigens BP180 and BP230 have been identified. Serum levels of autoantibodies against BP180 reflect the severity and activity of the disease. T cells are memory CD4+ cells that produce both Th1 and Th2 cytokines, primarily IL-4, IL-5, and IL-13. IL-5 and eotaxin are abundantly found in blister fluid. IL-5 production is indeed associated with significant eosinophil infiltration in the skin of patients with BP and blood eosinophilia. Eosinophils are believed to be crucially involved in blister formation by releasing toxic granule proteins (ESP, MBP) and proteolytic enzymes.

[0036] As used herein, the term "eosinophilic esophagitis" (EoE) refers to an allergic inflammatory condition of the esophagus involving eosinophils. Symptoms are difficulty swallowing, food impaction, and heartburn. EoE is characterized by dense infiltration of the epithelial lining of the esophagus by white blood cells of the eosinophil type. Based on the important role that eosinophils play in allergic reactions, EoE is considered an allergic reaction to ingested food. An EoE diagnostic panel can be used to diagnose EoE. EoE can also be diagnosed if gastroesophageal reflux does not respond to a 6-week trial of high-dose proton pump inhibitors (PPIs) twice a day, or if negative dynamic pH values ​​rule out gastroesophageal reflux disease (GERD). Endoscopically, ridges, grooves, or rings can be seen on the esophageal wall. Sometimes, multiple rings can appear in the esophagus, leading to the terms "corrugated esophagus" or "cat esophagus" due to the rings' similarity to a cat's esophagus. The presence of white exudate in the esophagus is also suggestive of the diagnosis. When a biopsy is taken during endoscopy, numerous eosinophils are usually seen in the superficial epithelium. A minimum of 15 eosinophils per high-power field is required to make the diagnosis. The eosinophilic inflammation is not limited to the esophagus alone but extends throughout the gastrointestinal tract. There may also be prominent degranulated eosinophils, as may microabscesses and dilated basal lamina. Radiologically, the term "annular esophagus" has been used to describe the appearance of eosinophilic esophagitis on barium swallow studies, in contrast to the transient transverse folds sometimes seen with esophageal reflux (called "cat esophagus").

[0037] Treatment with a composition comprising mepolizumab, or a composition of the disclosure comprising mepolizumab, can be used to treat COPD according to the methods of the disclosure.

[0038] Subjects with "chronic obstructive pulmonary disease" (COPD) may meet one or more of the following criteria: a) Previous COPD diagnosis: Subjects with a clinically documented history of COPD of at least 1 year, as defined by the American Thoracic Society / European Respiratory Society; b) COPD severity: Subjects may present with the following: a measured forced expiratory volume in one second / forced vital capacity (FEV1 / FVC) ratio <0.70 before and after albuterol administration to confirm the diagnosis of COPD; a measured post-albuterol FEV1 >20 percentile and <=80 percentile of predicted normal values, calculated using the National Health and Nutrition Examination Survey (NHANES) III reference equation; c) Exacerbation history: A well-documented history of the following within 12 months (as verified by medical records): at least two moderate COPD exacerbations. Moderate is defined as the use of systemic corticosteroids (IM, intravenous, or oral) and / or treatment with antibiotics, or at least one severe COPD exacerbation. Severe is defined as requiring hospitalization. NOTE: Subjects must have experienced at least one exacerbation while taking an inhaled corticosteroid (ICS) plus a long-acting beta2 agonist (LABA) plus a long-acting muscarinic antagonist (LAMA). NOTE: Prior use of antibiotics alone does not qualify as a moderate exacerbation unless used specifically to treat exacerbations of COPD; and d) Concomitant COPD therapy: Well-documented need for optimized standard of care (SoC) background therapy, which includes ICS plus two additional COPD medications (i.e., triple therapy) for the prior 12 months, and meets the following criteria: Minimum 3 months of inhaled corticosteroid use (at a dose of >= 500 micrograms (mcg) / day of fluticasone propionate equivalents plus) immediately prior to visit to a healthcare provider; or LABA and LAMA.

[0039] Treatment with a composition comprising mepolizumab, or a composition of the disclosure comprising mepolizumab, can be used to treat COPD according to the methods of the disclosure.

[0040] As used herein, the term "eosinophilic granulomatosis with polyangiitis" (EGPA) refers to an autoimmune condition that causes inflammation of small and medium-sized blood vessels (vasculitis) in people with a history of allergic hypersensitivity (atopy) of the airways. EGPA may also be called Churg-Strauss Syndrome (CSS) or allergic granulomatosis. EGPA typically manifests in three stages. The early (prodromal) stage is marked by airway inflammation; almost all patients experience asthma and / or allergic rhinitis. The second stage is characterized by abnormally high numbers of eosinophils (hypereosinophilia), which cause tissue damage, most commonly to the lungs and digestive tract. The third stage consists of vasculitis, which can ultimately lead to cell death and can be life-threatening.

[0041] A subject with EGPA may meet one or more of the following criteria: a) asthma; b) a blood eosinophil level greater than 10% of a differential white blood cell count; c) the presence of mononeuropathy or polyneuropathy; d) unfixed pulmonary infiltrates; e) the presence of paranasal sinus abnormalities; and e) histological evidence of extravascular eosinophils. For classification purposes, a patient should be said to have EGPA if at least four of the six aforementioned criteria are positive.

[0042] Treatment with a composition comprising mepolizumab or a composition of the present disclosure comprising mepolizumab can be used to treat EGPA according to the methods of the present disclosure. The composition of the present disclosure can be administered to pediatric EGPA patients in an amount of 300 mg once every 4 weeks.

[0043] As used herein, the term "hypereosinophilic syndrome" (HES) refers to a disease characterized by persistently elevated eosinophil counts (≥ 1500 eosinophils / mm³) in the blood for at least six months without any identifiable cause, accompanied by involvement of the heart, nervous system, or bone marrow.

[0044] A subject with hypereosinophilic syndrome may meet one or more of the following criteria: a) a documented history of hypereosinophilic syndrome; b) a blood eosinophil count greater than 1500 cells for six months; c) signs and symptoms of organ system involvement; and d) no evidence of parasitic, allergic, or other causes of eosinophilia after a comprehensive evaluation.

[0045] Treatment with a composition comprising mepolizumab, or a composition of the disclosure comprising mepolizumab, can be used to treat hypereosinophilic syndrome according to the methods of the disclosure.

[0046] As used herein, the term "nasal polyps" means a disease characterized by the presence of nasal polyps, such polyps may be in the upper nasal cavity and / or may arise within the ostiomeatal complex.

[0047] A subject with nasal polyps may meet one or more of the following criteria: a) a documented history of nasal polyps; or b) nasal polyps evident on examination (eg, endoscopy).

[0048] Treatment with a composition comprising mepolizumab, or a composition of the disclosure comprising mepolizumab, can be used to treat nasal polyps according to the methods of the disclosure.

[0049] As used herein, the term "atopic dermatitis" means an inflammatory skin condition characterized by chronic itching, lichenification, xerosis, erythematous papules and plaques.

[0050] In the methods of the present disclosure, “atopic dermatitis” may be “moderate to severe atopic dermatitis.” A subject with moderate to severe atopic dermatitis may meet one or more of the criteria described in Table 9.

[0051] Subjects with moderate to severe atopic dermatitis can be children under 18 years of age, adults at least 18 years of age or older, adults between 18 and 70 years of age (inclusive), or pediatric subjects between 11 and 6 years of age (inclusive). Subjects can be male or female. Preferably, female subjects to be treated are not pregnant, not breastfeeding, and / or are unlikely to become pregnant.

[0052] The diagnosis of atopic dermatitis was based on the Hanifin and Rajka criteria modified by Eichenfield. See Table 10 and Eichenfield et al., 70 J Am Acad Dermatol 338 (2014).

[0053] The Health Care Professional's Global Assessment (HGA) is a clinical tool used to assess the current state / severity of a subject's atopic dermatitis. See Rehal et al., 6 PLos ONE e17520 (2011) and Table 11. It is a static 5-point morphological assessment of overall disease severity, as determined by a trained health care professional using the clinical features of erythema, infiltrate, papule, exudate, and crusting as a guide. The HGA is performed without reference to previous scores. Each assessment should be performed as a visual 'average' of the severity of all affected areas at the time of assessment.

[0054] The evaluation of body surface area percentage (% BSA) is the estimated amount of the total skin percentage involved about atopic dermatitis.Referring to Table 12.% BSA evaluation can be performed by checking the inflamed area from each of 4 body surface areas respectively: head and neck, upper limbs, trunk and lower limbs, and each in these body areas can potentially have up to 100% involvement.Assessor (for example, health care professional) estimates the skin percentage involved about each area for % BSA area scoring, and then multiplies it by appropriate ratio multiplier, to draw % BSA area involvement value (for the experimenter of ≥8 years old, head 0.1, upper limb 0.2, trunk 0.3 and lower limb 0.4).Regional % BSA involvement value is summed, to generate total involvement % BSA.Regional % BSA area scoring is also used as the part of matrix, to calculate EASI scoring.

[0055] The EASI scoring system is a standardized clinical tool for evaluating atopic dermatitis that takes into account the overall extent of the affected % body surface area (% BSA) and the severity score of each clinical sign: erythema, induration / papule, exfoliation, and lichenification. See Hanifin et al., 10 Exp Dermatol 11 (2001); Rullo et al., 36 Allergol et Immunopathol 201 (2008) and Table 13. The % BSA area score from the % BSA evaluation will be used as part of the matrix to calculate the EASI score. For each of the 4 body regions (head and neck, upper limbs, lower limbs, and trunk), a severity score for each clinical sign (erythema, induration / papule, exfoliation, and lichenification) is rated on a 4-point scale (0 to 3). The severity scores for each sign for each region are summed and then multiplied by the % BSA area score and the appropriate scale multiplier (0.1 for the head, 0.2 for the upper extremities, 0.3 for the trunk, and 0.4 for the lower extremities for subjects ≥8 years of age) to generate the regional EASI score. The regional EASI scores are then summed to arrive at the final EASI score. The EASI score is a static assessment made without reference to previous scores.

[0056] A therapeutically effective amount of mepolizumab or a composition comprising mepolizumab of the present disclosure can be used to treat patients suffering from atopic dermatitis or to reduce the absolute blood eosinophil count in such patients. Such atopic dermatitis can be moderate atopic dermatitis or severe atopic dermatitis.

[0057] According to the methods of the present disclosure, treating atopic dermatitis, such as moderate atopic dermatitis or severe atopic dermatitis, with mepolizumab or a composition comprising mepolizumab of the present disclosure can produce at least one result selected from the following: a) an HGA score of 0 or 1, and at least a 2-grade improvement in HGA (e.g., relative to the starting HGA score); b) a decrease in the Eczema Area and Severity Index (EASI) score (e.g., relative to the starting EASI score); c) a decrease in the percentage of total body surface area (% BSA) affected (e.g., relative to the starting % BSA); and / or d) Subjects were determined by a healthcare professional not to have atopic dermatitis according to the Hanifin and Rajka criteria modified by Eichenfield (Eichenfield et al., 70 J Am Acad Dermatol 338 (2014)).

[0058] The EASI score after treatment according to the methods of the present disclosure can be less than 16, for example, from about 0 to less than 16. The EASI score after treatment can also be from about 0 to about 15, from about 0 to about 14, from about 0 to about 13, from about 0 to about 12, from about 0 to about 11, from about 0 to about 10, from about 0 to about 9, from about 0 to about 8, from about 0 to about 7, from about 0 to about 6, from about 0 to about 5, from about 0 to about 4, from about 0 to about 3, from about 0 to about 2, from about 0 to about 1, from about 1 to less than 16, from about 2 to less than 16, from about 3 to less than 16, from about 4 to less than 16, from about 5 to less than 16, from about 6 to less than 16, from about 7 to less than 16, about 8 to less than 16, about 9 to less than 16, about 10 to less than 16, about 11 to less than 16, about 12 to less than 16, about 13 to less than 16, about 14 to less than 16, about 15 to less than 16, about 2 to about 15, about 3 to about 14, about 4 to 13, about 5 to about 12, about 6 to about 11, about 7 to about 10, about 8 to about 9, about 0 to about 8, about 8 to less than 16, about 0 to about 4, about 4 to about 8, about 8 to about 12, and about 12 to less than 16.

[0059] The % BSA after treatment according to the methods of the present disclosure can be less than about 10%, for example, from about 0% to less than 10%. The % BSA after treatment can also be from about 1% to less than 10%, from about 2% to less than 10%, from about 3% to less than 10%, from about 4% to less than 10%, from about 5% to less than 10%, from about 6% to less than 10%, from about 7% to less than 10%, from about 8% to less than 10%, from about 9% to less than 10%, from about 0% to about 9%, from about 0% to about 8%, from about 0% to about 7%, from about 0% to about 6%, from about 0% to about 5%, from about 0% to about 4%, from about 0% to about 3%, from about 0% to about 3%, from about 0% to about 2%, from about 0% to about 1%, from about 0% to about 5%, from about 5% to less than 10%, from about 0% to about 2.5%, from about 2.5% to about 5%, from about 5% to about 7.5%, and from about 7.5% to less than 10%.

[0060] As used herein, the term "antigen binding protein" refers to isolated antibodies, antibody fragments (e.g., Fab, etc.) and other antibody-derived protein constructs, such as those containing antibody domains (e.g., domain antibodies, etc.), which are capable of binding to human IL-5 (SEQ ID NO: 11).

[0061] As used herein, the term "antibody" refers to a molecule with an immunoglobulin-like domain (e.g., IgG, IgM, IgA, IgD or IgE), and includes monoclonal, recombinant, polyclonal, monoclonal, recombinant, polyclonal, chimeric, people and humanized molecules of this type. Monoclonal antibodies can be produced by eukaryotic cell clones expressing the antibody. Monoclonal antibodies can also be produced by eukaryotic cell lines, which can recombinantly express the heavy and light chains of the antibody due to the nucleic acid sequences encoding these being introduced into the cell. The method for producing antibodies from different eukaryotic cell lines, such as Chinese hamster ovary cells, hybridomas or immortalized antibody cells derived from animals (e.g., people) is well known.

[0062] Antibodies can be derived from rat, mouse, primate (eg, cynomolgus monkey, Old World monkey, or Great Ape), human, or other sources, for example, using molecular biology techniques to generate nucleic acid encoding the antibody molecule.

[0063] The antibody may comprise a constant region, which may be of any isotype or subclass. The constant region may be of an IgG isotype, such as IgG1, IgG2, IgG3, IgG4, or a variant thereof. The antigen binding protein constant region may be IgG1.

[0064] The antigen binding protein may comprise one or more modifications selected from mutated constant domains such that the antibody has enhanced effector function / ADCC and / or complement activation.

[0065] The antibody may be capable of binding to a target antigen. Examples of such target antigens include human IL-5, which comprises the amino acid sequence shown in SEQ ID NO: 11.

[0066] Mepolizumab, which comprises the heavy chain amino acid sequence shown in SEQ ID NO: 1 and the light chain amino acid sequence shown in SEQ ID NO: 2, is an example of such an antibody. Mepolizumab or a composition comprising mepolizumab of the present disclosure binds to human IL-5 and antagonizes its activity.

[0067] Mepolizumab is a recombinant humanized monoclonal antibody (IgG1, kappa) with two light chains and two heavy chains.

[0068] The heavy and light chains of mepolizumab are covalently linked by a single disulfide bond, and the heavy chains are linked to each other by two disulfide bonds, resulting in a typical IgG molecule.

[0069] Mepolizumab or a composition comprising mepolizumab of the present disclosure can be provided as a lyophilized powder containing an antibody and an excipient, which can be reconstituted with a pharmaceutically acceptable carrier (e.g., sterile water). This reconstituted pharmaceutical composition can then be administered subcutaneously or intravenously (e.g., with further dilution). Mepolizumab or a composition comprising mepolizumab of the present disclosure can also be provided as a liquid formulation containing an antibody, an excipient, and a pharmaceutically acceptable carrier. This liquid pharmaceutical composition can then be administered subcutaneously or intravenously (e.g., with further dilution).

[0070] As used herein, the term "antibody variant" means an antibody that differs from a parent antibody due to at least one amino acid modification (e.g., by having a different amino acid side chain), post-translational modification, or other modification in at least one heavy chain, light chain, or combination thereof, which results in a structural change relative to the parent antibody (e.g., a different amino acid side chain, a different post-translational modification, or other modification). Mepolizumab is an example of such a parent antibody. Structural changes can be determined directly by various methods well known in the art, such as LC-MS, direct sequencing, or indirectly via methods such as isoelectric focusing. Such methods are well known to those of ordinary skill in the art.

[0071] As used herein, the term "IL-5" means human IL-5 comprising the amino acid sequence shown in SEQ ID NO:11.

[0072] As used herein, the term "specifically binds" in relation to antigen binding proteins means that the antigen binding protein binds to the target antigen and discrete domains or discrete amino acid sequences within the target antigen, without binding or not significantly binding to other (e.g., unrelated) proteins. However, this term does not exclude the fact that the antigen binding protein may also cross-react with closely related molecules (e.g., those with a high degree of sequence identity or from another genus or species). The antigen binding proteins described herein can bind to human IL-5 or human IL-5 receptor with an affinity that is at least 2, 5, 10, 50, 100, or 1000 times greater than their affinity for closely related molecules.

[0073] The binding affinity (K) of the antigen binding protein-target antigen interaction D ) can be 1 mM or less, 100 nM or less, 10 nM or less, 2 nM or less, or 1 nM or less. Alternatively, K D It can be 5 to 10 nM; or 1 to 2 nM. D Can be 1 pM to 500 pM; Or 500 pM to 1 nM. The binding affinity of an antigen-binding protein is determined by the association constant (Ka) and the dissociation constant (Kd) (KD = Kd / Ka). Binding affinity can be measured by BIACORE™, for example, by capturing the test antibody on a protein A-coated sensor surface and flowing the target antigen over the surface. Alternatively, binding affinity can be measured by FORTEBIO, for example, by capturing the test antibody receptor on a protein A-coated needle and flowing the target antigen over the surface.

[0074] K d Can be 1x10 -3 Ms -1 or less, 1x10 -4 Ms -1 or less, or 1x10 -5 Ms -1 or less. K d Can be 1x10 -5 Ms -1 Up to 1x10 -4 Ms -1 ; or 1x10 -4 Ms -1 Up to 1x10 -3 Ms -1 . Slow K d It can lead to slow dissociation of the antigen binding protein-target antigen complex and improve the neutralization of the target antigen.

[0075] As used herein, the term "specific antigen binding activity" means antigen binding activity as measured by surface plasmon resonance (SPR). IL-5 specific binding activity can be measured using BIACORE TM The instrument was determined by SPR, the BIACORE TM For example, if it is performed in binding mode, it is the binding activity divided by the total protein content in the sample.

[0076] As used herein, the term "FcRn binding activity" means neonatal Fc (FcRn) receptor binding activity as measured by surface plasmon resonance (SPR). FcRn binding can be measured using BIACORE TM The activity is determined by the instrument and is the binding activity to the FcRn receptor divided by the total protein concentration of the sample.

[0077] The SPR method for specific antigen binding and FcRn binding uses a reference standard of mepolizumab. The mepolizumab reference standard can be used in the assay to obtain system suitability and sample comparability data to ensure that the method is performing appropriately. The reference standard allows for the establishment of a calibration curve and the interpolation of sample concentrations from the curve.

[0078] "Isolating" contemplates removing a molecule, such as an antigen-binding protein or nucleic acid, from the environment in which it may be found in nature. For example, a molecule can be purified from the substances with which it normally occurs in nature. For example, the mass of the molecules in a sample can be 95% of the total mass. Importantly, mepolizumab and antigen-binding proteins of the present disclosure are typically provided as compositions that can include any combination of nucleic acids of the present disclosure, buffer, residual buffer, salt, counterions, water, alcohol, or carrier, among others.

[0079] The term "V H ” and “V L " are used herein to refer to the heavy chain variable region and light chain variable region, respectively, of an antigen binding protein.

[0080] "CDR" is defined as the complementary determining region amino acid sequence of an antigen binding protein. These are the hypervariable regions of the heavy and light chains of immunoglobulins. There are three heavy chain and three light chain CDRs (or CDR regions) in the variable portion of an immunoglobulin. Therefore, as used herein, "CDR" refers to all three heavy chain CDRs, all three light chain CDRs, all heavy chain and light chain CDRs, or at least one CDR, and wherein the at least one CDR is CDRH3. The framework region follows each of these CDR regions. Acceptable heavy chain variable region and light chain variable region framework 1, framework 2, and framework 3 regions are easily identified by those of ordinary skill in the art. Acceptable heavy chain constant region (including hinge region) and light chain constant region are also easily identified by those of ordinary skill in the art. Acceptable antibody isotypes are similarly easily identified by those of ordinary skill in the art.

[0081] Throughout the specification, the amino acid residues in the variable domain sequences and full-length antibody sequences are numbered according to the Kabat numbering convention. Similarly, the terms "CDR", "CDRL1", "CDRL2", "CDRL3", "CDRH1", "CDRH2", "CDRH3" used in the specification follow the Kabat numbering convention.

[0082] It will be apparent to those skilled in the art that there are alternative numbering conventions for amino acid residues in variable domain sequences and full-length antibody sequences. There are also alternative numbering conventions for CDR sequences, such as those set forth according to the Chothia numbering convention. The structure and protein folding of the antibody may mean that other residues are considered part of the CDR sequence, and should be understood by the skilled artisan as such.

[0083] Other numbering conventions for CDR sequences available to the skilled artisan include the "AbM" (University of Bath) and "contact" (University College London) methods. The minimum region of overlap can be determined using at least two of the Kabat, Chothia, AbM, and contact methods to provide a "minimum binding unit." The minimum binding unit can be a subpart of a CDR.

[0084] Table 14 below shows one definition using each numbering convention for each CDR or binding unit. The Kabat numbering scheme is used to number the variable domain amino acid sequences in Table 14. It should be noted that some CDR definitions may vary depending on the individual publications used.

[0085] " identity percentage " between query nucleic acid sequence and the subject nucleic acid sequence is " identity " value expressed as percentage, after execution pairing BLASTN is compared, when subject nucleic acid sequence and query nucleic acid sequence have 100% query coverage, by BLASTN algorithm calculation described " identity " value.This type of pairing BLASTN between query nucleic acid sequence and the subject nucleic acid sequence is compared, and is carried out by the default setting of the obtainable BLASTN algorithm on the website of U.S. National Center for Biotechnology Information (National Center forBiotechnology Institute), wherein closes the filter about low complexity zone.Importantly, query sequence can be described by the nucleotide sequence identified in one or more claims of this paper.

[0086] Nucleic acid sequences that may be useful and included in the compositions and related methods of the present disclosure can have from about 85% to about 100%, from about 90% to about 100%, from about 95% to about 100%, from about 91%, from about 92%, from about 93%, from about 94%, from about 95%, from about 96%, from about 97%, from about 98%, from about 99%, and from about 100% identity to the nucleic acid sequences identified in the present disclosure (e.g., nucleic acids encoding antibody heavy chains or antibody light chains). In the present disclosure, the percent identity between the described nucleic acid sequences can include any discrete subrange of the percent identity ranges recited above (e.g., any range of integer values ​​within a particular range or discrete subvalues ​​within a particular range).

[0087] "Percent identity" between a query amino acid sequence and a subject amino acid sequence is an "identity" value expressed as a percentage, calculated by the BLASTP algorithm after performing a paired BLASTP comparison where the subject amino acid sequence and the query amino acid sequence have 100% query coverage. Such a paired BLASTP comparison between the query amino acid sequence and the subject amino acid sequence is performed using the default settings of the BLASTP algorithm available on the website of the National Center for Biotechnology Information, with filters for low complexity regions turned off. Importantly, the query sequence can be described by the amino acid sequence identified in one or more claims herein.

[0088] Amino acid sequences that may be useful and included in the compositions and related methods of the present disclosure can have from about 85% to about 100%, from about 90% to about 100%, from about 95% to about 100%, from about 91%, from about 92%, from about 93%, from about 94%, from about 95%, from about 96%, from about 97%, from about 98%, from about 99%, and about 100% identity to an amino acid sequence identified in the present disclosure (e.g., an antibody heavy chain or an antibody light chain). In the present disclosure, the percent identity between the described amino acid sequences can include any discrete subrange of the percent identity ranges recited above (e.g., any range of integer values ​​within a particular range or discrete subvalues ​​within a particular range).

[0089] The terms "peptide," "polypeptide," "protein," and "peptide chain" each refer to a molecule comprising two or more amino acid residues. Peptides can be monomeric or polymeric.

[0090] It is well recognized in the art that certain amino acid substitutions are considered "conservative." Amino acids are grouped based on common side chain properties and substitutions, and substitutions within a group that maintain all or substantially all binding affinity of the antigen binding protein are considered conservative substitutions. See Table 15. The antigen binding proteins disclosed herein may contain such "conservative" amino acid substitutions.

[0091] Table 15. Sidechain member hydrophobic met, ala, val, leu, ile Neutral hydrophilic cys、ser、thr acidic asp、glu Alkaline asn, gln, his, lys, arg Residues that affect chain orientation gly、pro Aromatic trp、tyr、phe

[0092] As used herein, the term "pharmaceutical composition" means a composition suitable for administration to a patient.

[0093] The pharmaceutical compositions described herein may comprise purified preparations of antibodies as described herein.

[0094] For example, a pharmaceutical preparation may comprise a purified preparation of an antibody as described herein in combination with a pharmaceutically acceptable carrier.

[0095] Typically, such pharmaceutical compositions comprise a pharmaceutically acceptable carrier, as known and required by accepted pharmaceutical practice. Examples of such carriers include sterile carriers such as saline, Ringer's solution or dextrose solution, optionally buffered to a pH in the range of 5 to 8 with a suitable buffer.

[0096] The pharmaceutical composition can be administered by injection or infusion (e.g., intravenously, intraperitoneally, intradermally, subcutaneously, intramuscularly, or intraportally). Such compositions are suitably free of visible particulate matter. The pharmaceutical composition can comprise 1 mg to 10 g of antigen-binding protein, e.g., 5 mg to 1 g of antigen-binding protein. Alternatively, the composition can comprise 5 mg to 500 mg, e.g., 5 mg to 50 mg of antigen-binding protein.

[0097] Methods for preparing such pharmaceutical compositions are well known to those skilled in the art. The pharmaceutical composition may comprise 1 mg to 10 g of the antigen-binding protein in a unit dosage form, optionally together with instructions for use. The pharmaceutical composition may be lyophilized (freeze-dried) according to methods well known or apparent to those skilled in the art for reconstitution prior to administration. When the antibody has an IgG1 isotype, a copper chelator such as citrate (e.g., sodium citrate) or EDTA or histidine may be added to the pharmaceutical composition to reduce the degree of copper-mediated degradation of this isotype antibody. The pharmaceutical composition may also comprise a solubilizing agent such as arginine, a surfactant / anti-aggregating agent such as polysorbate 80, and an inert gas such as nitrogen to replace the oxygen in the vial headspace.

[0098] As used herein, the term "therapeutically effective amount" means an amount of an agent (e.g., an antibody or pharmaceutical composition) that provides a therapeutic benefit in the treatment or management of one or more symptoms of the condition to be treated (e.g., asthma, mild asthma, moderate asthma, severe asthma, mild eosinophilic asthma, moderate eosinophilic asthma, severe eosinophilic asthma, uncontrolled eosinophilic asthma, eosinophilic asthma, subeosinophilic asthma, chronic obstructive pulmonary disease, eosinophilic granulomatosis with polyangiitis, hypereosinophilic syndrome, nasal polyps, bullous pemphigoid, eosinophilic esophagitis, atopic dermatitis, moderate atopic dermatitis, and severe atopic dermatitis). Examples of such treatment or management of one or more symptoms of asthma, including asthma, mild asthma, moderate asthma, severe asthma, mild eosinophilic asthma, moderate eosinophilic asthma, severe eosinophilic asthma, uncontrolled eosinophilic asthma, eosinophilic asthma, and subeosinophilic asthma, include 1) reducing the frequency of asthma exacerbations; 2) reducing the time to the first clinically significant exacerbation requiring oral or systemic corticosteroids, hospitalization, and / or emergency department (ED) visit; 3) reducing the frequency of exacerbations requiring hospitalization (including intubation and intensive care unit admission) or ED visit; 4) reducing the time to the first exacerbation requiring hospitalization or ED visit; 5) change from baseline in clinical pre-bronchodilator FEV1; 6) change from baseline in clinical post-bronchodilator FEV1; 7) Asthma Control Questionnaire (ACPQ) questionnaire. 8) change from baseline in the ACQ score; 9) improved lung function as assessed by spirometry (e.g., vital capacity (VC), forced vital capacity (FVC), forced expiratory volume (FEV) at intervals of 0.5, 1.0 (FEV1), 2.0, and 3.0 seconds, total lung capacity at forced expiratory flow 25–75% (FEF 25–75), and maximal ventilation (MVV), tidal volume, residual volume, expiratory reserve volume, inspiratory reserve volume, inspiratory capacity, inspired vital capacity, vital capacity, functional residual capacity, residual volume expressed as a percentage of total lung capacity, alveolar gas volume, actual lung capacity including conducting airway capacity, forced vital capacity, etc.); and 10) reduction in acute asthma exacerbations requiring steroids for control (e.g., oral steroids or steroids administered by any route—e.g., prednisone, prednisolone, etc.). Such a reduction in asthma exacerbations requiring steroids for control can be about a 50% reduction in exacerbations requiring steroids (eg, oral steroids).

[0099] Therapeutically effective amounts and treatment regimens can generally be determined empirically and may depend on factors such as the age, weight, and health of the patient, and the disease or condition being treated. Such factors are within the purview of the attending physician.

[0100] The dosage of the antigen-binding protein administered to a subject is generally 1 μg / kg to 150 mg / kg, 0.1 mg / kg to 100 mg / kg, 0.5 mg / kg to 50 mg / kg, 1 to 25 mg / kg, about 0.3 mg / kg to about 3 mg / kg, or 1 to 10 mg / kg of the subject's body weight. For example, the dosage can be 10 mg / kg, 30 mg / kg, or 60 mg / kg. The dosage can also be 10 mg / kg to 110 mg / mg, 15 mg / kg to 25 mg / kg, or 15 mg / kg to 100 mg / kg. The antigen-binding protein can be administered, for example, parenterally, subcutaneously, intravenously, or intramuscularly. The dosage can also be administered on a per-subject basis, for example, from about 20 mg / subject to about 750 mg / subject, from about 75 mg / subject to about 750 mg / subject, from about 20 mg / subject to about 200 mg / subject. The dosage can be any discrete subrange of these dosage ranges. For example, the dose can also be administered subcutaneously on a per-subject basis, e.g., about 100 mg / subject (e.g., once every four weeks), or 300 mg / subject (or other doses can be administered subcutaneously so long as they provide about the same, or comparable, bioavailability as achieved with intravenous administration—e.g., three doses of 100 mg / subject to achieve a total dose of 300 mg / subject administered subcutaneously).

[0101] Preferably, pediatric subjects weighing less than 40 kg receive a dose of 40 mg, and pediatric subjects weighing greater than or equal to 40 kg receive a dose of 100 mg of mepolizumab. It is further preferred that such doses be administered subcutaneously.

[0102] Any type of range provided herein includes all values ​​within the particular range as well as values ​​about the endpoints of the particular range.

[0103] If desired, the effective daily dose of an antibody or antigen-binding protein of the present disclosure (e.g., as a pharmaceutical composition) can be administered as two, three, four, five, six or more doses administered separately at appropriate intervals throughout the day, optionally in unit dosage form. For example, a 100 mg dose to be administered to a pediatric subject weighing greater than or equal to 40 kg can be administered as two separate doses of 0.5 ml (e.g., subcutaneous injections), each containing 50 mg of mepolizumab.

[0104] The dosage can be administered by slow continuous infusion over a period of 2 to 24 hours, for example 2 to 12 hours, or 2 to 6 hours. Such administration can result in reduced side effects.

[0105] The administration of dosage can be repeated once or many times as needed, for example three times a day, once a day, once every 2 days, once a week, once every 14 days, once a month, once every 3 months, once every 4 months, once every 6 months or once every 12 months. Antigen-binding proteins can be administered by maintenance therapy, for example once a week for a total of 6 months or longer period. Antigen-binding proteins can be administered by intermittent therapy for a period of, for example, 3 to 6 months in a cycle, followed by a 3 to 6 month no-dose period, followed by another 3 to 6 month antigen-binding protein administration, and the rest can be deduced by analogy.

[0106] For example, the dose may be administered subcutaneously once every 14 or 28 days, in the form of multiple doses administered per day. In one embodiment, the dose of the composition is 100 mg once every 4 weeks (28 days).

[0107] The antigen binding protein can be administered to a subject in such a way as to target therapy to a particular site (eg, upper arm or thigh, etc.).

[0108] The antigen binding proteins of the methods of the present disclosure can be used in combination with one or more other therapeutically active agents, such as antibodies or small molecule inhibitors.

[0109] As used herein, the term "treat" and its grammatical variations refer to therapeutic therapy. With reference to a particular condition, treatment means: (1) ameliorating the condition or one or more biological manifestations of the condition, (2) interfering with a) one or more points in the biological cascade that leads to or is responsible for the condition, or b) one or more biological manifestations of the condition, (3) alleviating one or more symptoms, effects, or side effects associated with the condition or its treatment, (4) slowing the progression of the condition or one or more biological manifestations of the condition, or (5) preventing the onset of one or more biological manifestations of the condition. Prophylactic therapy is also contemplated. The skilled artisan will appreciate that "prevention" is not an absolute term. In medicine, "prevention" is understood to refer to the prophylactic administration of a drug to substantially reduce the likelihood or severity of a condition or its biological manifestations, or to delay the onset of such a condition or its biological manifestations.

[0110] The terms "individual," "subject," and "patient" are used interchangeably herein. The subject is typically a human. The subject may also be a mammal, such as a mouse, rat, or primate (e.g., a marmoset or monkey). The subject may be a non-human animal. The antigen-binding proteins, compositions, and methods of the present disclosure also have veterinary applications. The subject to be treated may be a farm animal, such as a cow or bull, sheep, pig, steer, goat, or horse, or may be a domesticated animal, such as a dog or cat. The animal may be of any age, or a mature adult animal.

[0111] Treatment may be therapeutic, prophylactic or preventative. The subject will be a subject in need thereof. Those in need of treatment may also include individuals already suffering from a particular medical disease, in addition to those who may develop the disease in the future.

[0112] Thus, if indicated, the methods, antigen-binding proteins, and compositions of the present disclosure described herein can be used for prophylactic or preventative treatment. In this case, the methods, antigen-binding proteins, and compositions of the present disclosure can be used to prevent or delay the onset of one or more aspects or symptoms of a disease. The subject can be asymptomatic. The subject can have a genetic predisposition to the disease. A prophylactic effective amount of the antigen-binding protein is administered to such an individual. A prophylactic effective amount is an amount that prevents or delays the onset of one or more aspects or symptoms of a disease described herein.

[0113] The methods, antigen-binding proteins and compositions of the present disclosure do not need to achieve a complete cure, or eradicate every symptom or manifestation of a disease, to constitute a viable therapeutic treatment. As is recognized in the art, drugs used as therapeutic agents in therapeutic methods can reduce the severity of a given disease state, but do not need to eliminate every manifestation of the disease to be considered a useful therapeutic agent. Similarly, treatments administered prophylactically do not need to be completely effective in preventing the onset of the disease in order to constitute a viable prophylactic agent. It is sufficient to simply reduce the impact of the disease in the subject (e.g., by reducing the number or severity of its symptoms, or by increasing the effectiveness of another treatment, or by producing another beneficial effect), or to reduce the likelihood of the disease occurring (e.g., by delaying the onset of the disease) or worsening.

[0114] As used herein, "area under the curve" or "AUC" is the area under the curve in a plot of the concentration of a substance in plasma against time. AUC can be a measure of the integral of the instantaneous concentrations over a time interval and has the units: mass*time / volume. AUC is typically calculated by the trapezoidal method (e.g., linear, linear logarithmic). AUC is typically given for a time interval from zero to infinity, with other time intervals indicated (e.g., AUC(t1, t2), where t1 and t2 are the start and end times of the interval). Thus, as used herein, "AUC" 0-24”refers to the AUC over a 24-hour period, whereas AUC(0-inf) refers to the AUC over an infinite period of time.

[0115] As used herein, "Tmax" refers to the observed time to reach a maximum concentration of a substance in the plasma of a mammal following administration of the substance to the mammal.

[0116] As used herein, "maximum plasma concentration" or "Cmax" means the highest concentration of a substance (eg, an antibody) observed in the plasma of a mammal following administration of the substance to the mammal.

[0117] As used herein, "maximum plasma concentration at steady state" or "Cmax SS" means the maximum concentration in plasma at steady state, as determined by pharmacokinetic modeling, and is a population estimate of Cmax at steady state obtained by population PK methods. Steady state refers to a condition in which the overall uptake of a drug is in proper dynamic equilibrium with its elimination. Generally, steady state is generally considered to be achieved when the time is 4 to 5 times the half-life of the drug after the start of regular dosing of the drug.

[0118] As used herein, "serum or plasma half-life" refers to the time required for half the amount of a substance administered to a mammal to be metabolized or eliminated from the serum or plasma of the mammal by normal biological processes.

[0119] One aspect of the present disclosure is a method of treating a disease in a pediatric subject comprising the steps of: a) identifying a pediatric subject weighing less than 40 kg having a disease selected from the group consisting of asthma, mild asthma, moderate asthma, severe asthma, mild eosinophilic asthma, moderate eosinophilic asthma, severe eosinophilic asthma, uncontrolled eosinophilic asthma, eosinophilic asthma, subeosinophilic asthma, chronic obstructive pulmonary disease, eosinophilic granulomatosis with polyangiitis, hypereosinophilic syndrome, nasal polyposis, bullous pemphigoid, eosinophilic esophagitis, atopic dermatitis, moderate atopic dermatitis, and severe atopic dermatitis; and b) subcutaneously administering to the pediatric subject a therapeutically effective amount of an antibody comprising the heavy chain amino acid sequence set forth in SEQ ID NO: 1 and the light chain amino acid sequence set forth in SEQ ID NO: 2, wherein the therapeutically effective amount of the antibody provides about 10.1960 ± 10.1960 % of the total dose. A maximum plasma concentration (Cmax) of the antibody of 0.3345 μg / mL, and an area under the curve [0-infinity] value of approximately 454.39 ± 15.8876 μg*day / mL; thereby treating the disease in pediatric subjects.

[0120] In one embodiment of the methods of the disclosure, the therapeutically effective amount of the antibody is about 40 mg.

[0121] In another embodiment of the methods of the disclosure, the antibody is administered about once a month.

[0122] In another embodiment of the methods of the present disclosure, the pediatric subject has an absolute blood eosinophil count selected from: greater than or equal to 150 cells / μL and greater than or equal to 300 cells / μL.

[0123] In another embodiment of the methods of the present disclosure, the pediatric subject is selected from premature newborns, full-term newborns, children 28 days to 11 months, children 12 months to 23 months, children 2 years to 6 years, children 16 years to 12 years, children 6 years to less than 18 years, and children 12 to 18 years.

[0124] In another embodiment of the methods of the present disclosure, the pediatric subject is between about 6 years of age and about 12 years of age, inclusive.

[0125] In another embodiment of the methods of the present disclosure, the pediatric subject has severe eosinophilic asthma.

[0126] In another embodiment of the methods of the disclosure, the pediatric subject has a treatment outcome selected from the group consisting of: a decrease in absolute blood eosinophil count relative to the absolute blood eosinophil count before antibody administration, a decrease in the score on the Asthma Control Questionnaire-7 scale relative to the score on the Asthma Control Questionnaire-7 scale before antibody administration, and a decrease in the score on the Childhood Asthma Control Test relative to the score on the Childhood Asthma Control Test before antibody administration.

[0127] In another embodiment of the methods of the disclosure, the antibody is administered by a safety syringe or autoinjector.

[0128] Another aspect of the present disclosure is a method of treating a disease in a pediatric subject comprising the steps of: a) identifying a pediatric subject weighing greater than or equal to 40 kg who has a disease selected from the group consisting of asthma, mild asthma, moderate asthma, severe asthma, mild eosinophilic asthma, moderate eosinophilic asthma, severe eosinophilic asthma, uncontrolled eosinophilic asthma, eosinophilic asthma, subeosinophilic asthma, chronic obstructive pulmonary disease, eosinophilic granulomatosis with polyangiitis, hypereosinophilic syndrome, nasal polyposis, bullous pemphigoid, eosinophilic esophagitis, atopic dermatitis, moderate atopic dermatitis, and severe atopic dermatitis; and b) subcutaneously administering to the pediatric subject a therapeutically effective amount of an antibody comprising the heavy chain amino acid sequence set forth in SEQ ID NO: 1 and the light chain amino acid sequence set forth in SEQ ID NO: 2, wherein the therapeutically effective amount of the antibody provides approximately 16.3412 ± 0.6364 A maximum plasma concentration (Cmax) of the antibody of about 675.20 ± 35.8980 μg / mL, and an area under the curve [0-infinity] value of about 675.20 ± 35.8980 μg*day / mL; thereby treating the disease in pediatric subjects.

[0129] In another embodiment of the methods of the disclosure, the therapeutically effective amount of the antibody is about 100 mg.

[0130] Another aspect of the present disclosure is a method of reducing the absolute blood eosinophil count in a pediatric subject comprising the steps of: a) identifying a pediatric subject weighing less than 40 kg who has a condition selected from the group consisting of asthma, mild asthma, moderate asthma, severe asthma, mild eosinophilic asthma, moderate eosinophilic asthma, severe eosinophilic asthma, uncontrolled eosinophilic asthma, eosinophilic asthma, subeosinophilic asthma, chronic obstructive pulmonary disease, eosinophilic granulomatosis with polyangiitis, hypereosinophilic syndrome, nasal polyposis, bullous pemphigoid, eosinophilic esophagitis, and atopic dermatitis; and b) subcutaneously administering to the pediatric subject a therapeutically effective amount of an antibody comprising the heavy chain amino acid sequence set forth in SEQ ID NO: 1 and the light chain amino acid sequence set forth in SEQ ID NO: 2, wherein the therapeutically effective amount of the antibody provides approximately 10.1960 ± 0.3345 The invention provides a maximum plasma concentration (Cmax) of the antibody of about 1.5 μg / mL and an area under the curve [0-infinity] value of about 454.39 ± 15.8876 μg*day / mL; thereby reducing the absolute blood eosinophil count in pediatric subjects.

[0131] Another aspect of the present disclosure is a method of reducing the absolute blood eosinophil count in a pediatric subject comprising the steps of: a) identifying a pediatric subject weighing greater than or equal to 40 kg who has a condition selected from the group consisting of asthma, mild asthma, moderate asthma, severe asthma, mild eosinophilic asthma, moderate eosinophilic asthma, severe eosinophilic asthma, uncontrolled eosinophilic asthma, eosinophilic asthma, subeosinophilic asthma, chronic obstructive pulmonary disease, eosinophilic granulomatosis with polyangiitis, hypereosinophilic syndrome, nasal polyposis, bullous pemphigoid, eosinophilic esophagitis, and atopic dermatitis; and b) subcutaneously administering to the pediatric subject a therapeutically effective amount of an antibody comprising the heavy chain amino acid sequence set forth in SEQ ID NO: 1 and the light chain amino acid sequence set forth in SEQ ID NO: 2, wherein the therapeutically effective amount of the antibody provides about 16.3412 ± 16.3412 eosinophils. A maximum plasma concentration (Cmax) of the antibody of 0.6364 μg / mL, and an area under the curve [0-infinity] value of approximately 675.20 ± 35.8980 μg*day / mL; thereby reducing absolute blood eosinophil counts in pediatric subjects.

[0132] Another embodiment of the present disclosure is a composition according to the present disclosure and / or a method of the present disclosure for use in therapy.

[0133] Another embodiment of the present disclosure is a composition according to the present disclosure and / or a method of the present disclosure for use in the treatment of asthma, mild asthma, moderate asthma, severe asthma, mild eosinophilic asthma, moderate eosinophilic asthma, severe eosinophilic asthma, uncontrolled eosinophilic asthma, eosinophilic asthma, subeosinophilic asthma, chronic obstructive pulmonary disease, eosinophilic granulomatosis with polyangiitis, hypereosinophilic syndrome, nasal polyps, bullous pemphigoid, eosinophilic esophagitis, atopic dermatitis, moderate atopic dermatitis, and severe atopic dermatitis.

[0134] Another aspect of the disclosure is a composition for treating a disease selected from the group consisting of asthma, mild asthma, moderate asthma, severe asthma, mild eosinophilic asthma, moderate eosinophilic asthma, severe eosinophilic asthma, uncontrolled eosinophilic asthma, eosinophilic asthma, subeosinophilic asthma, chronic obstructive pulmonary disease, eosinophilic granulomatosis with polyangiitis, hypereosinophilic syndrome, nasal polyposis, bullous pemphigoid, eosinophilic esophagitis, atopic dermatitis, moderate atopic dermatitis, and severe atopic dermatitis in a pediatric subject weighing less than 40 kg, wherein the composition comprises an antibody comprising the heavy chain amino acid sequence set forth in SEQ ID NO: 1 and the light chain amino acid sequence set forth in SEQ ID NO: 2, and a therapeutically effective amount of the antibody administered subcutaneously to the pediatric subject provides approximately 10.1960 ± 0.3345 The maximum plasma concentration (Cmax) of the antibody was approximately 454.39 ± 15.8876 μg*day / mL, and the area under the curve [0-infinity] value was approximately 454.39 ± 15.8876 μg*day / mL.

[0135] In another embodiment of the composition of the present disclosure, the therapeutically effective amount of the antibody is about 40 mg.

[0136] Another aspect of the disclosure is a composition for treating a disease selected from the group consisting of asthma, mild asthma, moderate asthma, severe asthma, mild eosinophilic asthma, moderate eosinophilic asthma, severe eosinophilic asthma, uncontrolled eosinophilic asthma, eosinophilic asthma, subeosinophilic asthma, chronic obstructive pulmonary disease, eosinophilic granulomatosis with polyangiitis, hypereosinophilic syndrome, nasal polyposis, bullous pemphigoid, eosinophilic esophagitis, atopic dermatitis, moderate atopic dermatitis, and severe atopic dermatitis in a pediatric subject weighing greater than or equal to 40 kg, wherein the composition comprises an antibody comprising the heavy chain amino acid sequence set forth in SEQ ID NO: 1 and the light chain amino acid sequence set forth in SEQ ID NO: 2, and a therapeutically effective amount of the antibody administered subcutaneously to the pediatric subject provides approximately 16.3412 ± 0.6364 The maximum plasma concentration (Cmax) of the antibody was 1.5 μg / mL, and the area under the curve [0-infinity] value was approximately 675.20±35.8980 μg*day / mL.

[0137] In another embodiment of the composition of the present disclosure, the therapeutically effective amount of the antibody is about 100 mg.

[0138] Another aspect of the present disclosure is a composition for reducing the absolute blood eosinophil count in a pediatric subject weighing less than 40 kg, wherein the pediatric subject has a disease selected from the group consisting of asthma, mild asthma, moderate asthma, severe asthma, mild eosinophilic asthma, moderate eosinophilic asthma, severe eosinophilic asthma, uncontrolled eosinophilic asthma, eosinophilic asthma, subeosinophilic asthma, chronic obstructive pulmonary disease, eosinophilic granulomatosis with polyangiitis, hypereosinophilic syndrome, nasal polyposis, bullous pemphigoid, eosinophilic esophagitis, atopic dermatitis, moderate atopic dermatitis, and severe atopic dermatitis, and the composition comprises an antibody comprising the heavy chain amino acid sequence set forth in SEQ ID NO: 1 and the light chain amino acid sequence set forth in SEQ ID NO: 2, and a therapeutically effective amount of the antibody administered subcutaneously to the pediatric subject provides approximately 10.1960 ± 0.3345 The maximum plasma concentration (Cmax) of the antibody was approximately 454.39 ± 15.8876 μg*day / mL, and the area under the curve [0-infinity] value was approximately 454.39 ± 15.8876 μg*day / mL.

[0139] Another aspect of the present disclosure is a composition for reducing the absolute blood eosinophil count in a pediatric subject weighing greater than or equal to 40 kg, wherein the pediatric subject has a disease selected from the group consisting of asthma, mild asthma, moderate asthma, severe asthma, mild eosinophilic asthma, moderate eosinophilic asthma, severe eosinophilic asthma, uncontrolled eosinophilic asthma, eosinophilic asthma, subeosinophilic asthma, chronic obstructive pulmonary disease, eosinophilic granulomatosis with polyangiitis, hypereosinophilic syndrome, nasal polyposis, bullous pemphigoid, eosinophilic esophagitis, atopic dermatitis, moderate atopic dermatitis, and severe atopic dermatitis, and the composition comprises an antibody comprising the heavy chain amino acid sequence set forth in SEQ ID NO: 1 and the light chain amino acid sequence set forth in SEQ ID NO: 2, and a therapeutically effective amount of the antibody administered subcutaneously to the pediatric subject provides approximately 16.3412 ± 0.6364 The maximum plasma concentration (Cmax) of the antibody was 1.5 μg / mL, and the area under the curve [0-infinity] value was approximately 675.20±35.8980 μg*day / mL.

[0140] In one embodiment of the methods of the present disclosure, the amount of antigen binding protein is from about 2 mg to about 600 mg. For example, the amount of antigen binding protein (e.g., antibody) can be a 2 mg, 10 mg, 30 mg, 100 mg, 300 mg, or 600 mg dose.

[0141] In one embodiment of the methods of the present disclosure, the antigen binding protein is administered once every 3 months or once every 6 months.

[0142] One embodiment is the method of the present disclosure, wherein the pharmaceutically effective carrier comprises an aqueous liquid formulation at about pH 5.5 to about pH 6.0 containing about 40 mM histidine, about 180 mM trehalose, about 100 mM arginine, about 8 mM methionine, about 0.02% weight / volume polysorbate 80, and about 0.05 mM EDTA.

[0143] The compositions of the present disclosure and those comprising the antibodies or antigen-binding proteins of the present disclosure may further comprise a buffer selected from sodium phosphate dibasic heptahydrate, phosphate, citric acid, citrate, sodium phosphate, potassium phosphate, sodium citrate and histidine, which provides a pH of 6.8 to 7.2 or a pH of pH 6.2 to pH 6.6, with a pH of 6.3 being preferred. The buffer in the compositions of the present disclosure may be present in a range of about 10-30 mM, about 10-20 mM, about 20 mM or about 15.5 mM. For example, the buffer in the compositions of the present disclosure is present in about 20 mM or about 15.5 mM sodium phosphate dibasic heptahydrate. These compositions can be used in the methods of the present disclosure.

[0144] The compositions of the present disclosure may comprise sodium phosphate dibasic heptahydrate and citric acid buffer, which provide a pH of 6.2 to 6.6 (inclusive), with a pH of 6.3 being preferred. The sodium phosphate dibasic heptahydrate buffer may be present in a range of about 15-16.4 mM, and the citric acid buffer may be present in a range of about 3.8-4.9 mM. For example, the compositions of the present disclosure may comprise about 15.5 mM sodium phosphate dibasic heptahydrate and about 4.5 mM citric acid monohydrate.

[0145] The compositions of the present disclosure may further comprise sugar. The compositions of the present disclosure may further comprise sucrose. The sucrose may be present in the compositions of the present disclosure in a range of about 5-20%, about 10-15%, about 11-13%, or about 12% weight / volume.

[0146] The composition of the present disclosure may further comprise polysorbate 80. Polysorbate 80 may be present in the range of about 0.01-0.1% weight / volume. For example, polysorbate 80 may be present in the composition of the present disclosure at about 0.02% weight / volume, or about 0.05% weight / volume.

[0147] The compositions of the present disclosure may further comprise EDTA. EDTA may be present in a range of about 0.01-0.1 mM. For example, EDTA may be present in about 0.05 mM.

[0148] In one embodiment, the composition of the present disclosure further comprises 20 mM sodium phosphate dibasic heptahydrate, 12% weight / volume sucrose, and 0.05% weight / volume polysorbate 80.

[0149] In another embodiment, the composition of the present disclosure further comprises 15.5 mM sodium phosphate dibasic, 3.9 mM citric acid monohydrate, 12% weight / volume sucrose, 0.02% weight / volume polysorbate 80, and 0.05 mM EDTA.

[0150] The composition of the present disclosure may comprise an aqueous liquid formulation at pH 6.2 containing 16.1 mM sodium phosphate dibasic heptahydrate, 3.9 mM citric acid monohydrate, 12% weight / volume sucrose, 0.02% weight / volume polysorbate 80, and 0.05 mM EDTA.

[0151] The composition of the present disclosure may comprise an aqueous liquid formulation at pH 6.2 containing 15.2 mM sodium phosphate dibasic heptahydrate, 4.8 mM citric acid monohydrate, 12% weight / volume sucrose, 0.02% weight / volume polysorbate 80, and 0.05 mM EDTA.

[0152] The composition of the present disclosure may comprise an aqueous liquid formulation at pH 6.4 containing 15.8 mM sodium phosphate dibasic heptahydrate, 4.2 mM citric acid monohydrate, 12% weight / volume sucrose, 0.02% weight / volume polysorbate 80, and 0.05 mM EDTA.

[0153] The composition of the present disclosure may comprise an aqueous liquid formulation at pH 6.6 containing 16.3 mM sodium phosphate dibasic heptahydrate, 3.7 mM citric acid monohydrate, 12% weight / volume sucrose, 0.02% weight / volume polysorbate 80, and 0.05 mM EDTA.

[0154] The composition of the present disclosure may comprise an aqueous liquid formulation at pH 6.3 containing 15.5 mM sodium phosphate dibasic heptahydrate, 4.5 mM citric acid monohydrate, 12% weight / volume sucrose, 0.02% weight / volume polysorbate 80, and 0.05 mM EDTA. Importantly, the tangential filtration and ultrafiltration exchange steps of the production process can be adjusted to produce a composition of the present disclosure, such as a composition of the present disclosure at pH 6.3 containing 15.5 mM sodium phosphate dibasic heptahydrate, 4.5 mM citric acid monohydrate, 12% weight / volume sucrose, 0.02% weight / volume polysorbate 80, 0.05 mM EDTA—or other such liquid formulations.

[0155] The compositions of the present disclosure may comprise a purified preparation of a monoclonal antibody and a buffer, wherein the composition is at a pH of 6.8 to 7.2, wherein the buffer is histidine, phosphate, citric acid, citrate, or a salt thereof.

[0156] In the composition of the present disclosure, the buffer may be at least one selected from the group consisting of disodium hydrogen phosphate heptahydrate, phosphate, citric acid, and citrate.

[0157] In the compositions of the present disclosure, the buffering agent may be sodium phosphate, potassium phosphate, or sodium citrate.

[0158] The compositions of the present disclosure may contain sugars, carbohydrates and / or salts.

[0159] The compositions of the present disclosure may further comprise sucrose or trehalose.

[0160] The compositions of the present disclosure may also comprise a purified preparation of a monoclonal antibody and a buffer, wherein the composition is at a pH of 6.8 to 7.2, wherein the buffer is phosphate or a salt thereof.

[0161] The composition of the present disclosure may further comprise one selected from the group consisting of: a first formulation having 20 mM disodium phosphate heptahydrate, 12% weight / volume sucrose, and 0.05% weight / volume polysorbate 80; a second formulation having 15.5 mM disodium phosphate heptahydrate, 3.9 mM citric acid monohydrate, 12% weight / volume sucrose, 0.02% weight / volume polysorbate 80, and 0.05 mM EDTA; and a third formulation having 26 mM disodium phosphate heptahydrate, 15% weight / volume sucrose, and 0.065% weight / volume polysorbate 80. The composition may be at a pH of 6.8 to about 7.2, about 6.1 to about 6.5, or about 6 to about 6.6.

[0162] The compositions described herein can be produced by any number of conventional techniques. For example, the compositions can be expressed in a recombinant expression system and purified therefrom. In one embodiment, the compositions are produced by culturing a host cell under conditions suitable for expressing a polypeptide comprising SEQ ID NO: 1 and SEQ ID NO: 2, wherein the composition is expressed, optionally purified, and optionally formulated into a pharmaceutical composition.

[0163] Many different expression systems and purification schemes can be used to produce compositions. Generally, host cells are transformed with the recombinant expression vector encoding the antibody. A wide range of host cells can be used, including eukaryotic cell lines (e.g., CHO, Perc6, HEK293, HeLa, NSO) of mammalian origin. Suitable host cells include mammalian cells, such as CHO (e.g., CHOK1 and CHO-DG44).

[0164] The host cell can be an isolated host cell. The host cell is not typically part of a multicellular organism (e.g., a plant or animal). The host cell can be a non-human host cell.

[0165] Appropriate cloning and expression vectors and cloning methods for use with eukaryotic or mammalian cell hosts are known in the art.

[0166] Cells can be cultured under conditions that promote antibody expression. For example, a production bioreactor is used to culture cells. The volume of the production bioreactor can be: (i) about 20,000 liters, about 10,000 liters; about 5,000 liters; about 2,000 liters; about 1,000 liters; or about 500 liters; or (ii) 500 to 20,000 liters; 500 to 10,000 liters; 500 to 5,000 liters; 1,000 to 10,000 liters, or 2,000 to 10,000 liters. For example, cells can be cultured in a production bioreactor at a pH of about 6.75 to pH 7.00. Alternatively, cells can be cultured in a production bioreactor for about 12 to about 18 days. Alternatively, cells can be cultured in a production bioreactor at a pH of about 6.75 to pH 7.00 for about 12 to about 18 days. This incubation step can help control the levels of deamidated antibody variants, e.g., reduce the levels of deamidated antibody variants.

[0167] The composition can be recovered and purified by conventional protein purification procedures. For example, the composition can be harvested directly from the culture medium. The harvest of the cell culture medium can be clarified, for example, by centrifugation and / or depth filtration. The recovery of the composition is followed by purification to ensure sufficient purity.

[0168] One or more chromatography steps can be used in the purification, such as one or more chromatography resins; and / or one or more filtration steps. For example, affinity chromatography using resins such as Protein A, G, or L can be used to purify the composition. Alternatively, or in addition, ion exchange resins such as cation exchange resins can be used to purify the composition. Alternatively, or in addition, hydrophobic interaction chromatography resins can be used to purify the composition. Alternatively, the purification steps include: an affinity chromatography resin step, followed by a cation exchange resin step, followed by a hydrophobic interaction chromatography resin step.

[0169] For example, the harvest is contacted with a protein A resin. The solution comprising the composition can be eluted from the protein A resin and treated at a pH of 3.3 to 3.7 for 15 to 240 minutes. This protein A resin step can help control the level of aggregated antibody variants, for example, reduce the level of aggregated antibody variants.

[0170] The solution comprising the composition may then be further clarified by depth filtration and / or double filtration.

[0171] Alternatively, or in addition thereto, an anion exchange resin can be used. The solution comprising the composition can be contacted with an anion exchange resin (e.g., Q-SEPHAROSE™ Fast Flow anion exchange chromatography) at a loading pH of 8.3 to 8.7. The solution comprising the composition can be eluted from the anion exchange resin and maintained for 96 hours or less. This anion exchange resin step can help control the level of deamidated antibody variants, e.g., reduce the level of deamidated antibody variants.

[0172] Optionally, guanidine and / or ammonium sulfate may be added to the solution comprising the composition and maintained for 15 to 240 minutes.

[0173] Alternatively, or in addition thereto, a hydrophobic interaction chromatography resin can be used. The solution comprising the composition can be contacted with a hydrophobic interaction chromatography resin (e.g., phenyl SEPHAROSE™ fast flow chromatography) at a loading ratio of 12 to 27 g protein / L resin. For example, an elution gradient volume (bed volume; BV) of approximately 9 to approximately 11 can be used to elute the solution comprising the composition. In the process of eluting from the hydrophobic interaction chromatography resin, an elution peak cut-off of approximately 17 to approximately 23 can be used (cut stop) (% of maximum peak height). This hydrophobic interaction chromatography resin step can help control the level of the antibody variants of aggregation, for example, reducing the level of the antibody variants of aggregation.

[0174] The solution comprising the composition can then be filtered to remove the virus. The solution of the composition can then be prepared at an antibody concentration of about 76 g protein / L to about 82 g protein / L or about 100 g protein / L. The solution comprising the composition can be filled into a container and frozen. An aliquot of the solution comprising the composition can be lyophilized. The lyophilized product can be reconstituted by adding water to produce a composition comprising 75 mg / L of protein, monoclonal anti-IL-5 antibody, 20 mM sodium dihydrogen phosphate heptahydrate, 12% weight / volume sucrose, and 0.05% weight / volume polysorbate 80 at a pH of about 6.8 to about 7.2.

[0175] The methods of the present disclosure and the compositions of the present disclosure may also be used to achieve lower steroid doses or lower doses of other drugs—e.g. Figure 2 or those shown in Table 4.

[0176] In summary, this disclosure includes: In one aspect, a method of treating a disease in a pediatric subject comprises the steps of: a) identifying a pediatric subject weighing less than 40 kg having a disease selected from the group consisting of asthma, mild asthma, moderate asthma, severe asthma, mild eosinophilic asthma, moderate eosinophilic asthma, severe eosinophilic asthma, uncontrolled eosinophilic asthma, eosinophilic asthma, subeosinophilic asthma, chronic obstructive pulmonary disease, eosinophilic granulomatosis with polyangiitis, hypereosinophilic syndrome, nasal polyposis, bullous pemphigoid, eosinophilic esophagitis, atopic dermatitis, moderate atopic dermatitis, and severe atopic dermatitis; and b) subcutaneously administering to the pediatric subject a therapeutically effective amount of an antibody comprising the heavy chain amino acid sequence set forth in SEQ ID NO: 1 and the light chain amino acid sequence set forth in SEQ ID NO: 2, wherein the therapeutically effective amount of the antibody provides approximately 10.1960 ± 0.3345 A maximum plasma concentration (Cmax) of the antibody of about 1.5 μg / mL and an area under the curve [0-infinity] value of about 454.39 ± 15.8876 μg*day / mL; thereby treating the disease in pediatric subjects.

[0177] Another aspect of the present disclosure is a method of treating a disease in a pediatric subject comprising the steps of: a) identifying a pediatric subject weighing greater than or equal to 40 kg who has a disease selected from the group consisting of asthma, mild asthma, moderate asthma, severe asthma, mild eosinophilic asthma, moderate eosinophilic asthma, severe eosinophilic asthma, uncontrolled eosinophilic asthma, eosinophilic asthma, subeosinophilic asthma, chronic obstructive pulmonary disease, eosinophilic granulomatosis with polyangiitis, hypereosinophilic syndrome, nasal polyposis, bullous pemphigoid, eosinophilic esophagitis, atopic dermatitis, moderate atopic dermatitis, and severe atopic dermatitis; and b) subcutaneously administering to the pediatric subject a therapeutically effective amount of an antibody comprising the heavy chain amino acid sequence set forth in SEQ ID NO: 1 and the light chain amino acid sequence set forth in SEQ ID NO: 2, wherein the therapeutically effective amount of the antibody provides approximately 16.3412 ± 0.6364 A maximum plasma concentration (Cmax) of the antibody of about 675.20 ± 35.8980 μg / mL, and an area under the curve [0-infinity] value of about 675.20 ± 35.8980 μg*day / mL; thereby treating the disease in pediatric subjects.

[0178] Another aspect of the present disclosure is a method of reducing the absolute blood eosinophil count in a pediatric subject comprising the steps of: a) identifying a pediatric subject weighing less than 40 kg who has a condition selected from the group consisting of asthma, mild asthma, moderate asthma, severe asthma, mild eosinophilic asthma, moderate eosinophilic asthma, severe eosinophilic asthma, uncontrolled eosinophilic asthma, eosinophilic asthma, subeosinophilic asthma, chronic obstructive pulmonary disease, eosinophilic granulomatosis with polyangiitis, hypereosinophilic syndrome, nasal polyposis, bullous pemphigoid, eosinophilic esophagitis, and atopic dermatitis; and b) subcutaneously administering to the pediatric subject a therapeutically effective amount of an antibody comprising the heavy chain amino acid sequence set forth in SEQ ID NO: 1 and the light chain amino acid sequence set forth in SEQ ID NO: 2, wherein the therapeutically effective amount of the antibody provides approximately 10.1960 ± 0.3345 The invention provides a maximum plasma concentration (Cmax) of the antibody of about 1.5 μg / mL and an area under the curve [0-infinity] value of about 454.39 ± 15.8876 μg*day / mL; thereby reducing the absolute blood eosinophil count in pediatric subjects.

[0179] Another aspect of the present disclosure is a method of reducing the absolute blood eosinophil count in a pediatric subject comprising the steps of: a) identifying a pediatric subject weighing greater than or equal to 40 kg who has a condition selected from the group consisting of asthma, mild asthma, moderate asthma, severe asthma, mild eosinophilic asthma, moderate eosinophilic asthma, severe eosinophilic asthma, uncontrolled eosinophilic asthma, eosinophilic asthma, subeosinophilic asthma, chronic obstructive pulmonary disease, eosinophilic granulomatosis with polyangiitis, hypereosinophilic syndrome, nasal polyposis, bullous pemphigoid, eosinophilic esophagitis, and atopic dermatitis; and b) subcutaneously administering to the pediatric subject a therapeutically effective amount of an antibody comprising the heavy chain amino acid sequence set forth in SEQ ID NO: 1 and the light chain amino acid sequence set forth in SEQ ID NO: 2, wherein the therapeutically effective amount of the antibody provides about 16.3412 ± 16.3412 eosinophils. A maximum plasma concentration (Cmax) of the antibody of 0.6364 μg / mL, and an area under the curve [0-infinity] value of approximately 675.20 ± 35.8980 μg*day / mL; thereby reducing absolute blood eosinophil counts in pediatric subjects.

[0180] In one embodiment of the disclosed method, the therapeutically effective amount of the antibody is about 40 mg.

[0181] In another embodiment of the disclosed method, the antibody is administered about once a month.

[0182] In another embodiment of the disclosed method, the pediatric subject has an absolute blood eosinophil count selected from the group consisting of: greater than or equal to 150 cells / μL and greater than or equal to 300 cells / μL.

[0183] In another embodiment of the disclosed method, the pediatric subject is selected from premature newborns, full-term newborns, children aged 28 days to 11 months, children aged 12 months to 23 months, children aged 2 years to 6 years, children aged 16 years to 12 years, children aged 6 years to less than 18 years, and children aged 12 to 18 years.

[0184] In another embodiment of the disclosed method, the pediatric subject is between about 6 years of age and about 12 years of age, inclusive.

[0185] In another embodiment of the disclosed method, the pediatric subject has severe eosinophilic asthma.

[0186] In another embodiment of the disclosed methods, the pediatric subject has a treatment outcome selected from the group consisting of: a decrease in absolute blood eosinophil count relative to the absolute blood eosinophil count before antibody administration, a decrease in the score on the Asthma Control Questionnaire-7 scale relative to the score on the Asthma Control Questionnaire-7 scale before antibody administration, and a decrease in the score on the Childhood Asthma Control Test relative to the score on the Childhood Asthma Control Test before antibody administration.

[0187] In another embodiment of the disclosed method, the antibody is administered by a safety syringe or autoinjector.

[0188] In another embodiment of the disclosed method, the therapeutically effective amount of the antibody is about 100 mg.

[0189] Another embodiment of the invention is a composition according to any one aspect or embodiment of the method of the present disclosure for use in therapy.

[0190] Another embodiment of the invention is a composition according to any aspect or embodiment of the methods of the present disclosure for use in the treatment of asthma, mild asthma, moderate asthma, severe asthma, mild eosinophilic asthma, moderate eosinophilic asthma, severe eosinophilic asthma, uncontrolled eosinophilic asthma, eosinophilic asthma, subeosinophilic asthma, chronic obstructive pulmonary disease, eosinophilic granulomatosis with polyangiitis, hypereosinophilic syndrome, nasal polyps, bullous pemphigoid, eosinophilic esophagitis, atopic dermatitis, moderate atopic dermatitis, and severe atopic dermatitis.

[0191] Another aspect of the disclosure is a composition for treating a disease selected from the group consisting of asthma, mild asthma, moderate asthma, severe asthma, mild eosinophilic asthma, moderate eosinophilic asthma, severe eosinophilic asthma, uncontrolled eosinophilic asthma, eosinophilic asthma, subeosinophilic asthma, chronic obstructive pulmonary disease, eosinophilic granulomatosis with polyangiitis, hypereosinophilic syndrome, nasal polyposis, bullous pemphigoid, eosinophilic esophagitis, atopic dermatitis, moderate atopic dermatitis, and severe atopic dermatitis in a pediatric subject weighing less than 40 kg, wherein the composition comprises an antibody comprising the heavy chain amino acid sequence set forth in SEQ ID NO: 1 and the light chain amino acid sequence set forth in SEQ ID NO: 2, and a therapeutically effective amount of the antibody administered subcutaneously to the pediatric subject provides approximately 10.1960 ± 0.3345 The maximum plasma concentration (Cmax) of the antibody was approximately 454.39 ± 15.8876 μg*day / mL, and the area under the curve [0-infinity] value was approximately 454.39 ± 15.8876 μg*day / mL.

[0192] Another aspect of the disclosure is a composition for treating a disease selected from the group consisting of asthma, mild asthma, moderate asthma, severe asthma, mild eosinophilic asthma, moderate eosinophilic asthma, severe eosinophilic asthma, uncontrolled eosinophilic asthma, eosinophilic asthma, subeosinophilic asthma, chronic obstructive pulmonary disease, eosinophilic granulomatosis with polyangiitis, hypereosinophilic syndrome, nasal polyposis, bullous pemphigoid, eosinophilic esophagitis, atopic dermatitis, moderate atopic dermatitis, and severe atopic dermatitis in a pediatric subject weighing greater than or equal to 40 kg, wherein the composition comprises an antibody comprising the heavy chain amino acid sequence set forth in SEQ ID NO: 1 and the light chain amino acid sequence set forth in SEQ ID NO: 2, and a therapeutically effective amount of the antibody administered subcutaneously to the pediatric subject provides approximately 16.3412 ± 0.6364 The maximum plasma concentration (Cmax) of the antibody is about 1.5 μg / mL, and the area under the curve [0-infinity] value is about 675.20 ± 35.8980 μg*day / mL. Another embodiment is the composition of the present disclosure, wherein the therapeutically effective amount of the antibody is about 40 mg.

[0193] Another aspect of the present disclosure is a composition for reducing the absolute blood eosinophil count in a pediatric subject weighing less than 40 kg, wherein the pediatric subject has a disease selected from the group consisting of asthma, mild asthma, moderate asthma, severe asthma, mild eosinophilic asthma, moderate eosinophilic asthma, severe eosinophilic asthma, uncontrolled eosinophilic asthma, eosinophilic asthma, subeosinophilic asthma, chronic obstructive pulmonary disease, eosinophilic granulomatosis with polyangiitis, hypereosinophilic syndrome, nasal polyposis, bullous pemphigoid, eosinophilic esophagitis, atopic dermatitis, moderate atopic dermatitis, and severe atopic dermatitis, and the composition comprises an antibody comprising the heavy chain amino acid sequence set forth in SEQ ID NO: 1 and the light chain amino acid sequence set forth in SEQ ID NO: 2, and a therapeutically effective amount of the antibody administered subcutaneously to the pediatric subject provides approximately 10.1960 ± 0.3345 The maximum plasma concentration (Cmax) of the antibody was approximately 454.39 ± 15.8876 μg*day / mL, and the area under the curve [0-infinity] value was approximately 454.39 ± 15.8876 μg*day / mL.

[0194] A composition for reducing the absolute blood eosinophil count in a pediatric subject weighing greater than or equal to 40 kg, wherein the pediatric subject has a disease selected from the group consisting of asthma, mild asthma, moderate asthma, severe asthma, mild eosinophilic asthma, moderate eosinophilic asthma, severe eosinophilic asthma, uncontrolled eosinophilic asthma, eosinophilic asthma, subeosinophilic asthma, chronic obstructive pulmonary disease, eosinophilic granulomatosis with polyangiitis, hypereosinophilic syndrome, nasal polyposis, bullous pemphigoid, eosinophilic esophagitis, atopic dermatitis, moderate atopic dermatitis, and severe atopic dermatitis, and the composition comprises an antibody comprising the heavy chain amino acid sequence set forth in SEQ ID NO: 1 and the light chain amino acid sequence set forth in SEQ ID NO: 2, and a therapeutically effective amount of the antibody administered subcutaneously to the pediatric subject provides approximately 16.3412 ± 0.6364 The maximum plasma concentration (Cmax) of the antibody was 1.5 μg / mL, and the area under the curve [0-infinity] value was approximately 675.20±35.8980 μg*day / mL.

[0195] Another embodiment is the composition of the present disclosure, wherein said antibody is administered about once per month.

[0196] Another embodiment is the composition of the present disclosure, wherein the pediatric subject has an absolute blood eosinophil count selected from the group consisting of: greater than or equal to 150 cells / μL and greater than or equal to 300 cells / μL.

[0197] Another embodiment is the composition of the present disclosure, wherein the pediatric subject is selected from premature newborns, full-term newborns, children 28 days to 11 months, children 12 months to 23 months, children 2 years to 6 years, children 16 years to 12 years, children 6 years to less than 18 years, and children 12 to 18 years.

[0198] Another embodiment is the composition of the present disclosure, wherein the pediatric subject is between about 6 years of age and about 12 years of age, inclusive.

[0199] Another embodiment is the composition of the present disclosure, wherein the pediatric subject has severe eosinophilic asthma.

[0200] Another embodiment is the composition of the present disclosure, wherein the pediatric subject has a treatment outcome selected from the group consisting of: a decrease in absolute blood eosinophil count relative to the absolute blood eosinophil count before antibody administration, a decrease in the score on the Asthma Control Questionnaire-7 scale relative to the score on the Asthma Control Questionnaire-7 scale before antibody administration, and a decrease in the score on the Childhood Asthma Control Test relative to the score on the Childhood Asthma Control Test before antibody administration.

[0201] Another embodiment is the composition of the present disclosure, wherein the antibody is administered by a safety syringe or autoinjector.

[0202] Another embodiment is the composition of the present disclosure, wherein the therapeutically effective amount of the antibody is about 100 mg. Example

[0203] Example 1 Pharmacokinetics and pharmacodynamics of subcutaneously administered mepolizumab in children Mepolizumab is a humanized immunoglobulin G (IgG1) monoclonal antibody (mAb) that exhibits dose-proportional and time-dependent pharmacokinetics. As described herein, the pharmacokinetics (PK) and pharmacodynamics (PD) of mepolizumab in children aged 6 to 11 years with severe eosinophilic asthma were studied. In Part A of the study, the PK / PD of subcutaneously administered mepolizumab 40 milligrams (mg) or 100 mg, depending on the weight of the participant, were described. Part B of the study will be a long-term safety / pharmacodynamic phase, in which a further 52-week extended treatment will be provided on an optional basis to those subjects who qualify for continued treatment.

[0204] In the study, at Visit 2 (Week 0), pediatric subjects weighing <40 kilograms (kg) were dosed with mepolizumab 40 mg subcutaneously in the upper arm or thigh, while participants weighing >= 40 kg were dosed with mepolizumab 100 mg subcutaneously. Approximately 40 male or female participants aged 6 to 11 years were screened as described below to achieve approximately 28 eligible participants for the treatment phase to allow for the availability of 20 evaluable participants, with a minimum of 6 participants enrolled in the <40 kg weight group. The total duration of the study was 22 weeks and included a 1-2 week run-in phase, a 12-week treatment phase, and an 8-week follow-up phase. Participants were considered to have completed the study if they completed all phases of the study, including the follow-up phase (Week 20 [Visit 8]).

[0205] In the first arm of the study, participants were administered 40 mg of mepolizumab (this arm included patients who had completed Part A of the PK / PD study and were about to complete Part B). In this arm of the study, participants weighing <40 kg received 0.4 milliliters (mL) of reconstituted mepolizumab administered subcutaneously in the upper arm or thigh. Mepolizumab for this arm was provided as a 100 mg lyophilized cake in a sterile vial for subcutaneous administration in the upper arm or thigh. The vial was reconstituted with sterile water for injection prior to individual use.

[0206] In the second arm of the study, participants were administered 100 mg of mepolizumab (this arm included patients who had completed Part A of the PK / PD study and were about to complete Part B). In this arm of the study, participants weighing >= 40 kg received 1.0 mL of reconstituted mepolizumab subcutaneously in the upper arm or thigh. Mepolizumab for this second arm was provided and reconstituted as described above for the first arm of the study.

[0207] Pediatric patients in the study met the following inclusion criteria: • Aged 6 to 11 years (inclusive) at screening. • A diagnosis of severe asthma as defined by regional asthma guidelines (i.e., National Institute of Health (NIH), Global Initiative for Asthma (GINA), etc.) for at least 12 months prior to Visit 1. If the participant has never visited the study site, the participant / guardian must self-report a physician diagnosis of asthma, and the investigator must confirm this by reviewing the medical history with the participant / guardian. • Eosinophilic airway inflammation associated with asthma characterized as eosinophilic in nature as indicated by an elevated peripheral blood eosinophil count of >= 300 cells / microliter (cells / μL) demonstrated within the past 12 months or an elevated peripheral blood eosinophil count of >= 150 / μL at Visit 1. • Adequately documented need for regular treatment with inhaled corticosteroids (>200 μg / day fluticasone propionate powder inhaler [DPI] or daily equivalent), with or without maintenance oral corticosteroids (OCS), within the 12 months prior to visit 1. ICS doses should represent moderate or high doses in children aged 6–11 years [GINA]. • Current treatment with an alternative controller for at least 3 months, or documented failure of an alternative controller for at least 3 consecutive months within the past 12 months [e.g., long-acting beta-2-agonist (LABA), leukotriene receptor antagonist (LTRA), or theophylline.] • Forced expiratory volume in one second (FEV1): persistent airway obstruction at Visit 1 or Visit 2 (FEV1 performed before the first dose of study medication), as indicated by: pre-bronchodilator FEV1 <110% predicted (Quanjer, 2012) or FEV1:forced vital capacity (FVC) ratio <0.8. • History of two or more previously confirmed acute exacerbations requiring treatment with systemic corticosteroids (CS) (intramuscular [IM], intravenous, or oral) in the 12 months prior to Visit 1, despite the use of high-dose inhaled corticosteroids (ICS). For participants receiving maintenance CS, CS treatment for acute exacerbations must have involved a two-fold or greater increase in dose. • During the run-in period, there are no changes in the dose or regimen of baseline ICS and / or additional controller medication. • Males or females: Females of childbearing potential must commit to consistent and correct use of an acceptable method of contraception for the duration of the trial and for 4 months after the last dose of the investigational product. Females of childbearing potential require a urine pregnancy test. This test will be performed at the initial Screening Visit (Visit 1) and at each scheduled study visit prior to administration of the investigational product and during early withdrawal and follow-up visits. • Parents / guardians must be able to give written informed consent prior to participation in the research, which includes the ability to comply with the requirements and restrictions outlined in the consent form. Participants must be able and willing to give consent to participate in the research according to local requirements, if applicable. • For Part B: Subjects who completed all study assessments up to and including Visit 8 and received all 3 doses of Investigational Product (IP) in Part A • For Part B: The principal investigator (PI) has performed a benefit / risk evaluation, and this evaluation supports continued therapy with mepolizumab. • The subject's parent (or guardian) has given written consent and the subject has given consent for continued treatment.

[0208] The results for patients treated according to Part A of the study are as follows (Table 16); the outcome areas to be evaluated in Part B of the study are also as follows: The "50 kg" group identified above for certain rows corresponds to pediatric patients weighing greater than or equal to 40 kg, while the "27 kg" group identified above for certain rows corresponds to pediatric patients weighing less than 40 kg.

[0209] Example 2 Pharmacokinetic and pharmacodynamic analysis Pharmacokinetic (PK) and pharmacodynamic (PD) analyses of the study results were performed in sections as described herein. Plasma PK concentrations of mepolizumab collected in the first part of the study were analyzed by population PK methods using the NLMIXED procedure in SAS software (version 9.2). Because the number of blood draws was kept to a minimum, a two-compartment PK model with first-order absorption and first-order elimination, parameterized by macroconstants (i.e., A, B, α, β), was used, with distribution parameters fixed to adult values. Descriptive statistics were used to summarize the ratio of blood eosinophil counts to baseline. An indirect response PK / PD model was used to estimate the plasma concentrations that resulted in the maximal drug effect (maximal reduction in eosinophils) and 50% of the maximal drug effect. The derived exposure (area under the concentration-time curve to infinity) normalized to mean body weight in each subcutaneous dose group was 454 μg*day / mL (40 mg [subjects <40 kg]) and 675 μg*day / mL (100 mg [subjects ≥40 kg]). The estimated half-life (t ½ ) is about 23 days. PK data are shown in Table 17. In the methods or compositions of the present disclosure, Cmax SS values ​​may be described relative to those listed in Table 17 or including the indicated ranges shown in Table 17, rather than Cmax values.

[0210] In this 6- to 11-year-old population, significant and similar blood eosinophilia was observed in both treatment groups, which was evident from Week 4 (first post-dose assessment) and maintained throughout the treatment period, with recovery toward baseline by the 8-week post-treatment period ( Figure 3 At Week 12, reductions from baseline in blood eosinophils were 89% (40 mg SC), 83% (100 mg SC), and 87% (overall) ( Figure 3 Absolute blood eosinophil counts (geometric mean) showed similar decreases and trends in blood eosinophils. Figure 3 In the data presented, weeks 0–12 were considered as on-treatment, weeks 16–20 as post-treatment, and—when a zero result was recorded—a small value (0.005) was added before logarithmic transformation.

[0211] Example 3 Informal Sequence Listing The underlining below identifies the CDR sequences, or nucleic acid sequences encoding these CDR sequences, in the variable heavy and variable light chain portions of an antibody according to the Kabat definition of CDRs. For example, in SEQ ID NO: 1, the frameworks and CDRs are presented as plain text framework 1, underlined CDR1, plain text framework 2, underlined CDR2, plain text framework 3, underlined CDR3, and plain text framework 4, in order from the amino-proximal portion of the presented sequence to the carboxyl-terminal portion. Italics below identify signal sequences. An asterisk to the right of a character in a single-letter amino acid code indicates that the amino acid residue to the left is an N-glycosylation site. This scheme is used, for example, in SEQ ID NOs: 1-4, 11, 12, and 19-22. The amino-terminal methionine residues shown in these sequences can be cleaved. Therefore, sequences showing an amino-terminal methionine residue herein should also be considered to disclose cleaved forms of these proteins lacking such amino-terminal methionine residues. Nucleic acid sequences are presented as DNA nucleic acid sequences and include "t" nucleic acid residues. The corresponding RNA sequences should also be considered disclosed, such that the "t" nucleic acid residues can also be considered disclosed as "u" nucleic acid residues. In addition, the 5'-proximal "atg" start codon and the 3'-proximal "taa," "tag," and "tga" stop codons have been omitted from the cDNA nucleic acid sequences below. This is the case, for example, for SEQ ID NOs: 31-34, etc.

[0212] Mepolizumab full-length heavy chain Mepolizumab full-length light chain Mepolizumab VH Mepolizumab VL Mepolizumab CDRH1 Mepolizumab CDRH2 Mepolizumab CDRH3 Mepolizumab CDRL1 Mepolizumab CDRL2 Mepolizumab CDRL3 HUMANIL-5 (mature protein) Human IL-5 receptor subunit alpha isoform 1 (mature protein) Human IL-5 receptor common subunit β (mature protein)

[0213] The invention now being fully described, it will be apparent to those skilled in the art that many changes and modifications can be made therein without departing from the spirit or scope of the appended claims.

Claims

1. An aqueous liquid preparation comprising: (a) 40 mg of an antibody comprising a heavy chain amino acid sequence of SEQ ID NO: 1 and a light chain amino acid sequence of SEQ ID NO: 2; (b) a buffer selected from sodium hydrogen phosphate heptahydrate, phosphate, citric acid, citrate, sodium phosphate, potassium phosphate, sodium citrate, histidine or a combination thereof; (c) a pH between about 6.2 and about 6.6; (d) sugar; (e) polysorbate 80; and (f) EDTA.

2. The aqueous liquid formulation of claim 1, wherein the sugar is present in an amount ranging from about 5% to about 20% weight / volume.

3. The aqueous liquid formulation of claim 1, wherein polysorbate 80 is present in an amount ranging from about 0.01% to about 0.1% weight / volume.

4. The aqueous liquid formulation of claim 3, wherein polysorbate 80 is present in an amount of about 0.05% weight / volume.

5. The aqueous liquid preparation of claim 2, wherein the sugar is sucrose.

6. The aqueous liquid formulation of claim 5, wherein sucrose is present in an amount of about 12% weight / volume.

7. The aqueous liquid formulation of claim 1, wherein EDTA is present in an amount ranging from about 0.01 mM to about 0.1 mM.

8. The aqueous liquid formulation of claim 7, wherein EDTA is present in an amount of about 0.05 mM.

9. The aqueous liquid formulation of claim 1, wherein the buffer is selected from disodium hydrogen phosphate heptahydrate, citric acid monohydrate, or a combination thereof.

10. The aqueous liquid formulation of claim 9, wherein the sodium phosphate dibasic heptahydrate is present in an amount ranging from about 10 mM to about 30 mM.

11. The aqueous liquid formulation of claim 10, wherein the sodium phosphate dibasic heptahydrate is present in an amount ranging from about 15 mM to about 16.4 mM.

12. The aqueous liquid formulation of claim 9, wherein citric acid monohydrate is present in an amount ranging from about 3.8 mM to about 4.9 mM.

13. The aqueous liquid formulation of claim 9, wherein disodium hydrogen phosphate heptahydrate is present in an amount of about 15.5 mM and citric acid monohydrate is present in an amount of about 4.5 mM.

14. The aqueous liquid formulation of claim 1, wherein the pH is about 6.

3.

15. An aqueous liquid preparation comprising: (a) about 40 mg of an antibody comprising a heavy chain amino acid sequence of SEQ ID NO: 1 and a light chain amino acid sequence of SEQ ID NO: 2; (b) about 15 mM sodium phosphate dibasic heptahydrate and about 4.5 mM citric acid monohydrate, about pH 6.3; (c) about 12% weight / volume sugar; (d) about 0.02% weight / volume polysorbate 80; and (e) About 0.05 mM EDTA.

16. A safety syringe comprising the aqueous liquid preparation according to claim 15.

17. An automatic injector comprising the aqueous liquid formulation of claim 15.

Images