Methods for treating asthma using TSLP antibodies

Abstract

The present disclosure provides methods for treating asthma using TSLP antibodies. In particular, the present disclosure utilizes antibodies against thymic stromal lymphopoietin (TSLP) or antigen-binding fragments thereof to treat asthma and / or reduce the frequency and / or severity of asthma exacerbations and / or reduce the annualized asthma exacerbation rate and / or reduce a subject's ACQ-6 score.

Description

[Technical Field]

[0001] The present disclosure relates to methods for treating asthma, or reducing the frequency and / or severity of asthma exacerbations, or reducing the annualized asthma exacerbation rate, or reducing a subject's ACQ-6 score using antibodies or antigen-binding fragments thereof against thymic stromal lymphopoietin (TSLP). [Background technology]

[0002] Asthma is a serious chronic inflammatory disease of the airways that affects approximately 334 million people worldwide. Due to environmental degradation and increasing air pollution, more people may suffer from the disease, posing a serious threat to human life and health.

[0003] Thymic stromal lymphopoietin (TSLP) is a cytokine produced by epithelial cells in response to proinflammatory stimuli and promotes allergic inflammatory responses primarily through its effects on dendritic cells and mast cells. TSLP, a cytokine similar to interleukin-7 (IL-7), was first discovered in conditioned medium from mouse thymic stromal cells. TSLP is predominantly expressed in lung, skin, and intestinal epithelial cells. TSLP consists of four alpha-helices and two loops, AB and CD, with three pairs of disulfide bonds formed by six cysteine ​​residues, two N-glycosylation sites, and a molecular weight of approximately 15–20 kD. The TSLP receptor is a complex containing two components: TSLPR and IL-7Rα. TSLP first binds to TSLPR with relatively low affinity, then recruits IL-7Rα with high affinity, and finally activates signaling pathways such as STAT5, leading to dendritic cell (DC) maturation and T cell differentiation.

[0004] Myeloid dendritic cells (mDCs) are the primary effector cells of TSLP, and TSLP acts on immature mDCs to induce the secretion of cytokines IL-8, eotaxin-2, TARC, and MDC, and upregulate OX40L expression. In the absence of IL-12, OX40L binds to naive CD4+ T cells and promotes their differentiation into Th2 cells, which subsequently secrete Th2 cytokines such as IL-5, IL-4, IL-9, IL-13, and TNF, thus inducing a Th2-mediated inflammatory response. Furthermore, TSLP can induce DCs to produce IL-8, which recruits neutrophils, leading to neutrophil-mediated innate immune inflammation. TSLP can also induce DCs to produce eotaxin-2, which recruits eosinophils. Together with IL-5, this promotes rapid eosinophil infiltration and eosinophilic inflammation. TSLP also acts on mast cells and natural killer cells and mediates innate inflammation through the induction of IL-4, IL-6, IgE, and other factors. In summary, TSLP can simultaneously induce both innate and Th2 inflammation, leading to increased mucus production, airway remodeling and narrowing, severe cellular fibrosis, and the progressive development of asthma, atopic dermatitis, and allergic rhinitis—the three major allergic diseases. Therefore, blocking TSLP is a potentially effective strategy for treating asthma, atopic dermatitis, and related disorders.

[0005] Currently, only tezepelumab (TEZSPIRE), co-developed by AstraZeneca (AZ) and Amgen, is approved for the market. In the Phase II PATHWAY study conducted in patients with severe, uncontrolled asthma, the dosing protocol included a low-dose group (70 mg Q4W), a medium-dose group (210 mg Q4W), and a high-dose group (280 mg Q2W), with the final approved dosing regimen being 210 mg Q4W.

[0006] WO 2020 / 244544 discloses novel TSLP antibodies or antigen-binding fragments thereof, which are incorporated herein by reference. Summary of the Invention

[0007] The present disclosure provides methods and pharmaceutical uses for treating asthma or reducing the frequency and / or severity of asthma exacerbations and / or reducing the annualized asthma exacerbation rate and / or reducing the ACQ-6 score of a subject in need thereof, which methods are characterized by simple protocols, low administration frequency and small dosages, effectively reducing treatment burden and improving patient compliance.

[0008] The present disclosure provides any one or any combination of the following methods: 1) a method for treating a subject suffering from asthma, 2) a method for reducing the frequency and / or severity of asthma exacerbations in a subject, 3) a method for reducing the annualized asthma exacerbation rate in a subject, 4) a method for reducing the ACQ-6 score in a subject. Meanwhile, the present disclosure provides any one or any combination of the following uses: 1) Use of an anti-TSLP antibody or antigen-binding fragment thereof in the preparation of a medicament for treating a subject suffering from asthma; 2) Use of an anti-TSLP antibody or antigen-binding fragment thereof in the preparation of a medicament for reducing the frequency and / or severity of asthma exacerbations in a subject; 3) Use of an anti-TSLP antibody or antigen-binding fragment thereof in the preparation of a medicament for reducing the annualized asthma exacerbation rate in a subject; 4) Use of an anti-TSLP antibody or antigen-binding fragment thereof in the preparation of a medicament for reducing a subject's ACQ-6 score; 5) an anti-TSLP antibody or antigen-binding fragment thereof for treating a subject suffering from asthma; 6) an anti-TSLP antibody or antigen-binding fragment thereof for reducing the frequency and / or severity of asthma exacerbations in a subject; 7) An anti-TSLP antibody or antigen-binding fragment thereof for reducing the annualized asthma exacerbation rate in a subject; 8) An anti-TSLP antibody or antigen-binding fragment thereof for reducing a subject's ACQ-6 score.

[0009] Meanwhile, the present disclosure further provides any one of the following uses of an anti-TSLP antibody or antigen-binding fragment thereof in combination with other asthma medications (e.g., baseline therapies for asthma): 1) Use in the preparation of medicines, 2) Use in the preparation of a medicament for treating a subject suffering from asthma or reducing the frequency and / or severity of asthma exacerbations or reducing the ACQ-6 score of a subject; 3) Use for treating a subject suffering from asthma, or reducing the frequency and / or severity of asthma exacerbations, or reducing the annualized asthma exacerbation rate, or reducing the ACQ-6 score of a subject.

[0010] In some embodiments, the other asthma medication is: a) corticosteroids, such as inhaled corticosteroids (ICS) (e.g., fluticasone propionate, budesonide, beclomethasone) or oral corticosteroids (e.g., hydrocortisone, dexamethasone, prednisone), b) bronchodilators, such as long-acting beta-agonists (LABA) (e.g., vilanterol, formoterol), short-acting beta-agonists (SABA) (e.g., salbutamol, levalbuterol) and anticholinergics, such as ipratropium bromide, tiotropium bromide, aclidinium and glycopyrronium bromide; c) long-acting muscarinic receptor antagonists (LAMAs), such as tiotropium, umeclidinium, glycopyrronium, d) Leukotriene receptor antagonists (LTRAs), such as theophylline or its derivatives, montelukast, zafirlukast and pranlukast; e) Cromolyn, f) antihistamines, g) anti-leukotrienes, h PDE-4 inhibitors Includes.

[0011] In some embodiments, the other asthma medication is: 1) an inhaled corticosteroid (ICS) (e.g., fluticasone propionate, budesonide, beclomethasone); and 2) at least one additional medication selected from the group consisting of: a) oral corticosteroids (e.g., hydrocortisone, dexamethasone, prednisone), b) bronchodilators, such as long-acting beta-agonists (LABA) (e.g., vilanterol, formoterol), short-acting beta-agonists (SABA) (e.g., salbutamol, levalbuterol) and anticholinergics, such as ipratropium bromide, tiotropium bromide, aclidinium and glycopyrronium bromide; c) long-acting muscarinic receptor antagonists (LAMAs), such as tiotropium, umeclidinium, glycopyrronium, d) Leukotriene receptor antagonists (LTRAs), such as theophylline or its derivatives, montelukast, zafirlukast and pranlukast; e) Cromolyn, f) antihistamines, g) anti-leukotrienes, h) PDE-4 inhibitors Includes.

[0012] In some embodiments, the other asthma medication includes: a medium-dose ICS (e.g., fluticasone propionate, budesonide, beclomethasone), such as a DPI or pMDI fluticasone propionate or equivalent at a total daily dose of more than 250 μg and not more than 500 μg. In some embodiments, the other asthma medication includes: a high-dose ICS (e.g., fluticasone propionate, budesonide, beclomethasone), such as a DPI or pMDI fluticasone propionate or equivalent at a total daily dose of more than 500 μg. In some embodiments, the equivalent ICS dosage is based on the Guidelines for the Prevention and Treatment of Bronchial Asthma (2020 edition).

[0013] In some embodiments, the uses or methods disclosed herein delay the time to an asthma exacerbation compared to not administering the anti-TSLP antibody or antigen-binding fragment.

[0014] In some embodiments, the uses or methods disclosed herein reduce the dose or frequency of administration of a background therapy, in some embodiments, the background therapy comprises a medium- or high-dose inhaled corticosteroid (ICS) (e.g., fluticasone propionate, budesonide, beclomethasone).

[0015] The term "background therapy" refers to the standard or usual treatment for asthma known in the art.

[0016] In some embodiments, the method or use comprises: 1) administering at least one anti-TSLP antibody or antigen-binding fragment to a patient at intervals of more than 4 weeks between doses, wherein the dose interval is selected from 5, 6, 7, 8, 9, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32 weeks or more, or the dose interval is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 months or more, or 2) administering once every 10, 12, 14, 16, 18, 20, 22, 24, 26, 28 weeks, or once every 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 months. In some embodiments, the method comprises: 1) a dosing interval selected from 12, 16, 20, 24 weeks or more, or a dosing interval selected from 3 months, 4 months, 5 months, 6 months or more, or 2) administration once every 12, 16, 20, 24 weeks, or once every 3 months, 4 months, 5 months, 6 months. In some embodiments, the method comprises: 1) a dosing interval selected from 12 weeks, 24 weeks or more, or a dosing interval selected from 3 months, 6 months or more, or 2) administration once every 12 weeks, 24 weeks, or once every 3 months, 6 months.

[0017] In some embodiments, the method or use comprises administering to a patient at least one anti-TSLP antibody or antigen-binding fragment at a dosage of up to 600 mg per administration with an interval of 10 weeks or more between doses, wherein the interval between doses is selected from 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32 weeks or more, or the interval between doses is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 months or more. In some embodiments, the interval between doses is selected from 12, 16, 20, 24 weeks or more, or the interval between doses is selected from 3, 4, 5, 6 months or more. In some embodiments, the interval between doses is selected from 12 weeks, 24 weeks or more, or the interval between doses is selected from 3 months, 6 months or more.

[0018] In some embodiments, the dosage of an anti-TSLP antibody or antigen-binding fragment is up to about 600 mg, e.g., 50-600 mg, e.g., 50-550 mg, 50-500 mg, 50-450 mg, 50-400 mg, 100-550 mg, 100-500 mg, 100-450 mg, 100-400 mg, 150-500 mg, 150-450 mg, or 150-400 mg. In some embodiments, the dosage of an anti-TSLP antibody or antigen-binding fragment is 100-500 mg or 150-450 mg. In some embodiments, the dosage of the anti-TSLP antibody or antigen-binding fragment is about 50 mg, about 100 mg, about 150 mg, about 175 mg, about 200 mg, about 225 mg, about 250 mg, about 275 mg, about 300 mg, about 325 mg, about 350 mg, about 375 mg, about 400 mg, about 450 mg, or about 600 mg. In some embodiments, the dosage of the anti-TSLP antibody or antigen-binding fragment is selected from about 150 mg, about 200 mg, about 250 mg, about 300 mg, about 350 mg, about 400 mg, or about 450 mg. In some embodiments, the dosage of the anti-TSLP antibody or antigen-binding fragment is about 200 mg or about 400 mg.

[0019] In some embodiments, the dosage of the anti-TSLP antibody or antigen-binding fragment is 100-500 mg, and the dosing interval is selected from 12, 16, 20, or 24 weeks. In some embodiments, the dosage of the anti-TSLP antibody or antigen-binding fragment is 150-450 mg, and the dosing interval is 12 or 24 weeks. In some embodiments, the dosing regimen of the anti-TSLP antibody or antigen-binding fragment is selected from 200 mg Q24W, 400 mg Q24W, or 400 mg Q12W.

[0020] In some embodiments, the anti-TSLP antibody or antigen-binding fragment is administered for a period of at least 2 months, 3 months, 4 months, 5 months, 6 months, 8 months, 10 months, 12 months or more.

[0021] In some embodiments of the present disclosure, the anti-TSLP antibody or antigen-binding fragment is administered parenterally. In some embodiments, the TSLP antibody or antigen-binding fragment is administered intravenously. In some embodiments, the TSLP antibody or antigen-binding fragment is administered subcutaneously. In some embodiments, the TSLP antibody or antigen-binding fragment is administered by subcutaneous injection. For example, an injectable form of the anti-TSLP antibody or antigen-binding fragment is an injection solution or lyophilized powder injection containing the anti-TSLP antibody or antigen-binding fragment, a buffer, a stabilizer, and a surfactant. The buffer is a histidine-acetic acid system, and the stabilizer may be selected from one or more of sugars, amino acids, and EDTA, such as sugars, e.g., trehalose, sucrose, sorbitol, and mannitol; amino acids, e.g., histidine, tryptophan, and methionine; and EDTA. The surfactant is selected from polysorbates, e.g., polysorbate 80 or polysorbate 20, e.g., polysorbate 80. For example, an injectable formulation of a TSLP antibody or antigen-binding fragment includes an anti-TSLP antibody or antigen-binding fragment, histidine-acetate buffer, sucrose, and polysorbate 80. WO 2022 / 116858 relates to compositions of anti-TSLP antibodies, the entire contents of which are incorporated herein by reference.

[0022] In some embodiments, the subject suffers from allergic asthma or non-allergic asthma.

[0023] In some embodiments, the subject suffers from mild asthma.

[0024] In some embodiments, the subject has moderate to severe asthma or severe asthma.

[0025] In some embodiments, the subject has uncontrolled asthma. In some embodiments, the subject has severe uncontrolled asthma.

[0026] In some embodiments, the subject suffers from eosinophilic asthma or noneosinophilic asthma.

[0027] In some embodiments, the subject has a high eosinophil count at the start of treatment or at diagnosis, eg, 250 cells / μL or more, eg, 300 cells / μL or more.

[0028] In some embodiments, the subject has a low eosinophil count, eg, less than 300 cells / μL, eg, less than 250 cells / μL, at the start of treatment or at diagnosis.

[0029] In some embodiments, the subject is an adult, hi some embodiments, the subject is a child or adolescent.

[0030] In some embodiments, the subject exhibits a low Th2 profile, ie, an IgE level of 100 IU / mL or less or an eosinophil count of less than 140 cells / μL, particularly at the start of treatment or at the time of diagnosis.

[0031] In some embodiments, the subject has moderate to severe asthma or severe asthma with an incomplete or poorly controlled response to baseline asthma therapy.

[0032] In some embodiments, the subject continues on maintenance baseline therapy.

[0033] In some embodiments, the baseline therapy includes at least one of the following: a) corticosteroids, such as inhaled corticosteroids (ICS) (e.g., fluticasone propionate, budesonide, beclomethasone) or oral corticosteroids (e.g., hydrocortisone, dexamethasone, prednisone), b) bronchodilators, such as long-acting beta-agonists (LABA) (e.g., vilanterol, formoterol), short-acting beta-agonists (SABA) (e.g., salbutamol, levalbuterol) and anticholinergics, such as ipratropium bromide, tiotropium bromide, aclidinium and glycopyrronium bromide; c) long-acting muscarinic receptor antagonists (LAMAs), such as tiotropium, umeclidinium, glycopyrronium, d) Leukotriene receptor antagonists (LTRAs), such as theophylline or its derivatives, montelukast, zafirlukast and pranlukast; e) Cromolyn, f) antihistamines, g) anti-leukotrienes, h) PDE-4 inhibitors.

[0034] In some embodiments, the baseline therapy includes at least one of the following: 1) a medium or high dose inhaled corticosteroid (ICS) (e.g., fluticasone propionate, budesonide, beclomethasone), and 2) at least one additional agent selected from the following group: a) oral corticosteroids (e.g., hydrocortisone, dexamethasone, prednisone), b) bronchodilators, such as long-acting beta-agonists (LABA) (e.g., vilanterol, formoterol), short-acting beta-agonists (SABA) (e.g., salbutamol, levalbuterol) and anticholinergics, such as ipratropium bromide, tiotropium bromide, aclidinium and glycopyrronium bromide; c) long-acting muscarinic receptor antagonists (LAMAs), such as tiotropium, umeclidinium, glycopyrronium, d) Leukotriene receptor antagonists (LTRAs), such as theophylline or its derivatives, montelukast, zafirlukast and pranlukast; e) Cromolyn, f) antihistamines, g) anti-leukotrienes, h) PDE-4 inhibitors.

[0035] In some embodiments, the baseline therapy includes at least one of the following: 1) a medium or high dose inhaled corticosteroid (ICS) (e.g., fluticasone propionate, budesonide, beclomethasone), and 2) at least one additional agent selected from the following group: a) oral corticosteroids (e.g., hydrocortisone, dexamethasone, prednisone), b) bronchodilators, such as long-acting beta-agonists (LABA) (e.g., vilanterol, formoterol), short-acting beta-agonists (SABA) (e.g., salbutamol, levalbuterol) and anticholinergics, such as ipratropium bromide, tiotropium bromide, aclidinium and glycopyrronium bromide; c) long-acting muscarinic receptor antagonists (LAMAs), such as tiotropium, umeclidinium, glycopyrronium, d) Leukotriene receptor antagonists (LTRAs), such as theophylline or its derivatives, montelukast, zafirlukast and pranlukast; e) Cromolyn, f) antihistamines, g) anti-leukotrienes, h) PDE-4 inhibitors Including, High-dose inhaled corticosteroids are DPIs or pMDIs containing fluticasone propionate or equivalent at a total daily dose of more than 500 μg. The medium-dose inhaled corticosteroid is a DPI or pMDI fluticasone propionate or equivalent drug at a total daily dose of >250-500 μg. In some embodiments, the equivalent ICS dosage is based on the Guidelines for the Prevention and Treatment of Bronchial Asthma (2020 edition). In some embodiments, the baseline therapy is: a) corticosteroids, such as medium- or high-dose inhaled corticosteroids (ICS), b) In combination with at least one other asthma control medication selected from long-acting beta-2 receptor agonists (LABAs), long-acting cholinergic receptor antagonists (LAMAs), leukotriene receptor antagonists (LTRAs), or oral corticosteroids (OCSs) in accordance with the Guidelines for the Prevention and Treatment of Bronchial Asthma (2020 edition). Includes.

[0036] In some embodiments, the background therapy includes: a medium dose ICS (e.g., fluticasone propionate, budesonide, beclomethasone), such as a DPI or pMDI fluticasone propionate or equivalent drug at a total daily dose of more than 250 μg and less than or equal to 500 μg.

[0037] In some embodiments, the background therapy includes: high dose ICS (eg, fluticasone propionate, budesonide, beclomethasone), eg, a total daily dose of more than 500 μg DPI or pMDI fluticasone propionate or equivalent drug.

[0038] In some embodiments, the methods of the invention delay the time to an asthma exacerbation compared to without administration of an anti-TSLP antibody or antigen-binding fragment thereof.

[0039] In some embodiments, the methods of the present invention reduce the dosage or frequency of administration of a background therapy, in some embodiments, the background therapy comprises a medium- or high-dose inhaled corticosteroid (ICS) (e.g., fluticasone propionate, budesonide, beclomethasone).

[0040] In some embodiments, the subject is selected according to at least one of the following criteria: a) prior to administration of the antibody or antibody-binding fragment, the subject has a pre-bronchodilator FEV1 of 40% or greater and less than 80% predicted; b) Prior to administration of the antibody or antibody-binding fragment, the subject has an ACQ-6 score of 1.5 or greater.

[0041] In some embodiments, at 24 or 48 weeks of treatment, the subject achieves at least one of the following: a) Improvement in annualized asthma exacerbation rate (AAER), b) Improvement of forced vital capacity (FVC), c) Improvement in forced expiratory volume in 1 second (FEV1), d) improvement of peak expiratory flow (PEF), e) improvement in exhaled nitric oxide (FeNO), blood eosinophil count (EOS), and / or serum immunoglobulin (IgE); f) reduction in ACQ-6 score; g) improvement in Asthma Quality of Life Questionnaire (AQLQ) scores; h) improvement in asthma symptom diary scores; i) reduced use of rescue medications; j) a reduction in the average number of nighttime awakenings; k) reduction in AAERs leading to hospitalization at 48 weeks; l) improvement in time to first acute asthma attack; m) Improvement of SNOT-22, n)Improvement in the incidence of CompEx (composite exacerbation endpoint).

[0042] In some embodiments, the method reduces eosinophils in the subject's blood, sputum, bronchoalveolar lavage fluid, or lungs.

[0043] In some embodiments, the method reduces the number of Th2 cells, eg, converts the number of cells in a subject from a Th2-high to a Th2-low population.

[0044] In another aspect, the disclosure provides a method for treating asthma or reducing the frequency and / or severity of asthma exacerbations and / or reducing the annualized asthma exacerbation rate and / or reducing the ACQ-6 score in a subject, comprising administering a therapeutically effective amount of an anti-TSLP antibody or antigen-binding fragment thereof, wherein the subject has an eosinophilic or hypoeosinophilic profile.

[0045] In another aspect, the disclosure provides a method for treating asthma, or reducing the frequency and / or severity of asthma exacerbations, or reducing the annualized asthma exacerbation rate, or reducing the ACQ-6 score in a subject, comprising administering a therapeutically effective amount of an anti-TSLP antibody or antigen-binding fragment thereof, wherein the subject has a low Th2 profile.

[0046] In another aspect, the disclosure provides an anti-TSLP antibody or antigen-binding fragment thereof for use in treating asthma in a subject, and / or reducing the frequency and / or severity of asthma exacerbations, and / or reducing the annualized asthma exacerbation rate, and / or reducing the ACQ-6 score of the subject, wherein the subject has a low Th2 profile.

[0047] In some embodiments, the anti-TSLP antibody or antigen-binding fragment is any of the anti-TSLP antibodies or antigen-binding fragments disclosed in WO 2020 / 244544 or WO 2022 / 116858, which are incorporated by reference in their entireties.

[0048] In some embodiments, the anti-TSLP antibody or antigen-binding fragment comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 set forth in SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3, respectively; The light chain variable region comprises LCDR1, LCDR2 and LCDR3, which are shown in SEQ ID NO:4, SEQ ID NO:5 and SEQ ID NO:6, respectively.

[0049] The aforementioned CDR sequences are listed in the table below: [Table 1]

[0050] In some embodiments, the anti-TSLP antibody is a murine antibody, a chimeric antibody, or a humanized antibody.

[0051] In optional embodiments, the light and heavy chain framework region sequences in the variable regions of the humanized antibody light and heavy chains are human germline light and heavy chain framework region sequences or variants thereof.

[0052] In some embodiments, the anti-TSLP antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO:7 or an amino acid sequence having at least 90% identity thereto, and the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO:8 or an amino acid sequence having at least 90% identity thereto.

[0053] Heavy chain variable region [ka]

[0054] In some embodiments, the anti-TSLP antibody or antigen-binding fragment thereof further comprises an antibody constant region, e.g., the heavy chain constant region of the antibody constant region is selected from human IgG1, IgG2, IgG3, and IgG4 constant regions and conventional variants thereof, and the light chain constant region of the antibody constant region is selected from human antibody kappa and lambda chain constant regions and conventional variants thereof.

[0055] Light chain variable region [ka]

[0056] In some embodiments, the anti-TSLP antibody comprises a heavy chain and a light chain, wherein the heavy chain comprises the amino acid sequence set forth in SEQ ID NO:9 or an amino acid sequence having at least 90% identity thereto, and the light chain comprises the amino acid sequence set forth in SEQ ID NO:10 or an amino acid sequence having at least 90% identity thereto.

[0057] heavy chain [ka]

[0058] Light chain [ka]

[0059] In specific embodiments, an anti-TSLP antibody or antigen-binding fragment thereof comprises a heavy chain sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence of SEQ ID NO:9, and a light chain sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence of SEQ ID NO:10.

[0060] Unless otherwise defined, terms used in this disclosure have the following meanings:

[0061] Unless the context clearly requires otherwise, throughout the specification and claims, the terms "comprising," "having," "including," and the like, shall be understood to have an inclusive meaning rather than an exclusive or exhaustive meaning; that is, they shall be interpreted as meaning "including, but not limited to."

[0062] The term "and / or," e.g., "X and / or Y," shall be understood to mean "X and Y" or "X or Y," and is intended to clearly support an either / or interpretation.

[0063] The term "thymic stromal lymphopoietin (TSLP)" refers to a type I four-alpha-helical bundle cytokine, an epithelial cell-derived cytokine produced in response to proinflammatory stimuli. It is closely related to interleukin-7 (IL-7) and induces allergic responses by stimulating dendritic cells (DCs). TSLP is an important factor in regulating the human immune response. The term "TSLP" includes variants, isoforms, homologs, orthologs, and paralogs of TSLP.

[0064] "Buffer" refers to a buffer that can withstand pH changes due to the effect of its acid-base conjugate components. Examples of buffers that control pH within a suitable range include acetate, succinate, gluconate, histidine, oxalate, lactic acid, phosphate, citric acid, tartaric acid, fumarate, glycylglycine, and other organic acid buffers.

[0065] A "histidine buffer" is a buffer containing histidine ions. Examples of histidine buffers include histidine-hydrochloride, histidine-acetate, histidine-phosphate, and histidine-sulfate buffers, preferably histidine-acetate buffer or histidine-hydrochloride buffer, where histidine-acetate buffer is prepared from histidine and acetic acid, and histidine-hydrochloride buffer is prepared from histidine and hydrochloric acid.

[0066] "Stabilizer" refers to a component that helps maintain the structural integrity of a biopharmaceutical, particularly during freezing and / or lyophilization and / or storage (especially when exposed to stress). This stabilizing effect can occur for a variety of reasons, but typically such stabilizers act as osmolytes to mitigate protein denaturation. As used herein, stabilizers include sugars, amino acids, and EDTA, and amino acids included as stabilizers refer to amino acids added separately in addition to any amino acids in the buffer.

[0067] "Surfactant" refers to a surface active agent, preferably a nonionic surfactant. The use of a surfactant can reduce protein aggregation and / or particle formation in the formulation. The amount of surfactant added is an amount that can reduce protein aggregation and minimize particle formation in the formulation. Surfactants disclosed herein include, among others, polysorbates (including but not limited to polysorbate 20 and polysorbate 80), polyhydroxyalkanes, Triton, sodium dodecyl sulfate, sodium lauryl sulfate, sodium octyl glucoside, lauryl sulfobetaine, myristyl sulfobetaine, linoleyl sulfobetaine, stearyl sulfobetaine, lauroyl sarcosine salts, myristoyl sarcosine salts, linoleoyl sarcosine salts, stearoyl sarcosine salts, linoleyl betaine, myristyl betaine, cetyl betaine, and the like. The surfactant may be selected from the group consisting of methyl methyl taurate, lauramidopropyl betaine, cocamidopropyl betaine, linoleamidopropyl betaine, myristamidopropyl betaine, palmitamidopropyl betaine, isostearamidopropyl betaine, myristamidopropyl dimethylamine, palmitamidopropyl dimethylamine, isostearamidopropyl dimethylamine, sodium cocoyl methyl taurate, sodium oleoyl methyl taurate, polyethylene glycol, polypropylene glycol, and copolymers of ethylene glycol and propylene glycol. Preferred surfactants are polysorbate 80 or polysorbate 20, more preferably polysorbate 80.

[0068] The terms "about" and "approximately," as used herein, refer to values ​​within an acceptable range of error as determined by one of ordinary skill in the art, which may depend, in part, on how the value is measured or determined (i.e., the limitations of the measurement system). For example, in the art, "about" in each embodiment may mean within or more than one standard deviation. Alternatively, "about" or "substantially comprising" may mean within ±10% of the specified value. Furthermore, particularly in biological systems or processes, the term may mean up to an order of magnitude or up to five times the value. Unless otherwise specified, when a particular value appears in this application and claims, "about" or "substantially containing" should mean that the particular value is expected to be within an acceptable range of error.

[0069] Although the present disclosure provides content ranges or content values, one of ordinary skill in the art will understand that such content ranges or values ​​encompass an acceptable margin of error in the measurements.

[0070] A "pharmaceutical composition" refers to a mixture containing one or more antibody-drug conjugates described herein or physiologically / pharmaceutically acceptable salts or prodrugs thereof, together with other chemical components, such as physiologically / pharmaceutically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to maintain the stability of the active antibody component, facilitate administration to an organism, and promote absorption of the active components to allow them to exert their biological activity.

[0071] The term "combination," as used in this disclosure, refers to a method of administration and means administering at least one dose of an anti-TSLP antibody, or antigen-binding fragment thereof, and another asthma therapeutic within a specified period of time, during which both drugs exhibit pharmacological activity. The specified period of time may be within one administration cycle, e.g., within 12 weeks, 16 weeks, 20 weeks, or 24 weeks. The anti-TSLP antibody, or antigen-binding fragment thereof, and the other asthma therapeutic may be administered simultaneously or in no particular order. The period of time includes such treatments in which the anti-TSLP antibody, or antigen-binding fragment thereof, and the other asthma therapeutic are administered via the same or different routes of administration.

[0072] In this disclosure, "pharmaceutical composition" and "formulation" are not mutually exclusive.

[0073] Solution forms of the pharmaceutical compositions described in this disclosure contain water as a solvent, unless otherwise specified.

[0074] "Lyophilized formulation" refers to a formulation or pharmaceutical composition obtained from a liquid or solution form of a pharmaceutical composition, or a liquid or solution formulation, after a vacuum freeze-drying step.

[0075] The three-letter and one-letter amino acid codes used in this disclosure are as described in J. Biol. Chem, 243, p3558 (1968).

[0076] The term "antibody," as used in this disclosure, is used in its broadest sense and encompasses various antibody structures, including, but not limited to, monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), full-length antibodies, or antigen-binding fragments (also referred to as "antigen-binding portions") thereof, provided that they exhibit the desired antigen-binding activity. A full-length antibody is an immunoglobulin (Ig) comprising at least two heavy chains and two light chains linked by disulfide bonds. According to the amino acid composition and arrangement of the immunoglobulin heavy chain constant region, immunoglobulins can be classified into five types or isotypes: IgM, IgD, IgG, IgA, and IgE, which have corresponding heavy chains: μ, δ, γ, α, and ε. Each Ig class can be further subdivided into subclasses based on differences in the amino acid composition of the hinge region and the number and location of disulfide bonds in the heavy chain; for example, IgG can be divided into IgG1, IgG2, IgG3, and IgG4. Light chains are classified as kappa or lambda based on their constant region. Each of the five classes of Ig can contain either kappa or lambda light chains.

[0077] The term "variable region" or "variable domain" refers to the domain in an antibody heavy or light chain that is involved in antigen binding. VH and VL each contain four conserved framework regions (FR) and three complementarity-determining regions (CDR). The term "complementarity-determining region" or "CDR" refers to the region in the variable domain primarily involved in antigen binding, while "framework" or "FR" refers to the variable domain residues outside the CDRs. VH contains three CDRs: HCDR1, HCDR2, and HCDR3, and VL contains three CDRs: LCDR1, LCDR2, and LCDR3. Each VH and VL consists of three CDRs and four FRs, arranged from the amino terminus to the carboxy terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. A single VH or VL may be sufficient to confer antigen-binding specificity.

[0078] Various well-known methods can be used to determine the amino acid sequence range of a CDR, such as the "Kabat" numbering convention (see Kabat et al., (1991), "Sequences of Proteins of Immunological Interest," 5th ed., Public Health Service, National Institutes of Health, Bethesda, MD), the "Chothia" numbering convention, the "ABM" numbering convention, the "Contact" numbering convention (see Martin, ACR Protein Sequence and Structure Analysis of Antibody Variable Domains [J]. 2001), and the ImMunoGenTics (IMGT) numbering convention (Lefranc, MP et al., Dev Comp. Immunol., 27, 55-77 (2003); Front Immunol. 2018, October 16; 9:2278). The relationship between various numbering conventions is well known to those skilled in the art and is shown in Table 1 below.

[0079] [Table 2]

[0080] Unless otherwise specified, variable region and CDR sequences in embodiments of the present disclosure follow the "Kabat" numbering convention.

[0081] The terms "antigen-binding fragment," "functional fragment," or "antigen-binding portion" refer to one or more fragments of a full-length antibody that retain the ability to specifically bind to an antigen. It has been shown that fragments of a full-length antibody can be used to perform the antigen-binding function. Exemplary binding fragments encompassed by the term "antigen-binding fragment" include: (i) a Fab fragment, a monovalent fragment containing the VL, VH, CL, and CH1 domains; (ii) a F(ab')2 fragment, a bivalent fragment containing two Fab fragments linked via a disulfide bridge in the hinge region; (iii) a Fd fragment, containing the VH and CH1 domains; (iv) a Fv fragment, containing the VH and VL domains of a single antibody arm; (v) a dsFv, a stabilized antigen-binding fragment in which the VH and VL are linked by an interchain disulfide bond; (vi) a scFv; and (vii) diabodies, bispecific and multispecific antibodies, including scFv, dsFv, Fab, and other fragments.

[0082] The terms "single-chain antibody," "single-chain Fv," or "scFv" refer to a molecule comprising an antibody heavy chain variable domain (or region, VH) and an antibody light chain variable domain (or region, VL) linked by a linker. Such scFv molecules generally may have the structure: NH2-VL-linker-VH-COOH or NH2-VH-linker-VL-COOH. Numerous linkers suitable for linking antibody VH and VL are known in the art, such as linkers constructed using repeated GGGGS amino acid sequences or variants thereof, e.g., 1 to 4 repeats of such variants (Holliger et al., (1993), Proc. Natl. Acad. Sci. USA 90: 6444-6448). Other linkers useful in the present disclosure are described by Alfthan et al., (1995), Protein Eng. 8: 725-731, Choi et al., (2001), Eur. J. Immunol. 31: 94-106, Hu et al., (1996), Cancer Res. 56: 3055-3061, Kipriyanov et al., (1999), J. Mol. Biol. 293: 41-56 and Roovers et al., (2001), Cancer Immunol.

[0083] Diabodies are antibody fragments formed by dimerization of scFv or Fab fragments, resulting in bivalent antigen-binding activity, where the two antigens can be the same or different.

[0084] Bispecific and multispecific antibodies refer to antibodies capable of simultaneously binding to two or more antigens or epitopes, and include scFv or Fab fragments capable of binding TSLP.

[0085] "Conventional variants" of human antibody heavy chain constant regions and human antibody light chain constant regions, as described in this disclosure, refer to variants of human origin disclosed in the prior art that do not alter the structure and function of the antibody variable region; exemplary variants include IgG1, IgG2, IgG3, or IgG4 heavy chain constant region variants with point mutations and amino acid substitutions, such as the known YTE mutation, the L234A and / or L235A mutation, the S228P mutation, and / or mutations that result in knob-into-hole structures (such that the antibody heavy chain comprises knob-Fc and hole-Fc combinations), all of which have been shown to confer novel properties to antibodies without altering variable region function.

[0086] The terms "full length antibody," "intact antibody," "complete antibody," and "whole antibody" are used interchangeably herein to refer to an antibody in substantially its entire form, as distinguished from an antigen-binding fragment, as defined below. This term particularly refers to an antibody comprising constant regions in both the heavy and light chains.

[0087] Methods for producing and purifying antibodies and antigen-binding fragments are well known in the art, such as those described in Chapters 5-8 and 15 of the Cold Spring Harbor Antibody Laboratory Manual. For example, mice can be immunized with human TSLP or a fragment thereof, and the resulting antibodies can be refolded, purified, and sequenced using standard methods. Antigen-binding fragments can similarly be prepared by conventional methods. The antibodies or antigen-binding fragments described in this disclosure are engineered by adding one or more human framework (FR) regions to non-human CDR regions. Human germline FR sequences can be selected by aligning sequences in the ImMunoGeneTics (IMGT) human antibody variable region germline gene database using the MOE software on the ImMunoGeneTics (IMGT) website (http: / / imgt.cines.fr) or obtained from the Immunoglobulin journal (2001 ISBN 012441351).

[0088] The engineered antibodies or antigen-binding fragments of the present disclosure may be produced and purified by standard methods. For example, cDNA sequences encoding the heavy and light chains may be cloned and recombined into a GS expression vector. The recombinant immunoglobulin expression vector may be stably transfected into CHO cells. As a more preferred prior art, mammalian expression systems result in glycosylation of antibodies, particularly at the highly conserved N-terminal site within the Fc region. Stable clones are obtained by expressing antibodies that specifically bind to human TSLP. Positive clones are grown in serum-free medium in a bioreactor to produce the antibody. The culture medium containing the secreted antibody may be purified using conventional techniques. For example, purification may be performed using an A or G Sepharose FF column containing a conditioned buffer. Nonspecifically bound components are washed away. The bound antibody may then be eluted using a pH gradient, and antibody fragments may be detected and collected by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The antibody may be filtered and concentrated using conventional methods. Soluble compounds and polymers can be removed using conventional methods, such as molecular sieves and ion exchange. The resulting product should be immediately frozen, for example at -70°C, or lyophilized.

[0089] The terms "administration," "delivery," and "treatment," when applied to an animal, human, research subject, cell, tissue, organ, or bodily fluid, refer to the contact of an exogenous drug, therapeutic agent, diagnostic agent, or composition with an animal, human, subject, cell, tissue, organ, or bodily fluid. The terms "administration," "delivery," and "treatment" may refer, for example, to therapeutic, pharmacokinetic, diagnostic, research, and experimental methods. Treatment of a cell includes contact between a reagent and the cell, and between a reagent and a fluid when the fluid is in contact with the cell. The terms "administration," "delivery," and "treatment" also refer to the application of a drug, diagnostic agent, binding composition, or another type of ex vivo or in vitro manipulation, e.g., of cells. "Treatment," when applied to a human, veterinary, or research subject, refers to therapeutic treatment, preventative or prophylactic measures, research, and diagnostic applications.

[0090] "Treatment" refers to the administration of a therapeutic agent for internal or topical use, such as a composition containing any of the conjugate compounds disclosed herein, to a patient, where the patient has one or more disease symptoms and the therapeutic agent is known to have a therapeutic effect on those symptoms. Typically, the therapeutic agent is administered in an amount effective to alleviate one or more disease symptoms in the patient or population being treated, to induce regression of such symptoms, or to inhibit their progression to any clinically measurable extent. The amount of therapeutic agent that effectively alleviates any particular disease symptom (also referred to as a "therapeutically effective amount") can vary depending on several factors, such as the patient's disease state, age, and weight, and the drug's ability to produce the desired effect in the patient. Whether a disease symptom has been alleviated can be assessed by any clinical test method commonly used by physicians or other medical professionals to assess the severity or progression of symptoms. Although an embodiment (e.g., a method of treatment or product) in the present disclosure may be ineffective in alleviating each target disease symptom, any statistical testing method known in the art, such as Student's t-test, chi-square test, Mann-Whitney U test, Kruskal-Wallis test (H test), Jonckheere-Terpstra test, and Wilcoxon test, can determine that an embodiment alleviates a target disease symptom in a statistically significant number of patients.

[0091] The term "effective amount" or "effective dose" refers to the amount of a drug, compound, or pharmaceutical composition required to achieve any one or more beneficial or desired therapeutic outcomes. For prophylactic purposes, beneficial or desired outcomes include eliminating or reducing the risk, alleviating the severity, or delaying the onset of a condition, including symptoms of the condition, its complications, and biochemical, histological, and / or behavioral manifestations of intermediate pathological phenotypes that occur during disease progression. For therapeutic applications, beneficial or desired outcomes include clinical outcomes, such as reducing the incidence of a condition associated with a target antigen of the present disclosure, or ameliorating one or more symptoms of a condition, reducing the dosage of other drugs required to treat the condition, enhancing the effectiveness of another drug, and / or delaying the progression of a condition associated with a target antigen of the present disclosure in a patient.

[0092] "Homology" refers to the sequence similarity between two polynucleotide sequences or two polypeptides. If a position in the two sequences being compared is occupied by the same base or amino acid subunit monomer, for example, if both DNA molecules have adenine at the same position, the molecules are homologous at that position. The percentage of homology between two sequences is a function of the number of matching or homologous positions shared by the two sequences, divided by the number of positions compared, multiplied by 100. For example, if 6 out of 10 positions in the best alignment of two sequences are matching or homologous, the sequences are 60% homologous, and if 95 out of 100 positions are matching, they are 95% homologous. Typically, when comparing two sequences, the alignment that results in the highest percentage of homology is used. For example, comparison can be performed using the BLAST algorithm, where the algorithm parameters are selected to result in the maximum alignment across the entire length of each reference sequence. The following references relate to BLAST algorithms, which are commonly used for sequence analysis: BLAST ALGORITHMS: Altschul, S. F. et al., (1990) J. Mol. Biol. 215: 403-410; Gish, W. et al., (1993) Nature Genet. 3: 266-272; Madden, T. L. et al., (1996) Meth. Enzymol. 266: 131-141; Altschul, S. F. et al., (1997) Nucleic Acids Res. 25: 3389-3402; Zhang, J. et al., (1997) Genome Res. 7: 649-656. Other standard BLAST algorithms, such as the BLAST algorithm provided by NCBI BLAST, are also well known to those skilled in the art.

[0093] "Optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and the specification includes situations in which the event or circumstance occurs or does not occur.

[0094] " Asthma " refers to a common and potentially serious chronic disorder of the airways, characterized by inflammation and constriction of the airways, which leads to symptoms such as wheezing, shortness of breath, chest tightness, and coughing, and symptoms fluctuate over time in occurrence, frequency, intensity, and response to therapy.As used herein, the term "asthma" refers to all phenotypes and endotypes of asthma, including non-allergic asthma, allergic asthma, mild asthma, moderate asthma, severe asthma, moderate-severe asthma, uncontrolled asthma, non-eosinophilic asthma, hypoeosinophilic asthma, hypereosinophilic (eosinophilic) asthma, bronchial asthma, exercise-induced asthma, occupational asthma, and asthma caused by bacterial or viral infection, across all severity levels and regardless of disease control status.

[0095] An "acute asthma exacerbation" refers to a progressive worsening of asthma symptoms, such as shortness of breath, cough, wheezing, and chest tightness, accompanied by a progressive decline in lung function that requires a change in treatment. Subjects who die from worsening asthma symptoms are also included in acute asthma exacerbations. Such treatment changes may include any of the following: 1) treatment with systemic corticosteroids, or an increase in dose for subjects already receiving systemic corticosteroid maintenance therapy for three or more days; 2) an asthma event requiring treatment with systemic corticosteroids and emergency / urgent care or hospitalization. Other measures related to acute asthma attacks have also been studied to determine their impact, including, but not limited to, worsening of asthma symptoms, declines from baseline in lung function, or increased use of rescue medication. Annualized asthma exacerbation rate (AAER) = number of acute asthma attacks * 365.25 / length of follow-up (days).

[0096] An "asthma exacerbation" refers to an acute or subacute worsening of asthma symptoms and lung function from a patient's usual state, and may result in any of the following: the use of systemic corticosteroids for at least three consecutive days; a single long-acting injectable dose of corticosteroids considered equivalent to three days of systemic corticosteroids; a temporary doubling of the maintenance dose for at least three days in subjects receiving maintenance OCS; an asthma ED visit requiring systemic corticosteroids (as described above); or an asthma hospitalization. Other measures related to asthma exacerbations have also been studied to determine their impact. These measures include asthma exacerbation-related hospitalizations (i.e., severe asthma exacerbations), time to first asthma exacerbation, and the proportion of subjects experiencing one or more asthma exacerbation / severe asthma exacerbation events. A severe asthma exacerbation is typically defined as the need for systemic corticosteroids or an increase in the maintenance dose of oral corticosteroids for at least three days, and / or an asthma emergency department visit, hospitalization, or death. A moderate exacerbation is typically defined as an event requiring a change in treatment to prevent progression to a severe exacerbation and the presence of one or more of the following: an increase in asthma symptoms, an increase in rescue medication use, or a decline in lung function lasting 2 days or more but not severe enough to warrant systemic corticosteroids or hospitalization.

[0097] "Allergic asthma" refers to asthma caused by one or more inhaled allergens. Such patients have positive IgE fluorescent enzyme immunoassay (FEIA) levels for one or more allergens that induce an asthmatic response. Typically, most allergic asthma is associated with Th2-type inflammation.

[0098] The term "Th2-type inflammation" refers to subjects with a screening blood eosinophil count of 140 cells / μL or greater and a screening total serum IgE level of 100 IU / mL or greater (Corren et al., N Engl J Med. 22; 365(f2): 1088-98, 2011). A "high Th2" asthma population or profile refers to subjects with IgE levels of 100 IU / mL or greater and blood eosinophil counts of 140 cells / μL or greater. A "low Th2" asthma population refers to subjects with IgE levels of less than 100 IU / mL and blood eosinophil counts of less than 140 cells / μL.

[0099] "IgE" refers to immunoglobulin E, as known to those skilled in the art. IgE is measured as international units per milliliter (IU / mL), as disclosed, for example, in Seagroatt and Anderson (1981).

[0100] "Non-allergic asthma" refers to patients with low eosinophil counts, low Th2 or low IgE at the time of diagnosis. Typically, patients with "non-allergic asthma" have a negative IgE fluorescent enzyme immunoassay (FEIA) response to a panel of allergens, including area-specific allergens. In addition to low IgE, these patients generally have low or absent eosinophil counts and low Th2 counts at the time of diagnosis.

[0101] An eosinophil count is a blood test that measures the number of white blood cells called eosinophils. Typically, blood is drawn from a vein on the inside of the elbow or the back of the hand, or by puncturing the skin with a lancet-type device. The blood is placed in a small glass tube or on a glass slide or test strip. In the laboratory, the blood is placed on a microscope slide, and a dye is added to the sample. This causes the eosinophils to appear as orange-red granules, allowing the count of eosinophils present in a specific volume of blood (e.g., 1 microliter (μL)).

[0102] "Non-eosinophilic asthma" refers to asthma in which a patient has no eosinophil count. "Hypoeosinophilic asthma" refers to asthma in which a patient has a screening blood eosinophil count of less than 300 cells / μL. "Hypereosinophilic asthma" or "eosinophilic asthma" refers to asthma in which a patient has a screening blood eosinophil count of 300 cells / μL or greater.

[0103] "Mild asthma" refers to asthma that is adequately controlled by GINA Step 1 or Step 2 therapy, i.e., as-needed controller medication alone or with low-intensity maintenance controller therapy (e.g., low-dose ICS, leukotriene receptor antagonists, or chromones) (Global Strategy for Asthma Management and Prevention, GINA Report 2019, 35). "Moderate asthma" refers to asthma that is adequately controlled by GINA Step 3 therapy, e.g., low-dose ICS-LABA (Global Strategy for Asthma Management and Prevention, GINA Report 2019, 35). "Severe asthma" refers to asthma that requires GINA Step 4 or 5 therapy, e.g., high-dose ICS-LABA, to prevent the condition from becoming "uncontrolled," or that remains "uncontrolled" despite such therapy (Global Strategy for Asthma Management and Prevention, GINA Report 2019, 35). "Moderate-to-severe asthma" refers to both moderate and severe asthma.

[0104] "Uncontrolled asthma" refers to asthma that remains uncontrolled on GINA Step 4 or 5 treatment, eg, asthma with poor symptom control.

[0105] Asthma severity can be assessed retrospectively based on the level of treatment required to control symptoms and prevent exacerbations. Assessments can be made after several months of controller therapy, and, if appropriate, step-down therapy can be attempted to determine the patient's minimally effective level of treatment. Asthma severity is not a static characteristic and can change over months or years.

[0106] "Adult" refers to a subject aged 18 years or older. As used herein, the term "adolescent" refers to a subject aged 12 years or older but younger than 18 years. In some embodiments of the present invention, the subject is a child. As used herein, the term "child" refers to a subject under the age of 12 years.

[0107] The details of one or more embodiments of the present disclosure have been presented above. Although any methods and products similar or equivalent to those described herein can be used in practicing or testing the present disclosure, preferred methods and products are described below. Other features, objects, and advantages of the present disclosure will be apparent from the specification and claims. In the specification and claims, the singular forms "a," "an," and "the" include plural referents unless the context dictates otherwise. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this disclosure belongs. All patents and publications cited herein are incorporated by reference. The following examples are provided to more fully describe preferred embodiments of the present disclosure. These examples should not be construed as limiting the scope of the present disclosure, which is defined by the claims.

[0108] Abbreviation: FEV1:1 second amount ePRO: Electronic Patient-Reported Outcomes Pre / Post-BD: Before / after use of bronchodilators eGFR: Estimated glomerular filtration rate

[0109] Specific Embodiments The following specific embodiments or experimental examples are provided to further illustrate the present disclosure and are intended for illustrative purposes only and are not intended to limit the scope of the present disclosure.

[0110] Unless otherwise specified, experimental methods in the embodiments are performed under normal conditions or conditions recommended by the material or reagent manufacturer. Reagents for which a specific source is not indicated are commercially available conventional reagents. [Example]

[0111] Example 1: Pharmacokinetic study of anti-TSLP antibodies in cynomolgus monkeys 1) Test substance: an anti-TSLP antibody, prepared according to the method described in WO 2022 / 116858, designated hu179-33 antibody, having heavy and light chain sequences set forth in SEQ ID NOs: 9 and 10 in the present disclosure. 2) Test Method: Six cynomolgus monkeys (3 males and 3 females) were selected and administered a single subcutaneous injection of 3 mg / kg anti-TSLP antibody. Whole blood samples were collected before administration and at 2 h, 8 h, 24 h, 48 h, 72 h, 96 h, 120 h (D6), 168 h (D8), 336 h (D15), 504 h (D22), 672 h (D29), 840 h (D36), 1008 h (D43), 1176 h (D50), 1344 h (D57), 1512 h (D64), 1680 h (D71), 1848 h (D78), and 2016 h (D85) after administration, and serum was separated. Serum drug concentrations were determined using ELISA. t1 / 2 (half-life), T max (time to maximum concentration), C max Pharmacokinetic parameters, including (maximum concentration) and AUC (area under the concentration-time curve), were calculated using the WinNonLin noncompartmental analysis (NCA) model. Means and standard deviations were calculated using Waston. The experimental results are shown in Table 1, and data are presented as mean ± standard deviation.

[0112] [Table 3]

[0113] Example 2: Pharmacokinetic study of anti-TSLP antibody (reference antibody) in cynomolgus monkeys 1) Test substance: Tezepelumab was used as the reference antibody, prepared in-house. 2) Study Method: Three male cynomolgus monkeys were selected and administered a single subcutaneous injection of 3 mg / kg of the reference antibody tezepelumab. Whole blood was collected and serum was isolated before administration and at 2 h, 8 h, 24 h (D2), 48 h (D3), 72 h (D4), 96 h (D5), 120 h (D6), 168 h (D8), 336 h (D15), 504 h (D22), and 672 h (D29) after administration. The concentration of the reference antibody in the serum samples was determined by ELISA. Pharmacokinetic parameters: 1 / 2 , T max , C max and AUC were calculated for each animal using the WinNonlin non-compartmental analysis (NCA) model. The experimental results are shown in Table 2, and data are presented as mean ± standard deviation.

[0114] [Table 4]

[0115] Example 3: A single subcutaneous injection study of an anti-TSLP antibody in healthy subjects and patients with mild asthma to evaluate safety, tolerability, pharmacokinetics, and pharmacodynamics - A randomized, double-blind, dose-escalating, placebo-controlled Phase I trial 1. Test Pharmaceutical Product 1) An anti-TSLP antibody, prepared according to the methods described in WO 2022 / 116858, designated hu179-33 antibody. The heavy and light chain sequences of the anti-TSLP antibody are as disclosed in SEQ ID NOs:9 and 10, and formulated as an injection having a concentration of 1 mL:0.1 g. 2) Placebo, formulated as an injection with a concentration of 1 mL.

[0116] 2. Research method Eligible healthy adults (aged 18-55 years) were randomized in an 8:2 ratio to receive a single subcutaneous injection of anti-TSLP antibody (five cohorts: 50 mg, 100 mg, 200 mg, 400 mg, and 600 mg) or placebo, with follow-up ranging from 113 to 253 days for each cohort.

[0117] 3.Research results Fifty healthy subjects enrolled and completed treatment. All treatment-related adverse events (TRAEs) were mild to moderate in severity. The incidence of TRAEs was 45.0% (18 / 40) in the anti-TSLP antibody group and 20.0% (2 / 10) in the placebo group. The most common TRAEs (≥5%) in the anti-TSLP antibody group were headache (10.0%), injection site bruising (5.0%), and elevated troponin I (5.0%). Anti-TSLP antibody was absorbed slowly, reaching maximum serum concentrations (Cmax) between days 6.995 and 17.591. The area under the concentration-time curve increased with dose. Within the 50-600 mg dose range, the mean half-life was 69.7 to 84.9 days. The overall incidence of ADA-positive outcomes was similar between the anti-TSLP antibody and placebo groups, and no significant effect on pharmacokinetic properties was observed in ADA-positive subjects.

[0118] Conclusions: Single subcutaneous injections of anti-TSLP antibody at doses of 50 mg to 600 mg exhibited slightly greater dose-proportional pharmacokinetics and had acceptable safety and tolerability in healthy adult subjects. The overall safety and tolerability of anti-TSLP antibody was good within the 50-600 mg dose range.

[0119] Example 4: Efficacy and safety of anti-TSLP antibody injection in patients with severe, uncontrolled asthma - A multicenter, randomized, double-blind, placebo-controlled, phase II clinical study 1. Pharmaceutical Product Information Anti-TSLP antibody injection: the antibody sequences correspond to SEQ ID NO: 9 for the heavy chain and SEQ ID NO: 10 for the light chain, with a specification of 1 mL: 0.1 g, supplied by Jiangsu Hengrui Pharmaceuticals Co., Ltd. Placebo injection: specification 1mL, supplied by Jiangsu Hengrui Pharmaceuticals Co., Ltd.

[0120] 2. Study Design: This is a multicenter, randomized, double-blind, placebo-controlled, parallel-design phase II trial in patients with severe, uncontrolled asthma requiring medium- or high-dose ICS treatment and two or more acute asthma attacks (requiring oral or intravenous corticosteroids) in the 12 months prior to randomization. The study plans to enroll 248 subjects (50% receiving medium- or high-dose ICS before randomization, 50% each with baseline EOS <300 / μL and ≥300 / μL) and randomize them 1:1:1:1 to one of three treatment regimens or a placebo group, with 62 subjects per group. Subjects in the treatment groups will receive subcutaneous injections of an anti-TSLP antibody: Group A 200 mg Q24W, Group B 400 mg Q24W, or Group C 400 mg Q12W. Subjects in the placebo group will receive subcutaneous placebo every 12 weeks.

[0121] Research Procedure: The study consisted of five periods: screening, run-in, baseline, treatment, and follow-up.

[0122] Screening period (W-4~W-3): A total of 248 subjects with severe, uncontrolled asthma, regardless of sex, were screened. They had a predicted FEV1 of 40% to less than 80% and two or more acute exacerbations (requiring oral or intravenous corticosteroids) within the 12 months prior to randomization. Eligible subjects were trained in the use of a home peak flow meter with an electronic patient-reported outcomes (ePRO) system and were instructed accordingly, including on inhaled medication use.

[0123] Introductory period (W-2~W-1):Subjects must record a daily asthma diary in the ePRO system, complete a patient log, and measure PEF twice daily (morning and evening) using a home peak flow meter and record it in the ePRO system. The run-in visit will assess subject proficiency and compliance with the home peak flow meter and ePRO system. From W-1, subject compliance with the peak flow meter and ePRO system will be calculated and requires 70% or greater compliance within 7 days prior to randomization.

[0124] Baseline period: Subjects who pass screening will undergo further assessments and compliance checks during baseline. Subjects who meet all inclusion criteria and none of the exclusion criteria will receive a randomization and drug number during baseline and will be randomized 1:1:1:1 to treatment or placebo.

[0125] Treatment period (W1~W48): Treatment regimens for each group [Table 5]

[0126] Follow-up period (W49~W72): Subjects will enter a follow-up period after the end of the treatment period. During follow-up, subjects will be contacted prophylactically. Information collection must be conducted at least once every 4 weeks to record safety events in the trial.

[0127] Primary endpoints and key evaluands: The primary endpoint was the annualized asthma exacerbation rate (AAER) at 48 weeks. Annualized asthma exacerbation rate (AAER) = number of acute asthma attacks * 365.25 / duration of follow-up (days) The five attributes of a key estimand are defined as follows: 1. Treatment: The experimental group included anti-TSLP antibody (Group A 200mg Q24W, Group B 400mg Q24W, Group C 400mg Q12W) plus maintenance therapy, and the control group included placebo plus maintenance therapy; Maintenance therapy includes medium / high dose inhaled corticosteroids (ICS) plus at least one other asthma control medication, as defined by inclusion criteria 4 and 5. 2. Population: Patients with severe, uncontrolled asthma; 3. Variables (endpoints): Annualized Asthma Exacerbation Rate (AAER) at week 48; 4. Intercurrent Events and Strategies: [Table 6] 5. Population-level summary: ratio of AAER between treatment and control groups (estimated using a negative binomial regression model).

[0128] Secondary endpoints: Changes from baseline in pre- / post-bronchodilator (Pre- / Post-BD) forced expiratory volume in 1 second (FEV1), forced vital capacity (FVC), and peak expiratory flow (PEF) at each assessment time point. Changes from baseline in exhaled nitric oxide (FeNO), blood eosinophil count (EOS), and serum immunoglobulin E (IgE) at each evaluation time point. Change from baseline in Asthma Control Questionnaire (ACQ-6) scores at each assessment time point. Change from baseline in Asthma Quality of Life Questionnaire (AQLQ) scores at each assessment time point. Annualized rate of acute asthma exacerbations leading to hospitalization at 48 weeks. Change from baseline in mean weekly asthma symptom diary scores. Proportion of subjects experiencing an acute asthma attack. Change from baseline in weekly rescue medication use. Change from baseline in weekly mean morning and evening PEF. Change from baseline in the mean number of weekly nighttime awakenings. Time to first acute asthma attack. Change in 22-item Sino-Nasal Outcome Test (SNOT-22) scores at each assessment time point in patients with nasal polyps. Incidence of composite progression endpoint (CompEx).

[0129] Safety Endpoints: Physical examination, vital signs, ECG, laboratory tests and adverse events / serious adverse events.

[0130] Pharmacokinetic (PK) and immunogenicity endpoints: PK: serum drug concentration of anti-TSLP antibody. Immunogenicity: incidence, time of occurrence, duration and titer of anti-drug antibodies (ADA).

[0131] 3. Inclusion criteria To be eligible for this study, subjects must meet all of the following criteria: (1) Age 18 to 75 years old, male or female; (2) weight ≥40 kg; (3) diagnosed with asthma according to the 2020 edition of the Guidelines for the Prevention and Treatment of Bronchial Asthma and with a history of at least 1 year; (4) receiving medium / high dose inhaled corticosteroid (ICS) therapy for at least 6 months before screening and remaining stable for 3 months before randomization; a. High-dose ICS is defined as a total daily dose of >500 μg fluticasone propionate, or equivalent, delivered by dry powder inhaler (DPI) or pressurized metered-dose inhaler (pMDI); b. Medium-dose ICS is defined as a total daily dose of >250 μg to ≤500 μg fluticasone propionate, or equivalent, delivered via a DPI or pMDI; c. Equivalent ICS doses are determined according to the 2020 edition of the Asthma Treatment Guidelines; (5) Concomitant use of at least one other asthma control medication [including long-acting beta2 agonists (LABAs), long-acting muscarinic antagonists (LAMAs), leukotriene receptor antagonists (LTRAs), maintenance oral corticosteroids (OCSs; up to 10 mg daily or 20 mg every other day prednisone for maintenance) or equivalent, excluding theophylline], with stable use for 3 months prior to randomization; (6) Pre-bronchodilator FEV1 is ≥40% to <80% predicted during the screening and baseline periods; (7) ACQ-6 score ≥ 1.5 during screening; (8) At least two acute asthma exacerbation events within 12 months prior to randomization, with at least one occurring during medium / high-dose ICS treatment. An acute asthma exacerbation event is defined as the use of systemic corticosteroids or an increase in the dose of maintenance systemic corticosteroids for 3 days or more; (9) Female subjects of childbearing potential and male subjects with partners of childbearing potential must agree to have no plans for conception and to use suitable contraception from the time they sign the informed consent form until 19 months after the last dose; (10) Willingly sign the informed consent form to participate in this study.

[0132] 4. Exclusion criteria: Subjects will be excluded from the study if they meet any of the following criteria: 1. Clinically significant major pulmonary disease, including but not limited to tuberculosis (including old / latent TB), bronchiectasis, atelectasis, interstitial lung disease, allergic bronchopulmonary aspergillosis, chronic obstructive pulmonary disease (COPD), pulmonary vascular disease, tumor, etc. 2. Comorbidities other than asthma that may affect pulmonary function, including but not limited to clinically significant pleural disease, mediastinal disease, diaphragmatic disorders, myasthenia, and chest deformity; 3. Other eosinophil-related disorders, including but not limited to hypereosinophilic syndrome (HES), eosinophilic granulomatosis with polyangiitis (EGPA), eosinophilic esophagitis, etc.; 4. Immunodeficiency disorders; 5. Clinically significant, unstable or uncontrolled severe cardiovascular or cerebrovascular disease, including but not limited to myocardial infarction, unstable angina, heart failure, stroke, and subarachnoid hemorrhage; 6. Uncontrolled hypertension (systolic BP ≥ 180mmHg and / or diastolic BP ≥ 110mmHg during screening); 7. Uncontrolled diabetes (fasting blood glucose >11.1 mmol / L during screening); 8. Clinically significant infection within 4 weeks prior to randomization, including but not limited to upper or lower respiratory tract infection; 9. Major surgery within 3 months prior to randomization or scheduled surgery during the study, or any treatment considered by the investigator to be likely to affect subject assessments; 10. Known parasitic infection within 6 months prior to randomization; 11. Diagnosis of malignancy within 5 years prior to randomization (excluding malignancies with low risk of metastasis or death, e.g., adequately treated basal cell carcinoma of the skin or cervical carcinoma in situ); 12. Abnormal laboratory results during screening and baseline: White blood cell count (WBC) 3.0×10 9 / L or less; hemoglobin (Hb) 90 g / L or less; Platelet count (PLT) 100×10 9 / L or less; alanine aminotransferase (ALT) >3 × ULN (upper limit of normal); aspartate aminotransferase (AST) above 3 × ULN; Total bilirubin (TBIL) above ULN; Prothrombin time (PT) >ULN+3s; Creatinine (Cr) >1.5 × ULN; or glomerular filtration rate <50 mL / min (calculated using the modified MDRD formula for the Chinese population: eGFR [mL / min / 1.73 m 2 ]=175×Scr -1.234 ×[Age] -0.179× [female × 0.79], serum creatinine (Scr): (mg / dL), age: years, Scr1mg / dL=88.4μmol / L); 13. Active hepatitis B infection (peripheral HBV DNA 1×10 3 HBsAg positive with ≥ IU [or copies] / mL) or positive for hepatitis C antibody, HIV antibody, or syphilis antibody; 14. QTc prolongation on ECG (>450 ms for men, >470 ms for women) or any other clinically significant abnormal finding at screening or baseline that is considered to pose a significant safety risk to the subject; 15. Use of nonselective beta-blockers (e.g., propranolol, carvedilol) within 1 week prior to randomization; 16. Use of a 5-lipoxygenase inhibitor (e.g., zileuton) or a phosphodiesterase-4 inhibitor (e.g., roflumilast) within 1 week prior to randomization; 17. Live, live attenuated, or viral vector vaccine administered within 4 weeks prior to randomization; 18. Blood products or immunoglobulin infusion within 4 weeks prior to randomization; 19. Allergen immunotherapy within 8 weeks prior to randomization; 20. Use of systemic immunosuppressants (excluding systemic corticosteroids used to treat asthma), immunomodulators, biologics, or Th2 cytokine inhibitors within 12 weeks or 5 drug half-lives (as specified in the package insert) prior to randomization; if the half-life is unknown, this includes, but is not limited to, methotrexate, cyclosporine, interferon-α, anti-interleukin-5 (IL-5) monoclonal antibodies, anti-interleukin-4 receptor (IL-4R) monoclonal antibodies, anti-TSLP monoclonal antibodies, anti-IgE monoclonal antibodies, and pranlukast; 21. Bronchial thermoplasty within 1 year prior to randomization; 22. Current smoker or quit smoking less than 6 months prior to screening and / or positive urinary cotinine test at screening, or smoking history of 10 or more pack-years (pack-years = years smoked × packs per day); 23. History of substance abuse, drug abuse, and / or excessive alcohol use within 1 year prior to screening. Excessive alcohol use is defined as daily alcohol consumption of 15g or more for an adult, equivalent to 450mL of beer, 150mL of wine, 50mL of 38% spirits, or 30mL of high-alcohol spirits; 24. Known allergy to any biologic agent or any history of hypersensitivity that the investigator considers unsuitable for study participation; 25. Pregnancy (including a positive pregnancy test at screening or baseline), lactation, or planned pregnancy during the study period; 26. Participation in another clinical trial involving an investigational pharmaceutical product with an active ingredient within 4 weeks prior to screening or within 5 half-lives of the previous investigational product at the time of screening (whichever is longer); 27. Any other medical or social reason considered by the investigator to be unsuitable for study participation.

Claims

1. 1. A method for treating a subject suffering from asthma and / or reducing the frequency and / or severity of asthma exacerbations in the subject and / or reducing the annualized asthma exacerbation rate in the subject and / or reducing the ACQ-6 score in the subject, comprising the steps of: 1) administering to the patient at least one anti-TSLP antibody or antigen-binding fragment thereof with an interval between doses of 10 weeks or more; Preferably, the administration interval is selected from 10, 12, 14, 16, 18, 20, 22, 24, 26, 28 weeks or more; More preferably, the administration interval is selected from 12, 16, 20, 24 weeks or more; Most preferably, the administration interval is 12 weeks or 24 weeks. or 2) administering once every 10, 12, 14, 16, 18, 20, 22, 24, 26, or 28 weeks; More preferably, administering once every 12, 16, 20, or 24 weeks; Most preferably, once every 12 or 24 weeks. A method comprising:

2. the dosage of the anti-TSLP antibody or antigen-binding fragment thereof is up to about 600 mg; Preferably, it is 50 to 600 mg, 100 to 500 mg, or 150 to 450 mg. More preferably, it is selected from about 100 mg, about 150 mg, about 200 mg, about 250 mg, about 300 mg, about 350 mg, about 400 mg, about 450 mg or about 600 mg; Most preferably about 200 mg or about 400 mg. The method of claim 1.

3. 3. The method of claim 1 or 2, wherein the administration regimen of the anti-TSLP antibody or antigen-binding fragment thereof is selected from 200 mg Q24W, 400 mg Q24W, or 400 mg Q12W.

4. 4. The method of any one of claims 1 to 3, wherein the anti-TSLP antibody or antigen-binding fragment thereof is administered for a period of at least 2 months, 3 months, 4 months, 5 months, 6 months, 8 months, 10 months, 12 months or more.

5. The method of any one of claims 1 to 4, wherein the anti-TSLP antibody or antigen-binding fragment thereof is administered subcutaneously or intravenously, preferably by subcutaneous injection.

6. The method of any one of claims 1 to 5, wherein the subject has allergic asthma or non-allergic asthma.

7. The method of any one of claims 1 to 5, wherein the subject has mild asthma.

8. The method of any one of claims 1 to 5, wherein the subject has moderate asthma.

9. The method of any one of claims 1 to 5, wherein the subject has moderate to severe asthma or severe asthma.

10. 10. The method of claim 8 or 9, wherein the subject has uncontrolled asthma, preferably severe uncontrolled asthma.

11. The method of any one of claims 1 to 10, wherein the subject has eosinophilic asthma or non-eosinophilic asthma.

12. 12. The method of claim 11, wherein the subject has a high eosinophil count, preferably 250 cells / μL or more, more preferably 300 cells / μL or more, at the time of treatment initiation or diagnosis.

13. 12. The method of claim 11, wherein the subject has a low eosinophil count, preferably less than 300 cells / μL, more preferably less than 250 cells / μL, at the start of treatment or at diagnosis.

14. the anti-TSLP antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region; the heavy chain variable region comprises HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3, respectively; the light chain variable region comprises LCDR1, LCDR2 and LCDR3 set forth in SEQ ID NO:4, SEQ ID NO:5 and SEQ ID NO:6, respectively; The method according to any one of claims 1 to 13.

15. 15. The method of any one of claims 1 to 14, wherein the anti-TSLP antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 7 or has 90% identity thereto, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 8 or has 90% identity thereto.

16. the anti-TSLP antibody comprises an immunoglobulin constant region; Preferably, it comprises an Fc region derived from human IgG, Preferably comprising an Fc region from human IgG1, IgG2, IgG3 or IgG4, The method according to any one of claims 1 to 15.

17. 17. The method of any one of claims 1 to 16, wherein the anti-TSLP antibody comprises a heavy chain and a light chain, wherein the heavy chain comprises the amino acid sequence shown in SEQ ID NO: 9 or has 90% identity thereto, and the light chain comprises the amino acid sequence shown in SEQ ID NO: 10 or has 90% identity thereto.

18. The method of any one of claims 1 to 17, wherein the subject also maintains baseline therapy as maintenance therapy.

19. Basic therapy includes: a) a glucocorticoid, preferably an inhaled corticosteroid (ICS) or an oral glucocorticoid; b) bronchodilators, preferably long-acting beta2 agonists (LABAs), short-acting beta2 agonists (SABAs), anticholinergics; c) long-acting muscarinic receptor antagonists (LAMAs); d) leukotriene receptor antagonists (LTRAs); e) Cromolyn, f) antihistamines, g) anti-leukotriene agents; h) PDE-4 inhibitors 20. The method of claim 18, comprising at least one of:

20. 20. Use of an anti-TSLP antibody or antigen-binding fragment in combination with another asthma therapeutic agent in the preparation of a medicament for treating a subject suffering from asthma, or reducing the frequency and / or severity of asthma exacerbations in a subject, or reducing the annualized asthma exacerbation rate in a subject, or reducing the ACQ-6 score in a subject, wherein the anti-TSLP antibody or antigen-binding fragment thereof is as defined in any one of claims 1 to 17, and the other asthma therapeutic agent is selected from the background therapy agents defined in claim 18 or 19.

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