Pulmonary delivery of antibodies

Abstract

To provide a formulation system that is suitable for pulmonary delivery of antibodies and helps maintain antibody stability and activity upon aerosolization / nebulization. [Solution] An aerosol containing droplets containing a liquid formulation is provided, the liquid formulation comprising (i) an antibody or antibody derivative, (ii) a buffer selected from the group consisting of acetate, histidine, and combinations thereof, and (iii) an aqueous medium, wherein the liquid formulation has a pH of about 5.5 or less.

Description

[Technical Field]

[0001] The present invention relates to pulmonary delivery of antibodies or antibody derivatives. [Background technology]

[0002] Highly specific antigen targeting of therapeutic antibodies has received dramatically increased attention over the years, as it can provide highly effective treatments for a variety of diseases.

[0003] Pulmonary delivery of antibodies, such as monoclonal antibodies, may be an attractive, non-invasive alternative to parenteral delivery. The pulmonary administration route has proven effective for the local and systemic delivery of a variety of drugs and biopharmaceuticals to treat pulmonary and non-pulmonary diseases.

[0004] However, pulmonary administration of proteins such as antibodies presents many challenges, including the need for appropriate formulation of antibodies to overcome strong intermolecular / interparticle interactions and physicochemical degradation, which can lead to aggregation and potentially loss of biological / therapeutic activity and / or safety issues. For example, proteins can be sensitive to the shear stress and / or temperature increase associated with aerosolization and / or can exhibit reduced stability at the air-liquid interface in aerosols.

[0005] Therefore, an objective of the present invention was to identify a formulation system that is suitable for pulmonary delivery of antibodies and that helps maintain the stability and activity of the antibody upon aerosolization / nebulization. Summary of the Invention

[0006] In one aspect, the present invention provides an aerosol comprising droplets comprising a liquid formulation, the liquid formulation comprising: (i) an antibody or antibody derivative; (ii) a buffering agent selected from the group consisting of acetate, histidine, and combinations thereof; and (iii) Aqueous medium Including, The liquid formulation relates to an aerosol, having a pH of about 5.5 or less.

[0007] In one embodiment, the antibody or antibody derivative is an IgG1 antibody or antibody derivative.

[0008] In one embodiment, the antibody is a monoclonal antibody.

[0009] In one embodiment, the liquid formulation is citrate-free.

[0010] In one embodiment, the liquid formulation has a pH ranging from about 3.5 to about 5.5.

[0011] In one embodiment, the buffering agent is acetate and the liquid formulation has a pH of less than about 5.0 or less than about 4.5. In one embodiment, the buffering agent is acetate and the liquid formulation has a pH ranging from about 3.5 to less than about 4.5, or from about 3.7 to about 4.3, or from about 3.8 to about 4.2, or from about 3.9 to about 4.1. In one embodiment, the liquid formulation has a pH of about 4.0.

[0012] In one embodiment, the liquid formulation further comprises a surfactant. In one embodiment, the surfactant is a polysorbate, a poloxamer, a polyoxyethylene alkyl ether, an alkyl fluoride, or a hydroxypropyl methyl acrylate. The copolymer is selected from the group consisting of phenol polyoxyethylene and sodium dodecyl sulfate.

[0013] In one embodiment, the concentration of surfactant in the liquid formulation is about 0.1% (w / v) or less, or about 0.05% (w / v) or less.

[0014] In one embodiment, the liquid formulation does not contain any surfactants.

[0015] In one embodiment, the liquid formulation does not include NaCl.

[0016] In one embodiment, the liquid formulation does not include any non-buffering salts.

[0017] In one embodiment, the droplets have an average diameter ranging from about 0.5 μm to about 5 μm, from about 0.5 μm to about 4.5 μm, from about 0.5 μm to about 4 μm, from about 0.5 μm to about 3.5 μm, or from about 0.5 μm to about 3 μm.

[0018] In one embodiment, the droplets have an average diameter of less than 5 μm. In one embodiment, the droplets have an average diameter of less than 4.5 μm. In one embodiment, the droplets have an average diameter of less than 4.0 μm. In one embodiment, the droplets have an average diameter of less than 3.5 μm. In one embodiment, the droplets have an average diameter of less than 3.0 μm.

[0019] In another aspect, the present invention relates to a method for preparing an aerosol comprising droplets comprising a liquid formulation, the method comprising the steps of: (i) providing a liquid formulation as defined above; (ii) atomizing the liquid formulation provided in step (i) with a nebulizer, thereby preparing an aerosol; Includes:

[0020] In one embodiment, the nebulizer is a mesh nebulizer.

[0021] In one embodiment, the method further comprises, between steps (i) and (ii), the step of: (ia) lyophilizing the liquid formulation provided in step (i), thereby providing a lyophilized powder; and (ib) reconstituting the liquid formulation provided in step (i) by adding an appropriate amount of aqueous medium to the lyophilized powder provided in step (ia). Further includes:

[0022] In another aspect, the present invention relates to an aerosol comprising droplets comprising a liquid formulation, the aerosol being obtainable by a method as defined above. In one embodiment, the droplets have an average diameter ranging from about 0.5 μm to about 5 μm, or from about 0.5 μm to about 3 μm.

[0023] In another aspect, the invention relates to a liquid formulation as defined above or an aerosol as defined above for use in a method of delivering an antibody or antibody derivative to the lungs of a subject, wherein the aerosol is administered to the subject by inhalation and the liquid formulation is administered to the subject by inhalation via a nebulizer.

[0024] In one embodiment, the antibody or antibody derivative is an IgG1 antibody or antibody derivative.

[0025] In one embodiment, the nebulizer is a mesh nebulizer.

[0026] In another aspect, the present invention relates to a liquid formulation as defined above for use in a method of treating or preventing a disease in a subject, wherein the aerosol is administered to the subject by inhalation, or the liquid formulation is administered to the subject by inhalation via a nebulizer.

[0027] In one embodiment, the disease is a pulmonary disease.

[0028] In one embodiment, the nebulizer is a mesh nebulizer.

[0029] In another aspect, the invention relates to a method for delivering an antibody or antibody derivative to the lungs of a subject, the method comprising administering to the subject by inhalation an effective amount of an aerosol as defined above, or administering to the subject by inhalation via a nebulizer an effective amount of a liquid formulation as defined above.

[0030] In one embodiment, the antibody or antibody derivative is an IgG1 antibody or antibody derivative.

[0031] In one embodiment, the nebulizer is a mesh nebulizer.

[0032] In another aspect, the present invention relates to a method for treating or preventing a disease in a subject, said method comprising administering to the subject by inhalation an effective amount of an aerosol as defined above, or administering to the subject by inhalation via a nebulizer an effective amount of a liquid formulation as defined above.

[0033] In one embodiment, the disease is a pulmonary disease.

[0034] In one embodiment, the nebulizer is a mesh nebulizer.

[0035] In one embodiment, the above-mentioned pulmonary disease is selected from the group consisting of asthma, chronic obstructive pulmonary disease (COPD), lung cancer, interstitial lung diseases (ILDs) such as cystic fibrosis (CF), idiopathic pulmonary fibrosis, hypersensitivity pneumonitis, sarcoidosis and asbestosis, bacterial infections, viral infections, acute respiratory distress syndrome (ARDS), pulmonary alveolar proteinosis (PAP), acute bronchitis, bronchiolitis obliterans, and pulmonary hypertension.

[0036] In another aspect, the present invention relates to a nebulizer comprising a liquid formulation as defined above.

[0037] In one embodiment, the nebulizer is a mesh nebulizer.

[0038] In another aspect, the present invention provides a method for producing a pharmaceutical composition comprising: (i) a container containing the liquid formulation as defined above or a powder obtained by lyophilization of said liquid formulation, and (ii) Nebulizer The present invention relates to a kit comprising:

[0039] In one embodiment, the nebulizer is a mesh nebulizer.

[0040] In another aspect, the present invention relates to the use of a liquid formulation as defined above for preparing an aerosol by spraying with a nebulizer.

[0041] In one embodiment, the nebulizer is a mesh nebulizer.

[0042] In another aspect, the present invention relates to the use of a buffer selected from the group consisting of acetate, histidine, and combinations thereof, to enhance the stability of an antibody or antibody derivative when a liquid formulation containing the antibody or antibody derivative is nebulized through a nebulizer, wherein the buffer is included in the liquid formulation before nebulization.

[0043] In one embodiment, the antibody or antibody derivative is an IgG1 antibody or antibody derivative.

[0044] In one embodiment, the nebulizer is a mesh nebulizer.

[0045] In one embodiment, the liquid formulation has a pH of about 5.5 or less.

[0046] In one embodiment, the antibody is a monoclonal antibody.

[0047] In one embodiment, the liquid formulation is citrate-free.

[0048] In one embodiment, the liquid formulation has a pH ranging from about 3.5 to about 5.5.

[0049] In one embodiment, the buffering agent is acetate and the liquid formulation has a pH of less than about 5.0 or less than about 4.5. In one embodiment, the buffering agent is acetate and the liquid formulation has a pH ranging from about 3.5 to less than about 4.5, or from about 3.7 to about 4.3, or from about 3.8 to about 4.2, or from about 3.9 to about 4.1. In one embodiment, the liquid formulation has a pH of about 4.0.

[0050] In one embodiment, the liquid formulation further comprises a surfactant, hi one embodiment, the surfactant is selected from the group consisting of polysorbates, poloxamers, polyoxyethylene alkyl ethers, alkylphenol polyoxyethylenes, and sodium dodecyl sulfate.

[0051] In one embodiment, the concentration of surfactant in the liquid formulation is about 0.1% (w / v) or less, or about 0.05% (w / v) or less.

[0052] In one embodiment, the liquid formulation does not contain any surfactants.

[0053] In one embodiment, the liquid formulation does not include NaCl.

[0054] In one embodiment, the liquid formulation does not include any non-buffering salts. [Brief explanation of the drawings]

[0055] [Figure 1A] Figure 1 shows the analysis of the stability of two antibodies (mAb1 and mAb3) in formulations with different buffer systems for pulmonary delivery via nebulization by dynamic light scattering (DLS; A and D), SEC (B and E), flow cell microscopy (FCM; C and F), and visual inspection (B and E). [Figure 1B] Figure 1 shows the analysis of the stability of two antibodies (mAb1 and mAb3) in formulations with different buffer systems for pulmonary delivery via nebulization by dynamic light scattering (DLS; A and D), SEC (B and E), flow cell microscopy (FCM; C and F), and visual inspection (B and E). [Figure 1C] Figure 1 shows the analysis of the stability of two antibodies (mAb1 and mAb3) in formulations with different buffer systems for pulmonary delivery via nebulization by dynamic light scattering (DLS; A and D), SEC (B and E), flow cell microscopy (FCM; C and F), and visual inspection (B and E). [Figure 1D]Figure 1 shows the analysis of the stability of two antibodies (mAb1 and mAb3) in formulations with different buffer systems for pulmonary delivery via nebulization by dynamic light scattering (DLS; A and D), SEC (B and E), flow cell microscopy (FCM; C and F), and visual inspection (B and E). [Figure 1E] Figure 1 shows the analysis of the stability of two antibodies (mAb1 and mAb3) in formulations with different buffer systems for pulmonary delivery via nebulization by dynamic light scattering (DLS; A and D), SEC (B and E), flow cell microscopy (FCM; C and F), and visual inspection (B and E). [Figure 1F] Figure 1 shows the analysis of the stability of two antibodies (mAb1 and mAb3) in formulations with different buffer systems for pulmonary delivery via nebulization by dynamic light scattering (DLS; A and D), SEC (B and E), flow cell microscopy (FCM; C and F), and visual inspection (B and E). [Figure 2A] Analysis of the nebulization stability of two antibodies (mAb1 and mAb3) in a formulation containing PS80 for pulmonary delivery by DLS (A and D), SEC (B and E), FCM (C and F), and visual inspection (B and E). [Figure 2B] Analysis of the nebulization stability of two antibodies (mAb1 and mAb3) in a formulation containing PS80 for pulmonary delivery by DLS (A and D), SEC (B and E), FCM (C and F), and visual inspection (B and E). [Figure 2C] Analysis of the nebulization stability of two antibodies (mAb1 and mAb3) in a formulation containing PS80 for pulmonary delivery by DLS (A and D), SEC (B and E), FCM (C and F), and visual inspection (B and E). [Figure 2D] Analysis of the nebulization stability of two antibodies (mAb1 and mAb3) in a formulation containing PS80 for pulmonary delivery by DLS (A and D), SEC (B and E), FCM (C and F), and visual inspection (B and E). [Figure 2E]Analysis of the nebulization stability of two antibodies (mAb1 and mAb3) in a formulation containing PS80 for pulmonary delivery by DLS (A and D), SEC (B and E), FCM (C and F), and visual inspection (B and E). [Figure 2F] Analysis of the nebulization stability of two antibodies (mAb1 and mAb3) in a formulation containing PS80 for pulmonary delivery by DLS (A and D), SEC (B and E), FCM (C and F), and visual inspection (B and E). [Figure 3A] Analysis of the stability of an antibody (mAb1) in a pulmonary delivery formulation during nebulization using two different mesh nebulizers by DLS (A), SEC (B), FCM (C), and visual inspection (B). [Figure 3B] Analysis of the stability of an antibody (mAb1) in a pulmonary delivery formulation during nebulization using two different mesh nebulizers by DLS (A), SEC (B), FCM (C), and visual inspection (B). [Figure 3C] Analysis of the stability of an antibody (mAb1) in a pulmonary delivery formulation during nebulization using two different mesh nebulizers by DLS (A), SEC (B), FCM (C), and visual inspection (B). [Figure 4A] Figure 1 shows the analysis of the stability of antibodies of different isotypes (IgG1 and IgG4) in formulations for pulmonary delivery upon nebulization by DLS (A), SEC (B), FCM (C) and visual inspection (B). [Figure 4B] Figure 1 shows the analysis of the stability of antibodies of different isotypes (IgG1 and IgG4) in formulations for pulmonary delivery upon nebulization by DLS (A), SEC (B), FCM (C) and visual inspection (B). [Figure 4C] Figure 1 shows the analysis of the stability of antibodies of different isotypes (IgG1 and IgG4) in formulations for pulmonary delivery upon nebulization by DLS (A), SEC (B), FCM (C) and visual inspection (B). [Figure 5A]Figure 1 shows the analysis of the stability of an antibody (mAb1) in a formulation for pulmonary delivery upon nebulization by DLS and FCM (A), SEC and visual inspection (B), and additional analytical methods (C). [Figure 5B] Figure 1 shows the analysis of the stability of an antibody (mAb1) in a formulation for pulmonary delivery upon nebulization by DLS and FCM (A), SEC and visual inspection (B), and additional analytical methods (C). [Figure 6A-1] Figure 6 shows the analysis of the stability of two antibodies (mAb1 and mAb3) in formulations containing different concentrations of PS80 in citrate buffer during nebulization by DLS and FCM. Figure 6A shows the results for mAb1. [Figure 6A-2] Continuation of Figure 6A-1. [Figure 6A-3] Continuation of Figure 6A-2. [Figure 6A-4] Continuation of Figure 6A-3. [Figure 6B-1] Figure 6A shows the analysis of the stability of two antibodies (mAb1 and mAb3) in citrate buffer containing different concentrations of PS80 during nebulization by DLS and FCM. Figure 6B shows the results for mAb3. [Figure 6B-2] Continued from Figure 6B-1. [Figure 6B-3] Continued from Figure 6B-2. [Figure 6B-4] Continued from Figure 6B-3. [Figure 7-1] FIG. 1 shows the analysis of the stability of an antibody (mAb1) in a formulation containing different concentrations of PS80 in a histidine buffer for pulmonary delivery via nebulization by DLS and FCM. [Figure 7-2] Continued from Figure 7-1. [Figure 7-3] Continued from Figure 7-2. [Figure 7-4] Continued from Figure 7-3.

[0056] Detailed Description of the Invention Before describing the present invention in detail below, it is to be understood that the particular methodology, protocols, and reagents described herein may vary, and that the present invention is not limited thereto. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments and is not intended to limit the scope of the present invention, which is limited only by the appended claims. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. Preferably, terms used herein are defined in accordance with the following multilingual glossary of biotechnological terms: (IUPAC Recommendations), Leuenberger, H.G.W., Nagel, B. and Kolbl, H. (eds.) (1995), Helvetica The term "antibody" is defined as described in the Chimica Acta, CH-4010 Basel, Switzerland.

[0057] Several documents are cited throughout the text of this specification. Each document cited herein (including all patents, patent applications, scientific publications, manufacturer's specifications, instructions, etc.), whether supra or infra, is hereby incorporated by reference in its entirety. Nothing herein should be construed as an admission that the present invention is not entitled to antedate such disclosure by virtue of prior invention.

[0058] The following describes elements of the present invention. While these elements are described in conjunction with specific embodiments, it should be understood that they can be combined in any manner and in any number to create additional embodiments. The various described examples and preferred / specific embodiments should not be construed as limiting the invention to only the explicitly described embodiments. The description should be understood to support and encompass embodiments combining the explicitly described embodiments with any number of disclosed and / or preferred elements. Furthermore, any permutation and combination of all elements described herein shall be deemed to be disclosed by the description herein, unless otherwise indicated by context.

[0059] Throughout this specification and the appended claims, unless the context otherwise requires, the word "comprise," and variations such as "comprises" and "comprising," will be understood to mean the inclusion of a stated integer or step or group of integers or steps, and not the exclusion of other integers or steps or groups of integers or steps. As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the content clearly dictates otherwise.

[0060] In one aspect, the present invention provides an aerosol comprising droplets comprising a liquid formulation, the liquid formulation comprising: (i) an antibody or antibody derivative; (ii) a buffering agent selected from the group consisting of acetate, histidine, and combinations thereof; and (iii) Aqueous medium Including, The liquid formulation relates to an aerosol, having a pH of about 5.5 or less.

[0061] In one embodiment, the antibody or antibody derivative is an IgG1 antibody or antibody derivative.

[0062] The term "antibody" (or "immunoglobulin") generally refers to a glycoprotein comprising at least two heavy (H) chains and two light (L) chains interconnected by disulfide bonds. The term "antibody" includes monoclonal antibodies, recombinant antibodies, human antibodies, humanized antibodies, chimeric antibodies, and any combination of the above. Each heavy chain is composed of a heavy chain variable region (VH) and a heavy chain constant region (CH). Each light chain is composed of a light chain variable region (VL) and a light chain constant region (CL). Herein, the variable and constant regions are also referred to as variable and constant domains, respectively. The VH and VL regions can be further subdivided into regions of hypervariability called complementarity-determining regions (CDRs), interspersed with more conserved regions called framework regions (FRs). VH and VL each consist 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. The CDRs of VH are called HCDR1, HCDR2, and HCDR3, and the CDRs of VL are called LCDR1, LCDR2, and LCDR3. The variable regions of the heavy and light chains contain binding domains that interact with antigens. The constant region of an antibody includes a heavy chain constant region (CH) and a light chain constant region (CL), and the CH can be further subdivided into a constant domain CH1, a hinge region, and constant domains CH2 and CH3 (arranged from the amino terminus to the carboxy terminus in the following order: CH1, CH2, CH3). The constant region of an antibody can mediate the binding of immunoglobulins to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (Clq) of the classical complement system.

[0063] Antibodies may be derived from different species, including but not limited to mouse, rat, rabbit, guinea pig and human.

[0064] Antibodies described herein include antibodies of isotypes / classes such as IgA, such as IgA1 or IgA2, IgG1, IgG2, IgG3, IgG4, IgE, IgM, and IgD. In various embodiments, the antibody is an IgG1 antibody, more particularly an IgG1 kappa or IgG1 lambda isotype (i.e., IgG1, κ, λ), an IgG2a antibody (e.g., IgG2a, κ, λ), an IgG2b antibody (e.g., IgG2b, κ, λ), an IgG3 antibody (e.g., IgG3, κ, λ), or an IgG4 antibody (e.g., IgG4, κ, λ). In one embodiment, the antibody is an IgG1 antibody.

[0065] As used herein, the term "antibody derivative" refers to a molecule that does not have the entire structure of an antibody such as IgA, IgD, IgE, IgG, IgM, IgY, or IgW, particularly IgG1, but that contains at least a domain identified as containing it and that is still capable of binding a target molecule. Such derivatives may be, but are not limited to, functional (i.e., target-binding, particularly specific target-binding) antibody fragments thereof, such as Fab2, or combinations of such derivatives, such as bivalent Fabs. Also relevant are antibodies to which additional antibody domains, such as additional variable domains, have been added. In one embodiment, the term "antibody derivative" refers to single chain antibodies, e.g., as described in Spiess et al. (J Mol Imm. 2015, 67:95-106), WO2009 / 052081A2, Wu et al. (Nat. Biotechnol., 2007 25:1290-1297), Brinkmann & Kontermann (MAbs 2017, 9:182-212), and Fitzgerald et al. (Mol Cancer Ther. 2013, 13:410-25). Other non-limiting examples of antibody derivatives include, e.g., single chain antibodies described in Spiess et al. (J Mol Imm. 2015, 67:95-106). The term "IgG1 antibody derivative" refers to an antibody derivative that is recognizable as derived from an IgG1 antibody, for example, due to the presence of one or more IgG1-specific (sequence) elements.

[0066] In one embodiment, the antibody is a monoclonal antibody. As used herein, the term "monoclonal antibody" refers to a preparation of antibody molecules of a single molecular composition. A monoclonal antibody exhibits a single binding specificity and affinity. In one embodiment, a monoclonal antibody is produced by a hybridoma comprising a B cell obtained from a non-human animal, such as a mouse, fused to an immortalized cell.

[0067] In general, the terms "antibody" and "antibody derivative" as used herein include both monospecific and multispecific (e.g., bispecific, trispecific, or tetraspecific) antibodies and antibody derivatives.

[0068] In one embodiment, the antibody or antibody derivative is a pharmaceutically active antibody or antibody derivative. As used herein, the term "pharmaceutically active antibody or antibody derivative" refers to an antibody or antibody derivative that is suitable for therapy, i.e., that can be used to treat a disease or disorder. In one embodiment, the pharmaceutically active antibody or antibody derivative binds to a therapeutically relevant antigen.

[0069] Exemplary antibodies that can be used in accordance with the present invention include, but are not limited to, anti-respiratory syncytial virus (RSV) antibodies, anti-IL-4 antibodies, anti-IL-4 receptor antibodies, anti-IFN-beta antibodies, anti-IL-13 antibodies, anti-thymic stromal lymphopoietin (TSLP) antibodies, anti-IL-1 receptor antibodies, anti-IL-2 antibodies, and anti-TNF antibodies.

[0070] In one embodiment, the antibody concentration ranges from about 150 mg / ml to 1 mg / ml; or from about 150 mg / ml to about 5 mg / ml; or from about 150 mg / ml to about 10 mg / ml; or from about 150 mg / ml to about 20 mg / ml; or from about 150 mg / ml to about 50 mg / ml.

[0071] In one embodiment, the antibody concentration ranges from about 120 mg / ml to 1 mg / ml; or from about 120 mg / ml to about 5 mg / ml; or from about 120 mg / ml to about 10 mg / ml; or from about 120 mg / ml to about 20 mg / ml; or from about 120 mg / ml to about 50 mg / ml.

[0072] In one embodiment, the antibody concentration ranges from about 100 mg / ml to about 1 mg / ml; or from about 100 mg / ml to about 5 mg / ml; or from about 100 mg / ml to about 10 mg / ml; or from about 100 mg / ml to about 20 mg / ml; or from about 100 mg / ml to about 50 mg / ml.

[0073] In one embodiment, the antibody concentration ranges from about 80 mg / ml to 1 mg / ml; or from about 80 mg / ml to about 5 mg / ml; or from about 80 mg / ml to about 10 mg / ml; or from about 80 mg / ml to about 20 mg / ml; or from about 80 mg / ml to about 50 mg / ml.

[0074] In one embodiment, the antibody concentration ranges from about 50 mg / ml to about 5 mg / ml; or from about 400 mg / ml to about 5 mg / ml; or from about 30 mg / ml to about 5 mg / ml; or from about 20 mg / ml to about 5 mg / ml; or from about 10 mg / ml to about 5 mg / ml.

[0075] In one embodiment, the antibody concentration ranges from about 50 mg / ml to about 1 mg / ml; or from about 40 mg / ml to about 1 mg / ml; or from about 30 mg / ml to about 1 mg / ml; or from about 20 mg / ml to about 1 mg / ml; or from about 10 mg / ml to about 1 mg / ml.

[0076] In a preferred embodiment, the antibody concentration is about 150 mg / ml. In another preferred embodiment, the antibody concentration is about 140 mg / ml. In another preferred embodiment, the antibody concentration is about 130 mg / ml. In another preferred embodiment, the antibody concentration is about 120 mg / ml. In another preferred embodiment, the antibody concentration is about 110 mg / ml. In a preferred embodiment, the antibody concentration is about 100 mg / ml. In another preferred embodiment, the antibody concentration is about 90 mg / ml. In another preferred embodiment, the antibody concentration is about 80 mg / ml. In another preferred embodiment, the antibody concentration is about 70 mg / ml. In another preferred embodiment, the antibody concentration is about 60 mg / ml. In a preferred embodiment, the antibody concentration is about 50 mg / ml. In another preferred embodiment, the antibody concentration is about 40 mg / ml. In another preferred embodiment, the antibody concentration is about 30 mg / ml. In another preferred embodiment, the antibody concentration is about 20 mg / ml. In another preferred embodiment, the antibody concentration is about 10 mg / ml. In another preferred embodiment, the antibody concentration is about 5 mg / ml. In another preferred embodiment, the concentration of the antibody is about 1 mg / ml.

[0077] In one embodiment, the concentration of the buffering agent in the liquid formulation ranges from about 1 mM to about 200 mM, for example, from about 5 mM to about 150 mM, or from about 5 mM to about 100 mM, or from about 5 mM to about 50 mM. In one embodiment, the concentration of the buffering agent in the liquid formulation ranges from about 5 mM to about 25 mM, for example, from about 5 mM to about 20 mM, or from about 5 mM to about 15 mM, or from about 7.5 mM to about 12.5 mM. In one embodiment, the concentration of the buffering agent in the liquid formulation is about 10 mM. In one embodiment, the buffering agent is acetate, and the concentration of the acetate in the liquid formulation ranges from about 1 mM to about 200 mM, for example, from about 5 mM to about 150 mM, or from about 5 mM to about 100 mM, or from about 5 mM to about 50 mM. In one embodiment, the concentration of acetate in the liquid formulation ranges from about 5 mM to about 25 mM, e.g., from about 5 mM to about 20 mM, or from about 5 mM to about 15 mM, or from about 7.5 mM to about 12.5 mM. In one embodiment, the concentration of acetate in the liquid formulation is about 10 mM.

[0078] In one embodiment, the buffering agent is histidine, and the concentration of histidine in the liquid formulation is from about 1 mM to about 200 mM, e.g., from about 5 mM to about 150 mM, or from about 5 mM to about 200 mM. In one embodiment, the concentration of histidine in the liquid formulation is about 10 mM. In one embodiment, the concentration of histidine in the liquid formulation is about 20 mM. In one embodiment, the concentration of histidine in the liquid formulation is about 10 mM. In one embodiment, the concentration of histidine in the liquid formulation is about 20 mM.

[0079] In one embodiment, the acetate acting as a buffer is, for example, sodium acetate (or another suitable acetate, e.g., potassium acetate) in combination with acetic acid (i.e., in the form of an acetate buffer). Methods for preparing suitable acetate buffers are well known to those skilled in the art.

[0080] In one embodiment, the histidine that acts as a buffer is sodium L-histidine (or another suitable histidine salt), e.g., in the form of a histidine buffer. Methods for preparing suitable histidine buffers are well known to those skilled in the art.

[0081] In one embodiment, the liquid formulation is citrate-free.

[0082] As used herein, the term "aqueous medium" (or "aqueous solution") refers to a liquid medium or solution in which water is the solvent. In one embodiment, the aqueous medium is / consists of water, particularly purified water or water for injection (WFI). In one embodiment, the aqueous medium is sterile. In one embodiment, the liquid formulation is sterile.

[0083] In one embodiment, the liquid formulation has a pH ranging from about 3.5 to about 5.5.

[0084] In one embodiment, the buffering agent is acetate and the liquid formulation has a pH of less than about 5.0 or less than about 4.5. In one embodiment, the buffering agent is acetate and the liquid formulation has a pH ranging from about 3.5 to less than about 4.5, or from about 3.7 to about 4.3, or from about 3.8 to about 4.2, or from about 3.9 to about 4.1. In one embodiment, the liquid formulation has a pH of about 4.0.

[0085] In one embodiment, the buffering agent is acetate at a concentration of 10 mM, and the liquid formulation has a pH of less than about 5.0, or less than about 4.5. In one embodiment, the buffering agent is acetate at a concentration of 10 mM, and the liquid formulation has a pH ranging from about 3.5 to less than about 4.5, or from about 3.7 to about 4.3, or from about 3.8 to about 4.2, or from about 3.9 to about 4.1. In one embodiment, the liquid formulation has a pH of about 4.0, and the buffering agent is acetate at a concentration of 10 mM.

[0086] In one embodiment, the buffering agent is histidine and the liquid formulation has a pH of about 5.5 or less. In one embodiment, the buffering agent is histidine and the liquid formulation has a pH of less than about 5.0 or less than about 4.5. In one embodiment, the buffering agent is histidine and the liquid formulation has a pH ranging from about 3.5 to less than about 4.5, or from about 3.7 to about 4.3, or from about 3.8 to about 4.2, or from about 3.9 to about 4.1. In one embodiment, the liquid formulation has a pH of about 4.0.

[0087] In one embodiment, the buffering agent is histidine at a concentration of 10 mM, and the liquid formulation has a pH of about 5.5 or less. In one embodiment, the buffering agent is histidine at a concentration of 10 mM, and the liquid formulation has a pH of less than about 5.0 or less than about 4.5. In one embodiment, the buffering agent is histidine at a concentration of 10 mM, and the liquid formulation has a pH ranging from about 3.5 to less than about 4.5, or from about 3.7 to about 4.3, or from about 3.8 to about 4.2, or from about 3.9 to about 4.1. In one embodiment, The liquid formulation has a pH of about 4.0 and the buffering agent is histidine at a concentration of 10 mM.

[0088] In one embodiment, the buffering agent is histidine at a concentration of 20 mM, and the liquid formulation has a pH of about 5.5 or less. In one embodiment, the buffering agent is histidine at a concentration of 20 mM, and the liquid formulation has a pH of less than about 5.0 or less than about 4.5. In one embodiment, the buffering agent is histidine at a concentration of 20 mM, and the liquid formulation has a pH ranging from about 3.5 to less than about 4.5, or from about 3.7 to about 4.3, or from about 3.8 to about 4.2, or from about 3.9 to about 4.1. In one embodiment, the liquid formulation has a pH of about 4.0, and the buffering agent is histidine at a concentration of 20 mM.

[0089] The liquid formulation may contain one or more other excipients, so long as they are pharmaceutically acceptable and do not impair the suitability of the liquid formulation for administration by inhalation, particularly inhalation via a nebulizer. Suitable excipients are listed in the Pharmacopoeias or, for example, in REMINGTON'S PHARMACEUTICAL SCIENCES (18th ed., A.R. Gennaro, ed., Mack Publishing Company 1990) and its successors. As used herein, the term "pharmaceutically acceptable" refers to a substance that is non-toxic and, in one embodiment, does not interact with the action of the active agent in the liquid formulation.

[0090] In one embodiment, the liquid formulation further comprises a surfactant.

[0091] As used herein, the term "surfactant" (or "surface active agent") refers to a compound that reduces the surface tension (or interfacial tension) between two liquids, between a gas and a liquid, or between a liquid and a solid. In one embodiment, the compound reduces the surface tension (or interfacial tension) between a gas (e.g., air) and a liquid. In one embodiment, the surfactant is a non-ionic surfactant. In one embodiment, the surfactant is a polysorbate (e.g., polysorbate 20 or polysorbate 80), a poloxamer (e.g., poloxamer 188, commercially available as Pluronic® F68, or poloxamer 407, commercially available as Pluronic® F127), a polyoxyethylene alkyl ether (e.g., Brij™ surfactants), an alkylphenol polyoxyethylene (e.g., Triton® X100), and sodium dodecyl sulfate (SDS). In one embodiment, the surfactant is polysorbate 80 (PS80).

[0092] In one embodiment, the concentration of the surfactant in the liquid formulation is about 0.1% (w / v) or less, or about 0.05% (w / v) or less, for example, about 0.04% (w / v) or less, or about 0.03% (w / v) or less, or about 0.02% (w / v) or less, or about 0.01% (w / v) or less. In one embodiment, the concentration of the surfactant in the liquid formulation is less than about 0.01% (w / v).

[0093] In one embodiment, the concentration of polysorbate in the liquid formulation is about 0.1% (w / v) or less, or about 0.05% (w / v) or less, for example, about 0.04% (w / v) or less, or about 0.03% (w / v) or less, or about 0.02% (w / v) or less, about 0.01% (w / v) or less. In one embodiment, the concentration of polysorbate in the liquid formulation is less than about 0.01% (w / v).

[0094] In one embodiment, the concentration of PS80 in the liquid formulation is about 0.1% (w / v) or less, or about 0.05% (w / v) or less, for example, about 0.04% (w / v). ) or less, or about 0.03% (w / v) or less, about 0.02% (w / v) or less, or about 0.01% (w / v) or less. In one embodiment, the concentration of PS80 in the liquid formulation is less than about 0.01% (w / v).

[0095] In one embodiment, the liquid formulation contains only one surfactant, which in one embodiment is PS80.

[0096] In one embodiment, the liquid formulation does not contain any surfactants, eg, surfactants as defined above.

[0097] In one embodiment, the liquid formulation does not include NaCl.

[0098] In one embodiment, the liquid formulation does not contain any non-buffering salts. As used herein, the term "non-buffering salt" refers to a salt that does not contribute, or does not substantially contribute, to maintaining the pH of the liquid formulation upon addition of an acid or a base. In one embodiment, the non-buffering salt is a halogen salt (e.g., Cl). - or Br - In one embodiment, the non-buffering salt is sodium (Na + ), potassium (K + ), calcium (Ca 2+ ) or magnesium (Mg 2+ In one embodiment, the non-buffering salt is a halogen salt containing one or more cations of sodium (Na + ) or potassium (K + In yet another embodiment, the non-buffering salt is selected from the group consisting of NaCl, KCl, CaCl, and MgCl.

[0099] Generally, an aerosol is a suspension of fine solid particles or droplets in air or another gas. According to the present invention, the term "aerosol" means a suspension of droplets of a liquid formulation as defined above in a gas, e.g., air.

[0100] In one embodiment, the droplets have an average diameter of less than 5 μm. In one embodiment, the droplets have an average diameter of less than 4.5 μm. In one embodiment, the droplets have an average diameter of less than 4.0 μm. In one embodiment, the droplets have an average diameter of less than 3.5 μm. In one embodiment, the droplets have an average diameter of less than 3.0 μm.

[0101] In one embodiment, the droplets have an average diameter ranging from about 0.5 μm to about 5 μm. In one embodiment, the droplets have an average diameter ranging from about 0.5 μm to about 4.5 μm. In one embodiment, the droplets have an average diameter ranging from about 0.5 μm to about 4 μm. In one embodiment, the droplets have an average diameter ranging from about 0.5 μm to about 3.5 μm. In one embodiment, the droplets have an average diameter ranging from about 0.5 μm to about 3 μm. In one embodiment, the average diameter is the volume median diameter (VMD; also referred to as the Dv50 value). In one embodiment, the VMD is determined by laser diffraction, for example, as described in United States Pharmacopeia (USP) 429. Droplet size can also be measured, for example, by interferometric laser imaging. Results may vary depending on the measurement method used.

[0102] In one embodiment, the liquid formulation comprises an acetate buffer as a buffer, has a pH of about 5.0 or less than about 4.5, and comprises droplets having an average diameter of less than 5.0 μm. In one embodiment, the liquid formulation comprises an acetate buffer as a buffer, has a pH of about 5.0 or less than about 4.5, and comprises droplets having an average diameter ranging from about 0.5 μm to about 5 μm. In one embodiment, the liquid formulation comprises an acetate buffer as a buffer, has a pH ranging from about 3.5 to less than about 4.5, and comprises droplets having an average diameter of less than 5 μm. In one embodiment, the liquid formulation comprises an acetate buffer as a buffer, has a pH ranging from about 3.5 to less than about 4.5, and comprises droplets having an average diameter ranging from about 0.5 μm to about 5 μm. In one embodiment, the liquid formulation comprises an acetate buffer as a buffer, i.e., a pH between about 3.7 and about 4.3, and droplets having an average diameter of less than 5 μm. In one embodiment, the liquid formulation comprises an acetate buffer as a buffer, i.e., a pH between about 3.7 and about 4.3, and droplets having an average diameter ranging from about 0.5 μm to about 5 μm. In one embodiment, the liquid formulation comprises an acetate buffer as a buffer, i.e., a pH between about 3.8 and about 4.2, and droplets having an average diameter of less than 5 μm. In one embodiment, the liquid formulation comprises an acetate buffer as a buffer, i.e., a pH between about 3.8 and about 4.2, and droplets having an average diameter ranging from about 0.5 μm to about 5 μm. In one embodiment, the liquid formulation comprises an acetate buffer as a buffer, i.e., a pH between about 3.9 and about 4.1, and droplets having an average diameter of less than 5 μm. In one embodiment, the liquid formulation contains acetate buffer as a buffering agent, has a pH between about 3.9 and about 4.1, and contains droplets having an average diameter ranging from about 0.5 μm to about 5 μm. In another embodiment, the liquid formulation contains only one surfactant. In one embodiment, the surfactant is PS80.

[0103] In one embodiment, the liquid formulation comprises an acetate buffer as a buffer, has a pH of about 5.0 or less than about 4.5, and comprises droplets having an average diameter of less than 4.5 μm. In one embodiment, the liquid formulation comprises an acetate buffer as a buffer, has a pH of about 5.0 or less than about 4.5, and comprises droplets having an average diameter ranging from about 0.5 μm to about 4.5 μm. In one embodiment, the liquid formulation comprises an acetate buffer as a buffer, has a pH of about 3.5 to less than about 4.5, and comprises droplets having an average diameter of less than 4.5 μm. In one embodiment, the liquid formulation comprises an acetate buffer as a buffer, has a pH of about 3.5 to less than about 4.5, and comprises droplets having an average diameter ranging from about 0.5 μm to about 4.5 μm. In one embodiment, the liquid formulation comprises an acetate buffer as a buffer, has a pH between about 3.7 and about 4.3, and comprises droplets having an average diameter of less than 4.5 μm. In one embodiment, the liquid formulation comprises an acetate buffer as a buffer, i.e., a pH between about 3.7 and about 4.3, and droplets having an average diameter ranging from about 0.5 μm to about 4.5 μm. In one embodiment, the liquid formulation comprises an acetate buffer as a buffer, i.e., a pH between about 3.8 and about 4.2, and droplets having an average diameter less than 4.5 μm. In one embodiment, the liquid formulation comprises an acetate buffer as a buffer, i.e., a pH between about 3.8 and about 4.2, and droplets having an average diameter ranging from about 0.5 μm to about 4.5 μm. In one embodiment, the liquid formulation comprises an acetate buffer as a buffer, i.e., a pH between about 3.9 and about 4.1, and droplets having an average diameter less than 4.5 μm. In one embodiment, the liquid formulation contains acetate buffer as a buffering agent, has a pH between about 3.9 and about 4.1, and contains droplets having an average diameter ranging from about 0.5 μm to about 4.5 μm. In another embodiment, the liquid formulation contains only one surfactant. In one embodiment, the surfactant is PS80.

[0104] In one embodiment, the liquid formulation comprises an acetate buffer as a buffer, has a pH of about 5.0 or less than about 4.5, and comprises droplets having an average diameter of less than 4 μm. In one embodiment, the liquid formulation comprises an acetate buffer as a buffer, has a pH of about 5.0 or less than about 4.5, and comprises droplets having an average diameter ranging from about 0.5 μm to about 4 μm. In one embodiment, the liquid formulation comprises an acetate buffer as a buffer, has a pH ranging from about 3.5 to less than about 4.5, and comprises droplets having an average diameter of less than 4 μm. In one embodiment, the liquid formulation comprises an acetate buffer as a buffer, has a pH ranging from about 3.5 to less than about 4.5, and comprises droplets having an average diameter ranging from about 0.5 μm to about ...7 to about 4.3. In one embodiment, the liquid formulation comprises droplets having an average diameter of less than 4 μm, and a pH between about 3.7 and about 4.3, i.e., between about 0.5 μm and about 4 μm. In one embodiment, the liquid formulation comprises droplets having an average diameter of less than 4 μm, and a pH between about 3.8 and about 4.2, i.e., between about 3.8 and about 4.2, i.e., between about 3.8 and about 4.2. In one embodiment, the liquid formulation comprises droplets having an average diameter of less than 4 μm, and a pH between about 3.9 and about 4.1, i.e., between about 3.9 and about 4.1. In one embodiment, the liquid formulation contains acetate buffer as a buffering agent, has a pH between about 3.9 and about 4.1, and contains droplets having an average diameter ranging from about 0.5 μm to about 4 μm. In another embodiment, the liquid formulation contains only one surfactant. In one embodiment, the surfactant is PS80.

[0105] In one embodiment, the liquid formulation comprises an acetate buffer as a buffer, has a pH of about 5.0 or less than about 4.5, and comprises droplets having an average diameter of less than 3.5 μm. In one embodiment, the liquid formulation comprises an acetate buffer as a buffer, has a pH of about 5.0 or less than about 4.5, and comprises droplets having an average diameter ranging from about 0.5 μm to about 3.5 μm. In one embodiment, the liquid formulation comprises an acetate buffer as a buffer, has a pH of about 3.5 to less than about 4.5, and comprises droplets having an average diameter of less than 3.5 μm. In one embodiment, the liquid formulation comprises an acetate buffer as a buffer, has a pH of about 3.5 to less than about 4.5, and comprises droplets having an average diameter of about 0.5 μm to about 3.5 μm. In one embodiment, the liquid formulation comprises an acetate buffer as a buffer, has a pH between about 3.7 and less than about 4.3, and comprises droplets having an average diameter of less than 3.5 μm. In one embodiment, the liquid formulation comprises an acetate buffer as a buffer, i.e., a pH between about 3.7 and about 4.3, and droplets having an average diameter ranging from about 0.5 μm to about 3.5 μm. In one embodiment, the liquid formulation comprises an acetate buffer as a buffer, i.e., a pH between about 3.8 and about 4.2, and droplets having an average diameter less than 3.5 μm. In one embodiment, the liquid formulation comprises an acetate buffer as a buffer, i.e., a pH between about 3.8 and about 4.2, and droplets having an average diameter ranging from about 0.5 μm to about 3.5 μm. In one embodiment, the liquid formulation comprises an acetate buffer as a buffer, i.e., a pH between about 3.9 and about 4.1, and droplets having an average diameter less than 3.5 μm. In one embodiment, the liquid formulation contains acetate buffer as a buffering agent, has a pH between about 3.9 and about 4.1, and contains droplets having an average diameter ranging from about 0.5 μm to about 3.5 μm. In another embodiment, the liquid formulation contains only one surfactant. In one embodiment, the surfactant is PS80.

[0106] In one embodiment, the liquid formulation comprises an acetate buffer as a buffer, has a pH of about 5.0 or less than about 4.5, and comprises droplets having an average diameter of less than 3 μm. In one embodiment, the liquid formulation comprises an acetate buffer as a buffer, has a pH of about 5.0 or less than about 4.5, and comprises droplets having an average diameter ranging from about 0.5 μm to about 3 μm. In one embodiment, the liquid formulation comprises an acetate buffer as a buffer, has a pH ranging from about 3.5 to less than about 4.5, and comprises droplets having an average diameter of less than 3 μm. In one embodiment, the liquid formulation comprises an acetate buffer as a buffer, has a pH ranging from about 3.5 to less than about 4.5, and comprises droplets having an average diameter ranging from about 0.5 μm to about 3 μm. In one embodiment, the liquid formulation comprises an acetate buffer as a buffer, has a pH between about 3.7 and about 4.3, and comprises droplets having an average diameter of less than 3 μm. In one embodiment, the liquid formulation comprises an acetate buffer as a buffering agent, i.e., a pH between about 3.7 and about 4.3, and droplets having an average diameter ranging from about 0.5 μm to about 3 μm. In one embodiment, the liquid formulation comprises an acetate buffer as a buffering agent, i.e., a pH between about 3.8 and about 4.2, and droplets having an average diameter less than 3 μm. In one embodiment, the liquid formulation comprises an acetate buffer as a buffering agent, i.e., a pH between about 3.8 and about 4.2, and droplets having an average diameter ranging from about 0.5 μm to about 3 μm. In one embodiment, the liquid formulation comprises an acetate buffer as a buffering agent, i.e., a pH between about 3.9 and about 4.1, and droplets having an average diameter less than 3 μm. In one embodiment, the liquid formulation comprises an acetate buffer as a buffer, has a pH between about 3.9 and about 4.1, and comprises droplets having an average diameter ranging from about 0.5 μm to about 3 μm. In another embodiment, the liquid formulation comprises only one surfactant. In one embodiment, the surfactant is PS80.

[0107] In one embodiment, the liquid formulation comprises a histidine buffer as a buffer, has a pH of about 5.0 or less than about 4.5, and comprises droplets having an average diameter of less than 5 μm. In one embodiment, the liquid formulation comprises a histidine buffer as a buffer, has a pH of about 5.0 or less than about 4.5, and comprises droplets having an average diameter ranging from about 0.5 μm to about 5 μm. In one embodiment, the liquid formulation comprises a histidine buffer as a buffer, has a pH ranging from about 3.5 to less than about 4.5, and comprises droplets having an average diameter of less than 5 μm. In one embodiment, the liquid formulation comprises a histidine buffer as a buffer, has a pH ranging from about 3.5 to less than about 4.5, and comprises droplets having an average diameter ranging from about 0.5 μm to about 5 μm. In one embodiment, the liquid formulation comprises a histidine buffer as a buffer, has a pH between about 3.7 and about 4.3, and comprises droplets having an average diameter of less than 5 μm. In one embodiment, the liquid formulation comprises a histidine buffer as a buffering agent, i.e., a pH between about 3.7 and about 4.3, and droplets having an average diameter ranging from about 0.5 μm to about 5 μm. In one embodiment, the liquid formulation comprises a histidine buffer as a buffering agent, i.e., a pH between about 3.8 and about 4.2, and droplets having an average diameter less than 5 μm. In one embodiment, the liquid formulation comprises a histidine buffer as a buffering agent, i.e., a pH between about 3.8 and about 4.2, and droplets having an average diameter ranging from about 0.5 μm to about 5 μm. In one embodiment, the liquid formulation comprises a histidine buffer as a buffering agent, i.e., a pH between about 3.9 and about 4.1, and droplets having an average diameter less than 5 μm. In one embodiment, the liquid formulation contains a histidine buffer as a buffering agent, has a pH between about 3.9 and about 4.1, and contains droplets having an average diameter ranging from about 0.5 μm to about 5 μm. In another embodiment, the liquid formulation contains only one surfactant. In one embodiment, the surfactant is PS80.

[0108] In one embodiment, the liquid formulation comprises a histidine buffer as a buffer, has a pH of about 5.0 or less than about 4.5, and comprises droplets having an average diameter of less than 4.5 μm. In one embodiment, the liquid formulation comprises a histidine buffer as a buffer, has a pH of about 5.0 or less than about 4.5, and comprises droplets having an average diameter ranging from about 0.5 μm to about 4.5 μm. In one embodiment, the liquid formulation comprises a histidine buffer as a buffer, has a pH ranging from about 3.5 to less than about 4.5, and comprises droplets having an average diameter of less than 4.5 μm. In one embodiment, the liquid formulation comprises a histidine buffer as a buffer, has a pH ranging from about 3.5 to less than about 4.5, and comprises droplets having an average diameter ranging from about 0.5 μm to about 4.5 μm. In one embodiment, the liquid formulation comprises a histidine buffer as a buffer, has a pH ranging from about 3.5 to less than about 4.5, and comprises droplets having an average diameter ranging from about 0.5 μm to about 4.5 μm. In one embodiment, the liquid formulation comprises a histidine buffer as a buffer, has a pH between about 3.7 and about 4.3, and comprises droplets having an average diameter of less than 4.5 μm. In one embodiment, the liquid formulation contains histidine as a buffer. The liquid formulation comprises droplets containing an acetate buffer as a buffering agent, i.e., having a pH between about 3.7 and about 4.3, and having an average diameter ranging from about 0.5 μm to about 4.5 μm. In one embodiment, the liquid formulation comprises droplets containing an acetate buffer as a buffering agent, i.e., having a pH between about 3.8 and about 4.2, and having an average diameter less than 4.5 μm. In one embodiment, the liquid formulation comprises droplets containing an acetate buffer as a buffering agent, i.e., having a pH between about 3.8 and about 4.2, and having an average diameter ranging from about 0.5 μm to about 4.5 μm. In one embodiment, the liquid formulation comprises droplets containing a histidine buffer as a buffering agent, i.e., having a pH between about 3.9 and about 4.1, and having an average diameter less than 4.5 μm. In one embodiment, the liquid formulation contains a histidine buffer as a buffering agent, has a pH between about 3.9 and about 4.1, and contains droplets having an average diameter ranging from about 0.5 μm to about 4.5 μm. In another embodiment, the liquid formulation contains only one surfactant. In one embodiment, the surfactant is PS80.

[0109] In one embodiment, the liquid formulation comprises a histidine buffer as a buffer, has a pH of about 5.0 or less than about 4.5, and comprises droplets having an average diameter of less than 4 μm. In one embodiment, the liquid formulation comprises a histidine buffer as a buffer, has a pH of about 5.0 or less than about 4.5, and comprises droplets having an average diameter ranging from about 0.5 μm to about 4 μm. In one embodiment, the liquid formulation comprises a histidine buffer as a buffer, has a pH ranging from about 3.5 to less than about 4.5, and comprises droplets having an average diameter of less than 4 μm. In one embodiment, the liquid formulation comprises a histidine buffer as a buffer, has a pH ranging from about 3.5 to less than about 4.5, and comprises droplets having an average diameter ranging from about 0.5 μm to about 4 μm. In one embodiment, the liquid formulation comprises a histidine buffer as a buffer, has a pH between about 3.7 and about 4.3, and comprises droplets having an average diameter of less than 4 μm. In one embodiment, the liquid formulation comprises a histidine buffer as a buffering agent, i.e., a pH between about 3.7 and about 4.3, and droplets having an average diameter ranging from about 0.5 μm to about 4 μm. In one embodiment, the liquid formulation comprises a histidine buffer as a buffering agent, i.e., a pH between about 3.8 and about 4.2, and droplets having an average diameter less than 4 μm. In one embodiment, the liquid formulation comprises a histidine buffer as a buffering agent, i.e., a pH between about 3.8 and about 4.2, and droplets having an average diameter ranging from about 0.5 μm to about 4 μm. In one embodiment, the liquid formulation comprises a histidine buffer as a buffering agent, i.e., a pH between about 3.9 and about 4.1, and droplets having an average diameter less than 4 μm. In one embodiment, the liquid formulation comprises a histidine buffer as a buffering agent, has a pH between about 3.9 and about 4.1, and comprises droplets having an average diameter ranging from about 0.5 μm to about 4 μm. In another embodiment, the liquid formulation comprises only one surfactant. In one embodiment, the surfactant is PS80.

[0110] In one embodiment, the liquid formulation comprises a histidine buffer as a buffer, has a pH of about 5.0 or less than about 4.5, and comprises droplets having an average diameter of less than 3.5 μm. In one embodiment, the liquid formulation comprises a histidine buffer as a buffer, has a pH of about 5.0 or less than about 4.5, and comprises droplets having an average diameter ranging from about 0.5 μm to about 3.5 μm. In one embodiment, the liquid formulation comprises a histidine buffer as a buffer, has a pH ranging from about 3.5 to less than about 4.5, and comprises droplets having an average diameter of less than 3.5 μm. In one embodiment, the liquid formulation comprises a histidine buffer as a buffer, has a pH ranging from about 3.5 to less than about 4.5, and comprises droplets having an average diameter ranging from about 0.5 μm to about 3.5 μm. In one embodiment, the liquid formulation comprises a histidine buffer as a buffer, has a pH ranging from about 3.5 to less than about 4.5, and comprises droplets having an average diameter of less than 3.5 μm. In one embodiment, the liquid formulation comprises a histidine buffer as a buffer, has a pH between about 3.7 and about 4.3, i.e., between the pHs. In one embodiment, the liquid formulation contains hydroxybenzoates as a buffer. The liquid formulation comprises droplets containing a histidine buffer as a buffer, i.e., having a pH between about 3.7 and about 4.3, and having an average diameter ranging from about 0.5 μm to about 3.5 μm. In one embodiment, the liquid formulation comprises droplets containing a histidine buffer as a buffer, i.e., having a pH between about 3.8 and about 4.2, and having an average diameter of less than 3.5 μm. In one embodiment, the liquid formulation comprises droplets containing a histidine buffer as a buffer, i.e., having a pH between about 3.8 and about 4.2, and having an average diameter ranging from about 0.5 μm to about 3.5 μm. In one embodiment, the liquid formulation comprises droplets containing a histidine buffer as a buffer, i.e., having a pH between about 3.9 and about 4.1, and having an average diameter of less than 3.5 μm. In one embodiment, the liquid formulation contains a histidine buffer as a buffering agent, has a pH between about 3.9 and about 4.1, and contains droplets having an average diameter ranging from about 0.5 μm to about 3.5 μm. In another embodiment, the liquid formulation contains only one surfactant. In one embodiment, the surfactant is PS80.

[0111] In one embodiment, the liquid formulation comprises a histidine buffer as a buffer, has a pH of about 5.0 or less than about 4.5, and comprises droplets having an average diameter of less than 3 μm. In one embodiment, the liquid formulation comprises a histidine buffer as a buffer, has a pH of about 5.0 or less than about 4.5, and comprises droplets having an average diameter ranging from about 0.5 μm to about 3 μm. In one embodiment, the liquid formulation comprises a histidine buffer as a buffer, has a pH ranging from about 3.5 to less than about 4.5, and comprises droplets having an average diameter of less than 3 μm. In one embodiment, the liquid formulation comprises a histidine buffer as a buffer, has a pH ranging from about 3.5 to less than about 4.5, and comprises droplets having an average diameter ranging from about 0.5 μm to about 3 μm. In one embodiment, the liquid formulation comprises a histidine buffer as a buffer, has a pH between about 3.7 and about 4.3, and comprises droplets having an average diameter of less than 3 μm. In one embodiment, the liquid formulation comprises a histidine buffer as a buffering agent, i.e., a pH between about 3.7 and about 4.3, and droplets having an average diameter ranging from about 0.5 μm to about 3 μm. In one embodiment, the liquid formulation comprises a histidine buffer as a buffering agent, i.e., a pH between about 3.8 and about 4.2, and droplets having an average diameter less than 3 μm. In one embodiment, the liquid formulation comprises a histidine buffer as a buffering agent, i.e., a pH between about 3.8 and about 4.2, and droplets having an average diameter ranging from about 0.5 μm to about 3 μm. In one embodiment, the liquid formulation comprises a histidine buffer as a buffering agent, i.e., a pH between about 3.9 and about 4.1, and droplets having an average diameter less than 3 μm. In one embodiment, the liquid formulation contains a histidine buffer as a buffering agent, has a pH between about 3.9 and about 4.1, and contains droplets having an average diameter ranging from about 0.5 μm to about 3 μm. In another embodiment, the liquid formulation contains only one surfactant. In one embodiment, the surfactant is PS80.

[0112] According to the present invention, the antibodies or antibody derivatives present in the aerosol or liquid formulations described herein are characterized by reduced aggregation, for example, compared to the same antibodies or antibody derivatives formulated in a citrate-containing formulation.

[0113] In one embodiment, the antibody or antibody derivative present in the aerosol or liquid formulation described herein has one or more of the following properties: The polydispersity index (PDI) of the antibody or antibody derivative present in the aerosol or liquid formulation described herein is 0.5 or less, or 0.4 or less, or 0.3 or less, or 0.2 or less, or 0.1 or less, e.g., as determined by DLS (e.g., essentially as described in Example 1). The percent polydispersity of the antibody or antibody derivative monomers present in the aerosol or liquid formulations described herein is 30% or less, or 25% or less, or 20% or less, or 15% or less, e.g., as determined by DLS (e.g., essentially as described in Example 1). The mass percentage of antibody or antibody derivative monomer present in the aerosol or liquid formulations described herein is 99.7% or greater, or 99.8% or greater, or 99.8% or greater, e.g., as determined by DLS (e.g., essentially as described in Example 1). The percent intensity of antibody or antibody derivative monomer present in the aerosol or liquid formulations described herein is 80.0% or greater, or 85.0% or greater, or 90.0% or greater, or greater than 90.0%, e.g., as determined by DLS (e.g., essentially as described in Example 1). e.g., as determined by FCM (e.g., essentially as described in Example 1), the number of particles >2 μm is less than 10000 / mL, or less than 7500 / mL, or less than 5000 / mL, or less than 4000 / mL, or less than 3000 / mL, or less than 2000 / mL; the number of particles >10 μm is less than 500 / mL, or less than 400 / mL, or less than 300 / mL, or less than 200 / mL; and The number of particles >25 μm is less than 100 / mL, or less than 50 / mL, or less than 40 / mL, or less than 30 / mL, or less than 20 / mL.

[0114] In another aspect, the present invention relates to a method for preparing an aerosol comprising droplets comprising a liquid formulation, the method comprising the steps of: (i) providing a liquid formulation as defined above; (ii) nebulizing the liquid formulation provided in step (i) using a nebulizer, thereby preparing an aerosol. Includes:

[0115] In one embodiment, the nebulizer is a mesh nebulizer.

[0116] Nebulizers allow liquid formulations to be aerosolized and dispersed in gas for inhalation into the subject's respiratory tract. Examples of nebulizers include soft mist nebulizers, mesh nebulizers (e.g., vibrating mesh nebulizers), jet nebulizers, and ultrasonic nebulizers. Suitable nebulizer devices include Aerogen® Solo (Aerogen), Pari eFlow® (Pari GmbH), Philips I-neb™ (Philips), Pari These include the LC Sprint (Pari GmbH), the AERxRTM Pulmonary Delivery System (Aradigm), and the Pari LC Plus Reusable Nebulizer (Pari GmbH). In one embodiment, the nebulizer is a mesh nebulizer, particularly a vibrating mesh nebulizer. Nebulizers typically contain from about 1 mL to about 200 mL, more typically from 1 mL to 20 mL, of the liquid formulation.

[0117] In one embodiment, the method further comprises, between steps (i) and (ii), the step of: (ia) lyophilizing the liquid formulation provided in step (i), thereby providing a lyophilized powder; and (ib) reconstituting the liquid formulation provided in step (i) by adding an appropriate amount of aqueous medium to the lyophilized powder provided in step (ia). Further includes:

[0118] In another aspect, the present invention relates to an aerosol comprising droplets comprising a liquid formulation, the aerosol being obtainable by a method as defined above. In one embodiment, the droplets have an average diameter ranging from about 0.5 μm to about 5 μm, or from about 0.5 μm to about 3 μm.

[0119] In another aspect, the invention relates to a liquid formulation as defined above or an aerosol as defined above for use in a method of delivering an antibody or antibody derivative to the lungs of a subject, wherein the aerosol is administered to the subject by inhalation and the liquid formulation is administered to the subject by inhalation via a nebulizer.

[0120] In one embodiment, the antibody or antibody derivative is an IgG1 antibody or antibody derivative.

[0121] In one embodiment, the nebulizer is a mesh nebulizer.

[0122] The term "subject", according to the present invention, means a subject for treatment, particularly an affected subject (also called a "patient"), and includes humans, non-human primates or other animals, particularly mammals, such as cows, horses, pigs, sheep, goats, dogs, cats, rabbits or rodents, such as mice, rats, guinea pigs and hamsters. In one embodiment, the subject / patient is a human.

[0123] In another aspect, the present invention relates to a liquid formulation as defined above for use in a method for treating or preventing a disease in a subject, wherein the aerosol is administered to the subject by inhalation, or the liquid formulation is administered to the subject by inhalation via a nebulizer.

[0124] In one embodiment, the disease is a pulmonary disease.

[0125] In one embodiment, the nebulizer is a mesh nebulizer.

[0126] As used herein, the term "treating" refers to any procedure that improves the health and / or prolongs (increases) the lifespan of a patient.

[0127] According to the present invention, the term "disease" refers to any pathological condition, in particular cancer, infectious diseases, inflammatory diseases, metabolic diseases, autoimmune diseases, degenerative diseases, apoptosis-related diseases and transplant rejection.

[0128] The term "cancer" according to the present invention also includes cancer metastasis. "Metastasis" refers to the spread of cancer cells from their original site to another part of the body. The formation of metastasis is a highly complex process that relies on the detachment of malignant cells from the primary tumor, their invasion into the extracellular matrix, their penetration through the endothelial basement membrane to enter body cavities and blood vessels, and their subsequent transport by the blood followed by their invasion of the target organ. Ultimately, the growth of new tumors at the target site, i.e., secondary or metastatic tumors, depends on angiogenesis. Tumor metastasis often occurs even after removal of the primary tumor, as tumor cells or components may remain and develop metastatic potential. In one embodiment, the term "metastasis" according to the present invention refers to "distant metastasis," which refers to metastasis away from the primary tumor and the regional lymph node system.

[0129] The term "infectious disease" refers to any disease that can be transmitted from individual to individual or organism to organism and is caused by a microbial agent (e.g., the common cold). Examples of infectious diseases include viral infections, such as AIDS (HIV), hepatitis A, B, or C, herpes, shingles (chickenpox), rubella (rubella virus), yellow fever, dengue fever, flaviviruses, influenza viruses, respiratory syncytial virus (RSV), hemorrhagic infections ( Marburg virus or Ebola virus), and severe acute respiratory syndrome (SARS), bacterial infections, such as Legionnaires' disease (Legionella), sexually transmitted diseases (e.g., chlamydia, gonorrhea), stomach ulcers (Helicobacter), cholera (Vibrio), tuberculosis, diphtheria, infections with E. coli, Staphylococci, Salmonella or Streptococci (tetanus); infections with protozoan pathogens, such as malaria, sleeping sickness, leishmaniasis; toxoplasmosis, i.e., infections with Plasmodium, Trypanosoma, Leishmania and Toxoplasma; or, for example, Cryptococcus neoformans, Histoplasma capsulatum These include fungal infections caused by Coccidioides capsulatum, Coccidioides immitis, Blastomyces dermatitidis or Candida albicans.

[0130] The term "inflammatory disease" refers to any disease characterized by or associated with high levels of inflammation in tissues, especially connective tissues, or degeneration of these tissues. Chronic inflammatory diseases are medical conditions characterized by persistent inflammation. Examples of (chronic) inflammatory diseases include celiac disease, vasculitis, lupus, chronic obstructive pulmonary disease (COPD), asthma, irritable bowel disease, atherosclerosis, arthritis, ankylosing spondylitis, Crohn's disease, colitis, chronic active hepatitis, dermatitis, and psoriasis.

[0131] The term "metabolic disease" refers to any disease or disorder that interferes with normal metabolism. Examples include cystinosis, diabetes, dyslipidemia, hyperthyroidism, hypothyroidism, hyperlipidemia, hypolipidemia, galactosemia, Gaucher disease, obesity, and phenylketonuria.

[0132] The term "autoimmune disease" refers to any disease or disorder in which the body mounts an immunogenic (i.e., immune system) response against some component of the body's own tissues. In other words, the immune system loses the ability to recognize some tissue or system within the body as self and targets and attacks the self as if it were foreign. Autoimmune diseases are classified as those that primarily affect one organ (e.g., hemolytic anemia and antiimmune thyroiditis) or those in which the autoimmune disease process spreads through many tissues (e.g., systemic lupus erythematosus). For example, multiple sclerosis is thought to be caused by T cells attacking the sheaths surrounding nerve fibers in the brain and spinal cord. The results are loss of coordination, weakness, and blurred vision. Autoimmune diseases are known in the art and include, for example, Hashimoto's thyroiditis, Graves' disease, lupus, multiple sclerosis, rheumatoid arthritis, hemolytic anemia, anti-immune thyroiditis, systemic lupus erythematosus, celiac disease, Crohn's disease, colitis, diabetes, scleroderma, psoriasis, and the like.

[0133] The term "degenerative disease" refers to any disease in which the function or structure of the affected tissue or organ gradually deteriorates over time, including, for example, Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS), Huntington's disease, macular degeneration, multiple sclerosis, muscular dystrophy, Niemann-Pick disease, osteoporosis, and rheumatoid arthritis.

[0134] The term "apoptosis-related disease" refers to any disease that involves alterations in apoptosis. Examples include cancer, neurological diseases such as Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis (ALS) and stroke, heart diseases such as ischemia-reperfusion, chronic heart failure, infectious diseases and autoimmune diseases.

[0135] The term "transplant rejection" refers to the rejection of a transplanted tissue or organ by the recipient's immune system. This refers to a rejection reaction that occurs during transplantation and can ultimately destroy the transplanted tissue or organ.

[0136] The term "lung disease" (also referred to herein as "pulmonary disease") generally refers to a disease or disorder that affects the lungs of a subject. In one embodiment, the lung disease is selected from the group consisting of asthma, chronic obstructive pulmonary disease (COPD), lung cancer (e.g., non-small cell lung cancer), cystic fibrosis (CF), interstitial lung disease (ILD), such as idiopathic pulmonary fibrosis, hypersensitivity pneumonitis, sarcoidosis, and asbestosis, bacterial infection (e.g., mycobacterial infection), viral infection (e.g., influenza virus infection or respiratory syncytial virus (RSV) infection), acute respiratory distress syndrome (ARDS), pulmonary alveolar proteinosis (PAP), acute bronchitis, obstructive bronchitis, and pulmonary hypertension.

[0137] In another aspect, the invention relates to a method for delivering an antibody or antibody derivative to the lungs of a subject, the method comprising administering to the subject by inhalation an effective amount of an aerosol as defined above, or administering to the subject by inhalation via a nebulizer an effective amount of a liquid formulation as defined above.

[0138] In one embodiment, the antibody or antibody derivative is an IgG1 antibody or antibody derivative.

[0139] In one embodiment, the nebulizer is a mesh nebulizer.

[0140] The term "effective amount" as used herein particularly refers to a "therapeutically effective amount," which is an amount that achieves the desired therapeutic response or effect, either alone or together with further doses, without causing unacceptable side effects. In the case of the treatment of a specific disease or condition, the desired response particularly relates to inhibiting the course of the disease. This includes slowing the progression of the disease, particularly halting or reversing the progression of the disease. The desired response in the treatment of a disease or condition may also be delaying or preventing the onset of the disease or condition. The effective amount of the aerosol or liquid formulation described herein, and therefore the antibody or antibody derivative contained therein, will depend on the condition being treated, the severity of the disease, individual parameters of the subject, including age, physiological state, size and weight, the duration of treatment, the type of concomitant therapy (if any), the specific route of administration, and similar factors. Therefore, the administered dose of the aerosol or liquid formulation described herein may depend on several of these parameters. If the subject's response is insufficient with the initial dose, a higher dose may be used.

[0141] In another aspect, the present invention relates to a method for treating or preventing a disease in a subject, the method comprising administering to the subject by inhalation an effective amount of an aerosol as defined above, or administering to the subject by inhalation via a nebulizer an effective amount of a liquid formulation as defined above.

[0142] In one embodiment, the disease is a pulmonary disease.

[0143] In one embodiment, the nebulizer is a mesh nebulizer.

[0144] In another aspect, the present invention relates to a nebulizer comprising a liquid formulation as defined above.

[0145] In one embodiment, the nebulizer is a mesh nebulizer.

[0146] In another aspect, the present invention provides a method for producing a pharmaceutical composition comprising: (i) a container containing a liquid formulation as defined above or a powder obtainable by lyophilization of said liquid formulation, and (ii) Nebulizer The present invention relates to a kit comprising:

[0147] In one embodiment, the nebulizer is a mesh nebulizer.

[0148] As used herein, the term "kit of parts" (abbreviated kit) refers to an article of manufacture comprising one or more containers, a nebulizer (e.g., a mesh nebulizer), and optionally a data carrier. The one or more containers are filled with the liquid formulation defined above and / or a powder obtained by lyophilization of the liquid formulation. Additional containers may be included in the kit, including, for example, a diluent (e.g., an aqueous medium), a buffer, and further reagents as defined herein. The data carrier may be a non-electronic data carrier, e.g., a graphic data carrier such as an information leaflet, an information sheet, a barcode, or an access code, or an electronic data carrier, such as a compact disc (CD), a digital versatile disc (DVD), a microchip, or other semiconductor-based electronic data carrier. The access code may enable access to a database, e.g., an internet database, a centralized database, or a distributed database. The data carrier may contain instructions for using the kit in the methods and applications described herein.

[0149] In another aspect, the present invention relates to the use of a liquid formulation as defined above for preparing an aerosol by spraying with a nebulizer.

[0150] In one embodiment, the nebulizer is a mesh nebulizer.

[0151] In another aspect, the present invention relates to the use of a buffer selected from the group consisting of acetate, histidine, and combinations thereof, to enhance the stability of an antibody or antibody derivative when a liquid formulation containing the antibody or antibody derivative is nebulized using a nebulizer, wherein the buffer is included in the liquid formulation before nebulization.

[0152] In one embodiment, the antibody or antibody derivative is an IgG1 antibody or antibody derivative.

[0153] In one embodiment, the nebulizer is a mesh nebulizer.

[0154] In one embodiment, the term "enhancing stability" refers to preventing or reducing the extent of aggregation of an antibody or antibody derivative.

[0155] In one embodiment, the liquid formulation has a pH of about 5.5 or less.

[0156] In one embodiment, the antibody is a monoclonal antibody.

[0157] In one embodiment, the liquid formulation is citrate-free.

[0158] In one embodiment, the liquid formulation has a pH ranging from about 3.5 to about 5.5.

[0159] In one embodiment, the buffering agent is acetate and the liquid formulation has a pH of less than about 5.0, or less than about 4.5. In one embodiment, the buffering agent is acetate and the liquid formulation has a pH of between about 3.5 and less than about 4.5, or between about 3.7 and about 4.3, or between about 3.8 and about to 4.2, or in the range of about 3.9 to about 4.1. In one embodiment, the liquid formulation has a pH of about 4.0.

[0160] In one embodiment, the liquid formulation further comprises a surfactant, hi one embodiment, the surfactant is selected from the group consisting of polysorbates, poloxamers, polyoxyethylene alkyl ethers, alkylphenol polyoxyethylenes, and sodium dodecyl sulfate.

[0161] In one embodiment, the concentration of surfactant in the liquid formulation is about 0.1% (w / v) or less, or about 0.05% (w / v) or less.

[0162] In one embodiment, the liquid formulation does not contain any surfactants.

[0163] In one embodiment, the liquid formulation does not include NaCl.

[0164] In one embodiment, the liquid formulation does not contain any non-buffering salts.

[0165] The present invention will now be further described with reference to the following examples, which are intended to illustrate but not limit the scope of the invention. [Example]

[0166] Example 1: Materials and Methods The immunoglobulins used in the examples are shown in Table 1 below. [Table 1]

[0167] Dynamic Light Scattering (DLS) DLS measurements were performed using a DynaPro NanoStar (Wyatt Technology) instrument with a 663 nm laser. Each sample was placed in a disposable cuvette (Uvette, Eppendorf), and measurements were performed with 10 acquisitions of 7 seconds each. Data were analyzed using Dynamics 7.1.9 software (Wyatt Technology) to determine particle size distribution in the submicron range.

[0168] Results are expressed as Z-average, polydispersity index (PDI), monomer radius, percentage polydispersity of the monomer peak (pic), percentage intensity, and mass of the monomer.

[0169] Polydispersity is considered low if the PDI is less than 0.1 and high if the sample is diverse.

[0170] If the percent polydispersity of the monomers is less than 15%, the level of homogeneity is considered high. If the level of homogeneity is low (percent polydispersity greater than 30%), the particle population can be considered to contain significantly different sizes, i.e., polydisperse.

[0171] A monomer mass percentage greater than 99.8% is considered low cohesion, and a monomer mass percentage less than 99.6% is considered high cohesion. A monomer mass percentage greater than 90.0% is considered low cohesion, and a monomer mass percentage less than 80.0% is considered high cohesion.

[0172] Flow Cell Microscopy (FCM) Flow cell microscopy measurements were performed using a Flowcell FC200-IPAC (Occhio) instrument. Each analysis was performed on a 200 μL sample introduced into a disposable cone. Data were analyzed using Callisto software (Occhio) to determine particle size distribution in the subvisible region.

[0173] Results are expressed as particle concentrations (particles / mL) of >2 μm, >10 μm, and >25 μm.

[0174] Aggregation is considered low when particles >2 μm are less than 2000 / mL, particles >10 μm are less than 200 / mL, and particles >25 μm are less than 20 / mL. Aggregation is considered high when particles >2 μm are more than 10,000 / mL, particles >10 μm are more than 500 / mL, and particles >25 μm are more than 100 / mL.

[0175] Size Exclusion Chromatography (SEC) SEC measurements were carried out on an ultra-high performance chromatographic chain Agilent 1200. Results are expressed as percentage of high molecular weight (HMW).

[0176] For mAb1: Measurements were performed at 24 °C with a mobile phase consisting of 0.1 M phosphate buffer, pH 6.2, 0.3 M NaClO4, and acetonitrile at a flow rate of 0.3 mL / min. Samples were stored in vials, and 10 μL was injected onto a column series consisting of a guard column Prosec 300S 50 x 7.5 mm (Agilent) and two columns Prosec 300S 300 x 7.5 mm (Agilent). Detection was performed at 280 nm. Results were processed using Empower software.

[0177] For mAb3: SEC measurements were performed using an Agilent 1200 ultrafast chromatography chain. Measurements were performed at 30°C with a mobile phase consisting of 50 mM sodium phosphate, 300 mM sodium perchlorate, pH 7, at a flow rate of 0.3 mL / min. Samples were stored in vials, and 1 μL was injected onto an Aquity UPLC BEH200 SEC 200A, 1.7 μm, 4.6 mm x 300 mm column (Waters). Detection was performed using a 1.7 μm, 4.6 mm x 300 mm column (Waters). The results were processed using Empower software.

[0178] Visual inspection Visual inspection was performed on samples placed in glass vials and illuminated with an MLC-150 cold light source (Motic) on a black background.

[0179] Example 2: Effect of buffer and pH on IgG1 stability This example shows that acetate and histidine buffers at acidic pH promote the stability of an IgG1 antibody against aggregation upon nebulization compared to citrate buffer.

[0180] Two IgG1 immunoglobulins formulated in different buffers with different pH values ​​without PS80 The IgG1 and mAb3 were nebulized and their post-nebulization stability was evaluated. mAb1 and mAb3 were formulated at 10 mg / mL in six different buffer / pH systems: acetate pH 4.0, acetate pH 5.5, histidine pH 5.5, histidine pH 7.0, citrate pH 5.5, and citrate pH 6.5. The buffer concentration was 10 mM for each system. Nebulization was performed using a Solo (Aerogen) vibrating mesh nebulizer with 2 mL of IgG1 formulated in the different buffers. The degree of aggregation was measured by dynamic light scattering (DLS), flow cell microscopy (FCM), SEC, and visual inspection. The results are summarized in Figure 1A-C (mAb1) and Figure 1D-F (mAb3).

[0181] Visual inspection showed the presence of visible particles in the citrate formulation of mAb1. DLS results showed low aggregation to submicron particles in acetate and histidine buffers, especially at acidic pH, whereas the citrate formulation was not suitable for analysis without filtration (likely due to the high particle content in the sample). FCM results confirmed high aggregation in the citrate buffer.

[0182] Example 3: Effect of Polysorbate 80 (PS80) on IgG1 Stability This example shows that a large amount of PS80 (1000 ppm) helps to limit aggregation during nebulization of IgG1 in citrate buffer, but not completely, as submicron particles of mAb1 are beyond the level of quantification by DLS.

[0183] Two immunoglobulins (mAb1 and mAb3) were formulated in different buffers at different pH levels in the presence of polysorbate 80 (PS80). Acetate, histidine, and citrate buffers were used, with PS80 used as a surfactant. Two IgGs were formulated at 10 mg / mL in six different buffer / pH systems: acetate pH 4.0, acetate pH 5.5, histidine pH 5.5, histidine pH 7.0, citrate pH 5.5, and citrate pH 6.5. A 10 mM buffer concentration was used. PS80 was added to achieve a 1000 ppm concentration in the formulation. 2 mL of IgG1 formulated in the different buffers was nebulized using a Solo (Aerogen) vibrating mesh nebulizer. The degree of aggregation was measured by dynamic light scattering (DLS), flow cell microscopy (FCM), SEC, and visual inspection. The results are summarized in Figures 2A-C (mAb1) and Figures 2D-F (mAb3).

[0184] Visual inspection showed no visible particles in all formulations with PS80. DLS results indicated that the citrate formulation was not suitable for analysis without filtration, likely due to high particle content / large particles in the sample. FCM results demonstrated lower levels of aggregation compared to the same formulation without PS80 tested in Example 2.

[0185] Example 4: Nebulization with two different mesh vibrating nebulizers In this example, the stabilizing properties of acetate and histidine buffers are demonstrated in two different vibrating mesh nebulizers: the Solo nebulizer (see previous example) and the eFlow nebulizer from PARI. The stabilizing effect on IgG1 antibodies in acetate and histidine buffers (compared to citrate buffer) was also confirmed in the eFlow system.

[0186] Acetate, histidine, and citrate buffers were used. Two IgG1s were prepared at 10 mg / mL concentrations using six buffer / pH systems: acetate pH 4.0, acetate pH 5.5, histidine pH 5.5, histidine pH 7.0, citrate pH 5.5, and citrate pH 6.5. Two mL of IgG1 formulated in different buffers was subjected to nebulization stress using a customized eFlow (PARI) vibrating mesh nebulizer. The degree of aggregation was measured by dynamic light scattering (DLS), flow cell microscopy (FCM), SEC, and visual inspection. The results are summarized in Figure 3A-C (mAb1).

[0187] Example 5: Stability of different IgG isotypes (IgG1 and IgG4) in acetate buffer pH 5.5 This example demonstrates the stabilizing effect of acetate buffers at acidic pH on several IgG1 antibodies. For IgG4 antibodies, the stabilizing properties of these buffers are less pronounced.

[0188] Four IgG1s (mAb1, mAb2, mAb3, and mAb4) and one IgG4 (mAbX) were nebulized in the same formulation at a concentration of 10 mg / mL (pH 5.5 acetate buffer) using a Solo (Aerogen) vibrating mesh nebulizer at 2 mL. The degree of aggregation was measured using dynamic light scattering (DLS), flow cell microscopy (FCM), SEC, and visual inspection. The results are summarized in Figure 4A-C.

[0189] Visual inspection showed the presence of visible particles in the IgG4 X sample. DLS results indicated that the IgG4 X sample was not suitable for analysis without filtration, likely due to particles in the sample. FCM counting confirmed high aggregation in the IgG4 X sample and low aggregation in the four IgG1 samples.

[0190] Example 6: Stabilizing effect of histidine This example demonstrates the stabilizing properties of histidine buffer in formulations containing excipients (polyols and non-ionic surfactants), allowing for the manufacture and long-term storage of pharmaceutical products prior to nebulization.

[0191] An IgG1 antibody (mAb1) was formulated at 20 g / L in 20 mM histidine buffer (pH 6.0) with additional excipients, a nonionic surfactant, and a polyol, providing long-term storage stability. A 2 mL mAb1 formulation was subjected to nebulization using a Solo (Aerogen) vibrating mesh nebulizer. The aerosol droplet size was determined by laser diffraction measurement. The resulting VMD after nebulization was 4.4 μm, with most droplets having a diameter less than 5 μm. Approximately 30% of the droplets had diameters ranging from 0.5 μm to 3 μm. The degree of aggregation was assessed by dynamic light scattering (DLS), flow cell microscopy (FCM), SEC, and visual inspection. Additional analytical methods were performed. This formulation can be stored at 2-8°C for up to 24 months. The results are summarized in Figures 5A and 5B.

[0192] Example 7: Effect of PS80 concentration on IgG1 in citrate buffer The effect of PS80 concentration on the stability of nebulized IgG1 in citrate was evaluated. Nebulization stress was applied to two different IgG1s (mAb 1 and mAb 3) formulated in citrate pH 5.5 with different concentrations of PS80. PS80 concentrations of 0, 200, and 1000 ppm were used. The two IgG1s were prepared at 10 mg / mL in citrate buffer, and PS80 was added at the target concentration. Nebulization stress was applied to 2 mL of IgG1 formulated in citrate with different concentrations of PS80 using a vibrating mesh nebulizer. The degree of aggregation was measured using dynamic light scattering (DLS) and flow microscopy (FCM; Figure 6). Figure 6 shows that IgG1 in citrate buffer exhibited high aggregation levels at 0 ppm of PS80. A high concentration of PS80 (1000 ppm) was required to stabilize IgG1 in citrate. DLS results indicated that the citrate formulation was not suitable for analysis without filtration, likely due to particles in the sample. FCM results showed a high concentration of particles at 0 ppm PS80. As the PS80 concentration increased, the number of particles per mL decreased. 200 ppm PS80 was not sufficient to stabilize mAb 1. DLS results indicated that the citrate formulation was not suitable for analysis without filtration, likely due to particles in the sample at 0 ppm PS80 in most cases (n=5 / 6). 200 ppm PS80 At 1000 ppm of PS80, aggregation was observed. At 1000 ppm of PS80, aggregation was low. FCM results showed a high concentration of particles at 0 ppm of PS80. Addition of 200 ppm and 1000 ppm of PS80 reduced the particle concentration.

[0193] Example 8: Effect of PS80 concentration on IgG1 in histidine buffer The effect of PS80 concentration on the stability of IgG1 nebulized in histidine was evaluated. To this end, nebulization stress was applied to one IgG1 (mAb 1) formulated in histidine, pH 5.5, with different concentrations of PS80. PS80 concentrations of 0, 50, 200, and 1000 ppm were used. IgG1 was prepared at a concentration of 10 mg / mL in histidine buffer, and PS80 was added at the target concentration. Nebulization stress was applied to 2 mL of IgG1 formulated in histidine with different concentrations of PS80 using a vibrating mesh nebulizer. The degree of aggregation was measured using dynamic light scattering (DLS) and flow microscopy (FCM; Figure 7). Figure 7 shows that IgG1 in histidine buffer exhibited moderate aggregation at 0 ppm PS80. A low concentration of PS80 (50 ppm) appears sufficient to stabilize IgG1 in histidine. DLS results showed moderate aggregation at 0 ppm PS80. At 50, 200, and 100 ppm PS80, the level of aggregation was low and similar across different concentrations. FCM results showed moderate concentrations of particles at 0 ppm PS80. Addition of 50, 200, and 1000 ppm PS80 was able to reduce particle concentrations in a comparable manner.

Claims

1. an aerosol comprising droplets containing a liquid formulation; The liquid formulation comprises: (i) an antibody or antibody derivative; (ii) a buffering agent selected from the group consisting of acetate, histidine, and combinations thereof; and (iii) Aqueous medium wherein the liquid formulation has a pH that is about 5.5 or less.

2. The aerosol of claim 1 , wherein the liquid formulation is citrate-free.

3. 3. The aerosol of claim 1 or 2, wherein the buffer is acetate and the liquid formulation has a pH that is less than about 5.0 or less than about 4.

5.

4. The aerosol of any one of claims 1 to 3, wherein the liquid formulation further comprises a surfactant.

5. 5. The aerosol of claim 4, wherein the concentration of surfactant in the liquid formulation is about 0.1% (w / v) or less, or about 0.05% (w / v) or less.

6. 6. The aerosol of claim 4 or 5, wherein the surfactant is selected from the group consisting of polysorbate, poloxamer, polyoxyethylene alkyl ether, alkylphenol polyoxyethylene, and sodium dodecyl sulfate.

7. The aerosol according to any one of claims 1 to 3, wherein the liquid formulation does not contain a surfactant.

8. The aerosol according to any one of claims 1 to 7, wherein the antibody or antibody derivative is an IgG1 antibody or antibody derivative.

9. 9. The aerosol of any one of claims 1 to 8, wherein the droplets have an average diameter ranging from about 0.5 μm to about 5 μm, from about 0.5 μm to about 4.5 μm, from about 0.5 μm to about 4 μm, from about 0.5 μm to about 3.5 μm, or from about 0.5 μm to about 3 μm.

10. 1. A method for preparing an aerosol comprising droplets containing a liquid formulation, the method comprising the steps of: (i) providing a liquid formulation as defined in any one of claims 1 to 8, (ii) nebulizing the liquid formulation provided in step (i) using a nebulizer, thereby preparing an aerosol. A method comprising:

11. Optionally, between steps (i) and (ii), the following step: (ia) lyophilizing the liquid formulation provided in step (i), thereby providing a lyophilized powder; and (ib) reconstituting the liquid formulation provided in step (i) by adding an appropriate amount of aqueous medium to the lyophilized powder provided in step (ia). The method of claim 10 further comprising:

12. 12. An aerosol comprising droplets comprising a liquid formulation, the aerosol being obtainable by the method of claim 10 or 11.

13. 13. The aerosol of any one of claims 1 to 9 and 12 or the liquid formulation of any one of claims 1 to 8 for use in a method of delivering an antibody or antibody derivative to the lungs of a subject, wherein the aerosol is administered to the subject by inhalation, or the liquid formulation is administered to the subject by inhalation via a nebulizer.

14. 13. An aerosol according to any one of claims 1 to 9 and 12 or a liquid formulation according to any one of claims 1 to 8 for use in a method for treating or preventing a disease in a subject, wherein the aerosol is administered to the subject by inhalation or the liquid formulation is administered to the subject by inhalation via a nebulizer.

15. A nebulizer comprising a liquid formulation according to any one of claims 1 to 8.

16. A kit comprising: (i) a container containing the liquid formulation according to any one of claims 1 to 8 or a powder obtained by lyophilization of said liquid formulation; and (ii) Nebulizer Includes a kit.

17. Use of a liquid formulation according to any one of claims 1 to 8 for preparing an aerosol by spraying with a nebulizer.

18. 1. Use of a buffer selected from the group consisting of acetate, histidine, and combinations thereof, to enhance the stability of an antibody or antibody derivative when a liquid formulation containing the antibody or antibody derivative is nebulized using a nebulizer, wherein the buffer is included in the liquid formulation before nebulization.

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