Compositions and Methods for Inhalable Therapeutics

Abstract

Therapeutic inhaled antibodies and methods of delivering these therapeutic antibodies are described herein that may sustain the concentration of the therapeutic inhaled antibody in the upper respiratory tract (URT) and lower respiratory tract (LRT) as well as in the blood even after a single dose. The compositions and methods described herein can provide therapeutically relevant levels of an inhaled antibody delivered by inhalation in a single dose delivered at a frequency of once a day or less. These methods can result in concentrations in both the URT and LRT that are higher than a clinically relevant minimum threshold concentration.

Description

Technical Field

[0001] Claims of Priority

[0001] This application claims the benefit of U.S. Provisional Patent Application No. 63 / 345,019, filed May 23, 2022, entitled "COMPOSITIONS AND METHODS FOR INHALABLE THERAPEUTICS", and U.S. Provisional Patent Application No. 63 / 233,661, filed Aug. 16, 2021, entitled "METHODS AND APPARATUSES FOR DELIVERY OF AN AGENT TO THE LUNGS AND NASAL PASSAGES", and claims priority to U.S. Patent Application No. 17 / 889,141, filed Aug. 16, 2022, the entire contents of all of which are incorporated herein by reference in their entirety. Incorporation by Reference

[0002] All publications and patent applications cited herein are incorporated herein by reference in their entirety to the same extent as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference.

Background Art

[0002]

[0003] It is generally accepted that antibodies delivered to the lungs by inhalation, including those conjugated with Fc, are rapidly removed from the lungs. For example, proteins conjugated with Fc administered by inhalation typically have a Tmax (i.e., the time to reach Cmax) in serum in the range of 10 - 20 hours, and thus have a rather rapid clearance (on the order of hours or minutes) in the lungs. Bitonti and Durmont, "Pulmonary administration of therapeutic proteins using an immunoglobulin transport pathway," Advanced Drug Delivery Reviews, Volume 58, Issues 9 - 10, 31 October 2006, pages 1106 - 1118. In fact, the therapeutic efficacy of inhaled drugs has long been thought to be limited by their rapid clearance in the lungs. Small solutes delivered to the lungs rapidly diffuse into the lung epithelium and penetrate into the bloodstream within minutes. Peptides are similarly rapidly transported into the systemic circulation but are significantly metabolized locally. As summarized by Loira - Pastoriza et al., ("Delivery strategies for sustained drug release in the lungs," Advanced Drug Delivery Reviews, Volume 75, 30 August 2014, pages 81 - 91), "Macromolecules can be absorbed into the systemic circulation over several hours, but they may be rapidly taken up by alveolar macrophages, removed by the mucociliary escalator, or metabolized locally. For example, recombinant human deoxyribonuclease I is a 37 kDa glycoprotein that cleaves DNA in the respiratory secretions of patients with cystic fibrosis and thus reduces its viscosity. This glycoprotein is the most widely used mucolytic agent in the symptomatic treatment of cystic fibrosis. However, it is rapidly removed from the human lungs when a daily dose of 2.5 mg is inhaled and a concentration of 3 μg / ml is measured in sputum immediately after inhalation and reduced to 0.6 μg / ml after 2 hours."Therefore, once- or twice-daily dosing provides a restricted therapeutic application range for patients. Unfortunately, the short residence time of drugs in the lungs also requires more frequent dosing, which is thought to endanger patient compliance. For example, steps of inhaling corticosteroids at least twice a day and short-acting β2-agonists up to four times a day are recommended.

Summary of the Invention

[0003]

[0004] Therefore, it would be beneficial to provide a composition, particularly an mAb composition, that can remain in the lungs at clinically significant levels for an extended period without being removed. Such compositions and methods can provide numerous clinical and compliance benefits.

[0004]

[0005] The present invention relates to therapeutic inhaled antibodies and methods of delivering these therapeutic antibodies to maintain concentrations in the upper respiratory tract (URT), lower respiratory tract (LRT), and blood even after a single dose. Surprisingly, the compositions and methods described herein can provide therapeutically relevant levels of inhaled IgG antibodies delivered by inhalation in a single dose administered once a day or less frequently (e.g., between once a day and once every five days). These methods can result in concentrations above the minimum threshold concentration that is clinically relevant in both the URT and LRT.

[0005]

[0006] The persistence of therapeutic mAbs in the URT and LRT appears to be the result of interactions of the core Fc region of the IgG backbone common to the therapeutic antibodies described herein (including, for example, regdanvimab), regardless of the target-specific portion (variable region) of the individual mAbs. This could be because the Fc region is interacting with other components that drive the clearance of mAbs from mucus and the lung. The effects described herein are particularly relevant to compositions of mAbs in which the IgG Fc region is glycosylated in a manner that modulates mucin interactions. For example, these compositions can include an Fc region glycosylated with G0 glycosylation, including, for example, a biantennary core glycan structure of Manα1-6(Manα1-5)Manβ1-4GlcNAcβ1-4GlcNAcβ1 having terminal N-acetylglucosamine at each branch that enhances the capture efficacy of the recombinant antibody in mucus.

[0006]

[0007] Described herein are methods of treating a subject having or at risk of having a respiratory disorder, the method comprising administering by inhalation to the subject a formulation comprising a therapeutic antibody that binds to a respiratory virus in a dosing regimen that includes a once-daily or twice-daily dosing cycle.

[0007]

[0008] Accordingly, described herein is a method of treating a subject having or at risk of having a respiratory disorder, the method comprising administering by inhalation to the subject a formulation comprising a therapeutic human IgG monoclonal antibody (mAb) comprising a population of antibodies glycosylated with a G0 glycosylation pattern comprising at least 40% of a biantennary core glycan structure of Manα1-6(Manα1-3)Manβ1-4GlcNAcβ1-4GlcNAcβ1, the administering step comprising administering at a dose of 0.02 μmol or more of the therapeutic human mAb no more than twice daily such that a concentration higher than 20 ng / mL in the upper respiratory tract (URT) and higher than 100 ng / mL in the lower respiratory tract (LRT) is achieved for the therapeutic human mAb 12 hours or longer after dosing.

[0008]

[0009] In some examples, a method of treating a subject having or at risk of having a respiratory disorder comprises maintaining, for longer than 12 hours after dosing, a concentration of a therapeutic human IgG monoclonal antibody (mAb) that binds to a respiratory virus in the upper respiratory tract (URT) of the subject at higher than 20 ng / ml and in the lower respiratory tract (LRT) of the subject at higher than 100 ng / ml, by inhalation of a dose of a therapeutic human IgG monoclonal antibody (mAb) population that is glycosylated in a G0 glycosylation pattern comprising at least 40% of the branched core glycan structure of Manα1-6(Manα1-3)Manβ1-4GlcNAcβ1-4GlcNAcβ1, wherein the step of administering the dose comprises administering no more than twice a day a therapeutic human mAb of 0.02 μmol or more, and the maintaining step may be included.

[0009]

[0010] In any of these examples, the step of administering may comprise administering the dose no more than once a day.

[0011] In some examples, the therapeutic antibody may comprise at least 45% (e.g., at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, etc.) of the G0 glycosylation pattern.

[0010]

[0012] In some examples, the therapeutic antibody comprises an Fc sequence that is at least X% (e.g., 80%, 85%, 90%, 95%) identical to the sequence of SEQ ID NO: 1 (e.g., human IgG1). For example, the therapeutic antibody comprises an Fc sequence that is at least 85% identical to the sequence of SEQ ID NO: 1, including conservative peptide substitutions.

[0011]

[0013] The therapeutic antibody can be regdanvimab. The dosing regimen can include a twice-daily dosing cycle over a period of 2 to 7 days. The dosing regimen can include a dosing cycle every 2 days, every 3 days, or every 4 days. The dosage regimen can include the step of administering a dose of at least 10 mg of the therapeutic mAb. The dosage regimen can include the step of administering a dose of the therapeutic mAb between about 10 mg and 100 mg. In some examples, the step of administering includes the step of sustaining the release of the therapeutic mAb from the LRT into the blood over multiple days. The step of administering can include the step of sustaining the release of the mAb into the lungs and blood for at least 2 days.

[0012]

[0014] The formulation can also include a pharmaceutically acceptable diluent, excipient, and / or carrier. In some examples, the formulation further includes one or more of citric acid, arginine, mannitol, sorbitol, trehalose.

[0013]

[0015] The therapeutic antibody formulation can be administered to a subject via a nebulizer, such as a vibrating mesh nebulizer. In some examples, the therapeutic antibody formulation is administered via inhalation or via direct instillation into the upper airway. The therapeutic antibody formulation can be self-administered by the subject.

[0014]

[0016] Respiratory disorders can include lower airway disorders. Respiratory disorders can include upper airway disorders. In some examples, respiratory disorders include inflammatory disorders. Respiratory viruses can include coronaviruses. Respiratory viruses can include severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Respiratory viruses can include respiratory syncytial virus (RSV). Respiratory viruses can include one or more of influenza, metapneumovirus, parainfluenza, (certain coronaviruses). In some examples, respiratory viruses include paramyxovirus.

[0015]

[0017] The formulation may include a second or more therapeutic agents in addition to the therapeutic antibody. The formulation may include a therapeutic mAb and a second therapeutic antibody, and the first therapeutic antibody and the second therapeutic antibody bind to the same virus but do not compete for binding to the virus. In some examples, the formulation includes a second therapeutic antibody in addition to the first therapeutic antibody, and further, the first antibody and the second antibody bind to different viruses. The formulation includes a biologic in addition to the therapeutic mAb.

[0016]

[0018] For example, a method of treating a subject having or at risk of having a respiratory disorder comprises administering by inhalation to the subject a dose of a formulation comprising a therapeutic human IgG monoclonal antibody (mAb) that binds to a respiratory virus, and maintaining for more than 12 hours after dosing a concentration of the therapeutic human IgG monoclonal antibody (mAb) that binds to the respiratory virus that is greater than 25 ng / ml in the upper respiratory tract (URT) of the subject and greater than 25 ng / ml in the lower respiratory tract (LRT) of the subject, wherein the administering step comprises administering no more than twice a day a dose of 0.02 μmol or more of the therapeutic human mAb, and the maintaining step may be included.

[0017]

[0019] Also described herein is a method of treating a subject having or at risk of having a respiratory disorder, the method comprising administering by inhalation to the subject a formulation comprising a therapeutic human IgG monoclonal antibody (mAb) that binds to a respiratory virus, wherein the administering step comprises administering no more than once a day a dose of 0.02 μmol or more of the therapeutic human mAb to achieve a concentration of the therapeutic human mAb that is greater than 25 ng / ml in the upper respiratory tract (URT) and greater than 25 ng / ml in the lower respiratory tract (LRT) for more than 24 hours after dosing.

[0018]

[0020] Also described herein is a method of treating a subject having or at risk of having a respiratory disorder, the method comprising administering by inhalation a formulation comprising a therapeutic human IgG monoclonal antibody (mAb) glycosylated with a G0 glycosylation pattern comprising a bisecting core glycan structure of Manα1-6(Manα1-3)Manβ1-4GlcNAcβ1-4GlcNAcβ1, wherein the administering step comprises administering, no more than once a day, a dose of 0.02 μmol or more of the therapeutic human mAb to achieve a concentration of greater than 20 ng / mL in the upper respiratory tract (URT) and greater than 100 ng / mL in the lower respiratory tract (LRT) of the therapeutic human mAb for 24 hours or longer after dosing.

[0019]

[0021] For example, a method of treating a subject having or at risk of having a respiratory disorder may comprise maintaining, for longer than 24 hours after dosing, a concentration of greater than 25 ng / ml of a therapeutic human IgG monoclonal antibody (mAb) that binds to a respiratory virus in the upper respiratory tract (URT) of the subject and greater than 25 ng / ml in the lower respiratory tract (LRT) of the subject, the method comprising administering by inhalation a dose of a formulation comprising a therapeutic human mAb that binds to a respiratory virus, and the administering step comprises administering, no more than once a day, a dose of 0.02 μmol or more of the therapeutic human mAb.

[0020]

[0022] In one example, a method of treating a subject having or at risk of having a respiratory disorder comprises administering by inhalation to the subject a dose of a therapeutic human IgG monoclonal antibody (mAb) glycosylated with a G0 glycosylation pattern comprising a bisected core glycan structure of Manα1-6(Manα1-3)Manβ1-4GlcNAcβ1-4GlcNAcβ1, maintaining for longer than 24 hours after dosing a concentration of the therapeutic human IgG monoclonal antibody (mAb) that binds to a respiratory virus in the upper respiratory tract (URT) of the subject higher than 20 ng / ml and a concentration higher than 100 ng / ml in the lower respiratory tract (LRT) of the subject, and the step of administering the dose comprises administering 0.02 μmol or more of the therapeutic human mAb no more than once a day.

[0021]

[0023] In any of the methods described herein, the therapeutic antibody can be a therapeutic human IgG monoclonal antibody (mAb). In any of the methods described herein, the therapeutic human IgG monoclonal antibody (mAb) is a human IgG1 mAb. In any of the methods described herein, the therapeutic antibody comprises an Fc sequence that is at least X% (e.g., 80%, 85%, 90%, 95%) identical to the sequence of SEQ ID NO: 1 (e.g., human IgG G1). For example, the therapeutic antibody can comprise regdanvimab. Alternatively, in any of these methods and compositions, the Fc sequence can be at least X% identical to one or more of the sequences of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, and / or SEQ ID NO: 4.

[0022]

[0024] Generally, the subject can be any subject in need of therapy. In particular, the subject can be an adult subject and a young adult subject. As used herein, a young adult subject can refer to any individual 12 years of age or older.

[0023]

[0025] In any of the methods described herein, the therapeutic antibody may comprise an oligosaccharide that enhances the capture efficacy of the recombinant antibody in mucus. For example, the therapeutic antibody may comprise a population of mAbs that contain at least 40% oligosaccharides having a G0 glycosylation pattern that includes a bi-branched core glycan structure of Manα1-6(Manα1-5)Manβ1-4GlcNAcβ1-4GlcNAcβ1, with terminal N-acetylglucosamine on each branch that enhances the capture efficacy of the recombinant antibody in mucus.

[0024]

[0026] In any of the methods described herein, the dosing regimen may include a once-daily dosing cycle over a period of 2 to 7 days. The dosing regimen may include a dosing cycle every 2 days, every 3 days, or every 4 days. The dosing regimen may include a step of administering a total of 2, 3, or 4 doses. The dosing regimen may include a step of administering only a single dose. The dosing regimen may include a step of administering a dose of at least 30 mg of the therapeutic mAb. The dosing regimen may include a step of administering a dose of the therapeutic mAb between about 30 mg and 90 mg.

[0025]

[0027] In any of the methods described herein, the administering step may include a step of sustaining the release of the therapeutic mAb from the LRT into the blood over multiple days. The administering step may include a step of sustaining the release of the mAb into the lungs and blood over at least 2 days.

[0026]

[0028] In any of the methods described herein, the formulation further comprises a pharmaceutically acceptable diluent, excipient, and / or carrier. For example, the formulation may further comprise one or more of citric acid, arginine, mannitol, sorbitol, trehalose. The therapeutic antibody formulation may be administered to a subject via a nebulizer. The therapeutic antibody formulation may be administered to a subject via a vibrating mesh nebulizer. The therapeutic antibody formulation may be administered to a subject via a nebulizer. The therapeutic antibody formulation may be administered via inhalation or via direct instillation into the upper airway. The therapeutic antibody formulation may be self-administered by the subject.

[0027]

[0029] Respiratory disorders can include lower respiratory tract disorders. Respiratory disorders can include upper respiratory tract disorders. Respiratory disorders can include inflammatory disorders. Respiratory viruses can include coronaviruses. Respiratory viruses can include severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Respiratory viruses can include respiratory syncytial virus (RSV). Respiratory viruses can include one or more of influenza, metapneumovirus, parainfluenza, (certain coronaviruses). Respiratory viruses can include paramyxoviruses.

[0028]

[0030] In any of the methods described herein, the formulation can include a second or more therapeutic agents in addition to the therapeutic antibody. The formulation may include a therapeutic mAb and a second therapeutic antibody, where the first therapeutic antibody and the second therapeutic antibody bind to the same virus but do not compete for binding to the virus. The formulation includes a second therapeutic antibody in addition to the first therapeutic antibody, and further, the first antibody and the second antibody bind to different viruses. The formulation can include a biologic in addition to the therapeutic mAb.

[0029]

[0031] Also, a composition for use in a method of treating any of the respiratory disorders described herein by practicing any of the described methods (e.g., a therapeutic human IgG monoclonal antibody, particularly a therapeutic human IgG monoclonal antibody (mAb) comprising a population of antibodies glycosylated in a G0 glycosylation pattern comprising at least 40% of a bifurcated core glycan structure of Manα1-6(Manα1-3)Manβ1-4GlcNAcβ1-4GlcNAcβ1 is also described herein. For example, a therapeutic human IgG monoclonal antibody (mAb) comprising a population of antibodies glycosylated in a G0 glycosylation pattern comprising at least 40% of a bifurcated core glycan structure of Manα1-6(Manα1-3)Manβ1-4GlcNAcβ1-4GlcNAcβ1 for use in a method of treating a respiratory disorder by a step of administering the therapeutic human IgG monoclonal antibody (mAb) by inhalation is described herein, and the step of administering comprises administering at a dose of 0.02 μmol or more of the therapeutic human IgG mAb no more than twice a day so as to achieve a concentration higher than 20 ng / mL in the upper respiratory tract (URT) and higher than 100 ng / mL in the lower respiratory tract (LRT) of the subject for 12 hours or longer after dosing.

[0030]

[0032] Also, a therapeutic human IgG monoclonal antibody (mAb) comprising a population of antibodies glycosylated in a G0 glycosylation pattern comprising at least 40% of a bifurcated core glycan structure of Manα1-6(Manα1-3)Manβ1-4GlcNAcβ1-4GlcNAcβ1 for use in a method of treating a respiratory disorder by a step of maintaining, for longer than 12 hours after dosing, a concentration higher than 20 ng / ml in the upper respiratory tract (URT) of the subject and higher than 100 ng / ml in the lower respiratory tract (LRT) of the subject of the therapeutic human IgG mAb by a step of administering the dose of the therapeutic human IgG mAb by inhalation is described herein, and the step of administering the dose comprises administering at a dose of 0.02 μmol or more of the therapeutic human IgG mAb no more than twice a day.

[0031]

[0033] These methods, and in particular the dosing regimens described herein, are surprising and effective: Prior to this study, the predicted clearance of the therapeutic agent (mAb) from the lungs of such compositions was thought to be extremely rapid (e.g., less than 30 minutes), so relatively large and / or more frequent dosing was thought to be required.

[0032]

[0034] All of the methods and apparatuses described herein may be contemplated and used in any combination to achieve the benefits as described herein.

[0035] A better understanding of the features and advantages of the methods and apparatuses described herein is obtained by reference to the following detailed description which illustrates exemplary embodiments and the following accompanying drawings.

Brief Description of the Drawings

[0033]

Figure 1

[0036] Figure 1 is Table 1 depicting the demographics of the patients enrolled in the study described in Example 1, showing the persistence in the upper and lower airways (as would be seen at serum levels) of a therapeutic mAb having a human IgG Fc region that is glycosylated (e.g., more than 40% of the mAb is glycosylated).

Figure 2

[0037] Figure 2 is Table 2 summarizing the adverse events from the study described in Example 1. Side effects marked by (*) occurred within 2 hours of completion of inhaled administration; (cough, decrease in FEV1). Complications marked by (**) included use of contraceptive IUD.

Figure 3

[0038] Figure 3 (left) is a diagram showing an example of the process flow of the method as an example of the procedure described in Example 1. Figure 3 (right) is a diagram showing an example of the overview of the study used in Example 1 and the time points of sample collection.

Figure 4

[0039] Figures 4A to 4C are diagrams illustrating nasal fluid concentrations. Figure 4A shows the concentrations in the single-dose cohort. Figure 4B shows the concentrations in the multiple-doses-per-day cohort (e.g., 7 days of 90 mg). The arrows on the X-axis indicate the 7 administrations of the 90 mg dose in Figure 4B. Figure 4C shows the comparison of nasal concentrations between the single-dose and multiple-doses cohorts. The mean LLOQ for all nasal fluid samples is shown as 450 ng / g, but the LLOQ varied by sample depending on the mass of the nasal fluid collected by swab, and as a result, some detectable samples below the overall mean LLOQ were obtained. The fractions below each time point represent the number of samples that were below the LLOQ at that time.

Figure 5

[0040] Figures 5A to 5B show the serum IN-006 concentrations in the single-dose cohort (Figure 5A) and the multiple-doses cohort (Figure 5B, the last dose administered at 144 hours). The symbols plotted below the dotted line LLOQ of 25 ng / mL represent the number of samples in each group that were BLQ at each time point.

Figure 6

[0041] Figure 6 is a schematic diagram illustrating an example of the method as described herein.

DETAILED DESCRIPTION OF THE INVENTION

[0034]

[0042] Methods, compositions and devices (e.g., devices, systems, etc.) useful for treating a subject having or at risk of having a respiratory disorder are described herein. In some embodiments, provided is a method of administering a therapeutic antibody for treating a subject having or at risk of having a respiratory disorder that affects the upper respiratory tract (upper airway) or the lower respiratory tract (lower airway). The methods provided herein may be particularly useful for treating a subject having or at risk of having a respiratory disorder that affects both the upper respiratory tract (upper airway) and the lower respiratory tract (lower airway). Applicants have surprisingly and unexpectedly found the ability to achieve a long-term scope of application using therapeutic antibodies that allow for rare or sporadic dosing regimens (e.g., single delivery, once daily delivery, up to twice daily delivery) using the methods, compositions and devices described herein.

[0035]

[0043] The term "antibody" (Ab) refers to an immunoglobulin molecule that specifically binds to or immunologically reacts with a particular antigen. The basic antibody has a Y-shape with a stem region and two arm regions and can be classified into different categories called isotypes based on characteristics found in the antibody stem region. The basic antibody is a heterotetrameric glycoprotein composed of two identical light (L) chains and two identical heavy (H) chains. Each of the four chains has, at its amino terminus, a variable (V) region that contributes to the antigen-binding site and a constant (C) region that determines the isotype. Experimentally, an antibody can be cleaved with the proteolytic enzyme papain, which breaks each of the heavy chains and results in three separate subunits. Two of the subunits are composed of a light chain and a fragment of the broken heavy chain of approximately the same mass. Each of these two subunits can bind to an antigen separately and is called a Fab fragment (i.e., "fragment antigen-binding"). According to one estimate, humans have 10 18It may be possible to generate up to 100 quintillion distinct antibodies, each with a unique Fab fragment. The third of the three units is composed of two identical segments of the heavy chain. This third unit is not normally involved in antigen binding but is important in subsequent processes in the body that involve eliminating the antigen from the body. In contrast to the Fab fragment, the third unit from an antibody typically has only one of five physicochemical properties and is thus called the Fc fragment (i.e., the "crystallizable" fragment). The classes of human antibodies that contain one of the five types of Fc fragments are called IgA, IgD, IgE, IgG, and IgM isotypes. These isotypes also have several subclasses. For example, IgG antibodies can be further divided in humans into subclasses IgG1, IgG2, IgG3, and IgG4. IgG antibodies can be further subdivided in mice into subclasses IgG1, IgG2a, IgG2b, and IgG3. Antibody classes and modified forms can be generated by methods known in the art and include polyclonal, monoclonal, genetically engineered, bifunctional, chimeric antibodies, humanized antibodies, heteroconjugate antibodies (e.g., bispecific antibodies, diabodies, triabodies, and tetrabodies), and antigen-binding fragments of antibodies or single-chain antibodies containing, for example, Fab’, F(ab’) 2 2 including Fab, Fv, rIgG, and single-chain antibodies containing scFv fragments (e.g., single-chain Fv) fragments in which the VL domain is linked to the VH domain by a linker.

[0036]

[0044] A "blocking" antibody (also called an "antagonist" antibody) is an antibody that inhibits or reduces the biological activity of the antigen to which it binds. In some embodiments, the blocking or antagonist antibody substantially or completely inhibits the biological activity of the antigen.

[0037]

[0045] A "carrier" is generally designed to interact with an active pharmaceutical ingredient (API) (e.g., an antibody) and enhance its properties. A carrier is generally safe and non-toxic for the subjects and cells to which it is exposed at the dosages and concentrations used. An example of a physiologically acceptable carrier is an aqueous pH buffer solution, such as a physiological saline solution. Examples of physiologically acceptable carriers include buffers such as phosphoric acid, citric acid, and other organic acids; antioxidants including ascorbic acid; polypeptides of low molecular weight (less than about 10 residues); proteins such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrin; chelating agents such as EDTA; salt-forming counterions such as sodium and / or nonionic surfactants such as TWEEN™, polyethylene glycol (PEG), and PLURONICS™.

[0038]

[0046] The term "Cmax" refers to a standard pharmacokinetic measure used to determine drug dosing. Cmax is the peak (highest) concentration, the maximum (or peak) concentration, that a drug achieves in a designated compartment or test area of the body (e.g., blood, serum, nasal cavity, etc.) after the drug has been administered and prior to the administration of the subsequent (second) dose.

[0039]

[0047] The term "effective amount" (or "therapeutically effective amount") is at least the minimum dosage concentration necessary to cause a measurable improvement or prevent a particular disorder. The effective amount herein can vary depending on factors such as the particular disorder (e.g., disease state) of the subject, age, gender and weight, as well as the ability of the agent (e.g., antibody) to induce the desired response in an individual. The effective amount is also such that any toxic or detrimental effects of the treatment are outweighed by the therapeutically beneficial effects. For prophylactic use, beneficial or desired results include eliminating or reducing the risk, including biochemical, histological and / or behavioral symptoms of the disorder (disease), its complications and intermediate pathological phenotypes presented during the onset of the disorder (disease), reducing the severity of the disorder (disease) or delaying the onset of the disorder (disease). For therapeutic use, beneficial or desired results include reducing one or more symptoms resulting from the disorder (disease), enhancing the quality of life of a person suffering from the disorder (disease), reducing the dosage of other medications required to treat the disorder (disease), targeting and delaying the progression of the disease, enhancing the effect of another dosage regimen and / or extending survival, etc. Clinical outcomes such as these are included. The effective amount can be administered in one or more administrations. For the purposes herein, an effective amount of a drug, compound or pharmaceutical composition is an amount sufficient to effect a prophylactic or therapeutic treatment, either directly or indirectly. As understood in the clinical context, an effective amount of a drug, compound or pharmaceutical composition may or may not be achieved in combination with another drug, compound or pharmaceutical composition. Thus, an "effective amount" can be considered in the context of the administration of one or more therapeutic agents, where there is a possibility that the desired result is achieved in combination with one or more other agents, or where a single agent is considered to be administered in an effective amount.

[0040]

[0048] The term "excipient" refers to substances in a formulation other than the active ingredient(s) (e.g., antibody). Examples of excipients include antioxidants, buffers, emulsifiers, penetration enhancers, preservatives, release control reagents and viscosity modifiers.

[0041]

[0049] The term "humanized antibody" or "humanized" form of a non-human (e.g., mouse) antibody refers to a chimeric antibody containing minimal sequences derived from a non-human immunoglobulin. In some embodiments, a humanized antibody is a human immunoglobulin (recipient antibody) in which residues from the recipient's hypervariable regions have been replaced by residues from the hypervariable regions of a non-human species, such as a mouse, rat, rabbit, or non-human primate (donor antibody) having the desired specificity, affinity, and / or capacity. A humanized antibody may also typically include at least a portion of the Fc that is of a human immunoglobulin consensus sequence. Methods for antibody humanization are known in the art. In some examples, framework ("FR") residues of the human immunoglobulin are replaced by the corresponding non-human residues. Additionally, a humanized antibody may contain residues not found in the recipient antibody or in the donor antibody.

[0042]

[0050] The term "ka" (M -1 sec -1 ) is intended to refer to the association rate constant of a particular antibody-antigen interaction. The term "K A " (M) as used herein is intended to refer to the binding equilibrium constant of a particular antibody-antigen interaction.

[0043]

[0051] The term "kd" (sec -1 ) as used herein is intended to refer to the dissociation rate constant of a particular antibody-antigen interaction. This value is also referred to as the off-rate. The term "K D " (M -1 ) as used herein is intended to refer to the dissociation equilibrium constant of a particular antibody-antigen interaction.

[0044]

[0052] In certain embodiments, the antibodies of the present disclosure are monoclonal antibodies. The term "monoclonal antibody" as used herein includes, but is not limited to, antibodies produced by hybridoma technology. The term "monoclonal antibody" refers to an antibody derived from a single clone, including any eukaryotic, prokaryotic or phage clone, regardless of the method by which it is produced. Monoclonal antibodies useful in conjunction with the present disclosure can be prepared using a variety of techniques known in the art, including the use of hybridoma, recombinant and phage display technologies, or combinations thereof. The antibodies of the present disclosure include chimeric, primatized, humanized or human antibodies.

[0045]

[0053] The term "nebulizer" refers to a device configured to transform a medicament (formulation) from a liquid into an aerosol or a suspension of fine particles or droplets (also called a mist herein) and deliver the aerosol to a subject for inhalation into the lungs. Nebulizer devices include jet nebulizers, mesh nebulizers, and ultrasonic nebulizers. Nebulizers can also be heatable or refillable. A jet nebulizer (also sometimes called a compressor, nozzle, using compressed air, or a venturi nebulizer) uses compressed gas (e.g., air or oxygen) to form an aerosol. For example, a nebulizer reservoir can be filled with a medicament (formulation). The compressed gas is applied to the inlet of the reservoir, moves at high speed, exits through a narrow opening, and can create a low-pressure region at the outlet. The resulting pressure difference causes the fluid to be drawn up from the reservoir into and out of the reservoir. The fluid can then be broken into droplets of various sizes by the walls of the nebulizer or internal baffles. An ultrasonic nebulizer uses high-frequency vibrations such as 2 to 3 million times per second from a piezoelectric oscillator. The vibrations are transferred to the medicament (formulation) via a cooling water tank to form an aerosol. A mesh nebulizer uses an extremely fine mesh to form a mist. The vibrating element pushes the medicament (formulation) through extremely fine holes in a membrane (e.g., a mesh). This results in an aerosol of small droplets. The phrase "pharmaceutically acceptable" indicates that a substance or composition must be chemically and / or toxicologically compatible with the other components of the formulation, including the mammal being treated therewith.

[0046]

[0054] The term "peak level" refers to the highest concentration of a therapeutic agent (e.g., an antibody) in an individual body.

[0055] The term "pharmaceutically acceptable salt" refers to pharmaceutically acceptable organic or inorganic salts of the compounds of the present invention. Exemplary salts include, but are not limited to, acetate, bisulfate, bromide, chloride, citrate, iodide, nitrate, oleate, oxalate, pantothenate, sulfate, phosphate, acid phosphate, isonicotinate, lactate, salicylate, acid citrate, tartrate, tannate, bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucuronate, saccharate, formate, benzoate, glutamate, methanesulfonate ("mesylate"), ethanesulfonate, benzenesulfonate, p-toluenesulfonate, pamoate (i.e., 1,1'-methylene-bis-(2-hydroxy-3-naphthoate)) salts, alkali metal (e.g., sodium and potassium) salts, alkaline earth metal (e.g., magnesium) salts, and ammonium salts. Pharmaceutically acceptable salts can include those containing another molecule such as an acetate ion, succinate ion, or other counterion. The counterion can be any organic or inorganic moiety that stabilizes the charge of the parent compound. Further, pharmaceutically acceptable salts can have more than one charged atom in their structure. An example of a pharmaceutically acceptable salt that is part of a plurality of charged atoms can have a plurality of counterions. Thus, pharmaceutically acceptable salts can have one or more charged atoms and / or one or more counterions.

[0047]

[0056] The term "specific binding" of an antibody refers to antibody binding to a given antigen. Typically, an antibody binds with an affinity corresponding to a K of about 10 -8 M or less and binds to a given antigen with an affinity that is at least one-tenth, preferably at least one-hundredth, of its affinity for binding to a non-specific antigen (e.g., BSA, casein) other than the given antigen or an antigen closely related thereto (such as represented by K D ). Alternatively, an antibody binds to a given antigen with an affinity of about 10 D M or about 10 6 M -1 or about 10 7 M -1 or about 10 8 M-1 or 10 9 M -1 It can bind with an affinity corresponding to KA of 10 or higher, and binds to a predetermined antigen with an affinity that is at least 10-fold higher, preferably at least 100-fold higher, than its affinity for binding to non-specific antigens other than the predetermined antigen or an antigen closely related thereto (e.g., BSA, casein).

[0048]

[0057] The term "treatment" refers to a clinical intervention designed to alter the natural course of an individual or cell being treated during a clinical pathological process or to prevent the occurrence of a clinical pathological process. Desirable effects of treatment include reducing the rate of disease progression, restoring or alleviating a disease state, and improving remission or prognosis. For example, an individual is successfully "treated" when one or more symptoms associated with a respiratory disorder, such as pain, bronchitis, chill, confusion, cough, death, diarrhea, dyspnea, fatigue, fever, headache, inflammation, pale / grey / blue skin / lips / nail beds, pneumonia, rhinorrhea (congestion of the nasal cavity), shortness of breath, sneezing, pharyngalgia, vomiting, and weakness, are restored, reduced, eliminated, or prevented.

[0049]

[0058] The term "trough level" refers to the lowest concentration of a therapeutic agent in an individual's body at which the therapeutic agent is within the therapeutic range or the concentration of the therapeutic agent before administration of a further dose of the therapeutic agent.

[0059] The term "variable region" or "variable domain" of an antibody refers to the amino-terminal domain of the heavy or light chain of the antibody. The variable domains of the heavy and light chains may also be referred to as "VH" and "VL", respectively. These domains are generally the most variable parts of the antibody (relative to other antibodies of the same class) and contain the antigen-binding site. Therapeutic antibodies can be administered to the upper respiratory tract (also called the upper airway) and / or the lower respiratory tract (also called the lower airway). In some embodiments, the antibody is administered to both the upper and lower airways. The upper airway includes the nose and nasal cavity, paranasal sinuses, oral cavity, pharynx, and the part of the larynx above the vocal cords, while the lower airway is further divided into the conducting zone and the respiratory zone. The conducting zone consists of the part of the larynx below the vocal cords, the trachea, and the bronchi and bronchioles within the lungs. The respiratory zone is formed by the respiratory bronchioles, alveolar ducts, and alveoli. Therapeutic antibodies can be administered to the upper and / or lower airways by a dry powder inhaler (DPI), injection, metered-dose inhaler, nasal spray, or nebulizer. A nebulizer is a drug delivery device that converts a liquid drug, such as the antibody compositions described herein, into fine droplets (aerosol or mist) that are inhaled into the lungs of a subject via a face mask or mouthpiece. Nebulizers include jet nebulizers, ultrasonic nebulizers, and mesh nebulizers. Examples of nebulizers that can be used to administer therapeutic antibodies include the Acorn and Acorn II (registered trademark) nebulizers (Vital Signs), AERx nebulizer (Aradigm), AeroDose nebulizer (AeroGen Inc., Mountain View, CA), Respimat nebulizer (Boehringer, Germany), and UltraVent (trademark) nebulizer (Mallinckrodt). In some embodiments, the nebulizer used to perform the methods herein is a non-jet nebulizer and / or a non-ultrasonic nebulizer.In some embodiments, the nebulizer used to carry out the methods herein is a mesh nebulizer. Mesh nebulizers may be gentler and less disruptive to the antibody structure. Antibody potency is highly dependent on its higher order structure or conformation. Antibodies are proteins that undergo multiple stages of complex protein folding during formation to generate their complex higher order structure. These stages are primary, secondary, tertiary and quaternary. The primary stage is the sequence of amino acids held together by peptide bonds. The secondary stage is the protein beginning to fold (e.g., to form an alpha helix or a beta pleated sheet). Hydrogen bonds are formed between amino acids. The tertiary stage is the antibody tertiary structure when the protein folds into its 3D structure associated with its function. The quaternary structure is held together by various non-covalent interactions between side chains including ionic interactions, disulfide bridge formation, hydrophobic interactions, van der Waals forces and hydrogen bonds. The quaternary stage is when a single peptide binds to another peptide, e.g., when the heavy and light chains bind together. Antibodies may be sensitive to degradation by a number of physical and chemical stresses, such as freezing, heating, agitation, oxidation and pH change. Any of the compositions herein may contain a pharmaceutically acceptable diluent, excipient or carrier.

[0050]

[0060] The nebulizers described herein can be configured to produce a predetermined range of particle sizes. The particle size range can be within a predetermined range for deposition within both the lungs and nasal cavity using the methods described herein. Particles outside the desired range may not be delivered into the nasal cavity at the desired distribution pattern or level. For example, operating the nebulizer to continuously form particles containing a drug can include forming particles with an average particle or droplet size (commonly defined as the aerodynamic median particle diameter, MMAD) in the range of about 0.1 to about 200 microns (e.g., between about 1 and 10 microns, between about 2 and 7 microns, between about 2 and 20 microns, between about 10 and 40 microns, between about 20 and 60 microns, between about 30 and 70 microns, between about 40 and 80 microns, between about 50 and 90 microns, between about 60 and 100 microns, between about 70 and 110 microns, between about 80 and 120 microns, between about 90 and 130 microns, between about 100 and 150 microns, between about 125 and 200 microns, etc.). For example, operating the nebulizer to continuously form particles containing a drug can include forming particles with an average particle or droplet size in the range of about 2 to 7 microns. In some examples, the methods described herein can be used with two distributions of particle sizes that include smaller and larger particle sizes.

[0051]

[0061] In some variations, inhaled respiratory medications can be administered using a device called a metered-dose inhaler or MDI. An MDI is a pressurized canister of medication in a plastic holder with a mouthpiece. When sprayed, it can deliver a reliable, consistent dose of medication.

[0052]

[0062] The present disclosure provides a therapeutic regimen that includes administering one or more therapeutic agents, including one or more antibodies, to a subject having or at risk of having a disorder (respiratory disorder). The dosing regimen for administration (e.g., the therapeutic regimen) may vary depending on the age and size of the subject to be administered, the target disease, the antibody particles, the condition / health / disease state, and the route of administration. The dosing regimen may be more than once a day, but generally will be once a day or twice a day. In some variations, the dosing regimen may include more frequent therapeutic agent administrations, such as three times a day, four times a day, etc. In some embodiments, the dosing regimen is administered once a day for only one day (i.e., only one dose). In some embodiments, the dosing regimen can continue for one day to an indefinite period. In some embodiments, the dosing regimen continues for two days, three days, four days, five days, six days, seven days, etc. or longer. In some embodiments, the dosing regimen may have regular dosing intervals, such as daily, every two days, every three days, weekly, every two weeks, monthly, etc., or intervals therebetween. In some embodiments, the dosing regimen is a non-variable dose regimen (e.g., each dose is the same amount) or a variable dose regimen (different doses are different amounts, e.g., more antibody in the first dose and less, e.g., half, one-third, one-fourth, etc. in subsequent doses). A once-daily delivery regimen is convenient and can facilitate successful compliance with the regimen. A delivery regimen more frequent than once a day may not be as convenient, but there may be advantages to more frequent delivery. For example, for some therapeutic agents, such as expensive monoclonal antibodies, the total amount of the therapeutic agent delivered in two doses may be less than the amount of the therapeutic agent that would be delivered in a single once-daily dose, and a delivery regimen of two (or optionally, more) doses per day may result in lower costs. As illustrated in Example 4, either a single high dose once a day or substantially lower doses twice a day can be administered to minimize the trough level from becoming too low and losing efficacy. Each of these two doses may be quite low, compensating for the inconvenience of administering the dose twice a day.Accordingly, the step of providing once-daily versus twice-daily (or three times a day, etc.) dosing can balance the convenience and efficient use of the antibody, for example, according to antibody manufacturing costs, accessibility to delivery options (e.g., self-administration, availability of medical professionals for therapeutic drug administration, use of medical facilities for therapeutic drug administration, etc.).

[0053]

[0063] Specific antibodies and their EC50s Various pharmaceutical agents can benefit from the nebulized delivery methods and devices described herein. In particular, these methods and devices can benefit from pharmaceutical agents for treating respiratory disorders that affect at least one of the upper and lower airways, but can typically provide benefit (treatment) to both the upper and lower airways. In some embodiments, the methods and devices can be useful for reducing symptoms in the upper airway (e.g., from respiratory infections) and, typically, for treating the lower airway, which is more associated with hospitalization and other severe adverse outcomes. These methods and devices can be particularly effective in the delivery of pharmaceutical agents configured as mucosal binders and / or capture agents. For example, methods and devices as described herein can be particularly useful and / or effective when the pharmaceutical agent is a recombinant antibody containing an oligosaccharide having a G0 glycosylation pattern comprising a bisecting core glycan structure of Manα1-6(Manα1-3)Manβ1-4GlcNAcβ1-4GlcNAcβ1 having a terminal N-acetylglucosamine that enhances the capture efficacy of the recombinant antibody in mucus at each branch. In some examples, the pharmaceutical agent comprises a recombinant antibody comprising a human or humanized Fc region, and the recombinant antibody comprises a population of antibodies in which at least 20% (e.g., 20% or more, 25% or more, 30% or more, 35% or more, 40% or more, etc.) contains an oligosaccharide having a G0 glycosylation pattern comprising a bisecting core glycan structure of Manα1-6(Manα1-3)Manβ1-4GlcNAcβ1-4GlcNAcβ1 having a terminal N-acetylglucosamine that enhances the capture efficacy of the recombinant antibody in mucus at each branch.

[0054]

[0064] The methods and devices described herein can be used to treat a subject having or at risk of having a respiratory disorder by administering one or more therapeutic antibodies. Examples of antibodies that can be used with the methods and devices herein include antibodies or antigen-binding fragments thereof that can specifically bind to human surface antigen classification 39 (CD39), such as anti-surface antigen classification 39 (CD39) antibodies as described in US20210388105A1. The antibody or antigen-binding fragment thereof can specifically bind to human CD39 at an EC50 of ≧10 -8 M as measured by a fluorescence-activated cell sorting (FACS) assay. CD39 is involved in the pathogenesis of cigarette smoke-induced lung inflammation in patients and preclinical mouse models.

[0055]

[0065] Another antibody that can be used with the methods and devices herein is an anti-influenza B antibody as disclosed in US20210171612A1 (Regeneron Pharmaceuticals Inc., Tarrytown, NY). The anti-influenza B antibody can be an IgG1 or IgG4 antibody that confers increased protection from influenza B virus in animals (e.g., mammals) when administered either subcutaneously or intravenously and / or before or after infection with influenza B virus, and can reduce symptoms of headache, fever, pain, nasal congestion, chills, fatigue, weakness, sore throat, cough, shortness of breath, vomiting, diarrhea, pneumonia, bronchitis, and / or death. The anti-influenza B antibody binds to influenza B HA and has an EC50 of less than about 10 -9 M.

[0056]

[0066] Another antibody that can be used with the methods and devices described herein is an anti-PcrV antibody as disclosed in US20200392210A1 (Regeneron Pharmaceuticals Inc., Tarrytown, NY). The anti-PcrV antibody binds to the V-tip protein (PcrV) of Pseudomonas aeruginosa and can inhibit or neutralize the activity of the bacterial type III secretion system (T3SS) in Pseudomonas aeruginosa. The antibody is thought to be useful for blocking the translocation of toxins from bacteria to host cells and / or preventing host cell death. The anti-PcrV antibody may function by blocking pore-mediated membrane permeability in host cells. The anti-PcrV antibody can bind to full-length PcrV and has an EC50 of less than about 10 -8 μM. As disclosed in US20200392210A1, patients at greater risk of Pseudomonas aeruginosa infection may be patients with cystic fibrosis, with diabetes, on a ventilator, having surgery, having tuberculosis, having HIV, having a compromised immune system, having neutropenia, having an indwelling catheter, after physical trauma, having burns, being in the intensive care unit, being bedridden, having a malignant tumor, having chronic obstructive pulmonary disease, being in a long-term care facility, or being an intravenous drug user.

[0057]

[0067] As another antibody that can be used with the methods and devices herein, there is an anti-PD1 antibody as disclosed in US10981994B2 (Apollomics Inc., Foster City, CA). The anti-PD-1 antibody can be a humanized antibody having a PD-1 binding EC50 of about 200 ng / ml or less or about 150 ng / mL or less or about 100 ng / mL or less or about 80 ng / ml or less or about 60 ng / mL or less as measured by ELISA or FACS. The provided anti-PD-1 antibody and its fragments bind to PD-1 on T cells, disrupt the PD-1 / PD-L1 interaction, and as a result, increase T cell activation. US10981994B2 discloses that IgG1 and IgG4 versions of the humanized 7A4 and 13F1 antibodies were generated. The anti-PD-1 antibody can be useful for treating infectious diseases including respiratory diseases such as candidiasis, candidemia, aspergillosis, streptococcal pneumonia, streptococcal skin and pharyngeal conditions, gram-negative sepsis, tuberculosis, mononucleosis, influenza, respiratory syncytial virus-induced respiratory disease, malaria, schistosomiasis, and trypanosomiasis. As another antibody that can be used with the methods and devices herein, there is bamlanivimab / etesevimab (made by Eli Lilly). Bamlanivimab (LY-CoV555, also known as alias LY3819253), a monoclonal antibody of Eli Lilly, was originally derived from the blood of one of the first US patients to recover from COVID-19. It is a recombinant neutralizing monoclonal antibody against the SARS-CoV-2 spike protein. Eli Lilly's etesevimab (LY-CoV016, alias JS016, alias LY3832479) is a monoclonal antibody against the receptor binding domain of the SARS-CoV-2 surface spike protein. As another antibody that can be used with the methods and devices herein, there is bebtelovimab. The monoclonal antibody bebtelovimab (Eli Lilly, Indianapolis, IN) is used for the treatment of mild to moderate COVID-19.Vepolizumab binds to the SARS-CoV-2 spike protein. Vepolizumab was administered as a single 175 mg intravenous injection over at least 30 seconds. Vepolizumab is a human immunoglobulin G-1 (IgG1 variant) monoclonal antibody having two identical light chain polypeptides each consisting of 215 amino acids and two identical heavy chain polypeptides consisting of 449 amino acids. It is produced by Chinese hamster ovary (CHO) stable bulk cultures or cell lines and has a molecular weight of 144 kDa. Vepolizumab is a recombinant neutralizing human IgG1λ monoclonal antibody (mAb) against the spike protein of SARS-CoV-2 and is not modified in the Fc region. Vepolizumab binds to the spike protein, has a dissociation constant KD = 0.046 - 0.075 nM, blocks spike protein binding to the human ACE2 receptor, and has been reported to have an IC50 value of 0.39 nM (0.056 mcg / mL). Another antibody that can be used with the methods and devices herein is casirivimab / imdevimab (produced by Regeneron, trade name REGEN-COV). Regeneron's REGEN-COV (also still known as REGN-CoV2 or REGEN-CoV2) is two antibodies that bind to different regions of the SARS-CoV-2 spike protein receptor binding domain: casirivimab (REGN10933) and imdevimab (REGN10987). Another antibody that can be used with the methods and devices herein is regdanvimab or CT-P59 (Celltrion). Regdanvimab is a recombinant human monoclonal antibody targeted against the receptor binding domain (RBD) of the spike protein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). It is a recombinant monoclonal antibody expressed in CHO-K1 cells. Dose-dependent binding to RBD was found for the CT-P59 clinical batch, as shown by the fact that the half-maximal effective concentration (EC50) of CT-P59 against the SARS-CoV-2 RBD protein was 4.4 ng / ml.

[0058]

[0068] Another antibody that can be used with the methods and devices herein is sotrovimab (made by Vir Biotechnology / GSK). Sotrovimab (previously VIR-7831) has been reported to bind to a highly conserved epitope of the receptor-binding domain of the SARS-CoV-2 virus spike protein. Another antibody that can be used with the methods and devices herein is tixagevimab / cilgavimab (made by AstraZeneca, AZD7442, trade name Evusheld™). Evusheld™ was urgently authorized as pre-exposure prophylaxis against COVID-19 in immunocompromised individuals or those who cannot be vaccinated or initiate an immune response after vaccination. AZD7442 contains two monoclonal antibodies, tixagevimab (AZD8895) and cilgavimab (AZD1061), that target the receptor-binding domain of the SARS-CoV-2 spike protein.

[0059]

[0069] The methods and devices described herein may be particularly useful for the treatment of respiratory disorders. In some embodiments, the present disclosure provides a method of treating (improving, alleviating or reducing) the severity, duration or frequency of occurrence of at least one symptom of the disorder, or preventing all of the disorder. Symptoms that can be treated or prevented by the methods of the present disclosure can be one or more of headache, fever, pain, nasal discharge (congestion of the nasal cavity), chills, fatigue, weakness, sore throat, cough, shortness of breath, vomiting, diarrhea, pneumonia, bronchitis, inflammation and death. Inflammation and other symptoms can be acute or chronic. The causative agents of the disorder can include one or more of disease, environmental factors, genetic factors, illness, infection, pathogens, toxins and / or trauma. Pathogens can include archaea, eubacteria, fungi, protists and / or viruses. In some embodiments, the pathogen is a respiratory pathogen, such as bacteria (Haemophilus (Haemophilus influenzae, Haemophilus influenzae (type B), etc.), Moraxella (Morazella catarrhalis), Pseudomonas (Pseudomonas aeruginosa), Staphylococcus (Staphylococcus aureus), Streptococcus (Streptococcus pneumoniae; Streptococcus pyogenes)), fungi (e.g., Aspergillus, Blastomyces, Candida, Cryptococcus, Histoplasma, mold, yeast, zygomycetes) or viruses (adenovirus, coronavirus, influenza virus, metapneumovirus, Middle East respiratory syndrome coronavirus (MERS-CoV), parainfluenza virus, respiratory syncytial virus, severe acute respiratory syndrome coronavirus (SARS-CoV), rhinovirus, such as SARS-CoV-2).Other respiratory disorders that can be treated using the methods and devices described herein include asthma, rhinitis, pulmonary fibrosis, cystic fibrosis, and chronic obstructive pulmonary disease.

[0060]

[0070] For the treatment of respiratory disorders, one (or more) additional therapeutic agents can be used in combination with the antibodies described herein as appropriate. The additional therapeutic agent can be a short-acting beta-agonist, such as a catecholamine or non-catecholamine agent. By way of example, albuterol (ProAir HFA, Proventil HFA, Ventolin HFA), bitolterol, carbuterol, clenbuterol, epinephrine (Asthmanefrin, Primatene Mist), levalbuterol (Xopenex HFA), metaproterenol (Alupent), pirbuterol (Maxair), procaterol, terbutaline (Brethine), or other bronchodilators, among others, can be used. The additional therapeutic agent can be an anticholinergic agent, such as ipratropium (Atrovent), or other mucolytic agents. The additional therapeutic agent can be a corticosteroid, such as methylprednisolone and prednisone, or other swelling-reducing agents. For the treatment of respiratory disorders, anti-inflammatory agents can be used in combination with the antibodies of the present disclosure as appropriate. Anti-inflammatory agents include, but are not limited to, acetaminophen, aspirin, dexamethasone, diphenhydramine, meperidine, mesalazine, Asacol, codeine phosphate, benorylate, fenbufen, naprosyn, diclofenac, etodolac, indomethacin, and ibuprofen.

[0061]

[0071] In the example shown by the schematic diagram in FIG. 3, method 100 may include 101 instructing and / or guiding the patient to administer the inhaled dose, first (optionally) by sitting up straightening the back. The patient can then operate the nebulizer 103 to continuously provide the nebulized drug agent. For example, the method may include guiding the patient to press the on / off button on the nebulizer to start the treatment (e.g., in some examples, the button turns green and mist begins to appear at the mouthpiece and / or back of the nebulizer). The method can then guide the patient to hold the mouthpiece of the nebulizer between the lips, including holding the mouthpiece with the teeth and / or lips and closing the lips around the mouthpiece 105.

[0062]

[0072] Next, the method may include activating (e.g., starting) a first indicator to coach or guide a patient in the inhalation of an aerosolized pharmaceutical agent through the mouth 107. The first indicator can be, for example, light (LED(s)), sound, a message, a countdown, etc., that remains on while the patient inhales to guide the patient to inhale deeper and draw in the agent aerosolized through the mouth. The indicator can be a count (e.g., a count up or down). The indicator can be automatically activated, including based on the presence or absence of input from the patient by a controller. In some examples, the patient can manually initiate the start (activation) of the first indicator. Alternatively, in some examples, the first indicator can be activated upon sensing that the patient has initiated an inhalation through the mouth (e.g., in a nebulizer and / or in a dose guide device). The first indicator can remain on, for example, for an inhalation period of 4 seconds or more (e.g., 4 seconds, 4.5 seconds, 5 seconds, 6 seconds, 7 seconds, etc.). The inhalation period can be fixed or set (e.g., by a user, such as a physician, nurse, pharmacist, and / or patient), or can be variable. In some examples, the inhalation period can change to indicate that a minimum inhalation period (e.g., 4 seconds, 4.5 seconds, 5 seconds, etc.) has been reached but that continued inhalation is recommended. For example, the first indicator may be in an active state using a first sound, color, etc. for a 4-second minimum inhalation period and may remain on for another 2 - 3 seconds, but may change to a first optional / continuous indicator using a second sound, color, etc. For example, the nebulizer and / or dose guide device can change from red to yellow, or some other change, to indicate that inhalation can optionally continue.

[0063]

[0073] The first instruction (and / or the first optional / continuation indicator) can be automatically turned off, for example, after the patient has finished inhaling via the nebulizer and / or has started exhaling. The method and apparatus can include sensing inhalation and / or exhalation. For example, the nebulizer and / or dose guidance device can include one or more sensors for detecting or estimating the start / stop of inhalation and / or exhalation. For example, the nebulizer can include one or more sensors for detecting flow or pressure at the mouthpiece. A flow sensor can be used to determine the start and / or stop of inhalation through the mouthpiece. Any of these methods and apparatuses can include a controller (including one or more processors) capable of implementing these methods, including activating the first indicator, the second indicator, and the like. The controller can analyze sensor data for activating the first and / or second indicators.

[0064]

[0074] Generally, the methods described herein can include instructing or guiding the patient to breathe in such a way that each breath is slow and long, until the lungs are filled as much as possible (e.g., breathe as deeply as possible). Each breath within the body should last at least 4 seconds or more as described.

[0065]

[0075] A second indicator that guides the patient for a rapid (e.g., less than 3 seconds) exhalation can be automatically activated as described above (e.g., upon cessation of inhalation) or based on a pre-set and / or adjustable timer. Generally, the method can include turning off the first indicator and / or operating the second indicator to guide exhalation111. Since the exhalation phase is intended to be rapid and short in duration, the second indicator can include a "stop" indicator that warns the user to stop after a second (exhalation) period of less than 3 seconds (e.g., 2 seconds). For example, in some cases, the second indicator can include a first phase (2 - 3 seconds) from the start of exhalation to the end of the exhalation phase113, after which the second indicator can change, for example, by a change in volume, sound, intensity, color, persistence, etc. (e.g., emitting a flash) to emphasize that exhalation must be completed. The second indicator can then be turned off or otherwise may stop115.

[0066]

[0076] Accordingly, during inhalation, the patient can be instructed and / or guided to exhale through the nose and attempt to complete exhalation within about 3 seconds (within about 2 seconds, within about 2 - 3 seconds, etc.). As discussed herein, this can direct the nebulized pharmaceutical agent (e.g., mist) from the patient's lungs into the nose in a desired distribution, where it can be captured and treatment can be administered to this area.

[0067]

[0077] After completing a long inhalation / rapid exhalation, the patient may be instructed to rest, e.g., breathe normally without using the nebulizer for one or more breaths, or perform another cycle of long inhalation / rapid exhalation 117. For example, the patient may need to rest or may have a cough or an urge to cough. The patient may press the on / off button to stop the nebulizer. The treatment can be continued by pressing the on / off button on the nebulizer and / or the dose guide device again to initiate inhalation through the mouthpiece and exhalation through the nose (iterative steps 107 - 117 in FIG. 6). The patient can take as much rest as needed.

[0068]

[0078] The treatment can be continued until a desired (e.g., pre - set, user - set, etc.) dose is delivered. In some examples, the nebulizer and / or the dose guide device can continue a treatment that includes multiple cycles of long inhalation / rapid exhalation until it indicates that a complete treatment dose has been delivered. For example, the treatment can be continued until the nebulizer emits a warning (e.g., a beep and / or a flash of light) indicating that the treatment is complete. The device can automatically turn off. Any of the methods (including user interfaces) described herein can be implemented as software, hardware, or firmware, and may be described as a non - transitory computer - readable storage medium (e.g., a computer, a tablet, a smartphone, etc.) that stores a set of instructions executable by a processor, which, when executed by the processor, controls the processor to perform any of the steps including, but not limited to, displaying, exchanging information with the user, analyzing, modifying parameters (including timing, frequency, intensity, etc.), making decisions, warning, etc.

[0069]

[0079] As described above, the device can be configured to implement any of the methods described herein. For example, the device can be configured as a nebulizer integrated with (or forming) a dosage guidance device. The nebulizer can be configured to emit a first indicator, such as a sound (e.g., a beep sound, etc.) or the illumination of one or more LEDs (e.g., a countdown of LEDs), and a second indicator, such as a second sound or the illumination of a different color or set of LEDs, etc. As described above, the nebulizer includes one or more sensors for detecting and initiating inhalation and / or exhalation, and the device counts down and enables guiding the user as described herein during inhalation and exhalation.

[0070]

[0080] In some examples, a separate dosage guidance device can be used with the nebulizer. For example, the dosage guidance device can be software. In some examples, the software can be executed on a wearable or handheld computer device, such as a smartphone processor.

[0071]

[0081] These methods and devices can be used with any type of nebulizer. For example, these devices can be used with jet nebulizers that use compressed gas to create an aerosol, ultrasonic nebulizers that form an aerosol by high-frequency vibration, and / or mesh nebulizers in which a liquid passes through a very fine mesh to form an aerosol. In particular, these methods can be used with continuous nebulizers that continuously form particles when turned on. Alternatively, these methods can be used with on-demand nebulizers.

[0072]

[0082] Generally, the methods and apparatuses described herein can be adapted to aerosol particles of a specified or pre-determined size or size distribution. For example, the particles of the drug agent (MMAD) can be in the range of about 0.1 to about 200 microns (e.g., between about 1 and 10 microns, between about 2 and 7 microns, between about 2 and 20 microns, between about 10 and 40 microns, between about 20 and 60 microns, between about 30 and 70 microns, between about 40 and 80 microns, between about 50 and 90 microns, between about 60 and 100 microns, between about 70 and 110 microns, between about 80 and 120 microns, between about 90 and 130 microns, between about 100 and 150 microns, between about 125 and 200 microns, etc.). For example, the particles containing the drug agent can have a size of particles or droplets in the range of about 2 to 7 microns. In some examples, the methods described herein can be used with two particle size distributions including smaller and larger particle sizes.

[0073]

[0083] Any suitable drug agent can be used, including but not limited to drug agents that are mucosal capture drug agents and / or immunotherapy drugs. Generally, these drug agents can be drug agents for treating respiratory disorders / diseases, including those respiratory disorders / diseases caused by respiratory infections.

[0074]

[0084] In particular, the pharmaceutical agents described herein may include pharmaceutical agents that are trapped within mucus, as described, for example, in each of US 10,829,543, US 10,100,102, US 10,793,623, US Patent Application No. 16 / 982,682 (entitled "COMPOSITIONS AND METHODS FOR INHIBITING PATHOGEN INFECTION", filed on March 20, 2019), US Patent Application No. 17 / 063,122 (entitled "OPTIMIZED CROSSLINKERS FOR TRAPPING A TARGET ON A SUBSTRATE", filed on October 5, 2020), and US Patent Application No. 17 / 278,217 (entitled "SYNTHETIC BINDING AGENTS FOR LIMITING PERMEATION THROUGH MUCUS", filed on September 23, 2019), each of which is hereby incorporated by reference in its entirety.

[0075]

[0085] For example, the methods described herein may be particularly useful for delivering a dose of a pharmaceutical agent configured to have an enhanced mucus trapping efficacy, including, but not limited to, a protein (e.g., an antibody) having one or more glycosylation patterns that enhance trapping in mucus. In some examples, the pharmaceutical agent may be a recombinant antibody comprising an oligosaccharide having a G0 glycosylation pattern comprising a bisecting core glycan structure of Manα1-6(Manα1-3)Manβ1-4GlcNAcβ1-4GlcNAcβ1 having a terminal N-acetylglucosamine at each branch that enhances the trapping efficacy of the recombinant antibody in mucus. For example, the pharmaceutical agent may be a recombinant antibody comprising a human or humanized Fc region, wherein at least 40% of the antibody population comprises an oligosaccharide having a G0 glycosylation pattern comprising a bisecting core glycan structure of Manα1-6(Manα1-3)Manβ1-4GlcNAcβ1-4GlcNAcβ1 having a terminal N-acetylglucosamine at each branch that enhances the trapping efficacy of the recombinant antibody in mucus.

[0076]

[0086] The techniques and procedures described or referenced herein are, for example, Sambrook et al., Molecular Cloning: A Laboratory Manual 3d edition (2001) Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.; Current Protocols in Molecular Biology (F.M. Ausubel et al. (eds.), (2003)); the series Methods in Enzymology (Academic Press, Inc.): PCR 2: A Practical Approach (M.J. MacPherson, B.D. Hames and G.R. Taylor (eds.) (1995)), Harlow and Lane (eds.) (1988) Antibodies, A Laboratory Manual, and Animal Cell Culture (R.I. Freshney (ed.) (1987)); Oligonucleotide Synthesis (M.J. Gait et al. (eds.) 1984); Methods in Molecular Biology, Humana Press; Cell Biology: A Laboratory Notebook (J.E. Cellis (ed.) 1998) Academic Press; Animal Cell Culture (R.I. Freshney) (ed.), 1987); Current Protocols in Immunology (J.E. Coligan et al. (eds.), 1991); Short Protocols in Molecular Biology (Wiley and Sons, 1999); Immunobiology (C.A. Janeway and P. Travers, 1997); Antibodies (P. Finch, 1997); Antibodies: A Practical Approach (D. Catty (ed.), IRL Press, 1988 - 1989); Monoclonal Antibodies: A Practical Approach (P. Shepherd and C.It can be utilized using the methodologies described in Dean (ed.), Oxford University Press, 2000); Using Antibodies: A Laboratory Manual (E. Harlow and D. Lane (Cold Spring Harbor Laboratory Press, 1999); The Antibodies (M. Zanetti and J.D. Capra (eds.), Harwood Academic Publishers, 1995). Methods and techniques for identifying amino acid sequences in the constant and variable regions are well-known in the art and can be used to identify CDRs within the specific HCVR and / or LCVR amino acid sequences disclosed herein. Exemplary conventions that can be used to identify the boundaries of CDRs include, for example, the Kabat definition, the Chothia definition, and the AbM definition. Generally, the Kabat definition is based on sequence variability, the Chothia definition is based on the position of structurally looped regions, and the AbM definition is a compromise between the Kabat and Chothia approaches. See, for example, Kabat, "Sequences of Proteins of Immunological Interest," National Institutes of Health, Bethesda, Md, (1991); Al-Lazikani et al., J. Mol. Biol. 273:927-948 (1997); and Martin et al., Proc. Natl. Acad. Sci. USA 86:9268-9272 (1989). Public databases are also available for identifying CDR sequences within antibodies. For example, software packages and databases for determining antigenic fragments, leader sequences, protein folding, functional domains, glycosylation sites, and sequence alignments are available (e.g., GenBank, Vector NTI® suite (Informax, Inc, Bethesda, Md.); GCG Wisconsin package (Accelrys, Inc., San Diego, Calif.); DeCypher® (TimeLogic Corp., Crystal Bay, Nev.); see Menne et al. (2000) Bioinformatics 16:741-742; Menne et al. (2000) Bioinformatics Applications Note 16:741-742; Wren et al. (2002) Comput. Methods Programs Biomed. 68:177-181; von Heijne (1983) Eur. J. Biochem. 133:17-21; von Heijne (1986) Nucleic Acids Res. 14:4683-4690).

Example

[0077]

[0087] Using various mAbs with different variable regions along with the same general Fc region (e.g., SEQ ID NO: 1), surprisingly, contrary to the widely held belief that such proteins in the lung would be rapidly cleared (within minutes if not hours), instead, when inhaled in a safe and reasonable amount during single therapy as described herein, such mAb proteins, particularly those glycosylated to increase mucus capture, were shown to persist for several days at therapeutically appreciable levels.

[0078]

[0088] The results described herein are not limited to mAbs against pathogens. For example, since anti-inflammatory mAbs can use the same Fc region, they typically exhibit the same clearance profile as described herein. Anti-pathogen and anti-inflammatory mAbs can be co-administered to treat inpatients, and the methods and compositions described herein can be used for indications where anti-inflammatory mAbs are used alone.

[0079]

[0089] As shown in the following examples, the results described herein are particularly surprising. For example, considering that the increase in mucus adhesion due to having an mAb with a "mucus-capturing" Fc domain by concentrating on G0 glycosylation has long been thought to lead to faster clearance by the mucosa, surprisingly, the methods and compositions described herein may remain long enough to sustain significant concentrations at 24 hours in both the URT and LRT.

[0080] Example 1 Human IgG G1 Fc region

[0090] The human IgG G1 Fc region (IN-006, reformulation of legdanvimab) having an Fc region homologous to SEQ ID NO: 1 was investigated as part of a study conducted in Australia. Except for the pharmacy staff who prepared the study drug, the study staff and participants were not informed about the treatment assignment. The primary outcome was safety and tolerability. The exploratory outcome was the pharmacokinetic evaluation of IN-006 in nasal fluid and serum.

[0081]

[0091] Twenty-three participants were enrolled and randomized across two single-dose and one multiple-dose cohorts. There were no serious adverse events (SAEs). All enrolled participants completed the study without treatment interruption or discontinuation. Adverse events that occurred with all treatments were transient, were classified as mild to moderate in severity, and were not dose-dependent. Inhalation administration showed good tolerability and was completed in an average of 6 minutes for the high-dose group. The mean nasal fluid concentration of IN-006 in the multiple-dose cohort was 921 μg / mL at 30 minutes after dosing and 5.4 μg / mL at 22 hours. The mean serum level in the multiple-dose cohort reached a peak of 0.55 μg / mL three days after the final dose.

[0082]

[0092] IN-006 showed good tolerability and achieved concentrations in the airway that were orders of magnitude above its inhibitory concentration. These data support further clinical development of IN-006.

[0093] Similar to many viruses that cause acute respiratory infections (ARIs), SARS-CoV-2 almost invariably infects cells via the apical (luminal) side of the airway epithelium and buds out from infected cells mainly via the apical surface. Since the infection spreads from the upper respiratory tract (URT) to the lower respiratory tract (LRT) and into the deep lung, progeny viruses must move through airway mucus to reach uninfected epithelial cells. Thus, neutralizing monoclonal antibodies (mAbs) must reach the airway lumen in sufficient amounts to effectively neutralize the virus and halt infection.

[0083]

[0094] mAbs distribute very poorly and slowly from the blood into the airways, having concentrations in the airways that are orders of magnitude lower than those in serum after intravenous (IV) or intramuscular (IM) administration. Despite these limitations, clinical experience to date has shown that mAbs that neutralize SARS-CoV-2 can be effective in treating infected individuals at high risk of severe COVID when administered IV early in the course of infection. Nevertheless, high doses of mAbs are generally required to do so, reducing the available drug supply. The delay in lung distribution also limits the treatment window for preventing severe COVID.

[0084]

[0095] Inhalation administration is used to deliver protein therapeutics (e.g., Pulmozyme) directly to the lungs, enabling dosing within minutes. Importantly, direct inhalation delivery can achieve a considerably higher concentration of drug in the lungs than can be achieved by IV or IM administration and does so within minutes. Since the pattern of deposition along the airways is largely determined by the aerosol droplet size, it is possible to use a nebulizer that generates a wide aerosol size distribution to deliver the drug throughout the airways, from the nasal turbinates in the URT through the conducting airways in the LRT to the deep lung. Thus, nebulized delivery is likely to be the fastest way to achieve high inhibitory concentrations of mAb in the airway fluid. Inhalation administration also enables convenient home self-administration and reduces the burden on the patient and on the healthcare infrastructure associated with systemic delivery.

[0085]

[0096] IN-006 is a repurposing of regdanvimab, configured as specifically described herein for nebulized delivery, for the inhalation treatment of COVID-19. Regdanvimab, an IV-administered human IgG1 mAb against the SARS-COV-2 spike protein receptor binding domain (RBD), is approved in the European Union for adults with COVID-19 who do not require supplemental oxygen and who are not at increased risk of worsening severe COVID-19.

[0086]

[0097] A first-in-human, double-blind, placebo-controlled, escalating-dose pharmacokinetic and safety study was conducted in the Phase 1 unit in Melbourne, Australia. The study was conducted in accordance with the guidelines of the International Council for Harmonisation's Good Clinical Practice for the conduct of clinical trials of pharmaceuticals and complied with local regulatory requirements. It was approved by The Alfred Hospital Office of Ethics and Research Governance, Melbourne, Victoria, Australia. Informed consent was obtained prior to all study-related procedures. Eligible participants were sequentially enrolled in three cohorts: a single low-dose cohort (30 mg), a single high-dose cohort (90 mg), and a multiple high-dose cohort (seven 90-mg daily doses). For each single-dose cohort, a sentinel pair (with one active recipient and one placebo recipient) was dosed first, followed by a 2-day safety monitoring period prior to dosing the remainder of the cohort. Transition to subsequent cohorts was done after review of safety parameters 7 days after dosing of the preceding cohort. Figure 3 shows a diagram of the study structure and the timing of pharmacokinetic evaluations.

[0087]

[0098] Eligibility criteria required that participants be adults aged 18 to 55 years, in good health as determined by medical history, physical examination, clinical chemistry and hematological evaluations, electrocardiogram, forced expiratory volume in 1 second (FEV 1 ) ≥ 90% predicted value and negative serology for HBsAg, HCV, and HIV antibodies, and have a body mass index of 18 to 32 kg / m 2 . Participants were required to be non-smokers or light smokers. FEV 1The threshold was changed to ≥80% predicted value after the registration of the first 7 participants. Participants were excluded for known or suspected symptomatic viral infections or signs of pulmonary infectious or inflammatory conditions within 14 days of starting medication, airway hyperresponsiveness, angioedema, a history of anaphylaxis, or a positive alcohol breath test and / or urine drug screening for substance abuse. When replenishing the 7 participants including the first single-dose cohort, participants who had received the COVID-19 vaccine were excluded. However, due to the rapid increase in local vaccine availability and uptake, this criterion was modified to exclude only those who had received vaccination within 2 weeks of the first dose or those who had a scheduled vaccination within 2 weeks after the completion of dosing.

[0088]

[0099] The primary endpoint of the clinical trial was the safety and tolerability of IN-006. This was evaluated by monitoring adverse events that occurred due to the treatment, vital signs, ECG, FEV 1 , SpO 2 , hematological and chemical safety blood tests, and physical examinations before and after administration. The follow-up continued for 28 days and was evaluated on the days shown in Figure 3. The exploratory outcome was the drug levels in nasal fluid and serum at intervals before and after administration. The randomization schedule was prepared for each sentinel pair using software (SAS) verified by a statistical team member who had no responsibility for the monitoring and data management of this study, including one active and one saline placebo assignment, with the overall ratio of active to placebo assignment in each cohort being 3:1. The randomization code was held by the unblinded pharmacy staff, who prepared the doses in matching syringes with the same appearance for filling the nebulizer by the clinical staff.

[0089]

[0100] IN-006 was manufactured under Good Manufacturing Practices (GMP) and supplied by the manufacturer as a liquid formulation in glass vials. IN-006 was provided in syringes to be filled into the InnoSpire Go vibrating mesh nebulizer (Koninklijke Philips N.V.). Placebo participants received the same syringes containing saline instead of IN-006. Participants were instructed to breathe slowly through the nebulizer mouthpiece and exhale through their nose. Nasal fluid was obtained by rotating a flocked swab (Copans catalog number 56380CS01) at the middle turbinate depth (4 - 5 cm) for 10 - 15 seconds. Between subsequent sampling time points, sampling was alternated between the right and left nostrils. The amount of nasal fluid sample collected by each individual swab was determined by comparing the pre-weight and post-weight. This was achieved by weighing the sample-containing swab and sample tube before and after incubation in the extraction buffer, rinsing, and oven drying. The sampling times for nasal fluid and serum are shown in Figure 3. Vital signs and FEV 1 were measured before inhalation, 15 minutes and 30 minutes after completion. IN-006 concentrations were measured in human serum and nasal fluid. The sample size was selected according to the convention for the first phase in human studies. Formal calculation of the sample size and power was not performed. Continuous variables were summarized using descriptive statistics including the number of non-missing observations, mean, SD, median, minimum, and maximum values. Categorical variables were summarized using frequency counts and percentages. Placebo recipients from different cohorts were pooled. The safety analysis included all randomized participants who received any dose of the study drug. The pharmacokinetic population included all participants who received any dose of IN-006. Inferential statistical tests were not performed. The serum PK parameters of IN-006 were determined using Phoenix WinNonlin version 8.3.

[0090]

[0101] Twenty-three participants were sequentially assigned to one of three cohorts from 102 screened adults. The first participant was randomized on September 22, 2021, and the last participant visit was on December 29, 2021. Of these participants, 17 were randomly assigned to receive IN-006 and 6 were randomly assigned to receive placebo. All 23 participants received their assigned treatment as intended and completed their last study visit on study day 29. The study was completed on December 29, 2021. The participant flow is depicted in Figure 3 and the participant demographics are listed in Table 1 (Figure 1).

[0091]

[0102] Treatment-emergent adverse events (TEAEs) are listed in Table 2 (Figure 2). Inhaled administration of IN-006 was well tolerated and completed in an average of 6 minutes (range 4 - 9 minutes) for the 90 mg dose. Eight of 15 participants (53.3%) included in the single ascending dose cohort experienced at least one TEAE (6 received IN-006 and 2 received placebo). The most frequently reported TEAEs were headache (4 / 15; 26.7%) and pharyngolaryngeal pain (2 / 15; 13.3%). All TEAEs except one were mild. One participant receiving the low dose of IN-006 (30 mg) experienced a moderate event (increase in transaminases on day 29), which was considered by the study physician not to be related to the study drug. Three participants (3 / 15; 20.0%) experienced at least one TEAE considered by the study physician to be related to the study drug. These events included headache, cough, and pharyngolaryngeal pain. All three related TEAEs resolved and were mild. There was no evidence of dose-related effects.

[0092]

[0103] In the multiple-dose cohort, no TEAE was reported in participants receiving placebo. Among the 6 participants receiving IN-006, 4 (66.7%) participants experienced at least one TEAE. The most frequently reported TEAE was dizziness (2 / 6; 33.3%). All TEAEs except one were mild. One single-dose participant receiving IN-006 experienced a moderate event (severe pain). The event was considered unlikely to be related to the study drug by the investigator in charge of the clinical trial. Two (33.3%) participants experienced at least one TEAE considered by the investigator in charge of the clinical trial to be related to the study drug. These drug-related TEAEs were dizziness and FEV 1 decrease, the latter of which was noticed 15 minutes after inhalation administration, was not accompanied by symptoms or abnormal vital signs, resolved within 15 minutes, and did not recur with subsequent doses. Both events were mild.

[0093]

[0104] No serious TEAEs, SAEs or TEAEs leading to discontinuation were reported in either the single-dose or multiple-dose cohorts. There were no unexpected safety signals.

[0105] For the single-dose cohort, the mean nasal concentrations at 3 hours after dosing for the 30 mg and 90 mg doses, respectively, were 261 μg / g and 710 μg / g; these values are consistent with a three-fold increase in the dose administered. In the multiple-dose cohort, repeated dosing provided additional opportunities for more nasal concentration measurements over a greater number of time points. The nasal concentrations measured 30 minutes after dosing on Days 1, 2, and 3 were, on average, 773 μg / g, which is higher than the concentration measured 3 hours after dosing (405 μg / g). This indicates that peak exposure occurred immediately after dosing and that the nasal concentration decreased appreciably by 3 hours after administration. The concentration of IN-006 measured 22 hours after dosing was <2% of the concentration immediately after dosing, indicating minimal nasal accumulation (Figures 4A - 4C). The difference in nasal concentration between 30 minutes and 22 hours after dosing suggested that the interval spanned approximately 6 - 7 half-lives, and the difference between 30 minutes and 3 hours after dosing was approximately half. Both are consistent with a nasal half-life of approximately 3 - 4 hours, which is significantly longer than the time scale of the estimated mucociliary clearance transit time of approximately 5 - 15 minutes obtained from the saccharin transit time test.

[0094]

[0106] As shown in Figures 5A - 5B, the serum concentration of IN-006 became detectable by 12 hours after inhaled administration at the 90 mg dose and continued to rise until 120 hours after a single dose (cohorts 1 and 2) or until 216 hours after the first dose in those receiving multiple doses (cohort 3). The elimination half-life of IN-006 in serum was estimated to be approximately 253, 292, and 402 hours, equivalent to the previously estimated elimination half-life of regdanvimab from serum after intravenous administration (288 hours) in the cohorts receiving a single 30 mg dose, a single 90 mg dose, and 7 daily 90 mg doses, respectively. The serum concentration of IN-006 was significantly lower than that in nasal fluid (serum C of 0.52 μg / mL compared to 990 μg / mL in nasal fluid), but they were still significantly higher than the IC max ) of IN-006 (approximately 0.01 μg / mL). 50 (about 0.01 μg / mL).

[0095]

[0107] For many years, it has been very difficult to stably nebulize mAbs (see, for example, Respaud, R et al., Nebulization as a delivery method for mAbs in respiratory diseases. Expert Opin Drug Deliv, 2015. 12(6): 1027-1039; Mayor, A et al., Inhaled antibodies: formulations require specific development to overcome instability due to nebulization. Drug Deliv Transl Res, 2021. 11(4): 1625-1633; Bodier-Montagutelli, E et al., Protein stability during nebulization: Mind the collection step! Eur J Pharm Biopharm, 2020. 152: 23-34; and Bodier-Montagutelli, E et al., Designing inhaled protein therapeutics for topical lung delivery: what are the next steps? Expert Opin Drug Deliv, 2018. 15(8): 729-736), and biologics have been considered to be rapidly eliminated from the airways by either systemic absorption, physical mucociliary clearance, or degradation by alveolar macrophages, which would make it difficult to sustain therapeutic concentrations. See, for example, Loira-Pastoriza, C., J. Todoroff, and R. Vanbever, Delivery strategies for sustained drug release in the lungs. Adv Drug Deliv Rev, 2014. 75: 81-91 and Suri, R, The use of human deoxyribonuclease (rhDNase) in the management of cystic fibrosis. BioDrugs, 2005. 19(3): 135-144.

[0096]

[0108] Surprisingly, as described herein, the reformulation of IN-006, regdanvimab for aerosolized delivery, was safe in healthy adults, showed good tolerability, had minimal side effects, high concentrations of drug were recovered from nasal and serum samples, and therapeutic concentrations could be sustained. This may be, in part, due to the glycosylation of the therapeutic antibody as described herein, in combination with the administration technique. The treatment was easily self-administered by all participants and was completed within minutes. Encouragingly, the concentration of IN-006 measured in nasal secretions was well above its IC 50 even 22 - 24 hours after dosing. In the multiple-dose cohort receiving 7 daily doses of 90 mg, an average nasal fluid IN-006 concentration of 920 μg / mL was observed 30 minutes after the first dose, and an average concentration of 5.4 μg / mL was maintained 22 hours later, before receiving the second dose. The ability to maintain mAb concentrations in the range 3 - 7 orders of magnitude higher than the IC 50 (~4 - 20 ng / mL) of regdanvimab and other COVID mAbs against susceptible variants strongly supports the once-daily dosing regimen proposed by the inventors. Since SARS-CoV-2 infection and replication begin in the upper respiratory tract, the efficient delivery of IN-006 to the nasal cavity by the inventors has the potential to provide a more highly effective treatment for early COVID-19, suggesting that this could enable early resolution of infection and reduction of the risk of progression to severe COVID.

[0097]

[0109] mAbs have been proven to be effective therapeutics for COVID-19, but the need for administration via the IV, IM, or SC routes has limited their use in clinical practice. For intravenous administration, the need for infusion centers and post-dose observation severely limits the number of patients who can be treated and significantly increases costs. IM injections shorten the administration time but are limited by the volume that can be administered per injection (about 5 mL), which in turn limits the dose of mAb that can be administered per injection and can be painful when the maximum injection volume is used. In contrast, nebulized delivery using a handheld nebulizer enables home dosing convenience and takes only a few minutes to complete. Furthermore, the IV, IM, and SC routes deliver the mAb to the airway lining fluid only after a delay of one or multiple days, and even then, only achieve airway concentrations that are only a fraction of the plasma concentration. For example, in a recent clinical trial of the anti-influenza mAb CR6261 administered as a single 50 mg / kg dose IV, peak nasal concentrations were not achieved until 2 days after infusion, and despite the much lower total dose of IN-006 relative to CR6261 (90 mg IN-006 vs. about 2,000 - 4,000 mg of CR6261), the peak nasal concentration of 0.597 μg / mL was about one-tenth of the concentration observed for IN-006 at the trough of daily dosing by the inventors (about 5.4 μg / mL). Due to increased convenience, more efficient lung delivery, and superior pharmacokinetics, inhalation may be a more preferred route for mAb delivery for treating acute respiratory infections.

[0098]

[0110] COVID-19 is mainly a respiratory infection, but the currently available treatments are administered by systemic dosing. Methods and compositions of mAbs constructed using human IgG that binds to the spike protein of SARS-CoV-2 are described herein. The methods and compositions described herein may provide inhaled delivery of mucosal capture monoclonal antibodies (including an Fc region having G0 glycosylation), which may provide a more convenient and effective treatment for COVID-19. The results described in Example 1 demonstrate the safety, tolerability, and pharmacokinetics of an example of a human IgG G1 Fc region (IN-006, a reformulation of regdanvimab, an approved intravenous treatment for COVID-19) that can be used for nebulized delivery by a handheld nebulizer.

[0099]

[0111] Severe disease due to SARS-Cov-2 is associated with the spread of the virus from the initial upper respiratory tract infection site to the deep lungs. Unfortunately, the exact timing of such spread is highly likely to be highly variable among individuals. In fact, there is evidence suggesting that the virus has already reached the LRT even around the time symptoms appear during the early stages of the disease. Similarly, rapid spread of infection to the LRT is likely to be frequent for other viruses such as influenza. Therefore, in order to widen the treatment window and reduce the risk of COVID-induced pneumonia and hospitalization, it may be important to administer drugs not only to the URT (e.g., by nasal spray), but also to both the URT and LRT. The level of IN-006 in the LRT was not directly measured in this study, but the appreciable delay in serum concentration and serum Tmax both strongly suggest that IN-006 is efficiently delivered into the LRT and deep lungs. In fact, in multiple-dose inhalation administration studies of the pharmacokinetics of IN-006 in rats, the concentration of IN-006 in airway fluid exceeded the serum concentration by more than about 100-fold. Efficient delivery to the LRT is a direct result of the inventors' design requirements for the vibrating mesh nebulizer. To deliver mAb into the LRT and deep lungs, the droplet sizes (fine particle fraction, i.e., droplets <5 μm, especially those <2.5 μm) generated by the nebulizer were intentionally selected. Furthermore, the fact that a slow and steady increase in serum concentration was observed over about 4 days in the single-dose cohort and the peak serum concentration in the multiple-dose cohort at about 9 days (or 2 days after the last dose) means that high levels of IN-006 are sustained in the deep lungs for at least that period, i.e., for several days. Assuming a similar ratio of 100:1 airway fluid to serum concentration in humans as observed in rats, the mean human serum concentration of 200 ng / mL 2 days after the first dose and 550 ng / mL on day 9 would have to be converted to lung concentrations of about 50 μg / mL, which is more than 3 digits above the IC 50 and is equivalent to the serum concentrations achieved with some IV / IM-administered mAbs. The intrinsic activity of the mAb (IC 50) The extremely high mAb levels sustained against may potentially continue to provide effective treatment against variants, even in the presence of recognizable genetic drift, and may reduce risks including virus escape. It also suggests that a shorter course of therapy, perhaps as short as a single dose, may provide recognizable protection against hospitalization.

[0100]

[0112] Despite the significant extrapulmonary manifestations of severe COVID-19 and the frequent detection of SARS-CoV-2 RNA in the blood, infectious SARS-CoV-2 is rarely detected in the blood of affected patients, suggesting that extrapulmonary disorders may be caused by indirect factors such as inflammatory responses rather than extrapulmonary virus infections. Nevertheless, it is reassuring that the serum levels of IN-006 achieved after aerosolized delivery were observed to exceed the IC 50 by at least one order of magnitude.

[0101]

[0113] Regdanvimab (administered IV) has been shown to be highly effective for the prevention of severe COVID-19 in a global phase 3 study, leading to formal approvals (EMEA / H / C / 005854) in South Korea and the European Union for the prevention of severe disease in patients presenting with mild to moderate COVID-19, as well as emergency use authorizations (EUAs) or conditional manufacturing authorizations in several additional countries around the world. IN-006, in combination with a second potent neutralizing mAb, can generate an mAb cocktail with potent binding activity against any variant tested to date. The surprisingly long airway retention of IN-006 observed here can in fact be exploited for mAbs that include the Fc region (e.g., SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4). The reformulation of regdanvimab for inhaled delivery was found to be safe and well-tolerated in healthy participants at single doses of 30 mg and 90 mg, as well as 7 consecutive daily doses of 90 mg. Inhaled administration results in IN-006 levels in nasal fluid and presumably also in the lung, exceeding by orders of magnitude the inhibitory concentration of susceptible SARS-CoV-2 variants within 30 minutes, with serum concentrations continuing to rise for several days after dosing, meaning substantially persistent IN-006 levels in the lung.

[0102] Example 2

[0114] Using the methods described herein, sufficient levels of antibody can be provided in both the upper and lower respiratory tracts.

[0103]

[0115] Subjects receive a first dose of IN-006 or placebo via a nebulizer on day 1 of dosing. The placebo is 0.9% saline and is administered via a nebulizer in the same manner as IN-006. Subjects receive a second dose of IN-006 or placebo via a nebulizer on at least one of days 3 to 8 of dosing. The placebo is 0.9% saline and is administered via a nebulizer in the same manner as IN-006. Antibody measurements are performed using bronchoalveolar lavage (BAL) before treatment and 2 weeks after treatment, using bronchoscopy. Bronchoalveolar lavage is performed by injecting warmed sterile PBS into the segmental middle lobe bronchi using a bronchoscope. The fluid is recovered by gentle suction and collected in a sterile container. It is filtered through a sterile 100 μm mesh to remove mucus and cell debris and analyzed using the methods described herein.

[0104] Example 3

[0116] Using the methods described herein, sufficient levels of at least two therapeutic agents can be provided in both the upper and lower respiratory tracts. Subjects can receive a first dose of IN-006 and a second therapeutic agent (e.g., non-mAb) or placebo via a nebulizer on day 1 of dosing. The placebo is 0.9% saline and is administered via a nebulizer in the same manner as IN-006. Nasal swabs are taken on at least one of days 3 to 8 of dosing to measure antibody levels. Subjects receive a second dose of IN-006 or placebo via a nasal spray on at least one of days 3 to 8 of dosing. The placebo is 0.9% saline and is administered via a nebulizer in the same manner as IN-006.

[0105] Example 4

[0117] Using the methods described herein, sufficient levels of antibodies can be provided in at least one or both of the upper and lower airways using 1X (once) or 2X (twice) per day of dosing.

[0106]

[0118] Subjects receive a first dose of an antibody (e.g., IN-006 or others) or placebo at time 0 on day 1 of dosing via a nebulizer. The antibody can be an antibody glycosylated with a G0 glycosylation pattern. The placebo is 0.9% saline and is administered via a nebulizer in the same manner as the antibody (e.g., IN-006 or others). The first cohort of subjects receives a second dose of the antibody (e.g., IN-006 or others) or placebo via a nebulizer 12 hours after the first dose. The placebo is 0.9% saline and is administered via a nebulizer in the same manner as the antibody (e.g., IN-006 or others). Measurement of the antibody is performed using bronchoscopy with bronchoalveolar lavage fluid (BAL) before dosing, immediately after dosing, at 12 hours (before the second dose), and at 24 hours. Bronchoalveolar lavage is performed by injecting warmed sterile PBS into the segmental middle lobe bronchus using a bronchoscope. The fluid is recovered by gentle suction and collected in a sterile container. It is filtered through a sterile 100 μm mesh to remove mucus and cell debris and analyzed using the methods described herein.

[0107]

[0119]

[0108]

Table 1

[0109]

[0120] The methods and compositions described herein include those having a G0 glycosylation pattern and enable delivery of inhaled mAbs that, otherwise, would be predicted to be cleared within minutes based on published studies, achieving sustained high concentrations for over 24 hours, thereby enabling once-daily or twice-daily dosing using relatively low (and thus accessible) concentrations. Thus, the dose can deliver the therapeutic mAb such that sufficient levels (i.e., trough concentrations) are maintained until the next dose.

[0110]

[0121] As described herein, the peak concentration achieved in the upper airway corresponds to the amount of mAb inhaled (e.g., going from 30 mg to 90 mg inhalation resulted in an approximately 3x increase). Subsequently, the rate of clearance was, surprisingly, dose-independent, with equivalent clearance rates for single doses of 30 mg and 90 mg and equivalent clearance between a single 90 mg dose and repeated 90 mg doses on different days. "Mucus-trapping" mAbs (those with the G0 glycosylation pattern) are not cleared within minutes (e.g., faster than 30 minutes) as previously suggested for the turnover rate of nasal secretions in the nasal turbinates, but rather have a half-life in the range of 3.5 - 4 hours.

[0111]

[0122] This enables a dosing regimen that not only achieves a sufficiently excessive mAb dose that exceeds the IC50 immediately after dosing, but also allows sufficient mAb to be retained at the trough just prior to the next dosing. Thus, different mAb doses can be selected according to their characteristics to achieve a concentration that is sufficiently excessive relative to their intrinsic potency (e.g., maintaining 10 - 100x above the IC50 for an mAb with an IC50 of about 100 ng / mL). For example, the methods and compositions described herein show that a dose of 15 mg at each of two times per day provides a sufficient dose.

[0112]

[0123] Of course, all combinations of the above concepts and additional concepts discussed in more detail below (such concepts being non - conflicting with each other) are intended to be part of the subject matter of the invention discussed herein and can be used to achieve the benefits described herein.

[0113]

[0124] The sequences of process parameters and steps described and / or illustrated in this specification are provided by way of example only and may vary as appropriate. For example, the steps illustrated and / or described in this specification may be shown or discussed in a particular order, but these steps need not necessarily be performed in the order illustrated or discussed. The various example methods described and / or illustrated in this specification may also omit one or more of the steps described or illustrated in this specification, or may include additional steps in addition to those disclosed.

[0114]

[0125] When a feature or element is referred to in this specification as being "on" another feature or element, it may be directly on the other feature or element or there may also be intervening features and / or elements. In contrast, when a feature or element is referred to as being "directly on" another feature or element, there are no intervening features or elements. Also, when a feature or element is referred to as being "connected to", "attached to" or "coupled to" another feature or element, it will be understood that it may be directly connected, attached or coupled to the other feature or element or there may be intervening features or elements. In contrast, when a feature or element is referred to as being "directly connected to", "directly attached to" or "directly coupled to" another feature or element, there are no intervening features or elements. Although described or shown with respect to one embodiment, the features and elements so described or shown may apply to another embodiment. It will also be understood by those skilled in the art that a reference to a structure or feature being "adjacent" to another feature may mean that the adjacent feature overlaps or has a portion that underlies it.

[0115]

[0126] The technical terms used in this specification are for the sole purpose of describing specific embodiments and are not intended to be limiting of the present invention. For example, as used in this specification, the singular forms "a", "an", and "the" are to be construed to include the plural forms as well, unless the context clearly dictates otherwise. The terms "comprises" and / or "comprising", as used in this specification, specify the presence of the stated features, steps, operations, elements, and / or components, but it is further understood that they do not preclude the presence or addition of one or more other features, steps, operations, elements, components, and / or groups thereof. As used in this specification, the term "and / or" includes any and all combinations of one or more of the associated listed items and may sometimes be abbreviated as " / ".

[0116]

[0127] The terms "first" and "second" may be used in this specification to describe various features / elements (including steps), but these features / elements are not to be limited by these terms, unless otherwise specified. These terms may be used to distinguish one feature / element from another. Thus, a first feature / element discussed below may be referred to as a second feature / element, and similarly, a second feature / element discussed below may be referred to as a first feature / element without departing from the teachings of the present invention.

[0117]

[0128] Throughout this specification and the following claims, unless the context requires otherwise, the words "comprise", "comprises", and "comprising" and variations thereof are to be construed to mean that various components can be used jointly in methods and articles (e.g., apparatuses and methods including compositions and devices). For example, the term "comprising" is understood to mean including any recited element or step but not meaning the exclusion of any other element or step.

[0118]

[0129] In general, any of the devices and methods described in this specification should be understood to be inclusive, but all or subsets of components and / or steps may alternatively be exclusive and may be expressed as "consisting of" or alternatively "consisting essentially of" various components, steps, sub-components or sub-steps.

[0119]

[0130] In this specification, unless otherwise specified, all numbers, including when used in the specification, in the claims, and in the examples, can be read as if the word "about" or "approximately" were placed before them even if the term does not explicitly appear. The phrases "about" or "approximately" are used when describing a scale and / or a position to indicate that the recited value and / or position is within a reasonable predicted range of the value and / or position. For example, a numerical value may have a value that is + / -0.1% of the recited value (or range of values), + / -1% of the recited value (or range of values), + / -2% of the recited value (or range of values), + / -5% of the recited value (or range of values), + / -10% of the recited value (or range of values), etc. Any numerical value given in this specification should also be understood to include about or approximately that value, unless otherwise stated. For example, if the value "10" is disclosed, then "about 10" is also disclosed. Any numerical range recited in this specification is intended to include all sub-ranges incorporated within it. Also, as would be appropriately understood by those skilled in the art, it is understood that when a value is disclosed, the value "less than", the value "greater than", and the possible ranges between the values are also disclosed. For example, if the value "X" is disclosed, then "less than X" as well as "greater than X" (for example, if X is a numerical value) are also disclosed. Throughout this application, it is understood that data is provided in several different formats and that this data represents ranges of any combination of endpoints and starting points and data points. For example, if a particular data point "10" and a particular data point "15" are disclosed, then it is understood that values greater than, more than, less than, less than or equal to, and between 10 and 15 are considered and disclosed. It is also understood that each unit between two particular units is also disclosed. For example, if 10 and 15 are disclosed, then 11, 12, 13, and 14 are then also disclosed.

[0120]

[0131] Although various exemplary embodiments have been described above, any of several variations can be made to the various embodiments without departing from the scope of the invention as set forth in the claims. For example, in alternative embodiments, the order in which the various described method steps are performed can often be changed, and in other alternative embodiments, one or more method steps can be skipped together. Optional features of the various device and system embodiments may or may not be included in some embodiments. Accordingly, the foregoing description has been provided primarily for illustrative purposes and should not be construed as limiting the scope of the invention as set forth in the claims. The examples and illustrations included herein are for illustrative purposes, not limitations, and show specific embodiments in which the subject matter may be practiced. As noted above, other embodiments can be utilized and derived therefrom, so long as structural and logical substitutions and variations can be made without departing from the scope of the present disclosure. Such embodiments of the subject matter of the present invention, for convenience and without intending to limit the scope of the present application to any single invention or inventive concept if more than one is actually disclosed, may be referred to herein individually or collectively simply by the term "invention." Accordingly, specific embodiments have been illustrated and described herein, but any arrangement calculated to achieve the same purpose may be substituted for the specific embodiments shown. The present disclosure is intended to cover any and all adaptations or variations of the various embodiments. Combinations of the above embodiments and other embodiments not specifically described herein will be apparent to those skilled in the art upon review of the foregoing description. Sequence Listing SEQ ID NO:1 Human IgG G1 Fc region (CH2 and CH3 domains) Organism: Homo sapiens (Human) (CH2 is residues 1-113 and CH3 is residues 114-219) PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK Sequence number 2 Human IgG G2 Fc region (CH2 and CH3 domains) Organism: Homo sapiens (Human) (CH2 is residues 1 - 109 and CH3 is residues 110 - 216) APPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDISVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK Sequence number 3 Human IgG G3 Fc region (CH2 and CH3 domains) Organism: Homo sapiens (Human) (CH2 is residues 1 - 110 and CH3 is residues 111 - 216) APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFKWYVDGVEVHNAKTKPREEQYNSTFRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESSGQPENNYNTTPPMLDSDGSFFLYSKLTVDKSRWQQGNIFSCSVMHEALHNRFTQKSLSLSPG Sequence number 4 Human IgG G4 Fc region (CH2 and CH3 domains) Organism: Homo sapiens (Human) (CH2 consists of residues 1 - 110 and CH3 consists of residues 111 - 217) APEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK

Claims

1. A pharmaceutical composition for use in a method of treating a subject having or at risk of having respiratory impairment, comprising the step of administering to a subject by inhalation a pharmaceutical composition comprising a population of antibodies comprising a therapeutic human IgG monoclonal antibody (mAb) comprising antibodies glycosylated in a G0 glycosylation pattern comprising a branched coglycan structure of Manα1-6(Manα1-3)Manβ1-4GlcNAcβ1-4GlcNAcβ1, wherein the administration step comprises administering a dose of 0.02 μmol or more of therapeutic human mAb at least twice daily, such that concentrations of therapeutic human mAb are achieved at a higher concentration than 20 ng / mL in the upper respiratory tract (URT) and at a higher concentration than 100 ng / mL in the lower respiratory tract (LRT) 12 hours or longer after dose.

2. A pharmaceutical composition for use in a method of treating a subject having or at risk of having respiratory impairment, comprising the step of administering a dose of therapeutic human IgG monoclonal antibody (mAb) to a subject by inhalation, wherein the dose dose comprises the step of maintaining a concentration of therapeutic human IgG monoclonal antibody (mAb) that binds to respiratory viruses in the upper respiratory tract (URT) of the subject at a level higher than 20 ng / ml and in the lower respiratory tract (LRT) of the subject at a level higher than 100 ng / ml for a longer period than 12 hours after administration, wherein the dose administration step comprises administering 0.02 μmol or more of therapeutic human mAb twice a day or less.

3. The pharmaceutical composition according to claim 1 or claim 2, wherein the administration step includes administering a dose once a day or less.

4. The pharmaceutical composition according to claim 1, wherein at least 50% of the therapeutic antibody has a G0 glycosylation pattern.

5. The pharmaceutical composition according to claim 1 or 2, wherein the therapeutic antibody comprises an Fc sequence that is at least X% (e.g., 80%, 85%, 90%, 95%) homologous to the sequence of SEQ ID NO: 1 (e.g., human IgG1).

6. The pharmaceutical composition according to claim 1 or 2, wherein the target is an adult.

7. The pharmaceutical composition according to claim 1 or 2, wherein the therapeutic antibody comprises legdanvimab.

8. The pharmaceutical composition according to claim or 2, wherein the drug regimen comprises a drug cycle of two doses per day over a period of 2 to 7 days.

9. The pharmaceutical composition according to claim 1 or 2, wherein the drug regimen includes a drug cycle of every two days, every three days, or every four days.

10. The pharmaceutical composition according to claim 1 or 2, wherein the dosage regimen includes the step of administering a dose of at least 10 mg of therapeutic mAb.

11. The pharmaceutical composition according to claim 1 or 2, wherein the dosage regimen includes the step of administering a dose of therapeutic mAb between 10 mg and 100 mg.

12. The pharmaceutical composition according to claim 1 or 2, wherein the administration step includes a step of sustaining the release of therapeutic mAb from the LRT into the bloodstream over several days.

13. The pharmaceutical composition according to claim 1 or 2, wherein the administration step includes a step of sustaining the release of mAb into the lungs and blood for at least two days.

14. The pharmaceutical composition according to claim 1 or 2, wherein the formulation further comprises a pharmaceutically acceptable diluent, an excipient, and / or a carrier.

15. The pharmaceutical composition according to claim 1 or 2, wherein the formulation further comprises one or more of citric acid, arginine, mannitol, sorbitol, and trehalose.

16. The pharmaceutical composition according to claim 1 or 2, wherein the therapeutic antibody preparation is administered to a subject via a nebulizer.

17. The pharmaceutical composition according to claim 1 or 2, wherein the therapeutic antibody preparation is administered to a subject via a vibrating mesh nebulizer.

18. The pharmaceutical composition according to claim 1 or 2, wherein the therapeutic antibody preparation is administered by inhalation or by direct drip infusion into the upper respiratory tract.

19. The pharmaceutical composition according to claim 1 or 2, wherein the therapeutic antibody preparation is self-administered by the subject.

20. The pharmaceutical composition according to claim 1 or 2, wherein the respiratory disorder includes lower respiratory tract disorder.

21. The pharmaceutical composition according to claim 1 or 2, wherein the respiratory disorder includes upper respiratory tract disorders.

22. The pharmaceutical composition according to claim 1 or 2, wherein the respiratory disorder includes an inflammatory disorder.

23. The pharmaceutical composition according to claim 1 or 2, wherein the respiratory virus includes coronavirus.

24. The pharmaceutical composition according to claim 1 or 2, wherein the respiratory virus comprises severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).

25. The pharmaceutical composition according to claim 1 or 2, wherein the respiratory virus comprises respiratory syncytial virus (RSV).

26. The pharmaceutical composition according to claim 1 or 2, wherein the respiratory virus comprises one or more of influenza, metapneumovirus, parainfluenza, or coronavirus.

27. The pharmaceutical composition according to claim 1 or 2, wherein the respiratory virus comprises a paramyxovirus.

28. The pharmaceutical composition according to claim 1 or 2, wherein the formulation comprises a second or more therapeutic agents in addition to a therapeutic antibody.

29. The pharmaceutical composition according to claim 1 or 2, wherein the formulation comprises a therapeutic mAb and a second therapeutic antibody, and the first therapeutic antibody and the second therapeutic antibody bind to the same virus but do not compete for binding to the virus.

30. The pharmaceutical composition according to claim 1 or 2, wherein the formulation comprises a second therapeutic antibody in addition to a first therapeutic antibody, and further, the first antibody and the second antibody bind to different viruses.

31. The pharmaceutical composition according to claim 1 or 2, wherein the formulation comprises a biological agent in addition to a therapeutic mAb.

32. A pharmaceutical composition for use in a method of treating a subject having or at risk of having a respiratory disorder, comprising the step of administering a dose of the pharmaceutical composition containing a therapeutic human IgG monoclonal antibody (mAb) that binds to a respiratory virus to the subject by inhalation, thereby maintaining a concentration of the therapeutic human IgG monoclonal antibody (mAb) that binds to a respiratory virus in the subject's upper respiratory tract (URT) at a concentration higher than 25 ng / ml and in the subject's lower respiratory tract (LRT) at a concentration higher than 25 ng / ml for a period longer than 12 hours after administration, wherein the administration step comprises administering 0.02 μmol or more of therapeutic human mAb twice a day or less.

33. A therapeutic human IgG monoclonal antibody (mAb) comprising a population of antibodies in which at least 40% are glycosylated in a G0 glycosylation pattern containing a bibranched coreglycan structure of Manα1-6(Manα1-3)Manβ1-4GlcNAcβ1-4GlcNAcβ1, for use in a method of treating respiratory distress by administering the therapeutic human IgG monoclonal antibody (mAb) by inhalation, wherein the administration step comprises administering a dose of 0.02 μmol or more of the therapeutic human IgG monoclonal antibody (mAb) twice daily or less, such that concentrations of the therapeutic human mAb are achieved at a rate higher than 20 ng / mL in the upper respiratory tract (URT) and higher than 100 ng / mL in the lower respiratory tract (LRT) 12 hours or longer after dose.

34. A therapeutic human IgG monoclonal antibody (mAb) comprising a population of antibodies in which at least 40% are glycosylated in a G0 glycosylation pattern containing a branched coreglycan structure of Manα1-6(Manα1-3)Manβ1-4GlcNAcβ1-4GlcNAcβ1, for use in a method of treating respiratory distress, comprising the step of administering a dose of the therapeutic human IgG mAb by inhalation, thereby maintaining concentrations of the therapeutic human IgG mAb in the upper respiratory tract (URT) of the subject at a level higher than 20 ng / ml and in the lower respiratory tract (LRT) of the subject at a level higher than 100 ng / ml for more than 12 hours after administration, wherein the dose administration step comprises administering 0.02 μmol or more of the therapeutic human IgG mAb at a rate of no more than twice daily.

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