TW202529811A - Compositions of thymic stromal lymphopoietin (tslp) binding fragments and methods of use thereof - Google Patents

Abstract

The present disclosure relates to pharmaceutical compositions comprising antigen binding fragments specific for thymic stromal lymphopoietin (TSLP) suitable for inhalation, as well as a process for preparing a pharmaceutical composition for inhalation comprising said antigen binding fragment specific for TSLP. The present disclosure further relates to methods of treating TSLP-related conditions, such as asthma and COPD, using the pharmaceutical compositions. The pharmaceutical compositions include a mixture of leucine, trileucine, and histidine that results in a formulation that is suitable for delivering antigen binding fragments derived from anti-TSLP antibodies via inhalation.

Description

Compositions of thymic stromal lymphopoietin (TSLP) binding fragments and methods of use thereof

This disclosure relates to pharmaceutical compositions suitable for inhalation that contain antigen-binding fragments specific for thymic stromal lymphopoietin (TSLP), and processes for preparing pharmaceutical compositions suitable for inhalation that contain antigen-binding fragments specific for TSLP. The disclosure further relates to methods of using these pharmaceutical compositions to treat TSLP-related conditions, such as asthma and COPD. These pharmaceutical compositions include a mixture of leucine, trileucine, and histidine that produces a formulation suitable for delivering antigen-binding fragments derived from anti-TSLP antibodies via inhalation.

Asthma affects an estimated 300 million people worldwide, across all ages, and places a significant burden on health care systems and society through lost workplace productivity and disruption to families ("Pocket Guide for Asthma Management and Prevention," Global Initiative for Asthma; 2019). Asthma causes symptoms such as wheezing, shortness of breath, chest tightness, and cough, which vary in occurrence, frequency, and intensity over time. Symptoms are often associated with bronchoconstriction, thickening of the airway walls, and increased mucus production. Depending on the frequency and severity of attacks, asthma can be symptomatic and well or poorly controlled.

Thymic stromal lymphopoietin (TSLP) is an epithelial cell-derived interleukin produced in response to environmental and proinflammatory stimuli, leading to the activation of multiple inflammatory cells and downstream pathways. TSLP is elevated in the airways of patients with asthma and correlates with Th2 interleukin and proinflammatory cytokine expression, as well as disease severity. While TSLP is crucial for the regulation of Th2 immunity, it also plays a key role in other inflammatory pathways and, therefore, is associated with various asthma phenotypes.

Dry powder formulations containing anti-TSLP antibody fragments suitable for inhalation to treat asthma are described in WO 2021/083908. Anti-TSLP Fabs with improved stability are described in WO 2022/223514. WO 2017/042701 and WO 2021/152488 describe methods of using anti-TSLP antibodies or anti-TSLP antibody fragments to treat inflammatory or obstructive airway diseases such as asthma.

There remains a need for other compositions suitable for delivering antibodies against TSLP via inhalation. The present invention was designed with the above considerations in mind.

The present disclosure provides novel pharmaceutical compositions comprising antibody fragments of anti-TSLP antibodies that exhibit significant beneficial properties, including high manufacturing yield, low levels of subvisible particles, and minimal throat and device powder retention. Preclinical toxicity studies further confirmed that the novel compositions exhibit a favorable toxicity profile, with minimal immune-related toxicities. Therefore, the pharmaceutical compositions are expected to be safe and clinically beneficial when administered to patients for the treatment of TSLP-related diseases.

In a first aspect, the present disclosure provides a pharmaceutical composition comprising spray-dried particles comprising: a. about 5% (w/w) to about 15% (w/w) leucine; b. about 1% (w/w) to about 5% (w/w) trileucine; c. about 1% (w/w) to about 10% (w/w) histidine buffer, at a pH between about pH 5 and pH 6; d. about 1% (w/w) to about 80% (w/w) an antigen-binding fragment of an anti-thymic stromal lymphopoietin (TSLP) antibody; and e. a glass stabilizer.

In some cases, the pharmaceutical composition comprises about 1% (w/w) to about 3% (w/w) triceleucine.

In some cases, the pharmaceutical composition comprises about 2% (w/w) trileucine.

In some cases, the pharmaceutical composition comprises about 8% (w/w) to about 12% (w/w) leucine.

In some cases, the pharmaceutical composition comprises about 10.5% (w/w) leucine.

In some cases, the pharmaceutical composition comprises about 1% (w/w) to about 5% (w/w) histidine buffer.

In some cases, the pharmaceutical composition comprises about 2.5% (w/w) to about 3.5% (w/w) histidine buffer.

In some cases, the pharmaceutical composition comprises about 3.14% (w/w) histidine buffer.

In some cases, the pharmaceutical composition comprises about 0.55% (w/w) L-histidine and about 2.59% (w/w) histidine HCl.

In some cases, the antigen-binding fragment is present at a concentration of about 1% (w/w).

In some cases, the antigen-binding fragment is present at a concentration of about 2% (w/w).

In some cases, the antigen-binding fragment is present at a concentration of about 3% (w/w).

In some cases, the antigen-binding fragment is present at a concentration of about 10% (w/w).

In some cases, the antigen-binding fragment is present at a concentration of about 30% (w/w).

In some cases, the antigen-binding fragment is present at a concentration of about 40% (w/w).

In some cases, the antigen-binding fragment is present at a concentration of about 80% (w/w).

In some cases, the pharmaceutical composition has a total solid content of 20 mg, and the pharmaceutical composition comprises 0.2 mg, 0.4 mg, 0.6 mg, 2 mg, 6 mg, 8 mg, or 16 mg of the antigen-binding fragment. In such cases, the pharmaceutical composition may comprise 0.4 mg, 2 mg, or 8 mg of the antigen-binding fragment.

In some cases, the antigen-binding fragment comprises: a. HCDR1 comprising the amino acid sequence of SEQ ID NO: 1; b. HCDR2 comprising the amino acid sequence of SEQ ID NO: 2; c. HCDR3 comprising the amino acid sequence of SEQ ID NO: 3; d. LCDR1 comprising the amino acid sequence of SEQ ID NO: 5; e. LCDR2 comprising the amino acid sequence of SEQ ID NO: 6; and f. LCDR3 comprising the amino acid sequence of SEQ ID NO: 7.

In some cases, the antigen-binding fragment comprises: a. HCDR1 having the amino acid sequence of SEQ ID NO: 1; b. HCDR2 having the amino acid sequence of SEQ ID NO: 2; c. HCDR3 having the amino acid sequence of SEQ ID NO: 3; d. LCDR1 having the amino acid sequence of SEQ ID NO: 5; e. LCDR2 having the amino acid sequence of SEQ ID NO: 6; and f. LCDR3 having the amino acid sequence of SEQ ID NO: 7.

In some cases, the antigen-binding fragment comprises: a. HCDR1 consisting of the amino acid sequence of SEQ ID NO: 1; b. HCDR2 consisting of the amino acid sequence of SEQ ID NO: 2; c. HCDR3 consisting of the amino acid sequence of SEQ ID NO: 3; d. LCDR1 consisting of the amino acid sequence of SEQ ID NO: 5; e. LCDR2 consisting of the amino acid sequence of SEQ ID NO: 6; and f. LCDR3 consisting of the amino acid sequence of SEQ ID NO: 7.

In some cases, the antigen-binding fragment comprises a VH domain comprising a sequence at least 95%, 90%, 85%, or 80% identical to SEQ ID NO:4, and a VL domain comprising a sequence at least 95%, 90%, 85%, or 80% identical to SEQ ID NO:8.

In some cases, the antigen-binding fragment comprises a VH domain comprising the sequence of SEQ ID NO: 4; and a VL domain comprising the sequence of SEQ ID NO: 8.

In some cases, the antigen-binding fragment is Fab, Fab', F(ab')2, scFv, minibody, or diabody.

In some cases, the antigen binding fragment is Fab.

In some cases, the Fab is an IgG1 antibody.

In some cases, the antigen-binding fragment comprises a first subunit having the sequence shown in SEQ ID NO: 28 and a second subunit having the sequence shown in SEQ ID NO: 29.

In some cases, the glass stabilizer is selected from trehalose, sucrose, raffinose, inulin, dextran, mannitol, and cyclodextrin.

In some cases, the glass stabilizer is trehalose.

In some cases, the percentage (w/w) concentration of trehalose is made up to about 100%.

In some cases, the composition does not contain a surfactant.

In some cases, the pharmaceutical composition comprises: a. 2% (w/w) antigen-binding fragment ± 20%, 2% (w/w) trileucine ± 10%, 10.5% (w/w) leucine ± 10%, 84.2% (w/w) trehalose ± 10%, and 1.3% (w/w) histidine ± 10%, pH 5.5; b. 2% (w/w) antigen-binding fragment ± 20%, 2% (w/w) trileucine ± 10%, 10.5% (w/w) leucine ± 10%, 82.36% (w/w) trehalose ± 10%, and 3.14% (w/w) histidine ± 10%, pH 5.5; c. 2% (w/w) antigen-binding fragment ± 20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 80.5% (w/w) trehalose ±10%, and 5.0% (w/w) histidine ±10%, pH 5.5; d. 10% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 76.2% (w/w) trehalose ±10%, and 1.3% (w/w) histidine ±10%, pH 5.5; e. 10% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 74.36% (w/w) trehalose ±10%, and 3.14% (w/w) histidine ±10%, pH 5.5; f. 10% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 72.5% (w/w) trehalose ±10%, and 5.0% (w/w) histidine ±10%, pH 5.5; g. 40% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 44.36% (w/w) trehalose ±10%, and 3.14% (w/w) histidine ±10%, pH 5.5; h. 40% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 42.5% (w/w) trehalose ±10%, and 5.0% (w/w) histidine ±10%, pH 5.5; or i. 40% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 46.2% (w/w) trehalose ±10%, and 1.3% (w/w) histidine ±10%, pH 5.5.

In some cases, the pharmaceutical composition comprises: a. 0.4 mg of an antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 84.2% (w/w) trehalose ±10%, and 1.3% (w/w) histidine ±10%, pH 5.5; b. 0.4 mg of an antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 82.36% (w/w) trehalose ±10%, and 3.14% (w/w) histidine ±10%, pH 5.5; c. 0.4 mg of an antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 80.5% (w/w) trehalose ±10% and 5.0% (w/w) histidine ±10%, pH 5.5; d. 2 mg antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 76.2% (w/w) trehalose ±10%, and 1.3% (w/w) histidine ±10%, pH 5.5; e. 2 mg antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 74.36% (w/w) trehalose ±10%, and 3.14% (w/w) histidine ±10%, pH 5.5; f. 2 mg antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 72.5% (w/w) trehalose ±10%, and 5.0% (w/w) histidine ±10%, pH 5.5; g. 8 mg antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 44.36% (w/w) trehalose ±10%, and 3.14% (w/w) histidine ±10%, pH 5.5; h. 8 mg antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 42.5% (w/w) trehalose ±10%, and 5.0% (w/w) histidine ±10%, pH 5.5; or i. 8 mg antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 46.2% (w/w) trehalose ±10%, and 1.3% (w/w) histidine ±10%, pH 5.5.

When the pharmaceutical composition comprises approximately 3.14% (w/w) histidine, the pharmaceutical composition may comprise approximately 0.55% (w/w) L-histidine and approximately 2.59% (w/w) histidine HCl. In some instances, when the pharmaceutical composition comprises 3.14% (w/w) histidine, the pharmaceutical composition may comprise 0.55% (w/w) L-histidine and 2.59% (w/w) histidine HCl.

In some cases, after recovery, the number of subvisible particles between 5 μm and 200 μm was less than about 2.5 × 10 ⁴ particles/ml.

In some cases, after recovery, the number of subvisible particles between 5 μm and 200 μm was less than about 0.5×10 ⁴ /ml.

In some cases, after recovery, the number of subvisible particles between 10 μm and 200 μm was less than about 1 × 10 ⁴ particles/ml.

In some cases, after recovery, the number of subvisible particles between 10 μm and 200 μm was less than about 0.2 × 10 ⁴ particles/ml.

In some cases, after reconstitution, the number of subvisible particles between 25 μm and 200 μm is less than about 2×10 ³ /ml. For example, after reconstitution, the number of subvisible particles between 25 μm and 200 μm is less than about 0.2×10 ³ /ml.

In some cases, the number of subvisible particles was determined by dynamic flow imaging microscopy, optionally by microfluidic imaging (MFI).

In some cases, the number of subvisible particles was determined after reconstitution in water to a concentration of 2.5 mg/ml or 30 mg/ml of the antigen-binding fragment.

In a second aspect, the present disclosure provides a process for preparing a pharmaceutical composition for inhalation, comprising: a. providing an aqueous solution at about pH 5 to about pH 6, comprising leucine, trileucine, histidine, a glass stabilizer, and an antigen-binding fragment of an anti-thymic stromal lymphopoietin (TSLP) antibody; b. spray-drying the aqueous solution in (a) to produce dry powder particles; and c. collecting the dry powder particles; wherein the aqueous solution comprises about 5% (w/w) to about 15% (w/w) leucine, about 1% (w/w) to about 5% (w/w) trileucine, about 1% (w/w) to about 10% (w/w) histidine, about 5% (w/w) to about 50% (w/w) antigen-binding fragment, and a certain % (w/w) of glass stabilizer to achieve 100% total solids content.

In some cases, the aqueous solution has a pH of 5.5.

In some cases, the glass stabilizer is trehalose.

In some cases, the aqueous solution comprises: a. 2% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 84.2% (w/w) trehalose ±10%, and 1.3% (w/w) histidine ±10%, pH 5.5; b. 2% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 82.36% (w/w) trehalose ±10%, and 3.14% (w/w) histidine ±10%, pH 5.5; c. 2% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) (w/w) leucine ±10%, 80.5% (w/w) trehalose ±10%, and 5.0% (w/w) histidine ±10%, pH 5.5; d. 10% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 76.2% (w/w) trehalose ±10%, and 1.3% (w/w) histidine ±10%, pH 5.5; e. 10% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 74.36% (w/w) trehalose ±10%, and 3.14% (w/w) histidine ±10%, pH 5.5; f. 10% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 72.5% (w/w) trehalose ±10%, and 5.0% (w/w) histidine ±10%, pH 5.5; g. 40% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 44.36% (w/w) trehalose ±10%, and 3.14% (w/w) histidine ±10%, pH 5.5; h. 40% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 42.5% (w/w) trehalose ±10% and 5.0% (w/w) histidine ±10%, pH 5.5; or i. 40% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 46.2% (w/w) trehalose ±10%, and 1.3% (w/w) histidine ±10%, pH 5.5.

In a third aspect, the present disclosure provides a dry powder formulation obtained by the process of the second aspect of the present disclosure.

In a fourth aspect, the present disclosure provides a method for treating a TSLP-related disease in a subject in need thereof, comprising administering to the subject a pharmaceutical composition of the first aspect or a dry powder formulation of the third aspect of the present disclosure. The present disclosure also provides a pharmaceutical composition of the first aspect or a dry powder formulation of the third aspect for treating a TSLP-related disease. Furthermore, the present disclosure provides a use of the pharmaceutical composition of the first aspect or the dry powder formulation of the third aspect for manufacturing a medicament for treating a TSLP-related disease.

In some cases, the TSLP-associated condition is asthma, COPD, allergic rhinitis, allergic sinusitis, allergic conjunctivitis, eosinophilic esophagitis, chronic idiopathic urticaria, or chronic sinusitis.

In some cases, the TSLP-related condition is asthma.

In some cases, the TSLP-related condition is COPD.

In a fifth aspect, the present disclosure provides a method for improving lung function in a subject suffering from asthma or COPD, the method comprising administering to the subject a pharmaceutical composition of the first aspect or a dry powder formulation of the third aspect of the present disclosure. The present disclosure also provides the pharmaceutical composition of the first aspect or the dry powder formulation of the third aspect for use in a method for improving lung function in a subject suffering from asthma or COPD. Furthermore, the present disclosure provides the use of the pharmaceutical composition of the first aspect or the dry powder formulation of the third aspect for manufacturing a medicament for improving lung function in a subject suffering from asthma or COPD.

In some instances, improvement in lung function refers to improvement compared to baseline in one or more of the following parameters: (i) pre-bronchodilator (BD) FVC, (ii) post-BD FVC, (iii) pre-BD _FEV1 , (iv) post-BD _FEV1 , (v) mean morning PEF, and/or (vi) mean evening PEF.

In a sixth aspect, the present disclosure relates to a method for ameliorating symptoms of asthma or COPD in a subject in need thereof, the method comprising administering to the subject the pharmaceutical composition of the first aspect or the dry powder formulation of the third aspect of the present disclosure.

In some instances, improvement in asthma symptoms means an improvement compared to baseline in one of the following parameters: (i) mean Asthma Symptom Diary score, (ii) Asthma Control Questionnaire 6 (ACQ-6) score, (iii) Asthma Quality of Life Questionnaire (AQLQ) score, and/or (iv) St. George's Respiratory Questionnaire (SGRQ) score.

In some instances of any of the fourth, fifth, and sixth aspects of the disclosure, the asthma is moderate to severe asthma.

In some cases of any one of the fourth, fifth, and sixth aspects of the present disclosure, the pharmaceutical composition or dry powder formulation is administered by inhalation or intranasally. In some cases of any one of the fourth, fifth, and sixth aspects of the present disclosure, the pharmaceutical composition is administered by a dry powder inhaler.

In some aspects of any of the fourth, fifth, and sixth aspects of the present disclosure, the pharmaceutical composition or dry powder formulation to be administered to a subject comprises an antigen-binding fragment of an anti-TSLP antibody in an amount of about 0.2 mg to about 16 mg. In some aspects of any of the fourth, fifth, and sixth aspects of the present disclosure, the pharmaceutical composition or dry powder formulation to be administered to a subject comprises an antigen-binding fragment of an anti-TSLP antibody in an amount of about 0.4 mg to about 8 mg. In some aspects of any of the fourth, fifth, and sixth aspects of the present disclosure, the pharmaceutical composition or dry powder formulation administered or intended to be administered to a subject comprises an antigen-binding fragment of an anti-TSLP antibody in an amount of about 0.4 mg, about 2 mg, or about 8 mg.

In some instances of any of the fourth, fifth, and sixth aspects of the disclosure, the asthma is uncontrolled moderate to severe asthma.

In some cases of any of the fourth, fifth, and sixth aspects of the disclosure, the pharmaceutical composition or dry powder formulation is administered or is to be administered to a subject by inhalation once a day (Q1D).

This disclosure includes any combination of described aspects and features unless such combination is expressly disallowed or expressly avoided.

Aspects and embodiments of the present disclosure will now be discussed with reference to the accompanying drawings. Other aspects and embodiments will be apparent to those skilled in the art. All documents mentioned herein are incorporated herein by reference.

Pharmaceutical compositions As described herein, pharmaceutical compositions for stabilizing and delivering pharmaceutically active agents are provided. Suitably, the pharmaceutical compositions are formulated for pulmonary delivery, including via inhalation via a dry powder inhaler (DPI). Thus, the pharmaceutical compositions include "dry powder formulations."

The pharmaceutical compositions provided herein comprise spray-dried particles comprising: about 5% (w/w) to about 15% (w/w) leucine; about 1% (w/w) to about 5% (w/w) trileucine; about 1% (w/w) to about 10% (w/w) histidine buffer, at a pH between about pH 5 and pH 6; about 1% (w/w) to about 80% (w/w) an antigen-binding fragment of an anti-thymic stromal lymphopoietin (TSLP) antibody; and a glass stabilizer.

As used herein, a "pharmaceutical composition" refers to a preparation that is in a form that allows the biological activity of the active ingredient (e.g., an anti-TSLP Fab disclosed herein) to be effective and does not contain additional components that are unacceptably toxic to the subject to which the composition is administered. Such compositions may be sterile.

As used herein, "spray-dried particles" refers to particles produced in a process that uses an aerosol phase to spray-dry particles to form a desiccant-based formulation. Thus, spray-dried particles refer to a plurality of solid microparticles in a powder composition that suitably contains less than about 20% water, more suitably less than 10% water, less than about 5-6% water, or less than about 3% water. As described herein, the pharmaceutical composition can be used for delivery to a patient via inhalation. In other cases, the pharmaceutical composition can be reconstituted and administered orally, intravenously, parenterally, or the like in liquid form. An exemplary process for preparing spray-dried particles according to the examples herein can be performed as follows. An atomizer is used to atomize a liquid raw material containing the desired final components of the dry powder formulation into a fine mist. The mist is then dried as described herein. The atomized droplets contain the dissolved components, which are initially in the form of droplets. As the droplets dry, the different components of the formulation begin to saturate and precipitate at different rates. As described herein, a shell begins to form around the outer surface of the particles of the dry powder formulation. This shell suitably includes leucine and trileucine components located on the outer surface of the shell. It should be noted that leucine and trileucine become preferentially located on the outer surface of the particles, while smaller amounts of leucine and trileucine can also be found throughout the particles. In some cases, higher concentrations of leucine and trileucine are suitably found at or near the surface of the particles, rather than near the center of the particles. In some cases, the center of the particle contains a substantial amount of the active agent, as well as other excipient components as described herein, suitably in amorphous form. As used herein, "substantial amount" of the active agent means that at least about 60% of the active agent (i.e., the total active agent in the formulation) is located at or near the center of the microparticle, suitably at least about 70%, and more suitably at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, and in some cases about 95%-100% of the active agent is located at or near the center of the microparticle.

As used herein, "microparticles" refer to solid particles having a mass mean diameter (MMD) of less than 20 μm. The mass mean diameter is a measure of the average particle size of the microparticles, which is measured using a suitable method (including, for example, centrifugal sedimentation, electron microscopy, light scattering, laser diffraction, etc.).

Unless otherwise indicated, "active agent" refers to an antigen-binding fragment derived from an anti-TSLP antibody as defined herein.

Unless otherwise indicated, ratios described herein are expressed as weight % (w/w, also referred to as "weight ratios"). Unless otherwise indicated, the amounts of leucine, trileucine, histidine buffer, and antigen-binding fragments of anti-TSLP antibodies provided herein are provided as weight percent (wt%) of the formulation. Because the pharmaceutical compositions described herein contain substantially little, if any, water, the weight percentages of the pharmaceutical compositions are percentages based on the dry weight of the final formulation.

These ratios are achieved by providing the desired mg/mL concentrations of these components in the feedstock and then spray drying to remove the feedstock solvent to obtain aerosolized microparticles where the initial concentration ratios (expressed in mg/mL) are maintained as the final weight ratios.

As used herein, "leucine" refers to the amino acid leucine (C6H13NO2), whether present as a single amino acid or as an amino acid component of a peptide, which may be a racemic mixture or in its D or L form, as well as modified forms of leucine (i.e., in which one or more atoms of leucine have been substituted with another atom or functional group). The chemical structure of leucine is provided below:

As used herein, "trileucine" refers to a compound in which three leucine molecules are linked together in a peptide, such as leucine-leucine-leucine (Leu-Leu-Leu), C18H35N3O4. The chemical structure of trileucine is provided below:

Exemplary weight percentages of leucine and trileucine that can be used in pharmaceutical compositions to achieve a desired ratio are described herein. Suitably, the dry powder formulation comprises about 5% to about 15% leucine and about 1% to about 5% trileucine. The pharmaceutical compositions described herein comprise about 5% to about 15% leucine by weight, more suitably about 5% to about 14%, about 5% to about 13%, about 5% to about 12%, about 5% to about 11%, about 5% to about 10%, about 5% to about 9%, about 5% to about 8%, about 5% to about 7%, about 5% to about 6%, about 6% to about 15%, about 7% to about 15%, about 8% to about 15%, about 9% to about 15%, about 10% to about 15%, about 11% to about 15%, about 12% to about 15%, about 13% to about 15% by weight. to about 15%, about 14% to about 15%, about 5% to about 14%, about 6% to about 14%, about 7% to about 14%, about 8% to about 14%, about 9% to about 14%, about 10% to about 14%, about 11% to about 14%, about 12% to about 14%, about 13% to about 14%, about 5% to about 13%, about 6% to about 13%, about 7% to about 13%, about 8% to about 13%, about 9% to about 13%, about 10% to about 13%, about 11% to about 13%, about 11% to about 13%, about 12% to about 1 3%, about 5% to about 12%, about 6% to about 12%, about 7% to about 12%, about 8% to about 12%, about 9% to about 12%, about 10% to about 12%, about 11% to about 12%, about 5% to about 11%, about 6% to about 11%, about 7% to about 11%, about 8% to about 11%, about 9% to about 11%, about 10% to about 11%, about 5% to about 10%, about 6% to about 10%, about 7% to about 10%, about 8% to about 10%, about 9% to about 10%, about 5% to about 9%, about 6% to about 9% , about 7% to about 9%, about 8% to about 9%, about 5% to about 8%, about 6% to about 8%, about 7% to about 8%, about 5% to about 7%, about 6% to about 7%, about 5% to about 6%, about 5%, about 5.5%, about 6%, about 6.5%, about 7%, about 7.5%, about 8%, about 8.5%, about 9%, about 9.5%, about 10%, about 10.5%, about 11%, about 11.5%, about 12%, about 12.5%, about 13%, about 13.5%, about 14%, about 14.5%, about 15% leucine. In some instances, the pharmaceutical compositions described herein comprise about 10.5% leucine by weight. In some cases, the pharmaceutical compositions described herein comprise 10.5% leucine ± 1%, 10.5% leucine ± 2%, 10.5% leucine ± 3%, 10.5% leucine ± 4%, 10.5% leucine ± 5%, or 10.5% leucine ± 10% by weight.

The pharmaceutical composition comprises about 1% to about 5% trileucine by weight, more suitably about 1% to about 4%, about 1% to about 3%, about 1% to about 2%, about 2% to about 4%, about 2% to about 3%, about 3% to about 5%, about 3% to about 4%, about 4% to about 5%, about 1%, about 3%, about 4%, about 5% trileucine by weight. In some instances, the pharmaceutical compositions described herein comprise about 2% leucine by weight. In some instances, the pharmaceutical compositions described herein comprise 2% leucine ± 0.5%, 2% leucine ± 1%, 2% leucine ± 2%, 2% leucine ± 3%, 2% leucine ± 4%, 2% leucine ± 5%, 2% leucine ± 10% by weight.

In some cases, the pharmaceutical composition comprises about 8% to about 12% leucine and about 1% to about 3% trileucine, and in some cases, the pharmaceutical composition comprises about 10.5% leucine and about 2% trileucine.

In other cases, the particles contain leucine and trileucine located substantially throughout the particle, but with higher amounts at or near the particle surface. As used herein, "substantially throughout the particle" means that leucine and/or trileucine are located in a gradient from the outer surface of the particle toward the center of the particle, but preferably, the amount of leucine and/or trileucine decreases as one moves toward the center, and in some cases, no leucine or trileucine is found at the center of the particle where the active agent is located. In other cases, the amount of leucine and trileucine may be substantially uniform across the cross-section of the particle.

In some cases, substantially all of the particles of the pharmaceutical composition contain leucine and trileucine. That is, suitably, at least about 60% of the particles contain leucine and trileucine, or at least about 70%, and more suitably, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, and in some cases, about 95%-100% of the particles contain leucine and trileucine. In some cases, each of the particles of the dry powder formulation contains leucine and trileucine.

In some cases, leucine and/or trileucine may be present in the pharmaceutical composition but not within or complexed with the microparticles of the formulation. Thus, in some cases, free leucine and/or trileucine that is not complexed with the microparticles may be present in the dry powder formulation. However, generally, the amount of free leucine and/or trileucine (i.e., not complexed with the microparticles) is less than about 10%, less than about 5%, less than about 1%, and more preferably less than about 0.1% of the total amount of leucine and/or trileucine in the formulation.

"Histidine," whether present as a single amino acid or as an amino acid component of a peptide, refers to the amino acid histidine (C6H9N3O2), which may be a racemic mixture or its D- or L-form, as well as modified forms of histidine (i.e., in which one or more atoms of leucine have been replaced by another atom or functional group). The chemical structure of histidine is provided below.

As described herein, it has been surprisingly discovered that pharmaceutical compositions comprising a histidine buffer significantly reduce protein aggregation and device and throat deposits compared to a reference buffer, at least in part by reducing the level of subvisible particles. Accordingly, the pharmaceutical compositions described herein comprise a histidine buffer. Suitably, the histidine buffer is present at about 1% to about 10% by weight, for example, the histidine buffer may be present at about 1% to about 9% by weight, about 1% to about 8% by weight, about 1% to about 7% by weight, about 1% to about 6% by weight, about 1% to about 5% by weight, about 1% to about 4% by weight, about 1% to about 3% by weight, about 1% to about 2% by weight, about 2% to about 10% by weight, about 2% to about 9% by weight, about 2% to about 8% by weight, about 2% to about 7 ... % to about 6% by weight, about 2% to about 5% by weight, about 2% to about 4% by weight, about 2% to about 3% by weight, about 2.% to about 3.5% by weight, or about 1% by weight, about 1.5% by weight, about 2% by weight, about 2.5% by weight, about 3% by weight, about 3.5% by weight, about 4% by weight, about 4.5% by weight, about 5% by weight, about 5.5% by weight, or about 6%, about 6.5% by weight, about 7% by weight, about 7.5% by weight, about 8% by weight, about 8.5% by weight, about 9% by weight, about 9.5% by weight. In some instances, the pharmaceutical composition comprises about 3.14% by weight of a histidine buffer. In some instances, the pharmaceutical compositions described herein comprise 3.14% histidine buffer ± 0.5%, 3.14% histidine buffer ± 1%, 3.14% histidine buffer ± 2%, 3.14% histidine buffer ± 3%, 3.14% histidine buffer ± 4%, 3.14% histidine buffer ± 5%, 3.14% histidine buffer ± 10% by weight. In some instances, the pharmaceutical composition comprises approximately 3.14% histidine buffer ± 10% by weight (w/w). In some cases, the pharmaceutical composition comprises approximately 3.14% histidine buffer by weight (w/w). In some cases, the pharmaceutical composition comprises approximately 0.55% (w/w) L-histidine and 2.59% (w/w) histidine HCl.

As used herein, a "reference buffer" is a buffer that does not contain histidine in the ratios described herein. For example, the reference buffer may not contain any histidine buffer. In some cases, the reference buffer comprises citrate. In some cases, the reference buffer comprises trehalose, leucine, trileucine, and citrate (TLTC). In some cases, the reference buffer has a pH of about pH 6. In some cases, the reference buffer comprises a surfactant, such as polysorbate 80 (PS80).

The histidine buffer also provides control over the pH of the pharmaceutical composition, suitably maintaining a pH of about pH 5.5 to about pH 6 (e.g., about pH 5.5, about pH 5.6, about pH 5.7, about pH 5.8, about pH 5.9). In certain instances, the pH of the histidine buffer is 5.5.

The compositions described herein comprise antigen-binding fragments of anti-thymic stromal lymphopoietin (anti-TSLP) antibodies. Advantageously, the inventors have discovered that the formulations described herein enable direct delivery of the antigen-binding fragments to the lungs via inhalation. Delivery of therapeutically active antigen-binding fragments of anti-TSLP antibodies via inhalation allows for the use of biopharmaceuticals to treat asthma in primary care settings. Suitably, the antigen-binding fragment is present at about 1% to about 80% by weight (w/w), for example, the antigen-binding fragment may be present at about 1% to about 50% by weight (w/w), about 1% to about 45% by weight (w/w), about 1% to about 40% by weight (w/w), about 2% to about 50% by weight (w/w), 2% to about 45% by weight (w/w), or about 2% to about 40% by weight (w/w).

In some cases, the antigen-binding fragment is present in the pharmaceutical composition at a concentration of about 1% to about 10% by weight (w/w), about 1% to about 9% by weight (w/w), about 1% to about 8% by weight (w/w), about 1% to about 7% by weight (w/w), about 1% to about 6% by weight (w/w), about 1% to about 5% by weight (w/w), about 1% to about 4% by weight (w/w), about 1% to about 3% by weight (w/w), about 2% to about 10% by weight (w/w), about 2% to about 9% by weight (w/w), about 2% to about 8% by weight (w/w), about 2% to about 7% by weight (w/w), about 2% to about 6% by weight (w/w), about 2% to about 5% by weight (w/w), about 2% to about 4% by weight (w/w), about 2 wt % to about 3 wt % (w/w), about 3 wt % to about 10 wt % (w/w), about 3 wt % to about 9 wt % (w/w), about 3 wt % to about 8 wt % (w/w), about 3 wt % to about 7 wt % (w/w), about 3 wt % to about 6 wt % (w/w), about 3 wt % to about 5 wt % (w/w), about 3 wt % to about 4 wt % (w/w), about 4 wt % to about 10 wt % (w/w), about 4 wt % to about 9 wt % (w/w), about 4 wt % to about 8 wt % (w/w), about 4 wt % to about 7 wt % (w/w), about 4 wt % to about 6 wt % (w/w), about 4 wt % to about 5 wt % (w/w), about 5 wt % to about 10 wt % (w/w), about 5 wt % to about 9 wt % (w/w), about 5 wt % to about 8 wt % % to about 30 wt % (w/w), about 10 wt % to about 40 wt % (w/w), about 15 wt % to about 25 wt % (w/w), about 25 wt % to about 35 wt % (w/w), or about 35 wt % to about 45 wt % (w/w).

In some instances, the antigen-binding fragment is present at about 1 wt % (w/w), about 2 wt % (w/w), about 3 wt % (w/w), about 4 wt % (w/w), about 5 wt % (w/w), about 6 wt % (w/w), about 7 wt % (w/w), about 8 wt % (w/w), about 9 wt % (w/w), about 10 wt % (w/w), about 15 wt % (w/w), about 20 wt % (w/w), about 25 wt % (w/w), about 30 wt % (w/w), about 35 wt % (w/w), about 40 wt % (w/w), about 45 wt % (w/w), about 50 wt % (w/w), about 55 wt % (w/w), about 60 wt % (w/w), about 65 wt % (w/w), about 70 wt % (w/w), about 75 wt % (w/w), or about 80 wt %. In some cases, the antigen-binding fragment is present in the pharmaceutical composition at a concentration of about 1% (w/w) to about 80% (w/w) by weight (e.g., about 1% (w/w) to about 40% (w/w) by weight). In some cases, the antigen-binding fragment is present in the pharmaceutical composition at a concentration of about 1% (w/w), about 2% (w/w), about 3% (w/w), about 10% (w/w), about 15% (w/w), about 20% (w/w), about 25% (w/w), about 30% (w/w), about 35% (w/w), about 40% (w/w), about 45% (w/w), about 50% (w/w), about 55% (w/w), about 60% (w/w), about 65% (w/w), about 70% (w/w), about 75% (w/w), or about 80% (w/w).

The pharmaceutical compositions described herein further comprise a glass stabilizer to help stabilize the formulation, and in particular, to stabilize the active agent. A "glass stabilizer" refers to a preservative that stabilizes the active agent (suitably a polypeptide) in a dry powder formulation, preferably by displacing water at the active agent's surface during drying or otherwise hindering degradation processes, and forms an amorphous solid comprising the active agent. Examples of glass stabilizers include amorphous sugars, polymeric sugars, buffers, salts, or synthetic polymers (e.g., poly-L-glycolic acid), as well as mixtures of such components. In some cases, the glass stabilizer is an amorphous sugar. In additional cases, the glass stabilizer is a buffer. In other cases, the formulations described herein may include both an amorphous sugar and a buffering agent, which may together or individually act as glass stabilizers.

Exemplary amorphous sugars for use in the compositions described herein include, but are not limited to, trehalose, sucrose, raffinose, inulin, dextran, mannitol, and cyclodextrin. Suitably, the amorphous sugar is present in an amount of about 30% to about 70% (by weight) of the dry powder formulation. In other cases, the amorphous sugar is present in an amount of about 30% to about 65%, about 35% to about 65%, about 35% to about 60%, about 40% to about 60%, about 30% to about 50%, or about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, or about 60%. Suitably, the amorphous sugar is trehalose and is present in the formulation in an amount of about 30%-60%, more suitably about 35%-55%, or about 35%, about 40%, about 45%, or about 50% by weight of the dry powder formulation.

Compositions and formulations that consist essentially of the listed ingredients contain the listed ingredients plus any other ingredients that do not materially affect the basic and novel characteristics of the claimed formulation. Ingredients that do not materially affect the basic and novel characteristics of the claimed formulation are those that do not limit the ability of leucine and trileucine to stabilize the dry powder formulation. Suitably, compositions and formulations consisting essentially of the listed ingredients specifically exclude other amino acids or tripeptide amino acids but may include additional sugars, buffers, etc.

In an exemplary embodiment, a pharmaceutical composition is provided, comprising: a. 1% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 85.2% (w/w) trehalose ±10%, and 1.3% (w/w) histidine ±10%, pH 5.5; b. 1% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 83.36% (w/w) trehalose ±10%, and 3.14% (w/w) histidine ±10%, pH 5.5; c. 1% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 83.36% (w/w) trehalose ±10%, and 3.14% (w/w) histidine ±10%. (w/w) leucine ±10%, 81.5% (w/w) trehalose ±10%, and 5.0% (w/w) histidine ±10%, pH 5.5; d. 2% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 84.2% (w/w) trehalose ±10%, and 1.3% (w/w) histidine ±10%, pH 5.5; e. 2% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 82.36% (w/w) trehalose ±10%, and 3.14% (w/w) histidine ±10%, pH 5.5; f. 2% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 80.5% (w/w) trehalose ±10%, and 5.0% (w/w) histidine ±10%, pH 5.5; g. 3% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 83.2% (w/w) trehalose ±10%, and 1.3% (w/w) histidine ±10%, pH 5.5; h. 3% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 81.36% (w/w) trehalose ±10% and 3.14% (w/w) histidine ±10%, pH 5.5; i. 3% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 79.5% (w/w) trehalose ±10%, and 5.0% (w/w) histidine ±10%, pH 5.5; j. 10% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 76.2% (w/w) trehalose ±10%, and 1.3% (w/w) histidine ±10%, pH 5.5; k. 10% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 74.36% (w/w) trehalose ±10%, and 3.14% (w/w) histidine ±10%, pH 5.5; l. 10% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 72.5% (w/w) trehalose ±10%, and 5.0% (w/w) histidine ±10%, pH 5.5; m. 30% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 54.36% (w/w) trehalose ±10%, and 3.14% (w/w) histidine ±10%, pH 5.5; n. 30% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 52.5% (w/w) trehalose ±10%, and 5.0% (w/w) histidine ±10%, pH 5.5; o. 30% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 56.2% (w/w) trehalose ±10%, and 1.3% (w/w) histidine ±10%, pH 5.5; p. 40% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 44.36% (w/w) trehalose ±10%, and 3.14% (w/w) histidine ±10%, pH 5.5; q. 40% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 42.5% (w/w) trehalose ±10%, and 5.0% (w/w) histidine ±10%, pH 5.5; r. 40% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 46.2% (w/w) trehalose ±10%, and 1.3% (w/w) histidine ±10%, pH 5.5; s. 80% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 4.36% (w/w) trehalose ±10%, and 3.14% (w/w) histidine ±10%, pH 5.5; t. 80% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 2.5% (w/w) trehalose ±10%, and 5.0% (w/w) histidine ±10%, pH 5.5; or u. 80% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 6.2% (w/w) trehalose ±10% and 1.3% (w/w) histidine ±10%, pH 5.5.

In some cases, a pharmaceutical composition is provided comprising: a. 1% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 85.2% (w/w) trehalose, and 1.3% (w/w) histidine, pH 5.5; b. 1% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 83.36% (w/w) trehalose, and 3.14% (w/w) histidine, pH 5.5; c. 1% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 81.5% (w/w) trehalose, and 5.0% (w/w) histidine, pH 5.5; d. 2% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 84.2% (w/w) trehalose, and 1.3% (w/w) histidine, pH 5.5; e. 2% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 82.36% (w/w) trehalose, and 3.14% (w/w) histidine, pH 5.5; f. 2% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 80.5% (w/w) trehalose, and 5.0% (w/w) histidine, pH 5.5; g. 3% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 83.2% (w/w) trehalose, and 1.3% (w/w) histidine, pH 5.5; h. 3% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 81.36% (w/w) trehalose, and 3.14% (w/w) histidine, pH 5.5; i. 3% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 79.5% (w/w) trehalose, and 5.0% (w/w) histidine, pH 5.5; j. 10% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 76.2% (w/w) trehalose, and 1.3% (w/w) histidine, pH 5.5; k. 10% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 74.36% (w/w) trehalose, and 3.14% (w/w) histidine, pH 5.5; l. 10% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 72.5% (w/w) trehalose, and 5.0% (w/w) histidine, pH 5.5; m. 30% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 54.36% (w/w) (w/w) trehalose and 3.14% (w/w) histidine, pH 5.5; n. 30% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 52.5% (w/w) trehalose and 5.0% (w/w) histidine, pH 5.5; o. 30% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 56.2% (w/w) trehalose and 1.3% (w/w) histidine, pH 5.5; p. 40% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 44.36% (w/w) trehalose and 3.14% (w/w) histidine, pH 5.5; q. 40% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 42.5% (w/w) trehalose, and 5.0% (w/w) histidine, pH 5.5; r. 40% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 46.2% (w/w) trehalose, and 1.3% (w/w) histidine, pH 5.5; s. 80% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 4.36% (w/w) trehalose, and 3.14% (w/w) histidine, pH 5.5; t. 80% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 2.5% (w/w) trehalose, and 5.0% (w/w) histidine, pH 5.5; or u. 80% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 6.2% (w/w) trehalose, and 1.3% (w/w) histidine, pH 5.5.

In some cases, a pharmaceutical composition is provided that comprises 1% (w/w) antigen-binding fragment ± 20%, 2% (w/w) trileucine ± 10%, 10.5% (w/w) leucine ± 10%, 83.36% (w/w) trehalose ± 10%, and 3.14% (w/w) histidine ± 10%, pH 5.5. In some cases, the pharmaceutical composition comprises 1% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 83.36% (w/w) trehalose, and 3.14% (w/w) histidine, pH 5.5.

In some cases, a pharmaceutical composition is provided that comprises 10% (w/w) antigen-binding fragment ± 20%, 2% (w/w) trileucine ± 10%, 10.5% (w/w) leucine ± 10%, 74.36% (w/w) trehalose ± 10%, and 3.14% (w/w) histidine ± 10%, pH 5.5. In some cases, the pharmaceutical composition comprises 10% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 74.36% (w/w) trehalose, and 3.14% (w/w) histidine, pH 5.5.

In some cases, a pharmaceutical composition is provided that comprises 40% (w/w) antigen-binding fragment ± 20%, 2% (w/w) trileucine ± 10%, 10.5% (w/w) leucine ± 10%, 44.36% (w/w) trehalose ± 10%, and 3.14% (w/w) histidine ± 10%, pH 5.5. In some cases, the pharmaceutical composition comprises 40% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 42.5% (w/w) trehalose, and 5.0% (w/w) histidine, pH 5.5.

In some cases, a pharmaceutical composition is provided comprising: a. 0.2 mg (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 85.2% (w/w) trehalose ±10%, and 1.3% (w/w) histidine ±10%, pH 5.5; b. 0.2 mg (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 83.36% (w/w) trehalose ±10%, and 3.14% (w/w) histidine ±10%, pH 5.5; c. 0.2 mg (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 81.5% (w/w) trehalose ±10%, and 5.0% (w/w) histidine ±10%, pH 5.5; d. 0.4 mg antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 84.2% (w/w) trehalose ±10%, and 1.3% (w/w) histidine ±10%, pH 5.5; e. 0.4 mg antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 82.36% (w/w) trehalose ±10%, and 3.14% (w/w) histidine ±10%, pH 5.5; f. 0.4 mg antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 80.5% (w/w) trehalose ±10%, and 5.0% (w/w) histidine ±10%, pH 5.5; g. 0.6 mg (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 83.2% (w/w) trehalose, and 1.3% (w/w) histidine, pH 5.5; h. 0.6 mg (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 81.36% (w/w) trehalose, and 3.14% (w/w) histidine, pH 5.5; i. 0.6 mg (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 79.5% (w/w) trehalose, and 5.0% (w/w) histidine, pH 5.5; j. 2 mg antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 76.2% (w/w) trehalose ±10%, and 1.3% (w/w) histidine ±10%, pH 5.5; k. 2 mg antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 74.36% (w/w) trehalose ±10%, and 3.14% (w/w) histidine ±10%, pH 5.5; l. 2 mg antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 72.5% (w/w) trehalose ±10%, and 5.0% (w/w) histidine ±10%, pH 5.5; m. 6 mg (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 54.36% (w/w) trehalose, and 3.14% (w/w) histidine, pH 5.5; n. 6 mg (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 52.5% (w/w) trehalose, and 5.0% (w/w) histidine, pH 5.5; o. 6 mg (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 56.2% (w/w) trehalose, and 1.3% (w/w) histidine, pH 5.5; p. 8 mg antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 44.36% (w/w) trehalose ±10%, and 3.14% (w/w) histidine ±10%, pH 5.5; q. 8 mg antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 42.5% (w/w) trehalose ±10% and 5.0% (w/w) histidine ±10%, pH 5.5; r. 8 mg antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 46.2% (w/w) trehalose ±10%, and 1.3% (w/w) histidine ±10%, pH 5.5 s. 16 mg (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 4.36% (w/w) trehalose, and 3.14% (w/w) histidine, pH 5.5; t. 16 mg (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 2.5% (w/w) trehalose and 5.0% (w/w) histidine, pH 5.5; or u. 16 mg (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 6.2% (w/w) trehalose, and 1.3% (w/w) histidine, pH 5.5.

In these cases, the total mass solid content of the pharmaceutical composition is 20 mg.

In some cases, the pharmaceutical composition comprises: a. 0.2 mg (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 85.2% (w/w) trehalose, and 1.3% (w/w) histidine, pH 5.5; b. 0.2 mg (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 83.36% (w/w) trehalose, and 3.14% (w/w) histidine, pH 5.5; c. 0.2 mg (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 81.5% (w/w) trehalose, and 5.0% (w/w) histidine, pH 5.5; d. 0.4 mg antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 84.2% (w/w) trehalose, and 1.3% (w/w) histidine, pH 5.5; e. 0.4 mg antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 82.36% (w/w) trehalose, and 3.14% (w/w) histidine, pH 5.5; f. 0.4 mg antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 80.5% (w/w) trehalose, and 5.0% (w/w) histidine, pH 5.5; g. 0.6 mg (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 83.2% (w/w) trehalose, and 1.3% (w/w) histidine, pH 5.5; h. 0.6 mg (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 81.36% (w/w) trehalose, and 3.14% (w/w) histidine, pH 5.5; i. 0.6 mg (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 79.5% (w/w) trehalose, and 5.0% (w/w) histidine, pH 5.5; j. 2 mg antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 76.2% (w/w) trehalose, and 1.3% (w/w) histidine, pH 5.5; k. 2 mg antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 74.36% (w/w) trehalose, and 3.14% (w/w) histidine, pH 5.5; l. 2 mg antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 72.5% (w/w) trehalose, and 5.0% (w/w) histidine, pH 5.5; m. 6 mg (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 54.36% (w/w) trehalose, and 3.14% (w/w) histidine, pH 5.5; n. 6 mg (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 52.5% (w/w) trehalose, and 5.0% (w/w) histidine, pH 5.5; o. 6 mg (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 56.2% (w/w) trehalose, and 1.3% (w/w) histidine, pH 5.5; p. 8 mg antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 44.4% (w/w) trehalose, and 3.14% (w/w) histidine, pH 5.5; q. 8 mg antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 42.5% (w/w) trehalose, and 5.0% (w/w) histidine, pH 5.5; r. 8 mg antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 46.2% (w/w) trehalose, and 1.3% (w/w) histidine, pH 5.5; or s. 16 mg (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 4.36% (w/w) trehalose, and 3.14% (w/w) histidine, pH 5.5; t. 16 mg (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 2.5% (w/w) trehalose, and 5.0% (w/w) histidine, pH 5.5; or u. 16 mg (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 6.2% (w/w) trehalose, and 1.3% (w/w) histidine, pH 5.5.

In these cases, the total mass solid content of the pharmaceutical composition is 20 mg.

In some cases, a pharmaceutical composition is provided that comprises 0.4 mg ± 20% antigen-binding fragment ± 20%, 2% (w/w) trileucine ± 10%, 10.5% (w/w) leucine ± 10%, 83.36% (w/w) trehalose ± 10%, and 3.14% (w/w) histidine ± 10%, at pH 5.5. In some cases, the pharmaceutical composition comprises 0.4 mg antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 83.36% (w/w) trehalose, and 3.14% (w/w) histidine, at pH 5.5. In these cases, the total mass solid content of the pharmaceutical composition is 20 mg.

In some cases, a pharmaceutical composition is provided that comprises 2 mg ± 20% antigen-binding fragment ± 20%, 2% (w/w) trileucine ± 10%, 10.5% (w/w) leucine ± 10%, 74.36% (w/w) trehalose ± 10%, and 3.14% (w/w) histidine ± 10%, at a pH of 5.5. In some cases, the pharmaceutical composition comprises 2 mg of the antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 74.36% (w/w) trehalose, and 3.14% (w/w) histidine, at a pH of 5.5. In these cases, the total mass solid content of the pharmaceutical composition is 20 mg.

In some cases, a pharmaceutical composition is provided that comprises 8 mg ± 20% antigen-binding fragment ± 20%, 2% (w/w) trileucine ± 10%, 10.5% (w/w) leucine ± 10%, 44.36% (w/w) trehalose ± 10%, and 3.14% (w/w) histidine ± 10%, at a pH of 5.5. In some cases, the pharmaceutical composition comprises 8 mg antigen-binding fragment, 2% (w/w) trileucine ± 10%, 10.5% (w/w) leucine ± 10%, 42.5% (w/w) trehalose ± 10%, and 5.0% (w/w) histidine ± 10%, at a pH of 5.5. In these cases, the total mass solid content of the pharmaceutical composition is 20 mg. When the pharmaceutical composition disclosed herein comprises approximately 3.14% (w/w) histidine, the pharmaceutical composition may comprise approximately 0.55% (w/w) L-histidine and approximately 2.59% (w/w) histidine HCl. In some instances, when the pharmaceutical composition comprises 3.14% (w/w) histidine, the pharmaceutical composition may comprise 0.55% (w/w) L-histidine and 2.59% (w/w) histidine HCl.

The particles comprising the pharmaceutical compositions described herein, when provided in aerosol form, suitably have a specified mass median aerodynamic diameter (MMAD). The particles may also have a specified equivalent optical volume mean diameter (oVMD). The oVMD may also be referred to as the particle size distribution (PSD or pPSD).

As used herein, "mass median aerodynamic diameter" or "MMAD" is a measure of the aerodynamic size of dispersed particles. Aerodynamic diameter is used to describe atomized powders based on their settling behavior and is the diameter of a sphere of unit density that has the same settling velocity in air as the particle. Aerodynamic diameter encompasses the particle shape, density, and physical size of the particle. Unless otherwise indicated, MMAD, as used herein, refers to the midpoint or median of the aerodynamic particle size distribution of an atomized powder as determined by cascade impaction. Suitably, the microparticles of the dry powder formulations provided herein have a mass median aerodynamic diameter (MMAD) of about 1 μm to about 10 μm, more suitably about 2 μm to about 8 μm, about 2 μm to about 7 μm, about 2 μm to about 6 μm, about 2 μm to about 5 μm, about 2 μm to about 4 μm, about 3 μm to about 7 μm, about 4 μm to about 7 μm, about 3 μm to about 6 μm, or about 2 μm, about 3 μm, about 4 μm, about 5 μm, about 6 μm, or about 7 μm.

Suitably, the dry powder formulations described herein have a fine particle fraction (the fraction of particles having an aerodynamic particle size of less than 5 μm ejected from an inhalation device) of ≥ 50%, more preferably ≥ 60%. This fine particle fraction (FPF) can contribute to low device retention of less than 20%, preferably less than 15%, less than 10%, or less than 5% of the dry powder formulation remaining in the device after delivery to the patient.

In another embodiment, the spray-dried particles preferably have an equivalent optical volume mean diameter (oVMD) of about 0.5 μm to about 7 μm. The equivalent optical volume mean diameter (oVMD) is the average diameter of a sphere that best approximates the particle's specific optical interaction with light, when measured using suitable optical techniques, wherein half of the particles approximate an equivalent sphere smaller than the average and half of the particles approximate an equivalent sphere larger than the average. In exemplary embodiments, the microparticles have an equivalent optical volume mean diameter (oVMD) of about 0.5 μm to about 6 μm, or about 1 μm to about 5 μm, or about 1 μm to about 4 μm, or about 2 μm to about 4.5 μm, or about 2.5 μm to about 4 μm, or about 2 μm to about 4 μm, or about 2 μm to about 3 μm, or about 2 μm to about 3.5 μm, or about 1 μm, about 1.5 μm, about 2 μm, about 2.5 μm, about 3 μm, about 3.5 μm, about 4 μm, about 4.5 μm, or about 5 μm.

The use of leucine and trileucine in dry powder formulations also produces particles with a desired size (MMAD) as well as a desired specific surface area (SSA) and roughness, thereby producing particles that can flow properly and be delivered to the lungs using various inhalation platforms.

The specific surface area (SSA) of a particle is defined as the total surface area per unit mass of the particle (suitably, in ^m² /g). Methods for measuring SSA are known in the art and include, for example, the Brunauer–Emmett–Teller (BET) measurement, which uses nitrogen adsorption measured as a function of relative pressure to estimate the specific surface area of a material. The surface area is determined by calculating the amount of adsorbate gas corresponding to a monolayer on the particle surface. This technique measures the external area and any pore area estimate to determine the total specific surface area. Instruments for measuring BET are known in the art.

In some cases, the dry powder formulation has a specific surface area (SSA) of the particles of about 3 m ² /g to about 8 m ² /g. In certain cases, the SSA of the plurality of particles is about 3.5 m ² /g to 7.5 m ² /g, or about 4 m ² /g to 7 m 2 /g, or about 4.5 m ² /g to 7 m ² /g, or about 5 m ² /g to 7 m ² /g, or about 4.5 m ² /g to 6 m ² /g, or about 5 m ² /g to 6 m 2 /g, or about 4 m ² /g, about 4.5 m ² /g, about 5 m ² /g, about 5.5 m ² /g, about 6 m ² /g, about 6.5 m ² /g, or about 7 m ² /g.

In certain instances, the dry powder formulation has a compressed bulk density of about 0.4-1.0 g/cm ^3. Suitably, the dry powder formulation has a compressed bulk density of about 0.5-0.8 g/cm ^3. In some instances, the dry powder formulations described herein have a compressed bulk density of about 0.4-0.9 gm/cm 3, about 0.4-0.8 g/cm ³ , about 0.5-0.8 g/cm ³ , about 0.6-0.8 gm/cm ³ , or about 0.4 gm/cm ³ , about 0.5 gm/cm ³ , about 0.6 gm/cm ³ , about 0.7 gm/cm ³ , or about 0.8 gm/cm ³ . In certain instances, the dry powder formulations described herein have a compressed bulk density of about 0.4 gm/cm ³ to about 0.9 gm/cm ³ . In certain instances, the dry powder formulations described herein have a compressed bulk density of about 0.5 gm/cm ³ to about 0.8 gm/cm ³ .

In some cases, the pharmaceutical compositions described herein do not include surfactants, such as polysorbates, e.g., polysorbate-80 or polysorbate-20. While the addition of surfactants minimizes the formation of protein aggregates, they can increase device deposition, thereby reducing lung deposition and lowering dry aerosol yield. The inventors optimized the composition in such a way that omitting the surfactant has minimal effect on protein aggregation while advantageously minimizing powder device deposition ( Example 3 ).

It will be apparent to those skilled in the art that surfactants reduce the formation of subvisible particles (SVPs) after reconstitution of the formulation. In view of this, the minimal device deposition and protein aggregation observed for the compositions described herein that do not contain a surfactant is surprising.

As referred to herein, "subvisible particles" ("SVPs") are particles ranging from about 1 μm to about 200 μm that are not visible to the naked eye. Removing or reducing the formation of SVPs simplifies the analytical characterization of the formulation because it eliminates the burden of tracking the formation of SVPs during manufacturing. Analytical characterization of SVPs may involve the development of orthogonal techniques to identify and quantify SVPs for quality control purposes. Therefore, removing SVPs or reducing them to acceptable levels eliminates the need for this characterization step in the manufacturing process, thereby streamlining manufacturing. Removal of SVPs can also make the dosage range more predictable because the release kinetics of the drug from the SVPs are unknown. Furthermore, removing the SVP may increase the amount of active agent available to participate in pharmacological activity after recovery, which may mean not only that higher delivery doses can be achieved, but also that more accurate predictions of delivery doses can be calculated. Higher delivery doses may also benefit patients, for example, by potentially reducing the number or frequency of doses that must be delivered to extract a pharmacological benefit.

The presence of subvisible particles can be determined by reconstituting dry powder formulations and liquids with a turbid quality. The actual determination of the presence of SVPs can be confirmed using techniques such as dynamic flow imaging microscopy, such as microflow imaging (MFI). In MFI, which is also known as flow imaging microscopy (FIM) or dynamic imaging analysis (DIA), bright field images are captured in successive frames as a continuous sample stream passes through a flow cell located in the field of view of the microscope system. Digital images of the particles present in the sample are processed by image morphology analysis software, which allows quantification of their size and counting. MFI is an established technique for the analysis of subvisible particles. Dynamic flow imaging microscopy combines microfluidic flow microscopy with high-resolution imaging particle analysis to quantify SVP counts. MFI can bin these counts across a range of particle sizes, for example, by binning particle counts in the size ranges of about 1 to about 200 μm, about 2 μm to about 200 μm, about 5 μm to about 200 μm, about 10 μm to about 200 μm, and about 25 μm to about 200 μm. An alternative technique for measuring SVPs is background membrane imaging (BMI). Briefly, SVPs from a liquid sample are separated onto a filter surface for counting analysis by a microscope. The BMI software images the baseline before particle isolation and then subtracts the baseline from the isolated particles pixel by pixel, so that only the photographic information remains (Vargas et al., 2020). Optimization of the formulation modifier and other aspects of the composition significantly reduced SVP formation.

In some cases, after reconstitution, the number of SVPs between 5 μm and 200 μm is less than about 30,000 particles per milliliter, such as 25,000 particles per milliliter, 20,000 particles per milliliter, 15,000 particles per milliliter, 10,000 particles per milliliter, or 5,000 particles per milliliter. In some cases, the number of SVPs between 5 μm and 200 μm is less than 1,000 particles per milliliter. In some cases, the number of SVPs between 5 μm and 200 μm is less than 1,000 particles per milliliter. In some cases, the number of SVPs ranging in size from 5 μm to approximately 200 μm was less than 100 particles per milliliter.

In some cases, after reconstitution, the number of SVPs having a size of 10 μm to 200 μm is less than about 100,000 particles per milliliter, such as 90,000 particles per milliliter, 80,000 particles per milliliter, 70,000 particles per milliliter, 60,000 particles per milliliter, 50,000 particles per milliliter, 40,000 particles per milliliter, or 30,000 particles per milliliter. In some cases, the number of SVPs having a size of 10 μm to 200 μm is less than about 10,000 particles per milliliter. In some cases, the number of SVPs having a size of 10 μm to 200 μm is less than about 2,000 particles per milliliter. In some cases, the number of SVPs ranging in size from 10 μm to approximately 200 μm was less than 100 particles per milliliter.

In some cases, after reconstitution, the number of SVPs with a size of 25 μm to 200 μm is less than about 200,000 particles per milliliter, such as 180,000 particles per milliliter, 170,000 particles per milliliter, 160,000 particles per milliliter, 150,000 particles per milliliter, or 140,000 particles per milliliter. In some cases, the number of SVPs with a size of about 5 μm to about 200 μm is less than about 50,000 particles per milliliter. In some cases, the number of SVPs with a size of 5 μm to 200 μm is less than about 10,000 particles per milliliter. In some cases, the number of SVPs with a size of about 5 μm to about 200 μm is less than about 2,000 particles per milliliter. In some cases, the number of SVPs with a size of between 10 μm and about 200 μm is less than about 200 particles per milliliter.

In some cases, the number of SVPs was determined after reconstitution in water to an antigen-binding fragment concentration of 2.5 mg/ml or 30 mg/ml.

In other cases, provided herein is a process for preparing a pharmaceutical composition for inhalation, comprising: providing an aqueous solution at about pH 5 to about pH 6, the aqueous solution comprising leucine, trileucine, histidine, a glass stabilizer as described herein, and an antigen-binding fragment of an anti-thymic stromal lymphopoietin (TSLP) antibody; spray drying the aqueous solution of (a) to produce dry powder particles; and collecting the dry powder particles, wherein the aqueous solution comprises about 5% (w/w) to about 15% (w/w) leucine, about 1% (w/w) to about 5% (w/w) trileucine, about 1% (w/w) to about 10% (w/w) histidine, about 5% (w/w) to about 50% (w/w) antigen-binding fragment, and a certain % (w/w) glass stabilizer to achieve a total solids content of 100%. In some cases, the aqueous solution has a pH of 5.5.

In some cases, the aqueous solution comprises: a. 1% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 85.2% (w/w) trehalose ±10%, and 1.3% (w/w) histidine ±10%, pH 5.5; b. 1% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 83.36% (w/w) trehalose ±10%, and 3.14% (w/w) histidine ±10%, pH 5.5; c. 1% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 83.36% (w/w) trehalose ±10%, and 3.14% (w/w) histidine ±10%, pH 5.5. (w/w) leucine ±10%, 81.5% (w/w) trehalose ±10%, and 5.0% (w/w) histidine ±10%, pH 5.5; d. 2% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 84.2% (w/w) trehalose ±10%, and 1.3% (w/w) histidine ±10%, pH 5.5; e. 2% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 82.36% (w/w) trehalose ±10%, and 3.14% (w/w) histidine ±10%, pH 5.5; f. 2% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 80.5% (w/w) trehalose ±10%, and 5.0% (w/w) histidine ±10%, pH 5.5; g. 3% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 83.2% (w/w) trehalose ±10%, and 1.3% (w/w) histidine ±10%, pH 5.5; h. 3% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 81.36% (w/w) trehalose ±10% and 3.14% (w/w) histidine ±10%, pH 5.5; i. 3% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 79.5% (w/w) trehalose ±10%, and 5.0% (w/w) histidine ±10%, pH 5.5; j. 10% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 76.2% (w/w) trehalose ±10%, and 1.3% (w/w) histidine ±10%, pH 5.5; k. 10% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 74.36% (w/w) trehalose ±10%, and 3.14% (w/w) histidine ±10%, pH 5.5; l. 10% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 72.5% (w/w) trehalose ±10%, and 5.0% (w/w) histidine ±10%, pH 5.5; m. 30% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 54.36% (w/w) trehalose ±10%, and 3.14% (w/w) histidine ±10%, pH 5.5; n. 30% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 52.5% (w/w) trehalose ±10%, and 5.0% (w/w) histidine ±10%, pH 5.5; o. 30% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 56.2% (w/w) trehalose ±10%, and 1.3% (w/w) histidine ±10%, pH 5.5; p. 40% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 44.36% (w/w) trehalose ±10%, and 3.14% (w/w) histidine ±10%, pH 5.5; q. 40% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 42.5% (w/w) trehalose ±10%, and 5.0% (w/w) histidine ±10%, pH 5.5; r. 40% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 46.2% (w/w) trehalose ±10%, and 1.3% (w/w) histidine ±10%, pH 5.5; s. 80% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 4.36% (w/w) trehalose ±10%, and 3.14% (w/w) histidine ±10%, pH 5.5; t. 80% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 2.5% (w/w) trehalose ±10%, and 5.0% (w/w) histidine ±10%, pH 5.5; or u. 80% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 6.2% (w/w) trehalose ±10% and 1.3% (w/w) histidine ±10%, pH 5.5.

In some cases, the aqueous solution comprises: a. 1% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 85.2% (w/w) trehalose, and 1.3% (w/w) histidine, pH 5.5; b. 1% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 83.36% (w/w) trehalose, and 3.14% (w/w) histidine, pH 5.5; c. 1% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 81.5% (w/w) trehalose, and 5.0% (w/w) histidine, pH 5.5; d. 2% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 84.2% (w/w) trehalose, and 1.3% (w/w) histidine, pH 5.5; e. 2% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 82.36% (w/w) trehalose, and 3.14% (w/w) histidine, pH 5.5; f. 2% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 80.5% (w/w) trehalose, and 5.0% (w/w) histidine, pH 5.5; g. 3% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 83.2% (w/w) trehalose, and 1.3% (w/w) histidine, pH 5.5; h. 3% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 81.36% (w/w) trehalose, and 3.14% (w/w) histidine, pH 5.5; i. 3% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 79.5% (w/w) trehalose, and 5.0% (w/w) histidine, pH 5.5; j. 10% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 76.2% (w/w) trehalose, and 1.3% (w/w) histidine, pH 5.5; k. 10% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 74.36% (w/w) trehalose, and 3.14% (w/w) histidine, pH 5.5; l. 10% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 72.5% (w/w) trehalose, and 5.0% (w/w) histidine, pH 5.5; m. 30% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 54.36% (w/w) (w/w) trehalose and 3.14% (w/w) histidine, pH 5.5; n. 30% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 52.5% (w/w) trehalose and 5.0% (w/w) histidine, pH 5.5; o. 30% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 56.2% (w/w) trehalose and 1.3% (w/w) histidine, pH 5.5; p. 40% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 44.36% (w/w) trehalose and 3.14% (w/w) histidine, pH 5.5; q. 40% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 42.5% (w/w) trehalose, and 5.0% (w/w) histidine, pH 5.5; r. 40% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 46.2% (w/w) trehalose, and 1.3% (w/w) histidine, pH 5.5; s. 80% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 4.36% (w/w) trehalose, and 3.14% (w/w) histidine, pH 5.5; t. 80% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 2.5% (w/w) trehalose, and 5.0% (w/w) histidine, pH 5.5; or u. 80% (w/w) antigen-binding fragment, 2% (w/w) trileucine, 10.5% (w/w) leucine, 6.2% (w/w) trehalose, and 1.3% (w/w) histidine, pH 5.5.

When the aqueous solution contains about 3.14% (w/w) histidine, the pharmaceutical composition may contain about 0.55% (w/w) L-histidine and about 2.59% (w/w) histidine HCl. In some cases, when the pharmaceutical composition contains 3.14% (w/w) histidine, the pharmaceutical composition may contain 0.55% (w/w) L-histidine and 2.59% (w/w) histidine HCl.

An aqueous solution is prepared by combining these components in a liquid solvent to produce a raw material in which the components are dissolved. Temperature control may be added as desired or necessary to increase the solubility of the various components to form the aqueous solution. Exemplary liquid solvents include water, including deionized water, and dilute solutions of alcohol and water. In some cases, the active agent is suitably added to the aqueous solution after the remaining components of the raw materials have been added and dissolved. The aqueous solution may then be atomized. The aqueous solution may be filtered prior to atomization. In some cases, the liquid raw material may be filtered using a 0.22 micron filter. In some cases, the aqueous solution comprising leucine, trileucine, and histidine is filtered prior to the addition of the active agent. In some cases, the aqueous solution is filtered after the active agent has been added prior to atomization. Atomization refers to the conversion of a solution into fine droplets, suitably using a pressurized gas (such as an inert gas or compressed dry air). Exemplary apparatus for producing atomized solutions are known in the art and include the use of various atomizing nozzles having the desired size and flow characteristics. Exemplary parameters for atomization include an outlet temperature of about 50° C. to 90° C., suitably about 60° C. to 80° C., or about 70° C.; a raw material feed rate of about 8-15 ml/min, suitably about 9-14 ml/min, about 10-13 ml/min, or about 12 ml/min; an atomizer gas flow rate of about 9-15 kg/hour (hr. or h), suitably about 10-14 kg/hr, about 12-14 kg/hr, or about 13 kg/hr; and a drying gas flow rate of about 60-100 kg/hr, suitably about 60-90 kg/hr, about 70-90 kg/hr, or about 80 kg/hr.

The atomized solution can then be dried, suitably under heat and in combination with moving air to aid drying. Drying results in a plurality of microparticles. Drying temperatures are typically in the range of about 50°C to 100°C, or about 60°C to 100°C, or about 70°C to 90°C; and air flow rates can be about 10-40 ^m³ /hour.

Exemplary glass stabilizers (including amorphous sugars and buffers) and suitable amounts of the glass stabilizers are described herein. Suitable amounts of leucine and trileucine are also provided throughout. Since the final dry powder formulation should contain the listed amounts of leucine, trileucine, histidine (and other components), these amounts are also used in the liquid raw materials. The drying process after atomization results in the removal of any liquid solvent, and therefore the total amount of the original dry weight of the components corresponds to the final dry weight of the compound in the dry powder formulation. Exemplary active agents are also described herein.

The methods and formulations described herein allow for the production of capsules, blister packs, and other suitable containers for dry powder formulations. Such containers can be produced to hold 10-200 mg of dry powder, preferably 10-100 mg, 15-25 mg, or 20-75 mg, 20 mg, or 50 mg of dry powder formulation. Such containers can suitably deliver 0.1-10 mg of dry powder formulation to a patient's lungs.

In some cases, the methods described herein can be used to provide pharmaceuticals that reduce the total number of capsules required for use in an inhalation device. For example, the volume required to deliver 50-100 mg of active agent can be reduced from two larger 00 capsules to a single size 3 capsule.

Methods for generating dry powder particles in aerosol form are known in the art and include, for example, the use of an inhaler device, such as a dry powder inhaler (DPI) (e.g., Monodose RS01 DPI from PLASTIAPE (Osnago, Italy)). The pharmaceutical compositions described herein can be dispensed into an airstream by a passive or active inhalation device and maintained suspended in the air for a sufficient amount of time to allow at least a portion of the particles to be inhaled by the patient, such that a portion of the particles reaches the lungs.

Anti- TSLP Antibodies and Antigen-Binding Fragments Thereof Various methods, pharmaceutical compositions, and unit doses disclosed herein utilize antigen-binding fragments of anti-thymic stromal lymphopoietin (TSLP) antibodies. In some cases, the antigen-binding fragment is a Fab.

The sequence of the human TSLP polypeptide is provided below: Met Phe Pro Phe Ala Leu Leu Tyr Val Leu Ser Val Ser Phe Arg Lys Ile Phe Ile Leu Gln Leu Val Gly Leu Val Leu Thr Tyr Asp Phe Thr Asn Cys Asp Phe Glu Lys Ile Lys Ala Ala Tyr Leu Ser Thr Ile Ser Lys Asp Leu Ile Thr Tyr Met Ser Gly Thr Lys Ser Thr Glu Phe Asn Asn Thr Val Ser Cys Ser Asn Arg Pro His Cys Leu Thr Glu Ile Gln Ser Leu Thr Phe Asn Pro Thr Ala Gly Cys Ala Ser Leu Ala Lys Glu Met Phe Ala Met Lys Thr Lys Ala Ala Leu Ala Ile Trp Cys Pro Gly Tyr Ser Glu Thr Gln Ile Asn Ala Thr Gln Ala Met Lys Lys Arg Arg Lys Arg Lys Val Thr Thr Asn Lys Cys Leu Glu Gln Val Ser Gln Leu Gln Gly Leu Trp Arg Arg Phe Asn Arg Pro Leu Leu Lys Gln Gln (SEQ ID NO: 27). As used herein, the term "antibody" refers to a tetrameric glycoprotein composed of two heavy chains and two light chains, each comprising a variable region and a constant region. "Heavy chain" and "light chain" refer to substantially full-length typical immunoglobulin light and heavy chains (see, e.g., Immunobiology, 5th ed. (Janeway and Travers et al., eds., 2001). The term "antibody" includes naturally occurring antibodies as well as all recombinant forms of antibodies, such as humanized antibodies, fully human antibodies, and chimeric antibodies.

The term "antibody fragment" refers to a portion of an intact antibody. The term "antigen-binding fragment", "antigen-binding domain" or "antigen-binding region" of an antibody refers to the portion of an intact antibody that binds to the antigen. Antigen-binding fragments of an antibody include, inter alia, Fab, Fab', F(ab')2, Fv, domain antibodies (dAb), complementary determining region (CDR) fragments, CDR-grafted antibodies, single-chain antibodies (scFv), single-chain antibody fragments, chimeric antibodies, bibodies, triabodies, tetrabodies, minibodies, linear antibodies; recombinant antibodies, tribodies or bibodies, cytosolic antibodies, scFvs ... Intrabodies, nanobodies, small modular immunopharmaceuticals (SMIPs), antigen-binding domain immunoglobulin fusion proteins, single-domain antibodies (including camelized antibodies), VHH-containing antibodies, or variants or derivatives thereof, and polypeptides containing at least a portion of an immunoglobulin sufficient to confer specific antigen binding to the polypeptide, such as one, two, three, four, five, or six CDR sequences, as long as the antibody retains the desired biological activity. In some cases, the antigen-binding fragment of the antibody of the present disclosure is selected from Fab, Fab', F(ab')2, scFv, minibody, or bispecific antibody. In some cases, the antigen-binding fragment is Fab. In some cases, the antigen-binding fragment belongs to IgG, IgM, IgA, IgD, or IgE. In some cases, the anti-TSLP antibody fragment is IgG1.

"Fab" refers to an antibody fragment comprising the VH-CH1 and VL-CL pairings. The term encompasses Fabs containing atypical sequence variants, such as amino acid substitutions, deletions, or insertions outside of sequence regions typically associated with high sequence variability within Fab. For example, Fab variants include Fabs containing atypical amino acids or sequence variations in the VH or VL framework regions, or in the CH1 or CL domains. Such variations may include the presence of atypical cysteines or other derivatizable amino acids that can be used to conjugate the Fab variants to a heterologous moiety. Other such variations include the presence of atypical polypeptide linkers, which are polypeptide sequences that covalently bridge between the two domains. For example, a Fab variant may comprise a linker polypeptide covalently linking the CH1 domain to the VL domain or the CL domain to the VH domain, such that the Fab appears as a single polypeptide chain.

The light chain CDR (LCDR), light chain variable domain (VL), heavy chain CDR (HCDR), and heavy chain variable domain (VH) sequences of exemplary Fabs of the present disclosure (referred to herein as FABl) include those listed in Table 1 below.

Table 1 : Exemplary sequences of Fabs of the present disclosure (referred to herein as FABl). SEQ ID NO describe sequence 1 HCDR1 FAB1 Thr Tyr Gly Met His 2 HCDR2 FAB1 Val Ile Trp Tyr Asp Gly Ser Asn Lys His Tyr Ala Asp Ser Val Lys Gly 3 HCDR3 FAB1 Ala Pro Gln Trp Glu Leu Val His Glu Ala Phe Asp Ile 4 Heavy chain VH FAB1 Gln Met Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Arg Thr Tyr Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Val Ile Trp Tyr Asp Gly Ser Asn Lys His Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Thr Arg Asp Asn Ser Lys Asn Thr Leu Asn Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Ala Pro Gln Trp Glu Leu Val His Glu Ala Phe Asp Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser 5 LCDR1 FAB1 Gly Gly Asn Asn Leu Gly Ser Lys Ser Val His 6 LCDR2 FAB1 Asp Asp Ser Asp Arg Pro Ser 7 LCDR3 FAB1 Gln Val Trp Asp Ser Ser Ser Asp His Val Val 8 Light chain VL FAB1 Ser Tyr Val Leu Thr Gln Pro Pro Ser Val Ser Val Ala Pro Gly Gln Thr Ala Arg Ile Thr Cys Gly Gly Asn Asn Leu Gly Ser Lys Ser Val His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Val Leu Val Val Tyr Asp Asp Ser Asp Arg Pro Ser Trp Ile Pro Glu Arg Phe Ser Gly Ser Asn Ser Gly Asn Thr Ala Thr Leu Thr Ile Ser Arg Gly Glu Ala Gly Asp Glu Ala Asp Tyr Tyr Cys Gln Val Trp Asp Ser Ser Ser Asp His Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 9 FAB1 variable heavy chain VH (nucleic acid) cagatgcagt tggttgaatc tggtggcggc gtggtgcagc ctggcagatc tctgagactg 60 tcttgtgccg cctccggctt caccttcaga acctacggaa tgcactgggt ccgacaggcc 120 cctggcaaag gattggaatg ggtcgccgtg atttggtacg acggctccaa caagcactac 180 gccgactccg tgaagggcag attcaccatc accagagaca actccaagaa caccctgaac 240 ctgcagatga actccctgag agccgaggac accgccgtgt actattgtgc tagagcccct 300 cagtgggaac tcgtgcatga ggcctttgac atctggggcc agggaacaat ggtcaccgtc 360 tcctca 366 10 FAB1 variable light chain VL (nucleic acid) tcatatgttc ttacacaacc accgtcggtt tcggttgctc caggacaaac agctcgaatt 60 acatgcggag gaaacaacct cggatcgaag tcggttcact ggtatcaaca aaagccagga 120 caagctccag ttctcgtggt gtacgatgat tcagatcgac catcatggat cccagagcga 180 ttctcaggat caaactcggg aaatactgcc acgctcacaa tttcacgcgg agaagcggga 240 gatgaagctg attactattg ccaagtgtgg gactcgtcgt cagatcatgt tgttttcgga 300 ggtggaacaa agctcacagt gctc 324 28 FAB1 heavy chain (peptide) Gln Met Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Arg Thr Tyr Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Val Ile Trp Tyr Asp Gly Ser Asn Lys His Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Thr Arg Asp Asn Ser Lys Asn Thr Leu Asn Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Ala Pro Gln Trp Glu Leu Val His Glu Ala Phe Asp Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys 29 FAB1 light chain (peptide) Ser Tyr Val Leu Thr Gln Pro Pro Ser Val Ser Val Ala Pro Gly Gln Thr Ala Arg Ile Thr Cys Gly Gly Asn Asn Leu Gly Ser Lys Ser Val His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Val Leu Val Val Tyr Asp Asp Ser Asp Arg Pro Ser Trp Ile Pro Glu Arg Phe Ser Gly Ser Asn Ser Gly Asn Thr Ala Thr Leu Thr Ile Ser Arg Gly Glu Ala Gly Asp Glu Ala Asp Tyr Tyr Cys Gln Val Trp Asp Ser Ser Ser Asp His Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln Pro Lys Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Pro Ser Ser Glu Glu Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp Phe Tyr Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro Val Lys Ala Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn Lys Tyr Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser His Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu Lys Thr Val Ala Pro Thr Glu Cys Ser 30 FAB1 heavy chain (nucleic acid) cagatgcagt tggttgaatc tggtggcggc gtggtgcagc ctggcagatc tctgagactg 60 tcttgtgccg cctccggctt caccttcaga acctacggaa tgcactgggt ccgacaggcc 120 cctggcaaag gattggaatg ggtcgccgtg atttggtacg acggctccaa caagcactac 180 gccgactccg tgaagggcag attcaccatc accagagaca actccaagaa caccctgaac 240 ctgcagatga actccctgag agccgaggac accgccgtgt actattgtgc tagagcccct 300 cagtgggaac tcgtgcatga ggcctttgac atctggggcc agggaacaat ggtcaccgtc 360 tcctcagcct ccaccaaggg cccatcggtc ttccccctgg caccctcctc caagagcacc 420 tctgggggca cagcggccct gggctgcctg gtcaaggact acttccccga accggtgacg 480 gtgtcgtgga actcaggcgc cctgaccagc ggcgtgcaca ccttcccggc tgtcctacag 540 tcctcaggac tctactccct cagcagcgtg gtgacagtgc cctccagcag cttgggcacc 600 cagacctaca tctgcaacgt gaatcacaag cccagcaaca ccaaggtgga caagagagtt 660 gagcccaaat cttgtgacaa a 681 31 FAB1 light chain (nucleic acid) tcatatgttc ttacacaacc accgtcggtt tcggttgctc caggacaaac agctcgaatt 60 acatgcggag gaaacaacct cggatcgaag tcggttcact ggtatcaaca aaagccagga 120 caagctccag ttctcgtggt gtacgatgat tcagatcgac catcatggat cccagagcga 180 ttctcaggat caaactcggg aaatactgcc acgctcacaa tttcacgcgg agaagcggga 240 gatgaagctg attactattg ccaagtgtgg gactcgtcgt cagatcatgt tgttttcgga 300 ggtggaacaa agctcacagt gctcggtcag cccaaggctg ccccctcggt cactctgttc 360 ccgccctcct ctgaggagct tcaagccaac aaggccacac tggtgtgtct cataagtgac 420 ttctacccgg gagccgtgac agtggcctgg aaggcagata gcagccccgt caaggcggga 480 gtggagacca ccacaccctc caaacaaagc aacaacaagt acgcggccag cagctatctg 540 agcctgacgc ctgagcagtg gaagtcccac agaagctaca gctgccaggt cacgcatgaa 600 gggagcaccg tggagaagac agtggcccct acagaatgtt ca 642

In some cases, the antigen-binding fragment within the composition comprises: a heavy chain variable domain comprising: a heavy chain CDR1 sequence comprising the amino acid sequence shown in SEQ ID NO:1, a heavy chain CDR2 sequence comprising the amino acid sequence shown in SEQ ID NO:2, and a heavy chain CDR3 sequence comprising the amino acid sequence shown in SEQ ID NO:3, wherein any one of heavy chain CDR1, 2, or 3 optionally comprises a single amino acid substitution, and a light chain variable domain comprising: a light chain CDR1 sequence comprising the amino acid sequence shown in SEQ ID NO:5, a light chain CDR2 sequence comprising the amino acid sequence shown in SEQ ID NO:6, and a light chain CDR3 sequence comprising the amino acid sequence shown in SEQ ID NO:7, wherein any one of light chain CDRs 1, 2, or 3 optionally comprises a single amino acid substitution.

In some cases, the antigen-binding fragment within the composition comprises a heavy chain variable domain comprising a light chain CDR1 sequence having the amino acid sequence shown in SEQ ID NO: 1, a heavy chain CDR2 sequence having the amino acid sequence shown in SEQ ID NO: 2, and a heavy chain CDR3 sequence having the amino acid sequence shown in SEQ ID NO: 3, and a light chain CDR1 sequence having the amino acid sequence shown in SEQ ID NO: 5, a light chain CDR2 sequence having the amino acid sequence shown in SEQ ID NO: 6, and a light chain CDR3 sequence having the amino acid sequence shown in SEQ ID NO: 7.

In another embodiment, the antigen-binding fragment for use in the pharmaceutical composition comprises a heavy chain variable domain having an amino acid sequence that is at least 95%, 90%, 85% or 80% identical to SEQ ID NO: 4, and a light chain variable domain having an amino acid sequence that is at least 95%, 90%, 85% or 80% identical to SEQ ID NO: 8.

In another embodiment, the antigen-binding fragment used in the composition comprises (a) a heavy chain variable domain having an amino acid sequence at least 95%, 90%, 85% or 80% identical to SEQ ID NO: 4; or an amino acid sequence encoded by a polynucleotide sequence at least 80% identical to SEQ ID NO: 30, (b) a light chain variable domain having an amino acid sequence at least 95%, 90%, 85% or 80% identical to SEQ ID NO: 8; or an amino acid sequence encoded by a polynucleotide sequence at least 80% identical to SEQ ID NO: 31; or the heavy chain variable domain of (a) and the light chain variable domain of (b).

In another embodiment, the antigen-binding fragment for use in the pharmaceutical composition comprises a heavy chain variable domain comprising SEQ ID NO: 4 and a light chain variable domain comprising SEQ ID NO: 8. In another embodiment, the antigen-binding fragment for use in the pharmaceutical composition comprises a heavy chain comprising the sequence set forth in SEQ ID NO: 28 and a light chain comprising the sequence set forth in SEQ ID NO: 29.

Other light chain CDR (LCDR), light chain variable domain (VL), heavy chain CDR (HCDR) and heavy chain variable domain (VH) sequences of the antigen-binding fragments of the present disclosure include those listed in Table 2 below: Table 2 : Exemplary sequences of Fabs of the present disclosure (referred to herein as FAB2). SEQ ID NO describe sequence 11 LCDR1 FAB2 Gly Gly Asn Asn Ile Gly Ser Lys Ser Val His 12 Light chain VL FAB2 Ser Tyr Val Leu Thr Gln Pro Pro Ser Val Ser Val Ala Pro Gly Gln Thr Ala Arg Ile Thr Cys Gly Gly Asn Asn Ile Gly Ser Lys Ser Val His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Val Leu Val Val Tyr Asp Asp Ser Asp Arg Pro Ser Trp Ile Pro Glu Arg Phe Ser Gly Ser Asn Ser Gly Asn Thr Ala Thr Leu Thr Ile Ser Arg Gly Glu Ala Gly Asp Glu Ala Asp Tyr Tyr Cys Gln Val Trp Asp Ser Ser Ser Asp His Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 13 LCDR1 FAB3 Gly Gly Asn Asn Val Gly Ser Lys Ser Val His 14 Light chain VL FAB3 Ser Tyr Val Leu Thr Gln Pro Pro Ser Val Ser Val Ala Pro Gly Gln Thr Ala Arg Ile Thr Cys Gly Gly Asn Asn Val Gly Ser Lys Ser Val His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Val Leu Val Val Tyr Asp Asp Ser Asp Arg Pro Ser Trp Ile Pro Glu Arg Phe Ser Gly Ser Asn Ser Gly Asn Thr Ala Thr Leu Thr Ile Ser Arg Gly Glu Ala Gly Asp Glu Ala Asp Tyr Tyr Cys Gln Val Trp Asp Ser Ser Ser Asp His Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 15 HCDR2 FAB4 Val Ile Trp Tyr Asp Gly Ser Asn Lys His Tyr Ala Glu Ser Val Lys Gly 16 Heavy chain VH FAB4 Gln Met Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Arg Thr Tyr Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Val Ile Trp Tyr Asp Gly Ser Asn Lys His Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Thr Arg Asp Asn Ser Lys Asn Thr Leu Asn Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Ala Pro Gln Trp Glu Leu Val His Glu Ala Phe Asp Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser 17 HCDR2 FAB5 Val Ile Trp Tyr Asp Gly Ser Asn Lys His Tyr Ala Asp Ser Val Lys Ala 18 Heavy chain VH FAB5 Gln Met Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Arg Thr Tyr Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Val Ile Trp Tyr Asp Gly Ser Asn Lys His Tyr Ala Asp Ser Val Lys Ala Arg Phe Thr Ile Thr Arg Asp Asn Ser Lys Asn Thr Leu Asn Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Ala Pro Gln Trp Glu Leu Val His Glu Ala Phe Asp Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser 19 LCDR1 FAB6 Gly Gly Gln Asn Leu Gly Ser Lys Ser Val His 20 Light chain VL FAB6 Ser Tyr Val Leu Thr Gln Pro Pro Ser Val Ser Val Ala Pro Gly Gln Thr Ala Arg Ile Thr Cys Gly Gly Gln Asn Leu Gly Ser Lys Ser Val His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Val Leu Val Val Tyr Asp Asp Ser Asp Arg Pro Ser Trp Ile Pro Glu Arg Phe Ser Gly Ser Asn Ser Gly Asn Thr Ala Thr Leu Thr Ile Ser Arg Gly Glu Ala Gly Asp Glu Ala Asp Tyr Tyr Cys Gln Val Trp Asp Ser Ser Ser Asp His Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu twenty one LCDR1 FAB7 Gly Gly Asn Gln Leu Gly Ser Lys Ser Val His twenty two Light chain VL FAB7 Ser Tyr Val Leu Thr Gln Pro Pro Ser Val Ser Val Ala Pro Gly Gln Thr Ala Arg Ile Thr Cys Gly Gly Asn Gln Leu Gly Ser Lys Ser Val His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Val Leu Val Val Tyr Asp Asp Ser Asp Arg Pro Ser Trp Ile Pro Glu Arg Phe Ser Gly Ser Asn Ser Gly Asn Thr Ala Thr Leu Thr Ile Ser Arg Gly Glu Ala Gly Asp Glu Ala Asp Tyr Tyr Cys Gln Val Trp Asp Ser Ser Ser Asp His Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu twenty three LCDR3 FAB8 Gln Val Trp Asp Thr Ser Ser Ser Asp His Val Val twenty four Light chain VL FAB8 Ser Tyr Val Leu Thr Gln Pro Pro Ser Val Ser Val Ala Pro Gly Gln Thr Ala Arg Ile Thr Cys Gly Gly Asn Asn Leu Gly Ser Lys Ser Val His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Val Leu Val Val Tyr Asp Asp Ser Asp Arg Pro Ser Trp Ile Pro Glu Arg Phe Ser Gly Ser Asn Ser Gly Asn Thr Ala Thr Leu Thr Ile Ser Arg Gly Glu Ala Gly Asp Glu Ala Asp Tyr Tyr Cys Gln Val Trp Asp Thr Ser Ser Asp His Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 25 LCDR3 FAB9 Gln Val Trp Asp Ser Thr Ser Asp His Val Val 26 Light chain VL FAB9 Ser Tyr Val Leu Thr Gln Pro Pro Ser Val Ser Val Ala Pro Gly Gln Thr Ala Arg Ile Thr Cys Gly Gly Asn Asn Leu Gly Ser Lys Ser Val His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Val Leu Val Val Tyr Asp Asp Ser Asp Arg Pro Ser Trp Ile Pro Glu Arg Phe Ser Gly Ser Asn Ser Gly Asn Thr Ala Thr Leu Thr Ile Ser Arg Gly Glu Ala Gly Asp Glu Ala Asp Tyr Tyr Cys Gln Val Trp Asp Ser Thr Ser Asp His Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

In certain instances, the heavy and light variable chain domains of the antigen-binding fragments of the present disclosure comprise any combination of the CDR sequences listed in Table 3 below: Table 3 : CDR Combinations of Fabs of the Present Disclosure VH CDRs 1 , 2 , and 3 VL CDRs 1 , 2 , and 3 FAB1 SEQ ID NO: 1, 2 and 3 SEQ ID NO: 5, 6 and 7 Fab2 SEQ ID NO: 1, 2 and 3 SEQ ID NO: 11, 6 and 7 Fab3 SEQ ID NO: 1, 2 and 3 SEQ ID NO: 14, 6 and 7 Fab4 SEQ ID NO: 1, 15 and 3 SEQ ID NO: 5, 6 and 7 Fab5 SEQ ID NO: 1, 17 and 3 SEQ ID NO: 5, 6 and 7 Fab6 SEQ ID NO: 1, 2 and 3 SEQ ID NO: 19, 6 and 7 Fab7 SEQ ID NO: 1, 2 and 3 SEQ ID NO: 19, 6 and 7 Fab8 SEQ ID NO: 1, 2 and 3 SEQ ID NO: 5, 6 and 23 Fab9 SEQ ID NO: 1, 2 and 3 SEQ ID NO: 5, 6 and 25

In some cases, the antigen-binding fragment used in the pharmaceutical composition may be derived from WO2023098491A1, WO2021155634A1, WO2022166072A1, WO2021043221A1, WO2022184074A1, WO2021104053A1, WO2023116925A1, WO2021155861A1, WO2022116858A1, WO2022117079 A1, WO2020244544A1, WO2021152488A1, WO2022253147A1, WO2023070948A1, WO2023142309A1, WO2022095689A1, WO2021115240A1, WO2022166739A1 and WO2019100111A1, the disclosure of each of which is incorporated herein by reference.

In some cases, the antigen-binding fragment used in the pharmaceutical composition may be derived from one of those listed in Table 4 below (which includes sequence identifiers from the listed publications themselves): Table 4 : Alternative anti-TSLP antibodies that can be used in the formulations disclosed herein. WO2017/042701 An anti-TSLP antibody comprising a heavy chain (HC) CDR1 comprising the sequence of SEQ ID NO: 13, a HC CDR2 comprising the sequence of SEQ ID NO: 14, and a HC CDR3 comprising the sequence of SEQ ID NO: 15; an anti-TSLP antibody comprising a light chain (LC) CDR1 comprising the sequence of SEQ ID NO: 16, a LC CDR2 comprising the sequence of SEQ ID NO: 17, and a LC CDR3 comprising the sequence of SEQ ID NO: 18; an anti-TSLP antibody comprising a heavy chain (HC) CDR1 comprising the sequence of SEQ ID NO: 19, a HC CDR2 comprising the sequence of SEQ ID NO: 20, and a HC CDR3 comprising the sequence of SEQ ID NO: 15; an anti-TSLP antibody comprising a light chain (LC) CDR1 comprising the sequence of SEQ ID NO: 21, a LC CDR2 comprising the sequence of SEQ ID NO: 22, and a LC CDR3 comprising the sequence of SEQ ID NO: an anti-TSLP antibody comprising a HC variable region comprising a sequence of SEQ ID NO: 26 and/or a LC variable region comprising a sequence of SEQ ID NO: 27; an anti-TSLP antibody comprising a HC variable region comprising a sequence of SEQ ID NO: 28 and/or a LC variable region comprising a sequence of SEQ ID NO: 29; an anti-TSLP antibody comprising a complement of at least one of the following residues: Thr28, Asp31, Tyr32, Trp33, Asp56, Glu101, Ile102, Tyr103, Tyr104, Tyr105 of the heavy chain sequence of SEQ ID NO: 26, or a complement of at least one of the following residues: an anti-TSLP antibody that specifically binds to an antigenic determinant in human TSLP, wherein the antigenic determinant comprises at least one of the following residues: Lys38, Ala41, Leu44, Ser45, Thr46, Ser48, Lys49, Ile52, Thr53, Ser56, Gly57, Thr58, Lys59, Lys101, Gln145, and Arg149 of SEQ ID NO: 30; WO2016/142426 An anti-TSLP antibody comprising the amino acid sequence of SEQ ID NO: 31; an anti-TSLP antibody comprising a CDR1 comprising the sequence of SEQ ID NO: 32, a CDR2 comprising the sequence of SEQ ID NO: 33, and a CDR3 comprising the sequence of SEQ ID NO: 34; an anti-TSLP antibody comprising a CDR1 comprising the sequence of SEQ ID NO: 32, a CDR2 comprising the sequence of SEQ ID NO: 35, and a CDR3 comprising the sequence of SEQ ID NO: 34; an anti-TSLP antibody comprising a variant of CDR1 of SEQ ID NO: 31, wherein the residue corresponding to residue 28 in SEQ ID NO: 31 is Pro, the residue corresponding to residue 30 in SEQ ID NO: 31 is Arg, the residue corresponding to residue 31 in SEQ ID NO: 31 is Arg, The residue corresponding to residue 31 of SEQ ID NO: 31 is Asn, the residue corresponding to residue 32 of SEQ ID NO: 31 is Trp, and the residue corresponding to residue 34 of SEQ ID NO: 31 is Asp; An anti-TSLP antibody comprising a variant of CDR2 of SEQ ID NO: 31, wherein the residue corresponding to residue 50 of SEQ ID NO: 31 is Gly, the residue corresponding to residue 53 of SEQ ID NO: 31 is His, and the residue corresponding to residue 55 of SEQ ID NO: 31 is Gln; An anti-TSLP antibody comprising a variant of CDR3 of SEQ ID NO: 31, wherein the residue corresponding to residue 54 of SEQ ID NO: 31 is Asn, the residue corresponding to residue 32 of SEQ ID NO: 31 is Trp, and the residue corresponding to residue 34 of SEQ ID NO: 31 is Asp; The residue 91 in SEQ ID NO:31 is He, Leu, Val or Phe, the residue corresponding to residue 92 in SEQ ID NO:31 is Gly or Ala, the residue corresponding to residue 93 in SEQ ID NO:31 is Glu, Phe, Asp or Ser, and the residue corresponding to residue 94 in SEQ ID NO:31 is Asp. WO2010/017468 an anti-TSLP antibody (9B7) comprising a HC CDR3 comprising the sequence of SEQ ID NO: 38, wherein the other CDRs of the HC and LC comprise the sequences of SEQ ID NOs: 36, 37, and 39-41; an anti-TSLP antibody (6C5) comprising a HC CDR3 comprising the sequence of SEQ ID NO: 44, wherein the other CDRs of the HC and LC comprise the sequences of SEQ ID NOs: 42, 43, and 45-47; an anti-TSLP antibody (6A3) comprising a HC CDR3 comprising the sequence of SEQ ID NO: 50, wherein the other CDRs of the HC and LC comprise the sequences of SEQ ID NOs: 48, 49, and 51-53; an anti-TSLP antibody (1A11) comprising a HC CDR3 comprising the sequence of SEQ ID NO: 56, wherein the other CDRs of the HC and LC comprise the sequences of SEQ ID NOs: 54, 55, and 57-59; An anti-TSLP antibody comprising (i) a heavy chain variable region of SEQ ID NO: 60 and/or a light chain variable region of SEQ ID NO: 61; an anti-TSLP antibody comprising (i) a heavy chain variable region of SEQ ID NO: 62 and/or a light chain variable region of SEQ ID NO: 63; an anti-TSLP antibody comprising (i) a heavy chain variable region of SEQ ID NO: 64 and/or a light chain variable region of SEQ ID NO: 65; an anti-TSLP antibody comprising (i) a heavy chain variable region of SEQ ID NO: 66 and/or a light chain variable region of SEQ ID NO: 67; an anti-TSLP antibody comprising (i) a heavy chain variable region of SEQ ID NO: 68 and/or a light chain variable region of SEQ ID NO: 69; An anti-TSLP antibody comprising an HC CDR selected from the group consisting of SEQ ID NO: 38, SEQ ID NO: 44, SEQ ID NO: 50, and SEQ ID NO: 56, and analogs thereof; an anti-TSLP antibody comprising a heavy chain comprising the following CDRs or analogs thereof: CDRH1: RYNVH (SEQ ID NO: 36) [referred to herein as "SEQ ID NO: 32"], CDRH2: MIWDGGSTDYNSALKS (SEQ ID NO: 37) [referred to herein as "SEQ ID NO: 33"], CDRH3: NRYESG (SEQ ID NO: 38) [referred to herein as "SEQ ID NO: 34"], and a light chain comprising the following CDRs or analogs thereof: CDRL1: KSSQSLLNSGNRKNYLT (SEQ ID NO: 39) [referred to herein as "SEQ ID NO: 35"], CDRL2: WASTRES (SEQ ID NO: 40) [referred to herein as “SEQ ID NO: 36”], and CDRL3: QNDYTYPFTFGS (SEQ ID NO: 41) [referred to herein as “SEQ ID NO: 37”]; or an anti-TSLP antibody comprising a heavy chain comprising the following CDRs or their analogs: CRDH1: AYWMS (SEQ ID NO: 42) [referred to herein as “SEQ ID NO: 38”], CDRH2: EINPDSSTINCTPSLKD (SEQ ID NO: 43) [referred to herein as “SEQ ID NO: 39”], CDRH3: RLRPFWYFDVW (SEQ ID NO: 44) [referred to herein as “SEQ ID NO: 40”], and a light chain comprising the following CDRs or their analogs: CDRL1: RSSQSIVQSNGNTYLE (SEQ ID NO: 45) [referred to herein as “SEQ ID NO: 41”], CDRL2: KVSNRFS (SEQ ID NO: 46) [referred to herein as “SEQ ID NO: 47”]. NO: 46) [referred to herein as "SEQ ID NO: 42"], and CDRL3: FQGSHVPRT (SEQ ID NO: 47) [referred to herein as "SEQ ID NO: 43"]; an anti-TSLP antibody comprising a heavy chain comprising the following CDRs or their analogs: CRDH1: TDYAWN (SEQ ID NO: 48) [referred to herein as "SEQ ID NO: 44"], CDRH2: YIFYSGSTTYTPSLKS (SEQ ID NO: 49) [referred to herein as "SEQ ID NO: 45"], CDRH3: GGYDVNYF (SEQ ID NO: 50) [referred to herein as "SEQ ID NO: 46"], and a light chain comprising the following CDRs or their analogs: CDRL1: LASQTIGAWLA (SEQ ID NO: 51) [referred to herein as "SEQ ID NO: 47"], CDRL2: AATRLAD (SEQ ID NO: 52) [referred to herein as "SEQ ID NO: 53"] NO: 52) [referred to herein as "SEQ ID NO: 48"], and CDRL3: QQFFSTPWT (SEQ ID NO: 53) [referred to herein as "SEQ ID NO: 49"]; an anti-TSLP antibody comprising a heavy chain comprising the following CDRs or their analogs: CDRH1: GYTMN (SEQ ID NO: 54) [referred to herein as "SEQ ID NO: 50"], CDRH2: LINPYNGVTSYNQKFK [referred to herein as "SEQ ID NO: 51"] (SEQ ID NO: 55), CDRH3: GDGNYWYF (SEQ ID NO: 56) [referred to herein as "SEQ ID NO: 52"], and a light chain comprising the following CDRs or their analogs: CDRL1: SASSSVTYMHW (SEQ ID NO: 57) [referred to herein as "SEQ ID NO: 53"], CDRL2: EISKLAS (SEQ ID NO: 58) [referred to herein as "SEQ ID NO: 59"] NO: 58) [referred to herein as "SEQ ID NO: 54"], and CDRL3: QEWNYPYTF (SEQ ID NO: 59) [referred to herein as "SEQ ID NO: 55"]; an anti-TSLP antibody comprising a HC CDR1 comprising the sequence of SEQ ID NO: 70, a CDR2 comprising the sequence of SEQ ID NO: 71, and a CDR3 comprising the sequence of SEQ ID NO: 72; an anti-TSLP antibody comprising a LC CDR1 comprising the sequence of SEQ ID NO: 73, a CDR2 comprising the sequence of SEQ ID NO: 74, and a CDR3 comprising the sequence of SEQ ID NO: 75; US2012/0020988 An anti-TSLP antibody comprises a heavy chain variable domain comprising a CDR1 region of SEQ ID NO: 76, a CDR2 region of SEQ ID NO: 77, and a CDR3 region of SEQ ID NO: 78, and a light chain variable domain comprising a CDR1 region of SEQ ID NO: 79, a CDR2 region of SEQ ID NO: 80, and a CDR3 region of SEQ ID NO: 81. An anti-TSLP antibody comprising a heavy chain variable domain comprising SEQ ID NO: 82 and a light chain variable domain comprising SEQ ID NO: 83; an anti-TSLP antibody comprising a heavy chain variable domain comprising a CDR1 region of SEQ ID NO: 76 or 84, a CDR2 region of SEQ ID NO: 77 or 85, and a CDR3 region of SEQ ID NO: 78, and a light chain variable domain comprising a CDR1 region of SEQ ID NO: 79 or 86, a CDR2 region of SEQ ID NO: 80, 87 or 88, and a CDR3 region of SEQ ID NO: 81. An anti-TSLP antibody comprising a heavy chain variable domain comprising a CDR1 region of SEQ ID NO: 76, a CDR2 region of SEQ ID NO: 85, and a CDR3 region of SEQ ID NO: 78, and a light chain variable domain comprising a CDR1 region of SEQ ID NO: 86, a CDR2 region of SEQ ID NO: 87, and a CDR3 region of SEQ ID NO: 81; An anti-TSLP antibody comprising a heavy chain variable domain comprising a CDR1 region of SEQ ID NO: 76, a CDR2 region of SEQ ID NO: 85, and a CDR3 region of SEQ ID NO: 78, and a light chain variable domain comprising a CDR1 region of SEQ ID NO: 86, a CDR2 region of SEQ ID NO: 88, and a CDR3 region of SEQ ID NO: 81; An anti-TSLP antibody comprising a heavy chain variable domain comprising a CDR1 region of SEQ ID NO: 84, a CDR2 region of SEQ ID NO: 85, and a CDR3 region of SEQ ID NO: 78, and a light chain variable domain comprising a CDR1 region of SEQ ID NO: 86, a CDR2 region of SEQ ID NO: 88, and a CDR3 region of SEQ ID NO: 81; or an anti-TSLP antibody comprising a heavy chain variable domain comprising a CDR1 region of SEQ ID NO: 76, a CDR2 region of SEQ ID NO: 85, and a CDR3 region of SEQ ID NO: 78, and a light chain variable domain comprising a CDR1 region of SEQ ID NO: 86, a CDR2 region of SEQ ID NO: 80, and a CDR3 region of SEQ ID NO: 81. An anti-TSLP antibody comprising a heavy chain variable domain comprising SEQ ID NO: 89, and a light chain variable domain comprising SEQ ID NO: 90; an anti-TSLP antibody comprising a heavy chain variable domain comprising SEQ ID NO: 89, and a light chain variable domain comprising SEQ ID NO: 91; an anti-TSLP antibody comprising a heavy chain variable domain comprising SEQ ID NO: 92, and a light chain variable domain comprising SEQ ID NO: 93; an anti-TSLP antibody comprising a heavy chain variable domain comprising SEQ ID NO: 89, and a light chain variable domain comprising SEQ ID NO: 94; US8637019 An anti-TSLP antibody comprising a heavy chain variable region comprising a CDR-H1 sequence comprising SEQ ID NO: 95, a CDR-H2 sequence comprising SEQ ID NO: 96, and a CDR-H3 sequence comprising SEQ ID NO: 97; and/or an antibody light chain variable region or a TSLP-binding fragment thereof, wherein the light chain variable region comprises a CDR-L1 sequence comprising SEQ ID NO: 98, a CDR-L2 sequence comprising SEQ ID NO: 99, and a CDR-L3 sequence comprising SEQ ID NO: 100. The anti-TSLP antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 101 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 102. An anti-TSLP antibody comprising SEQ ID NO: 103 and SEQ ID NO: 104. WO2020244544 An anti-TSLP antibody comprises a heavy chain variable region and a light chain variable region, wherein: the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 26, SEQ ID NO: 94 and SEQ ID NO: 28, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 29, SEQ ID NO: 113 and SEQ ID NO: 31, respectively.

Treatment Methods Also provided herein are treatment methods comprising administering to a subject in need thereof a therapeutically effective amount of a pharmaceutical composition described herein. In some instances, the method is used to treat a TSLP-related disorder.

"Treat" or "treatment" refers to both therapeutic treatment and prophylactic or preventative measures, wherein the goal is to prevent or slow down (lessen) an undesirable physiological change or condition. Beneficial or desired clinical results include, but are not limited to, relief of symptoms, reduction in severity of disease, stabilization (i.e., non-worsening) of the disease state, delay or slowing of disease progression, improvement or palliation of the disease state, and remission (whether partial or complete), whether detectable or undetectable. "Treatment" may also mean prolonging survival as compared to expected survival if not receiving treatment. Those in need of treatment include those already with the disease or condition as well as those who are susceptible to it or those in whom it is to be prevented.

A "therapeutically effective amount" refers to an amount of an antigen-binding fragment of an anti-TSLP antibody or other drug disclosed herein that is effective to "treat" a disease or condition in a subject or mammal.

Unless the term "subject" is defined as a "healthy individual," the terms "subject," "individual," "animal," "patient," or "mammal" refer to any individual, particularly a mammalian individual, for whom diagnosis, prognosis, or treatment is desired. Mammalian individuals include humans, livestock, and farm animals, such as dogs, cats, guinea pigs, rabbits, rats, mice, horses, cattle, and cows. Preferably, the subject is a human. The patient may be an adult, child, or adolescent.

Also provided herein are pharmaceutical compositions for use in therapy. In some cases, the therapy is for treating a TSLP-related disorder.

Also provided herein are uses of antigen-binding fragments of anti-TSLP antibodies or pharmaceutical compositions comprising the antigen-binding fragments in the manufacture of a medicament for treating a disease. In some cases, the disease is a TSLP-related disease.

The present disclosure also provides for the use of an antigen-binding fragment of an anti-TSLP antibody or a pharmaceutical composition comprising the antigen-binding fragment in a therapy. In some cases, the therapy is for treating a TSLP-related disease.

In this section of the specification, any reference to a method of treatment should be construed as a reference to a corresponding disclosure of the use or pharmaceutical composition for use.

In some instances, the TSLP-related disease is a TSLP-related inflammatory disease. In some instances, the TSLP-related inflammatory disease is selected from asthma, sepsis, septic shock, atopic dermatitis, allergic rhinitis, allergic sinusitis, allergic conjunctivitis, eosinophilic esophagitis, rheumatoid arthritis, chronic obstructive pulmonary disease (COPD), asthma, COPD overlay syndrome (ACOS), chronic bronchitis, emphysema, chronic sinusitis with or without nasal polyps, vasculitis, GvHD, uveitis, chronic idiopathic urticaria, sinusitis, or pancreatitis.

In some cases, the TSLP-related condition is asthma.

In some cases, the TSLP-related condition is COPD.

In some instances, the asthma is moderate or severe asthma, or moderate to severe asthma. In some instances, a subject having moderate or severe asthma, or moderate to severe asthma, may mean that the subject has a diagnosis of moderate or severe asthma, or moderate to severe asthma. In some instances, the subject may have a documented history of moderate or severe asthma, or moderate to severe asthma, for at least one year.

Asthma is a chronic inflammatory disease of the airways that affects 1-18% of the population across different countries and is characterized by bronchial hyperresponsiveness and reversible airflow limitation. It is defined by a history of respiratory symptoms such as wheezing, shortness of breath, chest tightness, and cough. The etiology of asthma is believed to be multifactorial, with identifiable demographic, clinical, and/or pathophysiological phenotypes. In patients with more severe phenotypes, some phenotype-guided treatments are available. However, a close relationship between pathological symptoms and clinical manifestations and response to therapy has not been established.

Asthma can be diagnosed or assessed by a variety of different measures, including: Assessment of airway inflammation using the standardized single-breath fraction of exhaled nitric oxide (FeNO) (ATS, Am J Respir Crit Care Med. 171(8):912-30, 2005). FeNO has not been established for confirming a diagnosis of asthma, but elevated FeNO has been associated with asthma characterized by type 2 airway inflammation.

Determine the presence of atopy. This can be identified by skin prick testing for common environmental allergens or by measuring serum levels of specific IgE. Like FeNO, allergy testing cannot rule in or rule out a diagnosis of asthma, but the presence of an atopy increases the likelihood that a patient with respiratory symptoms has allergic asthma.

Bronchial provocation tests. These tests monitor variable airflow limitation to assess airway hyperresponsiveness (AHR). Individuals can be challenged with a chemical agent such as acetylcholine. These tests are moderately sensitive for diagnosing asthma.

The term "FENO" refers to fractional exhaled nitric oxide, a biomarker for bronchial or airway inflammation. FENO is produced by airway epithelial cells in response to inflammatory interleukins (such as TSLP, IL-4, and IL-13). FENO levels in healthy adults range from 2 to 30 parts per billion (ppb). An exemplary test for measuring FENO involves an individual inhaling to total vital capacity using a NIOX MINO® Airway Inflammation Monitor and then exhaling at 50 ml/sec for 10 seconds (assisted by visual and auditory cues).

Different subtypes of asthma have been identified, including allergic asthma, non-allergic asthma, late-onset asthma (which generally tends to be non-allergic), asthma with persistent airflow limitation (which is associated with airway wall remodeling, resulting in long-term, persistent, irreversible airflow limitation), and asthma associated with obesity (which is generally associated with a non-eosinophilic/low-eosinophilic mechanism of action). In some cases, individuals treated by the present disclosure may have asthma of any type or origin.

There are different levels of asthma severity, currently assessed retrospectively based on the level of treatment required to control symptoms and exacerbations. The severity index comprises three main groups: mild asthma, moderate asthma, and severe asthma. Asthma severity is defined on the GINA scale by the level of treatment required to achieve adequate symptom control. The GINA scale is defined in the "Asthma Management and Prevention Pocket Guide," Global Asthma Initiative; 2019. Unless otherwise specified herein, references to "moderate asthma" or "severe asthma" are based on the definition on the GINA scale. For example, moderate asthma is defined as asthma with a Global Initiative for Asthma (GINA) scale score of 3 or less, and appropriately a GINA scale score of 2 or 3 (i.e., GINA Step 2 or Step 3), and severe asthma is defined as asthma that requires high-intensity treatment (e.g., GINA Steps 4 and 5) to maintain good control or that is not well controlled despite high-intensity treatment (GINA, Global Strategy for Asthma Management and Prevention. Global Initiative for Asthma (GINA), December 2012).

In some cases, individuals have a history of ≥1 or ≥2 severe exacerbations in the last 12 months prior to treatment. Severe exacerbations are defined as those resulting in hospital admission, emergency room visit, and/or treatment with oral corticosteroids, as detailed below: Inpatient admission: admission to an inpatient facility and/or evaluation and treatment in a healthcare facility for asthma for ≥24 hours; Emergency room or urgent care visit: evaluation and treatment in an emergency room or urgent care center for asthma requiring systemic corticosteroids for <24 hours; and/or A temporary bolus/burst of systemic corticosteroids (or temporarily increase the stable OCS background dose) for at least 3 consecutive days to treat symptoms of worsening asthma; a single depo injectable dose of corticosteroids will be considered equivalent to a 3-day bolus/pulse of systemic corticosteroids.

In some cases, the asthma is moderate asthma, severe asthma, or moderate to severe asthma. In some cases, the asthma is not well controlled with standard of care therapy, which includes controllers or relievers, as defined in Steps 1 and 2 of the GINA scale. Thus, in some cases, the asthma to be treated by the present disclosure may be uncontrolled asthma. In some cases, the asthma is moderate asthma that is not controlled with standard of care therapy, severe asthma that is not controlled with standard of care therapy, or moderate to severe asthma that is not controlled with standard of care therapy. Standard of care (SOC) therapy is as defined in the GINA scale.

Chronic obstructive pulmonary disease (COPD) is a progressive disease and a significant cause of morbidity and mortality worldwide. Compared to other chronic diseases, COPD is increasingly prevalent and is projected to become the third leading cause of death and disability worldwide by 2020.

Acute exacerbations of COPD (AECOPD) contribute significantly to the economic burden of COPD. In addition to the significant economic burden, AECOPD also accounts for a significant portion of COPD morbidity and mortality. Compared to patients with less frequent exacerbations, patients with frequent AECOPD exhibit increased airway inflammation and accelerated lung function decline.

In some cases, the subject has a baseline blood eosinophil count of ≥ 150 cells/μl or ≥ 300 cells/μl. In some cases, baseline refers to a blood eosinophil count before starting treatment (e.g., within one month of starting treatment).

In some cases, the methods disclosed herein improve lung function in a subject with asthma. In some cases, improved lung function refers to an improvement compared to baseline in one or more of the following: pre-bronchodilator (BD) FVC, post-BD FVC, pre-BD FEV1, post-BD FEV1, mean morning PEF, or mean evening PEF.

In some instances, improvement means achieving a minimal clinically important difference (MCID) for each of pre-bronchodilator (BD) FVC, post-BD FVC, pre-BD FEV1, post-BD FEV1, mean morning PEF, or mean evening PEF.

The phrase "minimal clinically important difference," or "MCID," means the smallest change in a treatment outcome that an individual patient would identify as important and would indicate a change in patient management. Some MCIDs are experimentally validated, while others are study-specific.

The term "pre-BD FEV1,""pre-BDFEV1," or "pre-bronchodilator (BD) FEV1" refers to the pre-bronchodilator forced expiratory volume 1. This is a measure of the amount of air a person can exhale in 1 second before the administration of a bronchodilator. In some cases, the minimum clinically important difference in pre-BD _FEV1 is 100 ml.

In some cases, the increase in pre-BD _FEV1 compared to baseline (e.g., pre-BD _FEV1 value before starting treatment) is at least 5 ml, at least 10 ml, at least 15 ml, at least 20 ml, at least 25 ml, at least 30 ml, at least 35 ml, at least 40 ml, at least 45 ml, at least 50 ml, at least 55 ml, at least 60 ml, at least 65 ml, at least 70 ml, at least 75 ml, at least 80 ml, at least 85 ml, at least 90 ml, at least 95 ml, at least 100 ml, at least 105 ml, at least 110 ml, at least 115 ml, at least 120 ml, at least 125 ml, at least 130 ml, at least 135 ml, at least 140 ml, at least 145 ml, at least 150 ml, at least 160 ml, at least 170 ml, at least 180 ml, at least 190 ml, at least 200 ml, at least 210 ml, at least 220 ml, at least 230 ml ml, at least 240 ml, or at least 250 ml. In some instances, the increase in pre-BD _FEV1 compared to baseline (e.g., the pre-BD _FEV1 value before starting treatment) is at least 80 ml on day 2 after starting treatment, at least 45 ml or at least 100 ml on day 7 after starting treatment, at least 100 ml on day 14 after starting treatment, or at least 5 ml or at least 100 ml on day 28 after starting treatment.

The term "post-BD FEV1" or "post-bronchodilator (BD) FEV1" refers to the forced expiratory volume after bronchodilator 1. This is a measure of the amount of air a person can exhale in 1 second after the administration of a bronchodilator.

The term "pre-BD FVC" or "pre-bronchodilator (BD) forced vital capacity (FVC)" refers to the forced vital capacity after bronchodilators. This is the total volume of air exhaled during a forced expiratory volume test or FVC test before the administration of a bronchodilator.

The term "post-BD FVC" or "post-bronchodilator (BD) FVC" refers to forced vital capacity after bronchodilator administration. This is the total volume of air exhaled during a forced expiratory volume test or FVC test after the administration of a bronchodilator.

The term "bronchodilator" refers to a substance that dilates the bronchi and bronchioles, reduces airway resistance, and increases airflow to the lungs. Suitable bronchodilators include short-acting beta-agonists (SABAs), such as albuterol (90 1-1 g dose) or salbutamol (100 1-1 g dose), or equivalents (Sorkness et al., J Appl Physiol. 104(2): 394-403, 2008).

The term "peak expiratory flow (PEF)" indicates the fastest rate at which a person can force air out of the lungs during a forced expiratory volume test or FVC test, and is usually measured in liters per minute.

The term "forced expiratory volume (FEV)" refers to the volume of air exhaled during the first, second, and third seconds of an FVC test. As explained above, the term FEV1 refers to the volume of air exhaled during the first second of an FVC test.

The term "forced vital capacity" (FVC) or forced vital capacity test is a measurement of the total amount of air a person can forcefully and quickly exhale after taking as much air as possible.

In some cases, spirometry is performed according to the ATS/European Respiratory Society (ERS) guidelines (Miller et al., Eur Respir J. 26(1): 153-61, 2005) for _FEV1 before BD, _FEV1 after BD, FVC before BD, and FVC after BD. For example, multiple forced expiratory efforts (at least 3 but not more than 8) are performed during each spirometry, and the two best efforts that meet the ATS/ERS acceptability and reproducibility criteria are recorded. The best effort will be based on the highest FEV1. The maximum forced expiratory volume (FEV1) of the two best efforts will be used for analysis. Both absolute measurements (for FEV1 and forced vital capacity (FVC)) and percentages of predicted normal values will be recorded using appropriate reference values. The maximum FVC will be reported regardless of the effort with which it occurred (even if the effort did not produce the maximum FEV1).

Post-bronchodilator (post-BD) spirometry testing is assessed after the individual has performed pre-BD spirometry. Before administering an appropriate bronchodilator to the individual, the pre-BD FEV1 is measured using spirometry as defined above. To measure post-BD FEV1, a short-acting beta-agonist (SABA) (such as albuterol (90 1-1 g dose) or albuterol (100 1-1 g dose) or equivalent) is used to induce maximal bronchodilation using a spacer device for a total of up to 8 puffs (Sorkness et al., J Appl Physiol. 1 04(2): 394-403, 2008). Reversibility was determined using the highest pre-BD FEV1 and post-BD FEV1 achieved after 4, 6, or 8 puffs and was used for analysis. The reversibility algorithm was as follows: % reversibility = (post-BD FEV1 - pre-BD FEV1) × 1.00 / pre-BD FEV1. The Ph1b portion of the Investigating the Safety, Tolerability and Effects of FAB1 in Healthy Subjects and Subjects With Asthma on Inhaled Corticosteroids and Long-acting Beta-agonists study (NCT05110976) demonstrated that FAB1 resulted in numerical improvements in lung function (i.e., pre-BD _FEV1 ), as shown in Figure 26. Thus, in some cases, the methods disclosed herein improve pre-BD FEV ₁ in individuals with TSLP-related disorders, such as asthma.

In some cases, lung function improves within 0.5 hours of administering a first dose of a composition disclosed herein. In some cases, lung function improves within 1 hour of administering a first dose of a composition disclosed herein. In some cases, lung function improves within 6 hours of administering a first dose of a composition disclosed herein. In some cases, lung function improves within 24 hours of administering a first dose of a composition disclosed herein. In some cases, lung function improves within 7 days of administering a first dose of a composition disclosed herein. In some cases, lung function improves within 14 days of administering a first dose of a composition disclosed herein. In some cases, lung function improves within 28 days of administering a first dose of a composition disclosed herein. Figure 26B shows that subjects administered 8 mg of FAB1 in Part B of NCT05110976 showed a trend toward improvement in pre-BD FEV1 in the high-dose group, with an increase of 105 ml on day 28, and an early effect was observed 6 hours after the first dose.

In some cases, the methods disclosed herein improve asthma symptoms in a subject. In some cases, improving asthma symptoms means an improvement compared to baseline in one or more of the following: mean Asthma Symptom Diary score, ACQ-6 score, AQLQ score, or SGRQ score.

Asthma Control Questionnaire (ACQ)-6 The Asthma Control Questionnaire (ACQ-6) is a patient-reported questionnaire that assesses asthma symptoms (i.e., nocturnal awakenings, symptoms upon awakening, activity limitation, shortness of breath, wheezing), as well as daily rescue bronchodilator use and FEV1 (Juniper et al., October 1999). The ACQ-6 is a shortened version of the ACQ that omits the FEV1 measurement from the original ACQ score. Questions are equally weighted and scored from 0 (completely controlled) to 6 (very uncontrolled). The mean ACQ score is the average of the responses. An average score of 0.75 indicates well-controlled asthma, a score between 0.75 and 1.5 indicates partially controlled asthma, and a score > 1.5 indicates uncontrolled asthma (Juniper et al., Respir Med. 100(4):616-21, 2006). Individual changes of at least 0.5 are considered clinically meaningful (Juniper et al., Respir Med. 99(5):553-8, 2005). Therefore, in some cases, the minimum clinically important difference for the ACQ-6 is 0.5.

In some instances, the disclosure provides methods for improving lung function, wherein the method results in an improvement in ACQ-6 score of 0.5 points compared to baseline.

Standardized Asthma Quality of Life Questionnaire (AQLQ(S)+12) for individuals aged 12 years and older. The AQLQ(S)+12 (or "AQLQ") measures the health-related quality of life experienced by individuals with asthma. The questionnaire consists of four separate domains (symptoms, activity limitations, emotional function, and environmental stimulation). Individuals are asked to recall their experiences over the previous two weeks and rate each question on a 7-point scale ranging from 7 (no impairment) to 1 (severe impairment). The overall score is calculated as the average response to all questions. The scores for the four individual domains (symptoms, activity limitations, emotional function, and environmental stimulation) are the average of the responses to the questions in each domain. AQLQ+12 responders were defined as having an improvement of 0.5 points relative to baseline. Therefore, in some cases, the minimum clinically important difference in AQLQ was 0.5.

In some instances, the disclosure provides methods for improving lung function, wherein the method results in an improvement of 0.5 points in the AQLQ compared to baseline.

St. George's Respiratory Questionnaire (SGRQ) The SGRQ is a 50-item PRO instrument developed to measure the health status of patients with obstructive airway disease (Jones et al. 1991). The questionnaire consists of two parts: Part 1 consists of 8 items regarding the severity of respiratory symptoms in the previous 4 weeks; Part 2 consists of 42 items related to the daily activities and psychosocial impact of the individual's respiratory condition. The SGRQ produces a total score and three domain scores (symptoms, activities, and impact). The total score indicates the impact of the disease on overall health status. This total score is expressed as a percentage of overall impairment, with 100 indicating the worst possible health status and 0 indicating the best possible health status. Similarly, domain scores range from 0 to 100, with higher scores indicating greater impairment. Based on empirical data and patient interviews, a mean change score of 4 units is associated with the minimal clinically important difference (MCID). Specific details regarding the scoring algorithm are provided by the developer in the user manual (Jones et al. 2009). The SGRQ is a qualified biomarker, and the responder definition is typically an improvement of 4 points from baseline. Therefore, in some cases, the minimal clinically important difference for the SGRQ is 4.

In some instances, the disclosure provides methods for improving lung function, wherein the method results in an improvement of 4 points in SGRQ score compared to baseline.

In some cases, improvement in asthma symptoms occurs within 7 days of taking the first dose of a composition disclosed herein. In some cases, improvement in asthma symptoms occurs within 14 days of taking the first dose of a composition disclosed herein. In some cases, improvement in asthma symptoms occurs within 28 days of taking the first dose of a composition disclosed herein. Figure 27 shows that subjects administered 8 mg of FAB1 in Part B of NCT05110976 showed a trend toward improvement in ACQ-6 scores, as demonstrated by the mean change from baseline over time in the high dose group (8 mg).

Composite Exacerbation (CompEx) Event Rate CompEx asthma is a composite endpoint that allows evaluation of treatment effects on exacerbations and involves fewer participants than severe exacerbations. There are two main types of CompEx asthma events: • Severe exacerbations of asthma • Diary-based (objective worsening)

CompEx for asthma is described in more detail in Example 8.

In some aspects, the present disclosure provides methods for improving the time to first CompEx event in individuals with asthma.

In another aspect, the disclosure provides methods for improving lung function in an individual with asthma, wherein the improvement in lung function comprises an improvement in time to first CompEx event compared to placebo.

In another aspect, the present disclosure provides methods for improving lung function in an individual with asthma, wherein the improvement in lung function comprises an improvement in the time to first CompEx event. In some cases, the improved time is compared to a placebo. In some cases, the improved time is compared to a baseline. In some cases, the baseline is the time to first CompEx event in an individual who has not yet received a treatment described herein. As described herein, the ability to deliver antigen-binding fragments of anti-TSLP antibodies by inhalation provides a delivery mechanism that is more suitable for use in a primary care setting. Therefore, the pharmaceutical compositions described herein can be administered by inhalation or intranasally. Therefore, in some cases, the pharmaceutical compositions described herein can be inhalable pharmaceutical compositions. In some cases, the pharmaceutical compositions are administered by dry powder inhaler.

In the context of methods for treating asthma, the compositions can be administered more frequently and at lower doses than with systemically administered anti-TSLP drugs. In some cases, the formulation can be administered daily. This can be more convenient for the individual or patient. Furthermore, it can reduce side effects that can occur with systemic administration.

In some cases, after treatment with the pharmaceutical compositions or dry powder formulations disclosed herein, the prevalence of anti-drug antibodies (ADA) in a subject is less than 6%, less than 5%, or less than 4%, and/or the incidence of ADA is less than 4%, less than 3%, or less than 2%. ADA prevalence is the percentage of ADA-evaluable participants who are ADA+ at any time, while ADA incidence is the percentage of ADA-evaluable participants who have treatment-emergent anti-drug antibodies (TE-ADA+).

In some cases, the compositions provide the possibility of treating patients with moderate to severe asthma that can be managed in a primary care setting, or the possibility of treating patients with moderate to severe asthma that is difficult to treat through specialist care. For example, the compositions can be used to treat moderate to severe asthma patients with a Global Initiative for Asthma (GINA) scale of 4-5. Suitably, the compositions provide the possibility of treating uncontrolled moderate to severe asthma. Suitably, the compositions provide the possibility of treating moderate to severe asthma that is uncontrolled on moderate to high doses of ICS:LABA and has one or more exacerbations and frequent symptoms.

The pharmaceutical compositions disclosed herein can be administered in combination with any known treatment for asthma and/or COPD, including any agent or combination of agents known to be useful, used, or currently used to treat inflammatory diseases such as asthma or COPD. Exemplary active agents that can be administered in combination with the compositions described herein include, but are not limited to, inhaled corticosteroids (ICS), bronchodilators (including long-acting beta agonists (LABAs), long-acting antimuscarinic agonists (LAMAs), short-acting beta agonists (SABAs), and muscarinic beta2-agonists (MABAs)), antihistamines, antileukotrienes, PDE-4 inhibitors, janus kinase inhibitors, and phosphoinositide 3-kinase inhibitors.

Therefore, in some cases, background therapy is co-administered to an individual who is being treated or is currently being treated. In some cases, the individual is already receiving background therapy prior to treatment. In some instances, background therapy is selected from: inhaled corticosteroids; leukotriene modulators; long-acting beta agonists (LABAs); long-acting muscarinic antagonists (LAMAs); combination therapies such as fluticasone and salmeterol, adenine and formoterol, mometasone and formoterol, and fluticasone and vilanterol; theophylline; short-acting beta agonists (SABAs); ipratropium or a combination of ipratropium and albuterol, or oral corticosteroids. In some cases, background therapy includes a combination of medium- or high-dose ICS (as reported in GINA 2023) and LABA (GINA Step 4 or 5 therapy).

The term "combination" refers to a fixed combination in the form of a dosage unit, or a combined administration, wherein a pharmaceutical composition as described herein and a combination partner (e.g., another drug, also referred to as a "therapeutic agent" or "adjuvant") can be administered independently at the same time or separately within a time interval, especially where such time interval allows the combination partners to exhibit a synergistic effect (e.g., a synergistic effect). The individual components can be packaged in a kit or individually. One or both of the components (e.g., powders or liquids) can be reconstituted or diluted to the desired dose prior to administration. As used herein, the terms "co-administration" or "combination administration" or the like are intended to encompass administration of the selected combination partners to a single individual (e.g., a patient) in need thereof, and are intended to include treatment regimens in which the agents are not necessarily administered by the same route of administration or at the same time. As used herein, the term "pharmaceutical combination" refers to a product resulting from the admixture or combination of more than one therapeutic agent and includes both fixed and non-fixed combinations of therapeutic agents. The term "fixed combination" means that both therapeutic agents (e.g., anti-TSLP Fab and combination partner) are administered to a patient simultaneously in the form of a single entity or dosage. The term "non-fixed combination" means that both therapeutic agents (e.g., anti-TSLP Fab and a combination partner) are administered to a patient as separate entities simultaneously, concurrently, or sequentially, without specific time limits, wherein such administration provides therapeutically effective levels of both compounds in the patient. The latter also applies to cocktail therapies, e.g., administration of three or more therapeutic agents.

The term "combination therapy" refers to the administration of two or more therapeutic agents to treat the therapeutic conditions or disorders described in this disclosure. Such administration encompasses co-administration of these therapeutic agents in a substantially simultaneous manner (e.g., in a single capsule with a fixed ratio of active ingredients). Alternatively, such administration encompasses co-administration of each active ingredient in multiple or separate containers (e.g., tablets, capsules, powders, and liquids). Powders and/or liquids can be reconstituted or diluted to the desired dose prior to administration. Additionally, such administration encompasses the use of each type of therapeutic agent in a sequential manner, at approximately the same time or at different times. In either case, the treatment regimen will provide the beneficial effects of the drug combination in treating the conditions or disorders described herein.

In some cases, the pharmaceutical composition may be administered to humans or other animals in an amount sufficient to produce a therapeutic effect according to the above-mentioned treatment methods/medical uses.

In some cases, the pharmaceutical composition can be administered to such humans or other animals in a conventional dosage form prepared by combining anti-TSLP Fab with a conventional pharmaceutically acceptable carrier or diluent according to known techniques.

Those skilled in the art will recognize that the form and characteristics of the pharmaceutically acceptable carrier or diluent are determined by the amount of active ingredient to be combined with it, the route of administration and other well-known variables.

In some cases, the pharmaceutical composition is formulated to include a pharmaceutically acceptable nontoxic sterile carrier, such as physiological saline, nontoxic buffers, preservatives, and the like. In some cases, the pharmaceutical composition may include sterile aqueous or non-aqueous solutions, suspensions, and emulsions. Suitable formulations for use in the treatment methods disclosed herein are described in Remington's Pharmaceutical Sciences (Mack Publishing Co.), 16th edition (1980).

In some cases, the components described herein for preparing the pharmaceutical composition can be packaged and sold in a kit. In some cases, such a kit will have a label or package insert indicating that the pharmaceutical composition can be used to treat an individual suffering from or susceptible to a disease or condition.

Dosage Regimens In some instances, a pharmaceutical composition is administered or intended to be administered to a subject in need thereof at a dose of about 0.2 mg to about 16 mg of an anti-TSLP antigen-binding fragment. For example, a pharmaceutical composition is administered or intended to be administered to a subject in need thereof at a dose of about 0.2 mg, about 0.4 mg, about 0.6 mg, about 2 mg, about 6 mg, about 8 mg, or about 16 mg of an anti-TSLP antigen-binding fragment. In some instances, a pharmaceutical composition is administered or intended to be administered to a subject in need thereof at a dose of about 0.4 mg to about 8 mg of an anti-TSLP antigen-binding fragment. For example, a pharmaceutical composition is administered or intended to be administered to a subject in need thereof at a dose of about 0.4 mg, about 2 mg, or about 8 mg of an anti-TSLP antigen-binding fragment. In some cases, the pharmaceutical composition is administered or is to be administered to a subject in need thereof at a dose comprising about 0.5 mg, about 1 mg, about 2 mg, about 3 mg, about 4 mg, about 5 mg, about 6 mg, about 7 mg, about 8 mg, about 9 mg, about 10 mg, about 11 mg, about 12 mg, about 13 mg, about 14 mg, about 15 mg, or about 16 mg of an anti-TSLP antigen-binding fragment.

In some cases, a pharmaceutical composition is administered or intended to be administered to a subject in need thereof at a dosage of 0.2 mg to 16 mg of an anti-TSLP antigen-binding fragment. For example, a pharmaceutical composition is administered or intended to be administered to a subject in need thereof at a dosage of 0.2 mg, 0.4 mg, 0.6 mg, 2 mg, 6 mg, 8 mg, or 16 mg of an anti-TSLP antigen-binding fragment. In some cases, a pharmaceutical composition is administered or intended to be administered to a subject in need thereof at a dosage of 0.4 mg to 8 mg of an anti-TSLP antigen-binding fragment. For example, a pharmaceutical composition is administered or intended to be administered to a subject in need thereof at a dosage of 0.4 mg, 2 mg, or 8 mg of an anti-TSLP antigen-binding fragment. In some cases, the pharmaceutical composition is administered or is to be administered to a subject in need thereof at a dosage comprising 0.5 mg, 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, 10 mg, 11 mg, 12 mg, 13 mg, 14 mg, 15 mg, or 16 mg of an anti-TSLP antigen-binding fragment.

In some cases, a dose of an anti-TSLP antigen-binding fragment is administered or intended to be administered to a subject in need thereof once daily. In some cases, a dose of an anti-TSLP antigen-binding fragment is administered or intended to be administered to a subject in need thereof as a daily dose. In some cases, a dose of an anti-TSLP antigen-binding fragment is administered or intended to be administered to a subject in need thereof as a total daily dose. In some cases, a dose of an anti-TSLP antigen-binding fragment is administered or intended to be administered to a subject in need thereof every other day. In some cases, a dose of an anti-TSLP antigen-binding fragment is administered or intended to be administered to a subject in need thereof daily. In some cases, a dose of an anti-TSLP antigen-binding fragment is administered or intended to be administered to a subject in need thereof twice daily or once daily. In some cases, a dose of an anti-TSLP antigen-binding fragment is administered or is to be administered to a subject in need thereof once daily (Q1D).

In some cases, a pharmaceutical composition comprising an anti-TSLP antigen-binding fragment is administered or is intended to be administered to a subject in need thereof for at least 2 weeks, at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, or at least 12 months. In some cases, a pharmaceutical composition comprising an anti-TSLP antigen-binding fragment is administered or is intended to be administered to a subject in need thereof for at least 2 weeks, at least 4 weeks, at least 8 weeks, at least 12 weeks, at least 16 weeks, at least 20 weeks, at least 24 weeks, at least 28 weeks, at least 32 weeks, at least 36 weeks, at least 40 weeks, at least 44 weeks, at least 48 weeks, or at least 52 weeks. In some cases, a pharmaceutical composition comprising an anti-TSLP antigen-binding fragment is administered or intended to be administered to a subject in need thereof for about 12 to about 52 weeks, for example, for 12 to 52 weeks.

In some cases, a pharmaceutical composition comprising an anti-TSLP antigen-binding fragment is administered or intended to be administered to a subject orally or intranasally, e.g., in aerosolized form. In some cases, a pharmaceutical composition comprising an antigen-binding fragment is administered or intended to be administered to a subject by inhalation, e.g., using a dry powder inhaler. In some cases, a pharmaceutical composition comprising an antigen-binding fragment is administered or intended to be administered to a subject by oral inhalation, e.g., using a dry powder inhaler.

The dose of the anti-TSLP antigen-binding fragment to be administered to or to a patient is the target amount of the anti-TSLP antigen-binding fragment present in a pharmaceutical composition (e.g., a dry powder formulation) for administration to a patient. For example, it can be the amount present in an inhalation device for administration to a patient, e.g., the dose can be the nominal metered dose for addition to an inhalation device (e.g., a dry powder inhaler). In some cases, the dose can be the amount of the anti-TSLP antigen-binding fragment present in a capsule containing the pharmaceutical composition, e.g., as a dry powder formulation for addition to an inhalation device, e.g., a dry powder inhaler.

*** The features disclosed in the foregoing description, the accompanying patent claims, or the accompanying drawings (in their specific forms or as components for performing the disclosed functions or as methods or processes for obtaining the disclosed results) may, where appropriate, be used alone or in any combination to implement the present disclosure in its various forms.

Although the present disclosure has been described in conjunction with the above exemplary embodiments, many equivalent modifications and variations will be apparent to those skilled in the art upon receiving the present disclosure. Therefore, the exemplary embodiments of the present disclosure set forth above are to be considered illustrative rather than restrictive. Various modifications may be made to the described embodiments without departing from the spirit and scope of the present disclosure.

To avoid any doubt, any theoretical explanations provided herein are provided for the purpose of enhancing the reader's understanding. The inventors do not wish to be bound by any of such theoretical explanations.

Any section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.

Throughout this specification, including the claims below, unless the context requires otherwise, the words "comprise" and "include" and variations such as "comprises," "comprising," and "including" will be understood to imply the inclusion of a stated number or step or group of numbers or steps but not the exclusion of any other number or step or group of numbers or steps.

It is important to note that, as used in the specification and accompanying patent applications, the singular forms "a," "an," and "the" include plural referents unless the context clearly indicates otherwise. Ranges may be expressed herein as from "about" one particular value and/or to "about" another particular value. When such a range is expressed, another example includes from one particular value and/or to the other particular value. Similarly, when a value is expressed as an approximation by using the antecedent "about," it will be understood that the particular value forms another example. The term "about" with respect to a numerical value is contextual and means, for example, ± 20%. When the term "± x%" is used herein, it refers to a percentage of the preceding value, not to plus or minus the absolute percentage listed, for example, "20% ± 10%" means "18-22%," not "10-30%."

EXAMPLES These examples demonstrate the optimization of buffer conditions, composition and pH (Example 2), histidine strength and leucine/trileucine levels (Examples 3-4) to achieve a stable FAB1 powder formulation (Example 5) that exhibits minimal in vitro aggregation and a favorable toxicological profile (Example 6).

Example 1 - Background: FAB1 is an antibody fragment directed against the interleukin TSLP and can be used to treat patients with moderate to severe asthma. The route of administration is inhalation of a dry powder formulation via a dry powder inhaler (DPI). The selected Phase 1 formulation contains trehalose, leucine, trileucine, and citrate (TLTC) at pH 6, with polysorbate 80 (PS80) added to reduce protein aggregation and maintain low counts of subvisible particles. However, surfactants compromise powder properties, primarily by reducing manufacturing yield and significantly increasing powder retention in the device and throat. Therefore, alternative formulations for FAB1 were developed, and formulations with different buffers (citrate vs. histidine (trehalose, leucine, trileucine, and histidine (TLTH)), pH, histidine strength, and leucine/trileucine levels were tested to achieve the optimal formulation.

Three screening studies and one stability study were conducted: • Study 1: TLTC vs. TLTH at pH 5 and 6 • Study 2: TLTH with 1.3% histidine, added PS80, varying levels of trileucine, and leucine (e.g., 2x trileucine/leucine and homoleucine without trileucine). • Study 3: Titration of TLTH at optimal histidine levels • Study 4: Stability (1M accelerated) of TLTH

The selection criteria for drug products are based on aerosol performance, solid-state characterization, and protein aggregate formation after dry powder reconstitution.

Example 2 - Study 1 : TLTC vs. TLTH Experiment at pH 5 and 6 This study evaluated two factors of the formulation, namely, buffer (citrate vs. histidine) and pH (5 and 6).

To test the effect of histidine buffer at different pH on protein formulation and protein aggregation, six batches were produced as shown in Table 5. The composition of each batch is summarized in Table 6 . Batch number formulations FAB1 strength Batch size (g) FS volume (ml) 21-WS-016 TLTC, pH 5 10% 10 133 21-WS-017 TLTC, pH 5 40% 21-WS-018 TLTH, pH 6, 5% His 10% 21-WS-019 TLTH, pH 6, 5% His 40% 21-WS-023 TLTH, pH 5, 5% His 10% 21-WS-024 TLTH, pH 5, 5% His 40% 20 267 Table 5. Study 1 batches Components (w/w%) 10% FAB1 , TLTC , pH 5 40% FAB1 , TLTC , pH 5 10% FAB1 , TLTH , pH 6 40% FAB1 , TLTH , pH 6 10% FAB1 , TLTH , pH 5 40% FAB1 , TLTH , pH 5 batch 21-WS-016 21-WS-017 21-WS-018 21-WS-019 21-WS-023 21-WS-024 FAB1 10.0 40.0 10.0 40.0 10.0 40.0 Trehalose 69.55 39.55 72.70 42.70 72.05 42.05 Leucine 10.50 10.50 10.50 10.50 10.50 10.50 Trileucine 2.00 2.00 2.00 2.00 2.00 2.00 Trisodium citrate 5.50 5.50 - - - - Citric acid 2.45 2.45 - - - - L-histidine - - 2.00 2.00 0.35 0.35 Histidine-HCl - - 2.80 2.80 5.10 5.10 Table 6. Allocation targets for Study 1

result describe 10% TLTC , pH 5 (21-WS-016) 40% TLTC , pH 5 (21-WS-017) 10% TLTH , pH 6 (21-WS-018) 40% TLTH , pH 6 (21-WS-019) 10% TLTH , pH 5 (21-WS-023) 40% TLTH , pH 5 (21-WS-024) formulations Batch size (g) 10 10 10 10 10 20 Solid loading capacity (mg/ml) 75 75 75 75 75 75 Raw material (FS) volume (ml) 133.3 133.3 133.3 133.3 133.3 166.6 Yield ( based on batch size ) (%) 69.6 87.4 69.3 67.7 79 68.6 Yield ( based on FS quality ) (%) 75.0 94.2 81.5 79.6 85.3 71.1 FS pH 5.1 5.1 6.0 5.8 5.0 5.1 Table 7. Study 1 Processing Results. 1) Yield based on batch size was calculated as the net collector yield divided by the nominal batch size. 2) Yield based on FS mass was calculated as the net collector yield divided by the actual FS mass.

Table 6 shows that all batches were 10 g in size, with the exception of 40% TLTH (pH 5), which was 20 g. Despite the smaller batch sizes, the spray drying yields based on FS mass were high for all batches. Powders were produced using low-throughput processing parameters.

10% FAB1 40% FAB1 TLTC pH5 (21-WS-016) TLTH pH6 (21-WS-018) TLTH pH5 (21-WS-023) TLTC pH5 (21-WS-017) TLTH pH6 (21-WS-019) TLTH pH5 (21-WS-024) Drug Substance (DS) Protein mg/ml 42.8 44.7 41.7 42.8 44.7 41.8 Recovery rate % 85 85 84 85 85 84 FS protein mg/ml 7.1 7.0 7.3 28.9 27.5 29.0 Recovery rate % 94 94 97 97 92 97 Bulk Powder (BP) Protein w/w% 9.4 9.7 7.6* 35.5 37.0 36.4 Recovery rate % 93.9 96.6 75.7 88.8 92.6 91.0 DS High Performance Size Exclusion Chromatography (HP-SEC) Monomer main peak % (MPP) 99.9 100 99.8 99.9 100 99.8 Aggregate % 0.1 0.0 0.2 0.1 0.0 0.20 FS HP-SEC MPP% 99.9 99.9 99.8 99.9 99.9 99.8 Aggregate % 0.1 0.1 0.2 0.1 0.1 0.2 BP HP-SEC MPP% 99.9 99.9 99.7 99.9 99.9 99.7 Aggregate % 0.1 0.1 0.3 0.1 0.1 0.3 Primary particle size distribution (pPSD) d10 ( μm ) 0.5 0.5 0.5 0.5 0.5 0.5 d50 ( μm ) 1.5 1.5 1.8 1.7 1.8 1.5 d90 ( μm ) 3.5 3.5 3.8 3.8 3.9 3.4 span 2.0 2.0 1.9 1.9 1.9 2.0 Moisture content (%) 1.4 0.7 0.7 1.5 1.0 0.9 Glass transition temperature (Tg) (℃) 94 98 99 97 100 99 Compressed bulk density (cBD) (g/cm ³ ) 0.6584 0.7245 0.7498 0.6070 0.6479 0.6702 Specific surface area (SSA) (m ² /g) 5.8251 5.1879 4.9904 5.5543 5.5313 5.2441 Table 8. Solid-state testing for Study 1. *Values are lower than expected and inconsistent with FS protein concentration (97% recovery). Table 8 shows that the solid properties were similar between all formulations, with very high protein purity and low aggregation in DS, FS, and BP. The primary particle size distributions were similar for all formulations, with low moisture content, with the histidine formulation having a moisture content below 1%. The glass transition values were high and similar for all formulations. The high glass transition and low residual moisture content indicate room temperature stable powders. The compressed bulk density was in a favorable range, with slightly higher values for the TLTH formulation, indicating slightly more flowable powders, which were also reflected in a slightly increased surface area.

Scanning electron microscopy (SEM) analysis of the formulations confirmed the good morphology of both TLTC and TLTH formulations with similar appearance ( FIG1 ).

A key readout for dry powder formulations is protein aggregation after reconstitution. Particle counts were performed using mass flow imaging (MFI) in powders reconstituted to two concentrations: 2.5 mg/ml protein and 30 mg/ml stock.

Figure 2 shows the number of particles present in the powder formulations tested (TLTC pH 5 (21-WS-016); TLTH pH 6 (21-WS-018); and TLTH pH 5 (21-WS-023). The TLTC formulations had significantly higher particle counts compared to TLTH, which had "low" particle counts at 2.5 mg/ml and stock concentration.

Figure 3 shows the particle counts in the powder formulations tested: TLTC pH 5 (21-WS-017); TLTH pH 6 (21-WS-019); and TLTH pH 5 (21-WS-024). At the stock concentration, the TLTC formulation had higher particle counts than the TLTH formulation, but not at 2.5 mg/ml.

Figure 4 shows the particle counts in the tested formulations (API citrate pH 5; API histidine pH 6; API histidine pH 5). Notably, the particle counts for DS in TLTC were higher than those in TLTH, with pH 5 being lower than pH 6. The same trend was observed for FS (Figures 4B and 4C).

FIG5 compares the reconstituted powders with 10% and 40% FAB1 in different formulations, and the lowest particle counts were observed for TLTH (pH 5) (see FIG5C ).

In summary, Figures 2 to 5 show that particle counts in DS, FS, and reconstituted BP follow the same pattern, with counts being lowest in TLTH (pH 5), slightly higher in TLTH (pH 6), and significantly higher in TLTC (pH 5). Because protein aggregation levels were significantly higher in the citrate-containing buffer, the histidine buffer is considered superior.

Finally, the aerosol potency of the 10% and 40% batches was tested using a cascade impactor, a device used for pharmaceutical inhaler testing (see Figures 6 and 7 ). The deposition profile for FAB1 was collected using a next-generation impactor as described in United States Pharmacopeia (USP) <601> Apparatus 6 at a flow rate of 30.0 ± 1.0 L/min. The next-generation impactor consists of seven stages, or sample receptacles, each graded with a different mass median aerodynamic diameter value depending on the flow rate. The NGI is also equipped with a USP inlet port "throat" to accommodate the inhaler device. When the vacuum pump applies a given flow rate, the inhaler device is "activated," simulating a patient inhaling through the device's mouthpiece. After each actuation, the amount of active agent remaining in the device (including the capsule), as well as on the throat and in various fractions, was determined using appropriate analytical methods. Aerosol potency was comparable across formulations, with a very high (>80%) fine particle fraction (FPF) (<5 μm) and a favorable median mass aerodynamic diameter (MMAD) of 2.5 μm (± 0.5). Device deposition was low for all formulations, with the TLTH formulation exhibiting the lowest deposition.

These results indicate that histidine buffer is superior to citrate buffer in maintaining low protein aggregation.

Example 3 - Study 2 : TLTH Experiment with PS80 and Trileucine / Leucine. Study 2 was designed to explore the formulation space for histidine and encapsulating agents (leucine and trileucine). The primary objective was to investigate the effect of reduced histidine on the particle surface by either decreasing the total amount of histidine in the formulation (from 5% to 1.3%) or by increasing the leucine/trileucine ratio. The addition of small amounts of PS80 was also explored.

A design of experiments (DOE) was established to explore the design space for formulation components, focusing on subvisible particle counts and aerosol performance as key readout parameters, as well as device deposition. The factors evaluated were 1) histidine reduction, 2) increasing the amount of histidine on the surface by adding a shell-forming modifier, and 3) adding an optimized amount of PS80. The addition of PS80 resulted in increased device deposition, but it significantly reduced protein aggregation and maintained low particle counts (see WO2021/083908). The addition of the shell-forming components trileucine and leucine was explored to improve the moisture resistance of the powder. Finally, a formulation with very high leucine (37.5%) and no trileucine was added to study moisture resistance. Nine batches were produced, as shown in Table 9 . The composition of each batch is summarized in Table 10 . Batch number formulations FAB1 strength Batch size (g) FS volume (ml) 21-WS-039 TLTH, pH 5.5, 1.3% His 10% 15 200 21-WS-040 TLTH, pH 5.5, 1.3% His 40% 21-WS-041 TLTH, pH 5.5, 1.3% His, 2x TriLeu/Leu 10% 21-WS-042 TLTH, pH 5.5, 5 % His, 2x TriLeu/Leu 10% 21-WS-043 TLTH, pH 5.5, 1.3% His, 0.4% PS80 10% 21-WS-044 TLTH, pH 5.5, 1.3% His, 0.4% PS80 40% 21-WS-045 TLTH, pH 5.5, 5% His 10% 21-WS-046 TLTH, pH 5.5, 5% His 40% 21-WS-060 TLH, pH 5.5, 1.3% His, 37.5% Leu 10% Table 9. Study 2 batches Components (w/w%) 10% FAB1 1.3% His 40% FAB1 1.3% His 10% FAB1 1.3% His 2x Leu/trileu 10% FAB1 5% His 2× 3Leu/Leu , 0.4% PS80 10% FAB1 1.3% His 0.4% PS80 40% FAB1 1.3% His 0.4% PS80 10% FAB1 5% His 40% FAB1 5% His 10% FAB1 1.3% His 37.5% Leu batch 21-WS-039 21-WS-040 21-WS-041 21-WS-042 21-WS-043 21-WS-044 21-WS-045 21-WS-046 21-WS-060 FAB1 10.0 40.0 10.0 10.0 10.0 40.0 10.0 40.0 10.0 Trehalose 76.17 46.17 63.67 59.69 75.77 45.77 72.59 42.59 53 Leucine 10.50 10.50 21.0 21.0 10.5 10.5 10.5 10.5 37.5 Trileucine 2.0 2.0 4.0 4.0 2.0 2.0 2.0 2.0 - PS80 - - - 0.4 0.4 0.4 - - - L -histidine 0.25 0.25 0.25 0.93 0.25 0.25 0.93 0.93 0.23 Histidine -HCl 1.08 1.08 1.08 3.98 1.08 1.08 3.98 3.98 1.07 Table 10. Allocation targets for Study 2

Table 11 summarizes the treatment results, including actual treatment parameters and yields. batch 21-WS-039 21-WS-040 21-WS-041 21-WS-042 21-WS-043 21-WS-044 21-WS-045 21-WS-046 21-WS-060 describe 10% TLTH 1.3% His 40% TLTH 1.3% His 10% TLTH 1.3% His 2x Leu/Trileu 10% TLTH 5% His 2x Leu/trileu , 0.4% PS80 10% TLTH 1.3% His 0.4% PS80 40% TLTH 1.3% His 0.4% PS80 10% TLTH 5% His 40% TLTH 5% His 10% TLTH 1.3% His 37.5% Leu form . Batch size (g) 15 15 15 15 15 15 15 15 15 Solid loading capacity (mg/ml) 75 75 75 75 75 75 75 75 75 Raw material (FS) volume (ml) 200.00 200.00 200.00 200.00 200.00 200.00 200.00 200.00 200.00 Yield ( based on batch size ) (%) 80.7 70.7 63.3 60.1 87.0 82.3 87.6 83.2 84.6 Yield ( based on FS quality ) (%) 85.0 74.5 84.8 80.1 92.9 87.9 81.6 77.3 87.8 FS density (g/ml) 1.0238 1.0213 1.0214 1.0207 FS pH 5.3 5.5 5.5 5.5 5.4 5.4 5.4 5.3 5.4 Table 11. Study 2 Processing Results. 1) Yield based on batch size was calculated as the net collector yield divided by the nominal batch size. 2) Yield based on FS mass was calculated as the net collector yield divided by the actual FS mass.

10% FAB1 40% FAB1 TLTH 1.3% His (21-WS-039) TLTH 1.3% His 2x Leu/trileu (21-WS-041) TLTH 5% His 2x 3Leu/Leu , 0.4% PS80 (21-WS-42) TLTH 1.3% His 0.4% PS80 (21-WS-043) TLTH 5% His (21-WS-045) TLTH 1.3% His 37.5% Leu (21-WS-60) TLTH 1.3% His (21-ws-40) TLTH 1.3% His 0.4% PS80 (21-WS-044) TLTH 5% His (21-WS-046) Drug Substance (DS) Protein mg/ml 54.0 53.0 53.0 53.9 52.8 59.1 54.0 53.0 52.8 Recovery rate % 98 96 96 98 96 98 98 96 96 FS protein mg/ml 7.4 7.5 7.6 7.6 7.6 4.9 30.0 29.8 30.1 Recovery rate % 99 100 102 101 101 98.8 100 99 100 Bulk Powder (BP) Protein w/w% 9.9 10.2 10.2 9.8 10.0 9.7 38.8 39.8 39.3 Recovery rate % 99.5 101.6 101.6 98.3 99.6 97.4 97 99.4 98.3 DS High Performance Size Exclusion Chromatography (HP-SEC) MPP% 100 100 100 100 100 100 100 100 100 Aggregate % 0 0 0 0 0 0 0 0 0 FS HP-SEC MPP% 100 100 100 100 100 99.4 100 100 100 Aggregate % 0 0 0 0 0 0.6 0 0 0 BP HP-SEC MPP% 100.0 99.9 99.9 100.0 100.0 99.6 99.9 99.9 99.9 Aggregate % 0.0 0.1 0.1 0.0 0.0 0.4 0.1 0.1 0.1 Primary particle size distribution (pPSD) d10 ( μm ) 0.5 0.5 0.5 0.5 0.5 0.5/0.4 ¹ 0.5 0.5 0.5 d50 ( μm ) 1.5 1.6 1.7 1.5 1.6 1.7/1.3 ¹ 1.7 1.7 1.7 d90 ( μm ) 3.5 3.6 3.6 3.5 3.6 4.5/3.1 ¹ 3.9 3.9 3.9 span 2.0 1.9 1.9 2.0 1.9 2.4/2.0 ¹ 2.0 1.9 2.0 Moisture content (%) 0.9 0.9 0.7 0.7 0.6 0.6 1.0 0.9 1.1 Glass transition temperature (Tg) (℃) 100 100 98 102 100 108 104 81, 104 ² 100 Compressed bulk density (cBD) (g/cm ³ ) - - - - - 0.6969 - - - Specific surface area (SSA) (m ² /g) - - - - - 5.4687 - - - Table 12. Study 2 solid-state testing. ^1. Particle size was larger than expected in the initial test, so the pPSD test was repeated. ^2. Values refer to Tg onset and secondary Tg onset.

Scanning electron microscopy (SEM) analysis of the formulations confirmed the good morphology of both TLTC and TLTH formulations with similar appearance ( FIG. 8 and FIG. 9 ).

The MFI results for 10% FAB1 are shown in Figure 10. All formulations showed low particle counts compared to the TLTC formulation (21-WS-016 - see Figure 5). Adding PS80 further reduced particle counts (21-WS-042 and 043). Doubling the amount of leucine and trileucine had no effect on particle counts.

The MFI results for 40% FAB1 are shown in Figure 11. All formulations showed low particle counts compared to the TLTC formulation (21-WS-017). Adding PS80 further reduced particle counts (21-WS-044). Compared to 5% histidine, 1.3% histidine showed a trend toward lower particle counts. Similar results were observed for pH 5, pH 5.5, and pH 6.

Finally, the aerosol potency of the 10% and 40% batches was tested using a cascade impactor , which is used for testing pharmaceutical inhalers (see Figures 12 and 13 ).

When comparing TLTH formulations with 5% histidine and pH 5, 5.5, or 6, no significant differences were observed in solid-state properties ( Table 12 ), aerosol potency, or protein aggregation ( Figures 12 and 13 ) . In DOE analysis and modeling of histidine versus PS80, higher histidine and lower PS80 levels were found to reduce device deposition, while only PS80 affected protein aggregation. Addition of homoleucine/trileucine also had no significant effect on device deposition or protein aggregation. All formulations had similar water content and glass transition temperatures. Low levels (0.4%) of PS80 showed a stickier trend, with lower spray drying yields and higher device deposition ( Figures 12 and 13 ), but as expected, kept protein aggregation to a minimum ( Table 12 ) .

Conclusion: Together, these data demonstrate that higher concentrations of histidine combined with lower levels of PS80 reduce device deposits. A formulation containing TLTH at pH 5.5, maintaining trileucine at 2.0% w/w and leucine at 10.5% w/w, and without added PS80, exhibited favorable properties. Therefore, the inventors further optimized this formulation by investigating the optimal histidine concentration.

Example 4 - Study 3 : Optimal Histidine Level for TLTH . Study 3 investigated the optimal histidine concentration for TLTH formulations. Three histidine levels were explored to select the optimal concentration. Six batches were produced as shown in Table 13. The composition of each batch is summarized in Table 14 . Batch number formulations FAB1 strength Batch size (g) FS volume (ml) 21-WS-055 TLTH, pH 5.5, 1.3% His 10% 15 200 21-WS-056 TLTH, pH 5.5, 3.14% His 10% 21-WS-057 TLTH, pH 5.5, 5% His 10% 21-WS-058 TLTH, pH 5.5, 3.14% His 40% 21-WS-059 TLTH, pH 5.5, 5% His 40% 21-WS-061 TLTH, pH 5.5, 1.3% His 40% 21-WS-045 TLTH, pH 5.5, 5% His 10% 21-WS-046 TLTH, pH 5.5, 5% His 40% 21-WS-060 TLH, pH 5.5, 1.3% His, 37.5% Leu 10% Table 13. Study 3 batches Components (w/w%) 10% FAB1 1.3% His 10% FAB1 3.14% His 10% FAB1 5% His 40% FAB1 3.14% His 40% FAB1 5% His 40% FAB1 1.3% His batch 21-WS-055 21-WS-056 21-WS-057 21-WS-058 21-WS-059 21-WS-061 FAB1 10.0 10.0 10.0 40.0 40.0 40.0 Trehalose 76.20 74.36 72.52 44.36 42.52 46.16 Leucine 10.50 10.50 10.50 10.50 10.50 10.50 Trileucine 2.00 2.00 2.00 2.00 2.00 2.00 L -histidine 0.23 0.55 0.88 0.55 0.88 0.37 Histidine -HCl 1.07 2.59 4.10 2.59 4.10 0.97 Table 14. Allocation targets for Study 3

Results Table 15 shows a summary of the processing results for Study 3, including actual processing parameters and yields. Solid-state test results are shown in Table 16 and SEM images are shown in Figure 14 . batch 21-WS-055 21-WS-056 21-WS-057 21-WS-058 21-WS-059 21-WS-061 describe 10% TLTH , 1.3% His 10% TLTH , 3.14% His 10% TLTH , 5% His 40% TLTH , 3.14% His 40% TLTH , 5% His 40% TLTH, 1.3% His formulations Batch size (g) 15 15 15 15 15 15 Solid loading capacity (mg/ml) 75 75 75 75 75 75 Raw material (FS) volume (ml) 200.00 200.00 200.00 200.00 200.00 200.00 Yield ( based on batch size ) (%) 79.9 85.2 81.8 74.3 66.7 85.2 Yield ( based on FS quality ) (%) 83.4 88.9 85.4 77.4 69.6 89.2 FS pH 5.4 5.4 5.5 5.4 5.4 5.7 Table 15. Study treatment results 10% FAB1 40% FAB1 TLTH 1.3% His (21-WS-055) TLTH 3.14% His (21-WS-056) TLTH 5% His (21-WS-057) TLTH 1.3% His (21-WS-061) TLTH 3.14% His (21-WS-058) TLTH 5% His (21-WS-059) Drug Substance (DS) Protein mg/ml 59.1 59.1 59.1 52.4 59.1 59.1 Recovery rate % 98 98 98 96 98 98 FS protein mg/ml 7.4 7.4 7.4 28.6 29.5 29.4 Recovery rate % 99.1 98.4 98.9 95.5 98.1 98.1 Bulk Powder (BP) Protein w/w% 10.2 10.3 10.3 37.9 41.0 40.8 Recovery rate % 102.3 103.4 102.6 94.8 102.6 102.1 DS High Performance Size Exclusion Chromatography (HP-SEC) MPP% 100 100 100 100 100 100 Aggregate % 0 0 0 0 0 0 FS HP-SEC MPP% 100 100 100 100 100 100 Aggregate % 0 0 0 0 0 0 BP HP-SEC MPP% 100.0 100.0 100.0 99.9 99.9 99.9 Aggregate % 0.0 0.0 0.0 0.1 0.1 0.1 Primary particle size distribution (pPSD) d10 ( μm ) 0.5 0.5 0.5 0.6 / 0.4 0.5 0.5 d50 ( μm ) 1.6 1.6 1.6 2.1 / 1.7 1.7 1.6 d90 ( μm ) 3.6 3.6 3.7 5.0 / 3.9 4.1 3.9 span 2.0 2.0 2.0 2.2 / 2.0 2.1 2.1 Moisture content (%) 0.7 0.7 0.7 0.9 1.0 1.0 Glass transition temperature (Tg) (℃) 101, 103 100, 102 100, 101 116 99, 105 100, 102 Compressed bulk density (cBD) (g/cm ³ ) 0.6131 0.6901 0.6558 0.5529 0.6226 0.5378 Specific surface area (SSA) (m ² /g) 4.8447 5.2046 4.9938 5.7128 5.7473 5.8758 Table 16. Study 3 solid state tests

All TLTH formulations showed excellent solids properties based on the following criteria: D90 <5 μm (90% of particles smaller than 5 μm), % water <5%, and Tg >80°C at 2% water content.

For all TLTH formulations, subvisible particles detected by MFI were low (see Figures 15 and 16 ) .

The results for the 10% and 40% NGIs are plotted in Figures 17A and 17B and summarized in Figure 17 , and these data indicate that aerosol performance is excellent at all histidine concentrations.

Conclusions: Three histidine levels (1.3%, 3.14%, and 5%) were explored at 10% and 40% protein strengths to determine the optimal histidine concentration. No significant differences in solid or aerosol performance were observed between the formulations, and protein aggregation was low, with a trend toward lower numbers at higher histidine levels.

Example 5 - Study 4 : One -Month Stability Experiment of TLTH Study 4 was added to further understand the stability of the TLTH formulation with optimized histidine and pH. Powders and filled capsules previously produced in Studies 1-3 were used for stability testing, as shown in Table 17. High (5%) and low (1.3%) histidine formulations were placed at 40°C/75% RH (wrapped in foil and protected with desiccant) to support TLTH as the proposed Phase 2 formulation. Batches of no more than 2 g of powder were transferred to aluminum Tournaire containers, foil-wrapped with desiccant, and stability tested at 40°C/75% RH for one month. The capsules were filled and packaged in foil bags with desiccant and stored at 40°C/75% RH. Batch number formulations FAB1 strength 21-WS-039 TLTH, pH 5.5, 1.3% His 10% 21-WS-040 TLTH, pH 5.5, 1.3% His 40% 21-WS-045 TLTH, pH 5.5, 5% His 10% 21-WS-046 TLTH, pH 5.5, 5% His 40% 21-WS-056 TLTH, pH 5.5, 3.14% His 10% 21-WS-058 TLTH, pH 5.5, 3.14% His 40% Table 17. Study 4 Stability Batches

The solid state stability results are summarized in Tables 18 and 19 , and the SEM images are shown in FIG18 . formulations TLTH 1.3% His TLTH 3.14% His TLTH 5% His Batch number 21-WS-039 21-WS-056 21-WS-045 Time point ( month ) T=0 T=1 T=0 T=1 T=0 T=1 BP protein w/w% 9.9 9.43 10.3 9.81 10 9.59 Recovery rate % 99.5 94.3 103.6 98.1 99.6 95.9 BP HP-SEC MPP% 100 100 100 100 100 99.9 Aggregate % 0.0 0.0 0.0 0.0 0.0 0.1 pPSD d10 ( μm ) 0.5 0.4 0.5 0.5 0.5 0.4 d50 ( μm ) 1.5 1.5 1.6 1.6 1.6 1.6 d90 ( μm ) 3.5 3.5 3.6 3.6 3.6 3.6 span 2.0 2.1 2.0 2.0 1.9 2.0 Moisture content (%) 0.9 1.2, 1.6* 0.7 0.7 0.6 1.1.3* Table 18. Stability of 10% FAB1 TLTH Formulations. *KF was rerun to confirm the slight increase in water content in both formulations. formulations TLTH 1.3% His TLTH 3.14% His TLTH 5% His Batch number 21-WS-040 21-WS-058 21-WS-046 Time point ( month ) T=0 T=1 T=0 T=1 T=0 T=1 BP protein w/w% 38.3 36.4 41.0 39.3 39.3 37.8 Recovery rate % 97.0 88.1, 91.0* 103 98.4 98.3 90.1, 94.4* BP HP-SEC MPP% 99.9 99.9 99.9 99.9 99.9 99.9 Aggregate % 0.1 0.1 0.1 0.1 0.1 0.1 pPSD d10 ( μm ) 0.5 0.4 0.5 0.5. 0.5 0.4 d50 ( μm ) 1.7 1.7 1.7 1.6 1.7 1.7 d90 ( μm ) 3.9 3.9 4.1 3.7 3.9 3.9 span 2.0 2.1 2.1 2.0 2.0 2.1 Moisture content (%) 1.0 1.1, 1.2* 1.0 1.0 1.1 1.2, 1.3* Table 19. Stability of 40% FAB1 TLTH Formulations. *KF was rerun to confirm the slight increase in water content in both formulations.

The NGI results for Study 4 are shown in Figures 19 and 20 and summarized in Tables 20 and 21 . formulations TLTH 1.3% His TLTH 3.14% His TLTH 5% His Batch number 21-WS-039 21-WS-056 21-WS-045 Time point ( month ) T=0 T=1 T=0 T=1 T=0 T=1 FPF (＜5 μm ) (%) 78 83 78 81 84 78 FPM (＜5 μm ) (mg) 1.4 1.6 1.4 1.5 1.6 1.5 MMAD ( μm ) 2.7 2.6 2.7 2.7 2.7 2.9 Table 20. One -month accelerated stability results of NGI with 10% FAB1. formulations TLTH 1.3% His TLTH 3.14% His TLTH 5% His Batch number 21-WS-040 21-WS-058 21-WS-046 Time point ( month ) T=0 T=1 T=0 T=1 T=0 T=1 FPF (＜5 μm ) (%) 82 80 81 85 81 80 FPM (＜5 μm ) (mg) 5.8 5.6 6.1 6.3 5.8 5.5 MMAD ( μm ) 2.8 2.6 2.5 2.6 2.6 2.8 Table 21. One -month accelerated stability results of NGI for 40% FAB1.

Conclusions All formulations performed very well, and the formulation containing TLTH (pH 5.5) with 3.14% histidine (w/w) showed particularly good stability.

Example 6 - In Vivo Toxicity Study To test the in vivo toxicity of inhaled FAB1 Fab in a powder formulation using the new formulation described in Example 5 (TLTH, pH 5.5, with 3.14% histidine (w/w)), a 28-day good laboratory practice (GLP) toxicity study was established and compared to an earlier toxicity study using TLTC (pH 6) with PS80.

Group Dosage Exposure duration ( minutes ) Total aerosol dose level (mg/L) FAB1 aerosol dosage level (mg/L) Deposition dose level (mg/kg/d) animal Main research Recovery 1 Air comparison 60 LLOQ LLOQ 0 3M + 3F 2M + 2F 2 Placebo control 60 2.5 LLOQ 0 3M + 3F 3 Low dose 8 2.5 963 1.0 3M + 3F 4 Medium dose 20 2.5 958 2.3 3M + 3F 2M + 2F 5 High dose 60 2.5 893 7.1 3M + 3F 2M + 2F Table 22. Results of a 28- day cynomolgus monkey GLP toxicity study of FAB1 in TLTC (pH 6) with PS80 . LLOQ = lower limit of quantitation.

Table 22 shows the results of the first 28-day toxicity study in which cynomolgus monkeys were treated with FAB1 reconstituted in TLTC (pH 6) with PS80. No adverse effects were observed on food consumption, body weight, clinical observations, clinical pathology, pulmonary function tests, ECG, blood pressure, neurobehavioral assessments, or ophthalmology.

In animals treated with the low dose level (1.0 mg/kg/day sedimentation dose level), no adverse macroscopic or microscopic findings were observed in any tissue including the respiratory tract (i.e., lungs, trachea, larynx, esophagus, salivary glands, lymph nodes, tongue, tonsils, and soft palate).

However, at higher doses of 2.3 mg/kg/day (females only) and 7.1 mg/kg/day (both sexes), the FAB1-associated incidence and/or severity of pulmonary perivascular (PV)/peribronchial (PB) mononuclear inflammatory cell (MIC) infiltrates increased (see Figure 21 ). Therefore, in this first study using TLTC at pH 6 + PS80 as the adjuvant, the no-observed-adverse-effect-level (NOAEL) was 1 mg/kg/day.

Figure 22 further shows visible particles in the reconstituted formulation (FAB1 in TLTC (pH 6) with PS80) in water, a simple simulation of the expected reconstitution in the epithelial lining after lung sedimentation.

Group Dosage Exposure duration ( minutes ) Total aerosol dose level (mg/L) FAB1 aerosol concentration level (mg/L) Target delivery dose level (mg/kg/d) Target pulmonary sedimentation dose level (mg/kg/d) animal Main research 1 Air comparison 60 0 0 0 0 3M + 3F 2 Placebo control 60 0.724 LLOQ 0 0 3M + 3F 3 Low dose 60 NC (0.090) 36 1.2 0.3 3M + 3F 4 Medium dose 60 NC (0.303) 121 4 1 3M + 3F 5 High dose 60 0.72 278 9.2 2.3 3M + 3F Table 23. Results of a 28 -day GLP toxicity study of FAB1 in TLTH (pH 5.5) with 3.14% histidine in cynomolgus monkeys . LLOQ = lower limit of quantitation.

Table 23 shows the results of a second 28-day toxicity study in which cynomolgus monkeys were treated with TLTH (pH 5.5) with 3.14% histidine (w/w).

Figure 23 shows representative images of FAB1-related lung pathology. Mononuclear cell (MNC) infiltrates in the lungs were minimal at all doses tested. Microscopic findings suggest a local, low-level immune response to inhaled foreign proteins and are considered non-adverse ( Figure 24 ). The NOAEL for this study was 2.3 mg/kg sediment (compared to the 1 mg/kg sediment dose used in the first toxicity study).

In summary, these data indicate that a common microscopic finding associated with inhaled protein in cynomolgus monkeys is mononuclear inflammatory cell (MIC) infiltrates in the lungs. In the first 28-day toxicity study, PB/PB MIC infiltrates were observed, which were considered unfavorable in severity and limited the NOAEL to 1 mg/kg. The TLTH formulation with pH 5.5 and 3.14% histidine (w/w) was associated with reduced in vitro aggregation and exhibited a more favorable toxicology profile in cynomolgus monkeys compared to TLTC (pH 6) with PS80 (2.3 mg/kg NOAEL vs. 1 mg/kg NOAEL).

Example 7 - Phase I, Randomized, Blinded, Placebo-Controlled Study to Evaluate the Safety, Tolerability, Pharmacokinetics, and Pharmacodynamics of FAB1 in Healthy Adult Subjects ( Part A ) and Adults with Asthma Receiving Moderate- to High-Dose Inhaled Corticosteroids and Long-Acting Beta- Agonists ( Part B) Study Design Part A of the study was a randomized, single-blind, placebo-controlled study in healthy male and female volunteers to evaluate the safety, tolerability , PK, and immunogenicity of FAB1 administered by DPI (one cohort in Subpart A1 received IV FAB1).

Part A consists of 4 subparts (A1, A2, A3 and A4). The overall design of Part A is presented in Table 24.

Table 24 Study Design - Part A Sub-part group Groups and treatment received A1 Healthy Volunteers 5 groups: 0.2 mg, 0.6 mg, 2 mg, 6 mg and 16 mg of inhaled FAB1 or placebo SAD Group 1: single 5 mg IV dose of FAB1 or placebo A2 Chinese and Japanese health volunteers 2 groups (1 Chinese group and 1 Japanese group): a single 16 mg dose of inhaled FAB1 or placebo A3 Healthy Volunteers 3 groups: 2 mg, 6 mg, and 16 mg of inhaled FAB1 or placebo MAD once daily for 14 days A4 Chinese and Japanese health volunteers Two groups (one Chinese group and one Japanese group): The highest dose of inhaled FAB1 in subpart A3 or placebo, once daily for 14 days. MAD, multiple ascending dose; SAD, single ascending dose.

The primary objectives of Part A are the safety and tolerability of inhaled FAB1 and the PK and safety of IV FAB1. Secondary objectives are the PK of inhaled FAB1 (including Japanese and Chinese participants) and the immunogenicity of FAB1 after single and multiple doses.

Part B of the study was a randomized, double-blind, placebo-controlled study in male and female adults with asthma receiving a combination of moderate- to high-dose ICS plus LABA medication. In a parallel-group design, patients were randomly assigned to receive one of three inhaled doses of FAB1 (0.4 mg, 2 mg, and 8 mg) or placebo via a dry powder inhaler (DPI) once daily for 28 days.

The predicted human dose of FAB1 following inhaled administration is based on two factors: first, the predicted human PK profile of FAB1 in systemic and lung tissues; second, the target lung concentration identified based on clinical efficacy data for texas pembrolizumab. The clinical PK profile of FAB1 was predicted using PK parameters derived from isokinetic scaling in cynomolgus monkeys. Following inhaled administration, the average partitioning of FAB1 from the lungs to the systemic circulation was estimated to be 2500 based on bronchoalveolar lavage fluid data from cynomolgus monkeys.

Target C _trough concentrations in the lung were identified from therapeutically effective systemic exposure of a systemic TSLP-specific mAb (texepemab) and the assumed lung distribution coefficient from serum. A calculated lung deposition dose of 1 mg (once daily) produced a C _trough concentration above the target concentration in lung tissue, corresponding to the predicted mean lung concentration (C _mean ) of a 210 mg dose of a TSLP-inhibiting systemic mAb administered every 4 weeks, which has demonstrated efficacy in a Phase 3 study (Corren et al., N Engl J Med 2017: 377 : 936-946). Based on these assumptions, delivery doses of 0.4 mg, 2 mg, and 8 mg (once daily over 28 days) were proposed in Part B of the study, with reduction in FeNO as the primary outcome.

The primary objective was to assess the safety and tolerability of inhaled FAB1 in patients with asthma receiving moderate to high-dose ICS/LABAs. Secondary objectives were the PK and immunogenicity of inhaled FAB1, as well as its effects on FeNO compared to placebo after once-daily dosing for 28 days.

The exploratory objective was to evaluate the effects of various doses of FAB1 on lung function, as well as on asthma symptoms and rescue/reliever therapy use in asthmatics. Lung function measures were assessed by changes from baseline in _FEV1 and FVC before bronchodilator administration (pre-BD) and _FEV1 and FVC after bronchodilator administration (post-BD). Asthma symptom measures were assessed by weekly changes from baseline in ACQ-6 scores.

To be included in Part B of the study, patients had to meet the following criteria: 1. Aged 18 to 75 years (inclusive) with an appropriate intravenous line for intubation or repeated venous puncture. 2. BMI between 18 and 35 kg/ ^m² (inclusive) and weighing at least 45 kg. 3. A confirmed physician-led diagnosis of asthma for > 6 months prior to the screening visit. Patients had to be receiving a stable combination of a LABA and ICS at > 250 to 1000 μg fluticasone propionate DPI or equivalent for at least 1 month prior to the screening visit. If the patient is receiving asthma control medications in addition to ICS plus LABA, the dosage of other asthma control medications (xanthines, anticholinesterases, leukotriene modifiers (cromoglycates)) must be stable for at least 4 weeks prior to the screening visit. 4. Any of the following assessments (documented in their medical history) within the past 10 years to confirm variable airflow obstruction: (a) Variability between clinical visits: FEV1 > 12% and 200 mL. (b) Response to 4 weeks of anti-inflammatory therapy: FEV1 > 12% and 200 mL. (c) Exercise provocation test: FEV1 decrease > 10% and 200 mL. (d) Acetylcholine provocation test: FEV1 decrease of ≥ 20% at < 8 mg/mL. (e) Indirect provocation test (saline or mannitol): FEV1 decrease of ≥ 15%. Or during the screening period: (f) Inter-visit variability: FEV1 > 12% and 200 mL. (g) PEFR during the run-in period: Mean daily PEFR variability > 10% for 2 weeks. 5. Pre-bronchodilator FEV1 ≥ 40% predicted at the screening visit according to ATS/ERS guidelines. 6. FeNO ≥ 30 ppb at the screening visit and ≥ 30 ppb at randomization. 7. ACQ-6 score ≥ 0.75 and ≤ 3.0 at screening. 8. Demonstrated ≥ 65% compliance with each of the following for 7 consecutive days during the screening period (approximately 4.5 days): (a) Twice daily home spirometry and (b) Twice daily electronic diary entries (a compliant day includes an evening and subsequent morning diary entry).

Results Study Population Part A In Part A of the study, a total of 96 healthy volunteers were randomized and treated; 72 received FAB1 and 24 received placebo. All participants completed the study except for one participant in the 6 mg once daily FAB1 group who was lost to follow-up. The median age was 33.5 (range: 20 to 55) years, and the majority were male (94.4%). The majority were White (93.5%), 5.2% were Black or African American, and 1.3% were of other races. Baseline characteristics (height, weight, body mass index) were balanced between treatment groups.

Part B In Part B of the study, a total of 77 patients with asthma were randomized and treated; 51 received FAB1 and 26 received placebo. All patients completed the study, except for one patient in the 8 mg FAB1 group who withdrew consent. The median age was 52.0 (range: 21 to 75) years, and 49.4% were male. The majority were White (93.5%), 5.2% were Black or African American, and 1.3% were of other races. Baseline characteristics (height, weight, body mass index) were balanced between treatment groups.

Pharmacokinetics Section A: Following inhalation of a single dose (0.2 to 16 mg), the time to maximum serum concentration (tmax) of FAB1 was observed at a median time of 8.0 to 11.0 hours. Over the dose range of 2 mg to 16 mg, FAB1 serum concentrations decreased with geometric mean terminal half-lives (t1/2λz) ranging from 20.7 to 25.6 hours. Inter-participant variability was high, as determined by the geometric mean coefficient of variation (CV%) of the maximum plasma (peak) drug concentration (Cmax) after the number of doses (N) administered before steady-state was achieved, the area under the plasma concentration-time curve from time 0 to the last quantifiable concentration (AUClast), and the area under the plasma concentration-time curve from time 0 to infinity (AUCinfinity). No major differences were observed in the estimated PK parameters between Chinese/Japanese participants and non-Asian participants.

Table 25 Geometric Mean (CV%) PK Parameters of FAB1 ( Parts A1 and A2) Following Single DPI Dose Administration in Healthy Volunteers SAD (Section A1) Chinese and Japanese groups (Part A2) Parameter (unit) Group 2 FAB1 0.6 mg once (N = 6) Group 3 FAB1 2 mg once (N = 6) Group 4 FAB1 6 mg once (N = 6) Group 5 FAB1 16 mg once (N = 4) Chinese group: FAB1 16 mg once (N = 4) Japanese group FAB1 16 mg once (N = 6) AUC(0-24) (h*ng/mL) 17.66 (27.0) [n = 4] 26.88 (33.7) [n = 5] 95.17 (26.1) [n = 6] 273.0 (68.3) [n =4] 230.4 (28.6) [n = 4] 262.9 (68.4) [n = 6] AUC last (h*ng/mL) 11.02 (104.5) [n = 6] 31.94 (36.2) [n = 5] 158.2 (42.1) [n = 6] 568.2 (86.6) [n = 4] 412.9 (30.5) [n = 4] 501.3 (57.7) [n = 6] AUCinf (h*ng/mL) NC NC 192.5 (47.4) [n = 5 607.7 (82.4) [n = 4] 526.2 (21.7) [n = 3] 568.2 (57.8) [n = 6] Cmax (ng/mL) 0.9844 (24.4) [n = 6] 1.321 (49.3) [n = 6] 5.721 (36.6) [n = 6] 14.57 (67.3) [n = 4] 11.96 (31.7) [n = 4] 15.05 (70.8) [n = 6] tmax ^a (h) 11.07 (10.08- 12.10) [n = 6] 7.98 (5.98- 10.03) [n = 6] 8.88 (4.05- 24.00) [n = 6] 9.04 (8.00-10.13) [n =4] 8.96 (3.07- 10.35) [n = 4] 11.08 (4.00- 24.15) [n = 6] t1/2λz (h) NC 23.26 (46.7) [n = 4] 21.29 (46.7) [n = 5] 24.46 (27.4) [n = 4] 20.67 (24.6) [n = 3] 25.62 (25.3) [n = 6] Median (minimum-maximum). DPI, dry powder inhaler; N, number of participants in the treatment group; n, number of participants included in the analysis; NC, not calculated. CV%, coefficient of variation, percent; AUC(0-24), area under the plasma concentration-time curve from time 0 to 24 hours; AUClast, area under the plasma concentration-time curve from time 0 to the last quantifiable concentration; Cmax, maximum plasma (peak) drug concentration after a dose (N) before steady-state was achieved; t1/2λ, terminal elimination half-life; tmax, time to maximum concentration after FAB1 administration.

After 14 days of inhaled FAB1 at daily doses of 2 mg, 6 mg, and 16 mg, tmax of FAB1 was observed at median times of 3.1 to 10.1 hours. On day 14, serum concentrations decreased across the 2 mg to 16 mg dose range, with geometric mean t1/2λz ranging from 19.9 to 29.9 hours. Following repeat dosing, a 2- to 3-fold accumulation of both AUC and Cmax was observed, and systemic exposure generally increased in a dose-proportional manner. Inter-participant variability was high, as determined by the geometric mean CV% of Cmax, AUClast, and AUC(0-24). Estimated PK parameters did not differ significantly between Chinese/Japanese and non-Asian participants following repeat dosing.

Table 26 Geometric mean (CV%) PK parameters of FAB1 after multiple DPI doses in healthy volunteers , subsection A3 MAD (Section A3) Chinese and Japanese groups (Part A4) Parameter (unit) Group 1 FAB1 2 mg once daily (N = 6) Group 2 FAB1 6 mg once daily (N = 5) Group 3 FAB1 16 mg once daily (N = 6) Chinese population FAB1 16 mg once daily (N = 6) Japanese group FAB1 16 mg once a day (N = 5) Day 1 Day 14 Day 1 Day 14 Day 1 Day 14 Day 1 Day 14 Day 1 Day 14 AUC(0-24) (h*ng/mL) 37.24 (36.8) [n = 6] 77.58 (40.4) [n = 6] 100.2 (54.7) [n = 5] 280.9 (41.3) [n = 5] 265.2 (58.9) [n = 6] 773.4 (50.1) [n = 6] 246.7 (72.9) [n = 6] 600.9 (70.4) [n = 6] 247.5 (56.0) [n = 5] 484.2 59.0 [n = 5] AUC last (h*ng/mL) 36.84 (37.0) [n = 6] NC 99.59 (54.1) [n = 5] NC 264.6§ (58.3) [n = 6] NC 245.4 (73.3) [n = 6] NC 247.0 (55.8) [n = 5] NC AUCinf (h*ng/mL) NC NC NC NC NC NC NC NC NC NC Cmax (ng/mL) 2.161 (38.5) [n = 6] 4.413 (33.2) [n = 6] 5.816 65.5 [n = 5] 16.72 (46.0) [n = 5] 15.47 (57.7) [n = 6] 41.60 (47.7) [n = 6] 13.69 (70.9) [n = 6] 33.22 (73.4) [n = 6] 13.72 (57.9) [n = 5] 25.29 (41.2) [n = 5] C trough value (ng/mL) NC 3.517 (45.5) [n = 6] NC 12.25 (49.8) [n = 5] NC 34.98 (46.3) [n = 6] NC 23.19 (70.6) [n = 6] NC 18.61 (66.7) [n = 5] tmax ^a (h) 5.04 (3.95-12.03) [n = 6] 7.06 (2.88-10.12) [n = 6] 6.02 (3.00-10.05) [n = 5] 6.08 (3.98-10.28) [n = 5] 10.10 (8.00-23.70) [n = 6] 5.00 (0.00-24.00) [n = 6] 8.00 (3.00-11.97) [n = 6] 3.10 (1.50-9.83) [n = 6] 9.98 (3.10-23.70) [n = 5] 6.00 (3.00-12.02) [n = 5] t1/2λz (h) 36.78 (47.8) [n = 5] 22.00 (42.9) [n = 6] 19.92 (28.2) [n = 3] 27.68 (35.3) [n =5] NC 27.85 (39.9) [n = 6] NC 27.82 (24.0) [n = 6] NC 29.88 (11.7) [n = 5] Rac AUC NC 2.083 (33.1) [n = 6] NC 2.805 (44.0) [n = 5] NC 2.916 (30.5) [n = 6] NC 2.436 (57.9) [n = 6] NC 1.956 (54.2) [n = 5] Rac Cmax NC 2.042 (39.6) [n = 6] NC 2.874 (43.6) [n = 5] NC 2.689 (37.4) [n = 6] NC 2.427 (52.1) [n = 6] NC 1.843 (40.6) [n = 5] ▪ ^aMedian (minimum - maximum). ▪ DPI, dry powder inhaler; N, number of participants in treatment group; n, number of participants included in the analysis; NC, not calculated.

Part B The PK of FAB1 has been characterized in both healthy volunteers (Part A) and patients with asthma in Part B.

In healthy volunteers, after 14 days of inhaled FAB1 at daily doses of 2 mg, 6 mg, and 16 mg, and with a complete PK profile generated, the observed Tmax of FAB1 was a median time of 5-7 hours (range 3-24 hours), and the geometric mean t1/2λz for each dose was 22-28 hours (range 14-45 hours).

In patients who received inhaled FAB1 at daily doses of 0.4 mg, 2 mg, and 8 mg for 28 days, the observed Tmax of FAB1 was a median of 6 to 8 hours (range, 0.25-24 hours). After the last dose on day 28, serum concentrations in all subjects did not decrease sufficiently during the sampling period to characterize the t1/2λz using non-compartmental analysis; however, a geometric mean t1/2λz of 31 hours (range, 23-48 hours) could be estimated for 4 of the 24 subjects at the 8 mg once-daily dosing regimen.

After repeated dosing, a 1- to 2.5-fold accumulation of both AUC and Cmax was observed, and systemic exposure generally increased in a dose-proportional manner. Inter-patient variability was high, as determined by the geometric mean CV% of Cmax, AUClast, and AUC(0-24).

Based on the estimated PK parameters, it can be inferred that the PK of FAB1 is similar between healthy volunteers (Part A) and patients with asthma (Part B).

surface 27 Many times in patients with asthma DPI After dose administration FAB1 Geometric mean (CV%) PK Parameter, subpart B Parameters (Unit) Group 1 0.4 mg once daily (N = 13) Group 2 2 mg once daily (N = 13) Group 3 8 mg once daily (N = 24) Day 1 Day 28 Day 1 Day 28 Day 1 Day 28 AUC(0-24) (h*ng/mL) NC 24.05 (28.4) [n=7] 36.17 (56.8) [n=7] 75.66 (41.6) [n = 11] 86.49 (61.9) [n=22] 238.0 (54.3) [n = 21] AUC final (h*ng/mL) 10.70 (150.2) [n=3] NC 35.98 (58.0) [n=7] NC 86.95 (62.2) [n=22] NC AUCinf (h*ng/mL) NC NC NC NC NC NC Cmax (ng/mL) 1.138 (77.1) [n=4] 1.022 (33.8) [n=9] 1.514 (83.9) [n=10] 3.616 (62.6) [n = 12] 4.768 (64.5) [n=22] 12.09 (53.0) [n = 21] tmax ^a (h) ^a 3.51 (2.00-23.78) [n=4] 7.82 (0.25-24.00) [n=9] 8.00 (3.92-24.00) [n=10] 6.00 (0.25-24.00) [n = 12] 7.79 (2.00-26.50) [n=22] 7.58 (0.00-24.00) [n = 21] t1/2λz (h) NC NC NC NC 31.16 (60.1) [n=3] 30.73 (14.0) [n=4] Rac AUC NC 1.93 (86.2) NC 2.56 (70.6) NC 2.67 (40.3) Rac Cmax NC 0.8836 (40.5) [n=4] NC 2.554 (63.1) [n=10] NC 2.438 (39.9) [n=20]

Immunogenicity Part A In Part A, in the FAB1 group, the ADA prevalence (percentage of ADA-evaluable participants who were ADA+ at any time) was 5.6% (4 of 71 evaluable participants), and the ADA incidence (percentage of evaluable participants who were TE-ADA+) was 1.4% (1 of 71 evaluable participants).

Part B The prevalence and incidence of immunogenicity in Part B of the study were low (Table 28). One patient in the 2 mg group had a treatment-induced ADA-positive response on treatment day 28, and one patient in the 8 mg group had treatment-induced ADA-positive responses on treatment days 14 and 28.

Table 28 Anti-drug Antibody Results - Multidose DPI in Patients with Asthma ( Part B ) FAB1 0.4 mg once daily (N = 13) n (%) FAB1 2 mg once daily (N = 13) n (%) FAB1 8 mg once daily (N = 25) n (%) FAB1 (N = 51) n (%) ADA ^prevalence 0 1/13 (7.7) 1/25 (4.0) 2/51 (3.9) ADA incidence ^b 0 1/13 (7.7) 1/25 (4.0) 2/51 (3.9) ^a The percentage of ADA-evaluable patients who were ADA-positive at any time. ^b The percentage of ADA-evaluable patients who were ADA-positive after treatment induction or intensification. Percentages are based on the number of ADA-evaluable patients (those with at least one ADA assessment).

Pharmacodynamics: After 28 days of treatment, a numerical reduction in FeNO levels was observed in all FAB1 groups (Table 29). FAB1 treatment reduced FeNO levels as early as 6 hours after dosing in the 0.4 mg group and on day 7 in the 2 mg and 8 mg groups, and the reduction persisted throughout the 28 days (Figure 25). Although only the comparison between the FAB1 8 mg and placebo groups was statistically powered, statistically significant reductions in FeNO levels were observed in the FAB1 8 mg and 0.4 mg groups. Compared with placebo, a 23% reduction in FeNO levels was observed for the 8 mg and 0.4 mg doses (geometric mean ratio 0.77; one-sided p-value = 0.0369) and 46% reduction (geometric mean ratio 0.54; one-sided p-value = 0.0003), respectively.

surface 29 In the 28 Time FeNO Change in level relative to baseline – part B ( Pharmacodynamic Analysis Set ) treatment FAB1 vs. placebo n Geometric LS mean 80% CI Geometric mean ratio 80% CI of the ratio p-value FAB1 0.4 mg 13 0.4322 (0.3616, 0.5166) 0.5437 (0.4363, 0.6774) 0.0003 FAB1 2 mg 13 0.6429 (0.5376, 0.7689) 0.8087 (0.6488, 1.008) 0.1083 FAB1 8 mg twenty two 0.6124 (0.5354, 0.7005) 0.7703 (0.6396, 0.9278) 0.0369 Placebo 25 0.7950 (0.6991, 0.9040) NA

Changes in FeNO levels from baseline were analyzed using MMRM, with treatment group, baseline FeNO, visit, and treatment-by-visit interaction as fixed effects and patient as a random effect. Log-transformed FeNO data (change from baseline and percentage change) were analyzed to normalize the skewed distribution of this endpoint and back-transformed to a linear scale. Within-patient correlations were modeled using an unstructured covariate matrix. Denominator degrees of freedom were estimated using the Kenward-Roger approximation. Analyses were performed using only OC, without imputation of missing values. Estimation was performed using REML. Treatment effects were estimated using two-sided 80% confidence intervals (CIs) and one-sided tests of the LS mean for treatment-day interactions and p values for differences between treatment groups. One patient was excluded because of incompatible FeNO data in the CRF, and two patients were excluded because of major protocol deviations.

CI, confidence interval; FeNO, fractional exhaled nitric oxide; LS, least squares; n, number of patients in a given category.

There was also numerical improvement in lung function after 28 days of treatment, as demonstrated by clinically based pre-BD FEV ₁ (105 ml at the highest dose at week 4 - Table 30 and Figure 26) compared to placebo.

Table 30 Change in _FEV1 relative to baseline before clinical BD on Day 28 (ml) - Part B Change from baseline in FEV ₁ compared with placebo (1 h PSOI) Change from baseline in FEV ₁ compared with placebo (D2) Change from baseline in FEV ₁ compared with placebo (D7) Change from baseline in FEV ₁ compared with placebo (D14) Change from baseline in FEV ₁ compared with placebo (D28) FAB1 0.4 mg (n=13) - 18 ml [0.43] + 84 ml [0.20] + 47 ml [0.34] - 27 ml [0.38] - 6 ml [0.48] FAB1 2 mg (n=12) + 190 ml [0.03] + 193 ml [0.03] + 111 ml [0.17] + 120 ml [0.11] + 9 ml [0.47] FAB1 8 mg (n=21) + 152 ml [0.03] + 270 ml [0.0009] + 151 ml [0.06] + 113 ml [0.09] + 105 ml [0.14] PSOI = post-onset of inhalation; n = number of subjects (D28 numbers); numbers in square brackets indicate p-values

In addition, there was numerical improvement in ACQ-6 symptoms (Figure 27).

Example 8 - Population PK Model A population PK (popPK) model was developed to quantify the observed variability in the clinical PK data and to understand any population differences between the healthy adult volunteer population in Part 1A and the adult asthma patient population receiving moderate/high doses of inhaled corticosteroids/long-acting β2 agonists in Part 1B (NCT05110976). The popPK model had four compartments defined by the combined zero- and first-order absorption of the administered dose in the lungs and the observed value in FAB1 serum, defined by the dashed line (Figure 28). Briefly, intravenous (IV) and Part 1A single-ascending dose data were used to estimate bioavailability for each absorption type and then incorporated into a subsequent multiple-ascending dose popPK model that included subjects from Parts 1A and 1B. For patients in Part 1B who received 0.4, 2, and 8 mg once daily for 28 days, the popPK model ( n = 1000/dose) was simulated to obtain predicted serum concentrations and predicted intervals ( Figure 29 , dashed lines) and scaled to predicted lung concentrations ( Figure 29 , solid lines).

Phase 2 Dose Selection: The PK profile observed in Phase 1 (in both patients and healthy volunteers) was sufficiently consistent with the predicted clinical profile to establish confidence in the exposure hypothesis for the inhaled dose of FAB1. Based on the mean exposure associated with the effective dose of 210 mg QW4 of tespelumab, the predicted lung concentration after the intended therapeutic dose of 2 mg was expected to be above the target level (Figure 29). Further confirmation of the mechanism of the significant reduction in FeNO in asthmatic patients after inhaled administration of 8 mg QD FAB1 provides clinical relevance for TSLP inhibition in the lung and confidence in the dose range selected for Phase 2 (0.4-8 mg).

Example 9 – Phase 2b , Randomized, Double-Blind, Placebo-Controlled, Dose-Range-Finding Study to Evaluate the Efficacy and Safety of Three Dose Levels of Inhaled FAB1 This example describes a Phase 2b, randomized, double-blind, placebo-controlled, dose-ranging study to evaluate the efficacy and safety of three dose levels (8 mg, 2 mg, 0.4 mg) of inhaled FAB1 administered once daily by inhalation for 12 to 52 weeks in adults.

Patient Population: The study will include adults (N = approximately 516) with documented physician-diagnosed asthma of at least 12 months' duration and a history of ≥1 severe exacerbation in the last 12 months. All participants will remain symptomatic (Asthma Control Questionnaire [ACQ] score ≥ 1.5) on background asthma therapy with a moderate- or high-dose ICS (as reported in GINA 2023) plus LABA plus additional non-biologic controller therapy (GINA Step 4 or 5 therapy).

The target population includes severe asthma, similar to the tezemab clinical protocol but expanded to include moderate disease. Approximately 30% of patients will have one exacerbation (defined as worsening asthma leading to ≥3 days of OCS use, hospitalization leading to systemic CS use, or an ER visit) in the last 12 months, and approximately 70% of patients will have ≥2 severe exacerbations in the last 12 months.

Eligible patients will be randomized in a 1:1:1:1 ratio to receive FAB1 8 mg once daily, 2 mg once daily, 0.4 mg once daily, or placebo. The dose range in the Phase IIb study is based on results from the Phase Ib part study, in which these same three doses (8 mg, 2 mg, and 0.4 mg) were explored against placebo.

The study is of variable length, with a 12-week treatment period and an optional safety extension of up to 52 weeks of total dosing. The safety extension portion will end after the last enrolled patient completes 12 weeks of treatment. The study design is provided in Figure 30.

The primary and secondary endpoints are provided in Table 31.

surface 31 - Goals and End Points Target End main To evaluate the effect of FAB1 compared with placebo on the time to first CompEx asthma event in patients with uncontrolled moderate to severe asthma. Time of first CompEx asthma event secondary Evaluation of the effect of FAB1 on lung function compared with placebo Changes from baseline in the following: 1. FVC before BD: Weeks 1, 4, and 12 2. FVC after BD: Week 12 3. FEV1 before BD: Weeks 1, 4, and 12 4. FEV1 after BD: Week 12 5. Weekly mean morning PEF: Weeks 1, 4, 6, 8, 10, and 12 6. Weekly mean evening PEF: Weeks 1, 4, 6, 8, 10, and 12 To evaluate the effects of FAB1 compared with placebo on asthma symptoms and control. Change from Baseline: 1. Weekly Mean Asthma Symptom Diary Score, Weeks 1, 4, 6, 8, 10, and 12 2. ACQ-6: Baseline, Weeks 2, 4, 8, 12, and During Treatment 3. AQLQ: Baseline, Weeks 4, 8, and 12 4. SGRQ: Baseline and Week 12 To evaluate the effect of FAB1 on asthma-related biomarkers compared with placebo Changes from baseline: 1. FeNO: Week 1, Week 4, Week 8, and Week 12 2. Blood eosinophils: Week 1, Week 4, Week 8, and Week 12 3. IgE: Week 1, Week 4, Week 8, and Week 12 Evaluating the PK between FAB1 and ADA FAB1 and anti-drug antibody (ADA) plasma concentrations: before drug administration at baseline, week 4, week 8, and week 12

The primary endpoint , CompEx Asthma, is a composite endpoint that allows evaluation of treatment effects on exacerbations and involves fewer participants compared to severe exacerbations. There are two main types of CompEx Asthma events: • Severe exacerbations of asthma • Diary-based (objective worsening)

Severe Exacerbations of Asthma CompEx Events: Investigators will assess exacerbations of asthma at each visit. Severe exacerbations are defined as those resulting in hospital admission, emergency room visit, and/or treatment with oral corticosteroids, as detailed below: • Inpatient Admission: An inpatient admission to an inpatient facility and/or evaluation and treatment in a healthcare facility for asthma lasting ≥24 hours. • Emergency Room or Urgent Care Visit: An evaluation and treatment in an emergency room or urgent care center for asthma requiring systemic corticosteroids lasting <24 hours. • Use a temporary bolus/pulse of systemic corticosteroids (or a temporary increase in a stable OCS background dose) for at least 3 consecutive days to treat symptoms of worsening asthma; a single long-acting injectable dose of corticosteroids will be considered equivalent to a 3-day bolus/pulse of systemic corticosteroids.

Diary-Based CompEx Events Diary-based CompEx events are based on patient-reported deterioration of three electronic diary variables, captured twice daily (morning and evening). This combination yields six different electronic diary variables.

The following morning/evening electronic diary variables were used to define diary-based CompEx events using threshold and slope criteria: • Peak Expiratory Flow (PEF - morning [PEFm] and evening [PEFe]) PEF (L/min) was measured by home spirometry. PEF was captured following standardized procedures. During the data collection period, all required attempts (usually three) were recorded. Only the best of three attempts (maximum PEF) was included in the diary data set and used to calculate CompEx events. PEFm was measured by patients at home, except on field visit days. On field visit days, PEF was assessed on-site, and home PEFm data may not be available. PEFm cannot be estimated using on-site PEF measurements from the on-site visit day (this is because PEFm is patient-reported data, while on-site PEF is investigator-reported data, and these two data sources cannot be used interchangeably in CompEx calculations). • Symptom score (0-3) (Morning [Sm] and Night [Se])

Asthma symptoms during the night and day will be rated by the patient each morning and evening using the following rating system: 0: No asthma symptoms. 1: Asthma symptoms are noticed but tolerated with ease. 2: Asthma is causing sufficient discomfort to cause problems with normal activities (or sleeping). 3: Asthma prevents normal activities (or sleeping). • Rescue medication use (number of doses) (morning [Rm] and night [Re])

Rescue medication use was measured by the number of SABA puffs used during the study period.

The number of rescue medication doses was defined as the number of inhaler puffs recorded in the morning (for the previous night) and at night (for the previous day). If an inhaler was used in the study, the number of reliever doses was defined as the number of inhaler puffs plus twice the number of inhaler applications.

Determination of Log-Based ( Objective Deterioration ) CompEx Events Electronic log events are based on deterioration of the electronic log variables PEFm, PEFe, Sm, Se, Rm, and Re as defined above. Log-based CompEx breathlessness events can be of two types based on: • Threshold criteria • Threshold and slope criteria.

A participant will be considered to have a CompEx event during the planned treatment period if they have either or both of the following: • Objective worsening, defined as meeting the threshold criteria as defined below or • Slope criteria (or both) lasting ≥ 2 consecutive days.

For this purpose, "2 consecutive days" means 2 consecutive days with identical assessments of multiple requirements during those days. For electronic diary data (which are captured twice during a single day), a day will be defined using morning/evening pairings to align with published precedent for CompEx endpoints. (Note: Other electronic diary endpoints in this study will use evening/morning pairings to define a day.) Morning electronic diary records captured on the first day of treatment will not be included in the calculation of the CompEx endpoint.

Before assessing threshold and slope criteria for log-based variables , baseline values must be calculated for each of the six log-based variables: PEFm, PEFe, Sm, Se, Rm, and Re. For each individual patient, baseline values were calculated as the mean of the variable over the last ten days of the run-in period (Days -10 to -1, where Day -1 represents the day before randomization). If fewer than 10 days of data were available, at least five days of data were required to calculate baseline values.

CompEx asthma events could not be calculated for participants who were missing variables based on the baseline diary.

CompEx asthma events based on the following threshold criteria : a. A decrease of ≥15% from baseline in PEFm or PEFe in the morning or evening home-based PEF, and at least one of the following: b. An increase of ≥1.5 from baseline in the rescue medication Rm or Re dose in the morning (for the previous night) or evening (for the previous day) c. An increase of ≥1 from baseline in the symptom score Sm or Se score in the morning or evening or an absolute maximum symptom score (3). This means that the criterion is met even when the value is the highest symptom score of 3.

For (b), the number of rescue medication doses is defined as the number of puffs of the inhaler recorded in the morning and at night, respectively.

The assessment of the threshold criterion for any rolling 2-day period is based on the data available during that period. Non-missing values for fewer than the six variables specified above can meet the threshold criterion, provided that their non-missing values meet the criterion. In other words, this gives a total of eight variable combinations: PEFm-Rm, PEFm-Re, PEFe-Rm, PEFe-Re and PEFm-Sm, PEFm-Se, PEFe-Sm, and PEFe-Se, in which at least two variables in at least one combination must meet the deterioration criterion for at least two consecutive days.

CompEx Asthma Events Based on Threshold and Slope Criteria : A CompEx Asthma Event based on Threshold and Slope Criteria occurs when: At least two consecutive days meet the threshold criteria (a), (b), or (c) above, and the regression slope requirement is also met for the preceding five days. Note that CompEx events are never based solely on slope criteria.

The regression slope requirements for the first five days must meet all of the following conditions: • PEFm slope ≤ -3%/day • PEFe slope ≤ -3%/day • Rm slope ≥ 0.3 doses/day • Re slope ≥ 0.3 doses/day • Sm slope ≥ 0.2 points/day • Se slope ≥ 0.2 points/day

In all cases, the regression slopes are point estimates of the slopes obtained from linear regressions of the absolute values of each of the six variables against the number of days, without including the other variables in the model.

For PEFm and PEFe, the regression slopes thus obtained were also first divided by the baseline PEFm and PEFe values and multiplied by 100, respectively, before applying the above criteria.

The regression slope is calculated if there are at least two non-missing values within the required five days. If one or more of the six variables does not have at least two non-missing values within the required five days, the slope requirement is not met.

Duration of CompEx Asthma Events Based on Diary: The start date of a CompEx asthma event was defined as the earliest date of exacerbation or objective worsening that met the definition. The objective worsening start date was defined as the earliest of two consecutive days from any rolling series that were first qualified using the threshold or slope criteria.

The end date of a CompEx event is defined as the latest of the aggravation or objective deterioration end dates. The objective deterioration end date is defined as the latest of two consecutive days from any series of rolls that meet the threshold or slope criteria.

Evaluate the compliance of log-based CompEx criteria using a scrolling window, evaluating the compliance of each pair of consecutive days. This also applies if different types of criteria (thresholds only or thresholds and slopes) are met for different consecutive days.

Combining CompEx Asthma Events If the end date of the first CompEx event and the start date of the second CompEx event for any participant are less than 7 days apart, these events will be counted as one CompEx event.

Examples and experiments illustrating the principles of the present disclosure will now be discussed with reference to the accompanying figures, in which: FIG1 shows scanning electron microscopy (SEM) images of the formulations tested in Study 1. FIG2A and FIG2B show microfluidics imaging ( MFI) results for 10% bulk powder reconstituted in water for injection to ( FIG2A ) 2.5 mg/ml protein and ( FIG2B ) a stock concentration of 7.5 mg/ml protein. Results are expressed as particle counts per milliliter, as the average of three replicates. FIG3A and FIG3B show MFI results for 40% bulk powder ( BP) reconstituted in water for injection to ( FIG3A ) 2.5 mg/ml protein and ( FIG3B ) a stock concentration of 30 mg/ml protein. Results are expressed as particle counts per milliliter, as the average of three replicates. Figures 4A , 4B , and 4C show the MFI results for ( Figure 4A ) the drug substance (DS) and ( Figure 4B ) 10% and ( Figure 4C ) 40% FAB1 reconstituted BP at the stock protein concentrations (7.5 mg/ml for the 10% formulation and 30 mg/ml for the 40% formulation). Figures 5A , 5B , and 5C show the MFI results for 10% and 40% FAB1 BP at a protein concentration of 2.5 mg/ml for different formulations: ( Figure 5A ) TLTC pH 5 at a concentration of 2.5 mg/ml; ( Figure 5B ) TLTH pH 6 at a concentration of 2.5 mg/ml; and ( Figure 5C ) TLTH pH 5 at a concentration of 2.5 mg/ml. Figures 6A and 6B show the Next Generation Pharmaceutical Impactor (NGI) results for a 10% FAB1, plotted by stage ( Figure 6A ), and a summary of the results ( Figure 6B ). The NGI is described in the United States Pharmacopeia (USP) <601> Apparatus 6. Figures 7A and 7B show the Next Generation Pharmaceutical Impactor (NGI) results for a 10% FAB1, plotted by stage ( Figure 7A ), and a summary of the results ( Figure 7B ). The NGI is described in the USP <601> Apparatus 6. Figure 8 shows a SEM image of a 40% FAB1 from Study 2. Figure 9 shows a SEM image of a 10% FAB1 from Study 2. Figures 10A and 10B show the detection of visible particles (SVPs) in Study 2 for 10% FAB1 formulations reconstituted to a stock concentration of 2.5 mg/ml ( Figure 10A ) or 7.5 mg/ml ( Figure 10B ). Particle counts are expressed as particles/ml in BP of the listed sizes (no larger than (NLT) 2, 5, 10, 25 μm). Figures 11A and 11B show the MFI results for Study 2 for 40% FAB1 formulations reconstituted to a stock concentration of 2.5 mg/ml ( Figure 11A ) or 30 mg/ml ( Figure 11B ). Particle counts are expressed as particles/ml in BP of the listed sizes (no larger than (NLT) 2, 5, 10, 25 μm). Figures 12A and 12B show ( Figure 12A ) the Next Generation Pharmaceutical Impactor (NGI) results for Study 2, 10% FAB1, plotted by grade; and ( Figure 12B ) a summary of the results. The Next Generation Pharmaceutical Impactor (NGI) is as described in the United States Pharmacopeia (USP) <601> Apparatus 6. Figures 13A and 13B show ( Figure 13A ) the Next Generation Pharmaceutical Impactor (NGI) results for Study 2, 40% FAB1, plotted by grade; and ( Figure 13B ) a summary of the results. The Next Generation Pharmaceutical Impactor (NGI) is as described in the United States Pharmacopeia (USP) <601> Apparatus 6. Figure 14 shows the SEM images for Study 3. Figures 15A and 15B show the MFI results for Study 3 of 10% FAB1 reconstituted to a stock concentration of 2.5 mg/ml ( Figure 15A ) or 7.5 mg/ml ( Figure 15B ). Particle counts are expressed as particles/ml in BP of the listed sizes (no larger than (NLT) 2, 5, 10, 25 μm). Results are shown as the mean of triplicate samples. Figures 16A and 16B show the MFI detection of Study 3 SVPs for 40% FAB1 formulations reconstituted to a stock concentration of 2.5 mg/ml ( Figure 16A ) or 30 mg/ml ( Figure 16B ). Particle counts are expressed as particles/ml in BP of the listed sizes (no larger than (NLT) 2, 5, 10, 25 μm). Results are presented as the average of triplicate samples. Figures 17A and 17B show the NGI results for Study 3, plotted by grade ( Figure 17A ) for 10% FAB1 (batches 21-WS-055, 056, and 057), by grade (Figure 17B ) for 40% FAB1 (batches 21-WS-061, 058, and 059), and a summary of the results ( Figure 17C ). The Next Generation Pharmaceutical Impactor (NGI) is as described in United States Pharmacopeia (USP) <601> Apparatus 6. Figure 18 shows the stability SEM image (1-month accelerated conditions) for Study 4. Figures 19A and 19B show the results of the Study 4 NGI, plotted by grade, for 10% FAB1 ( Figure 19A ) with a total of 1.3% His, ( Figure 19B ) with a total of 3.15% His, and ( Figure 19C ) with a total of 5%. The Next Generation Pharmaceutical Impactor (NGI) is as described in the United States Pharmacopeia (USP) <601> Apparatus 6. Figures 20A and 20B show the results of the Study 4 NGI, plotted by grade, for 40% FAB1 ( Figure 20A ) with a total of 1.3% His, ( Figure 20B ) with a total of 3.14% His, and ( Figure 20C ) with a total of 5%. The Next Generation Pharmaceutical Impactor (NGI) is as described in the United States Pharmacopeia (USP) <601> Apparatus 6. Figure 21 shows perivascular/peribronchial mononuclear inflammatory cell infiltrates (arrows) with macrophage aggregates (triangles) in mice receiving 7.1 mg/kg/day of FAB1 in TLTC (pH 6) with PS80, compared to a placebo control labeled "2M 2001, placebo." Figure 22 shows visible particles in FAB1 reconstituted in TLTC (pH 6) containing PS80 in water. Figure 23 shows representative images of FAB1-related lung pathology from the second toxicity study at the indicated dose levels (placebo, 4 mg/kg, and 9.2 mg/kg). Arrows in lung images point to mononuclear cell (MNC) infiltrates. Figure 24 lists the pathology results from the second toxicity study. FIG25 Mean change in FeNO over time by dose relative to baseline, GLS mean (80% confidence interval (CI)) - results from part B of the study described in Example 7 FIG26A Pre - BD FEV1 (L) - mean change over time from baseline in clinical measures, LS mean - results from part B of the study described in Example 7 FIG26B Pre-BD FEV1 (L) - mean change over time from baseline in clinical measures, LS mean - results from part B of the study described in Example 7 High dose group (top-8 Figure 27 shows the change in ACQ-6 from baseline by dose over time, with least squares (LS) means (80% confidence intervals (CI)) for the high-dose group (top - 8 mg) or placebo (bottom) from Part B of the study described in Example 7. Figure 28 shows a schematic diagram of the FAB1 popPK model. ka = absorption rate constant, F1 and F2 = first- and zero-order bioavailability, respectively, D2 = duration of zero-order absorption, CL = clearance, Q1 and Q2 = compartmental clearance, Vc, V1, and V2 = volume of the central compartment, peripheral compartment 1, and peripheral compartment 2. Observations are represented by dashed lines. Figure 29 shows the serum (dashed line) and lung (solid line) concentrations predicted by FAB1 after inhaled 0.4 mg, 2 mg, and 8 mg QD administration. The gray shaded area is a visualization aid for separating the predicted serum and lung results. The horizontal dashed line shows the predicted mean lung C concentration of tezepelumab after SC 210 mg administration (Q4W). Figure 30 shows the Phase 2b protocol design for testing the efficacy of anti-TSLP Fab fragments.

TW202529811A_114103686_SEQL.xml

Claims (162)

A pharmaceutical composition comprising spray-dried particles comprising: a. about 5% (w/w) to about 15% (w/w) leucine; b. about 1% (w/w) to about 5% (w/w) trileucine; c. about 1% (w/w) to about 10% (w/w) histidine buffer, at a pH between about pH 5 and pH 6; d. about 1% (w/w) to about 80% (w/w) an antigen-binding fragment of an anti-thymic stromal lymphopoietin (TSLP) antibody; and e. a glass stabilizer. The pharmaceutical composition of claim 1, comprising about 1% (w/w) to about 3% (w/w) trileucine. The pharmaceutical composition of claim 2, comprising approximately 2% (w/w) trileucine. The pharmaceutical composition of any one of claims 1 to 3, comprising about 8% (w/w) to about 12% (w/w) leucine. The pharmaceutical composition of claim 4, comprising approximately 10.5% (w/w) leucine. The pharmaceutical composition of any one of claims 1 to 5, comprising about 1% (w/w) to about 5% (w/w) histidine buffer. The pharmaceutical composition of claim 6, comprising about 2.5% (w/w) to about 3.5% (w/w) histidine buffer. The pharmaceutical composition of claim 7, comprising approximately 3.14% (w/w) histidine buffer. The pharmaceutical composition of claim 1, wherein the antigen-binding fragment is present at a concentration of about 2% (w/w). The pharmaceutical composition of claim 1, wherein the antigen-binding fragment is present at a concentration of about 10% (w/w). The pharmaceutical composition of claim 1, wherein the antigen-binding fragment is present at a concentration of about 40% (w/w). The pharmaceutical composition of any one of claims 1 to 11, wherein the antigen-binding fragment comprises: a. HCDR1 comprising the amino acid sequence of SEQ ID NO: 1; b. HCDR2 comprising the amino acid sequence of SEQ ID NO: 2; c. HCDR3 comprising the amino acid sequence of SEQ ID NO: 3; d. LCDR1 comprising the amino acid sequence of SEQ ID NO: 5; e. LCDR2 comprising the amino acid sequence of SEQ ID NO: 6; and f. LCDR3 comprising the amino acid sequence of SEQ ID NO: 7. The pharmaceutical composition of any one of claims 1 to 11, wherein the antigen-binding fragment comprises: a. HCDR1 having the amino acid sequence of SEQ ID NO: 1; b. HCDR2 having the amino acid sequence of SEQ ID NO: 2; c. HCDR3 having the amino acid sequence of SEQ ID NO: 3; d. LCDR1 having the amino acid sequence of SEQ ID NO: 5; e. LCDR2 having the amino acid sequence of SEQ ID NO: 6; and f. LCDR3 having the amino acid sequence of SEQ ID NO: 7. The pharmaceutical composition of any one of claims 1 to 13, wherein the antigen-binding fragment comprises a VH domain comprising a sequence that is at least 95%, 90%, 85% or 80% identical to SEQ ID NO: 4, and a VL domain comprising a sequence that is at least 95%, 90%, 85% or 80% identical to SEQ ID NO: 8. The pharmaceutical composition of any one of claims 1 to 14, wherein the antigen-binding fragment comprises a VH domain comprising the sequence of SEQ ID NO: 4; and a VL domain comprising the sequence of SEQ ID NO: 8. The pharmaceutical composition of any one of claims 1 to 15, wherein the antigen-binding fragment is Fab, Fab', F(ab')2, scFv, minibody or diabody. The pharmaceutical composition of claim 16, wherein the antigen-binding fragment is Fab. The pharmaceutical composition of claim 17, wherein the Fab is an IgG1 antibody. The pharmaceutical composition of claim 18, wherein the antigen-binding fragment comprises a first unit having the sequence shown in SEQ ID NO: 28 and a second unit having the sequence shown in SEQ ID NO: 29. The pharmaceutical composition of any one of claims 1 to 19, wherein the glass stabilizer is selected from trehalose, sucrose, raffinose, inulin, dextran, mannitol and cyclodextrin. The pharmaceutical composition of claim 20, wherein the glass stabilizer is trehalose. The pharmaceutical composition of claim 21, wherein the percentage (w/w) concentration of trehalose is supplemented to about 100%. The pharmaceutical composition of any one of claims 1 to 22, wherein the composition does not contain a surfactant. The pharmaceutical composition of any one of claims 1 to 23, comprising: a. 2% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 84.2% (w/w) trehalose ±10%, and 1.3% (w/w) histidine ±10%, pH 5.5; b. 2% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 82.36% (w/w) trehalose ±10%, and 3.14% (w/w) histidine ±10%, pH 5.5; c. 2% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) (w/w) leucine ±10%, 80.5% (w/w) trehalose ±10%, and 5.0% (w/w) histidine ±10%, pH 5.5; d. 10% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 76.2% (w/w) trehalose ±10%, and 1.3% (w/w) histidine ±10%, pH 5.5; e. 10% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 74.36% (w/w) trehalose ±10%, and 3.14% (w/w) histidine ±10%, pH 5.5; f. 10% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 72.5% (w/w) trehalose ±10%, and 5.0% (w/w) histidine ±10%, pH 5.5; g. 40% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 44.36% (w/w) trehalose ±10%, and 3.14% (w/w) histidine ±10%, pH 5.5; h. 40% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 42.5% (w/w) trehalose ±10% and 5.0% (w/w) histidine ±10%, pH 5.5; or i. 40% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 46.2% (w/w) trehalose ±10%, and 1.3% (w/w) histidine ±10%, pH 5.5. The pharmaceutical composition of any one of claims 1 to 24, wherein after reconstitution, the number of subvisible particles between 5 μm and 200 μm is less than about 2.5×10 ⁴ particles/ml. The pharmaceutical composition of claim 25, wherein after reconstitution, the number of subvisible particles between 5 μm and 200 μm is less than about 0.5×10 ⁴ particles/ml. The pharmaceutical composition of any one of claims 1 to 26, wherein after reconstitution, the number of subvisible particles between 10 μm and 200 μm is less than about 1×10 ⁴ particles/ml. The pharmaceutical composition of claim 27, wherein after reconstitution, the number of subvisible particles between 10 μm and 200 μm is less than about 0.2×10 ⁴ particles/ml. The pharmaceutical composition of any one of claims 1 to 28, wherein after reconstitution, the number of subvisible particles between 25 μm and 200 μm is less than about 2×10 ³ particles/ml. The pharmaceutical composition of claim 29, wherein after reconstitution, the number of subvisible particles between 25 μm and 200 μm is less than about 0.2×10 ³ particles/ml. The pharmaceutical composition of any one of claims 24 to 30, wherein the number of subvisible particles is determined by dynamic flow imaging microscopy, optionally by microfluidic imaging (MFI). The pharmaceutical composition of any one of claims 24 to 31, wherein the number of subvisible particles is determined after reconstitution in water to a concentration of 2.5 mg/ml or 30 mg/ml of the antigen-binding fragment. A method for preparing a pharmaceutical composition for inhalation, comprising: a. providing an aqueous solution at about pH 5 to about pH 6, comprising leucine, trileucine, histidine, a glass stabilizer, and an antigen-binding fragment of an anti-thymic stromal lymphopoietin (TSLP) antibody; b. spray-drying the aqueous solution of (a) to produce dry powder particles; and c. collecting the dry powder particles; wherein the aqueous solution comprises about 5% (w/w) to about 15% (w/w) leucine, about 1% (w/w) to about 5% (w/w) trileucine, about 1% (w/w) to about 10% (w/w) histidine, about 5% (w/w) to about 50% (w/w) antigen-binding fragment, and a certain % (w/w) of glass stabilizer to achieve 100% total solids content. The method of claim 33, wherein the aqueous solution has a pH of 5.5. The method of any one of claims 33 to 34, wherein the glass stabilizer is trehalose. The method of claims 33 to 35, wherein the aqueous solution comprises: a. 2% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 84.2% (w/w) trehalose ±10%, and 1.3% (w/w) histidine ±10%, pH 5.5; b. 2% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 82.36% (w/w) trehalose ±10%, and 3.14% (w/w) histidine ±10%, pH 5.5; c. 2% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) (w/w) leucine ±10%, 80.5% (w/w) trehalose ±10%, and 5.0% (w/w) histidine ±10%, pH 5.5; d. 10% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 76.2% (w/w) trehalose ±10%, and 1.3% (w/w) histidine ±10%, pH 5.5; e. 10% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 74.36% (w/w) trehalose ±10%, and 3.14% (w/w) histidine ±10%, pH 5.5; f. 10% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 72.5% (w/w) trehalose ±10%, and 5.0% (w/w) histidine ±10%, pH 5.5; g. 40% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 44.36% (w/w) trehalose ±10%, and 3.14% (w/w) histidine ±10%, pH 5.5; h. 40% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 42.5% (w/w) trehalose ±10% and 5.0% (w/w) histidine ±10%, pH 5.5; or i. 40% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 46.2% (w/w) trehalose ±10%, and 1.3% (w/w) histidine ±10%, pH 5.5. A dry powder formulation obtained by the method of any one of claims 33 to 36. A method for treating a TSLP-related disease in a subject in need thereof, comprising administering to the subject the pharmaceutical composition of any one of claims 1 to 32 or the dry powder formulation of claim 37. The pharmaceutical composition of any one of claims 1 to 32 or the dry powder formulation of claim 36, for use in treating TSLP-related diseases. A use of the pharmaceutical composition of any one of claims 1 to 32 or the dry powder formulation of claim 37 for the manufacture of a medicament for treating TSLP-related diseases. The method of claim 38, the composition for use of claim 39, or the use of claim 40, wherein the TSLP-related disease is asthma, COPD, allergic rhinitis, allergic sinusitis, allergic conjunctivitis, eosinophilic esophagitis, chronic idiopathic urticaria, or chronic sinusitis. The method, use or composition for use of claim 41, wherein the TSLP-related disease is asthma. The method, use or composition for use of claim 41, wherein the TSLP-related disease is COPD. A method for improving lung function in a subject suffering from asthma or COPD, comprising administering to the subject the pharmaceutical composition of any one of claims 1 to 32 or the dry powder formulation of claim 37. A pharmaceutical composition according to any one of claims 1 to 32 or a dry powder formulation according to claim 37 for use in a method of improving lung function in a subject suffering from asthma or COPD. Use of the pharmaceutical composition of any one of claims 1 to 32 or the dry powder formulation of claim 37 for the manufacture of a medicament for improving lung function in a subject suffering from asthma or COPD. The method, use or composition for use of any one of claims 44 to 46, wherein improved lung function refers to an improvement compared to baseline in one or more of the following parameters: (i) pre-bronchodilator (BD) FVC, (ii) post-BD FVC, (iii) pre-BD _FEV1 , (iv) post-BD _FEV1 , (v) mean morning PEF and/or (vi) mean evening PEF. The method, use, or composition for use of claim 47, wherein improvement means achieving a minimal clinically important difference in each respective parameter. The method, use, or composition for use of claim 48, wherein the baseline is the value of the respective parameter before initiation of the treatment with the pharmaceutical composition or the dry powder formulation. In the method, use or composition for use of claim 49, before means within one month of commencing the treatment. The method, use or composition for use of claim 49 or 50, wherein the improvement in _FEV1 before BD compared to baseline is at least 5 ml, at least 10 ml, at least 15 ml, at least 20 ml, at least 25 ml, at least 30 ml, at least 35 ml, at least 40 ml, at least 45 ml, at least 50 ml, at least 55 ml, at least 60 ml, at least 65 ml, at least 70 ml, at least 75 ml, at least 80 ml, at least 85 ml, at least 90 ml, at least 95 ml, at least 100 ml, at least 105 ml, at least 110 ml, at least 115 ml, at least 120 ml, at least 125 ml, at least 130 ml, at least 135 ml, at least 140 ml, at least 145 ml, at least 150 ml, at least 160 ml, at least 170 ml, at least 180 ml, at least 190 ml, at least 200 ml, at least 210 ml, at least 220 ml, at least 230 ml ml, at least 240 ml or at least 250 ml. The method, use, or composition for use of claim 49 or 50, wherein the improvement in pre-BD _FEV1 compared to baseline is at least 80 ml on day 2 after initiation of treatment, at least 45 ml or at least 100 ml on day 7 after initiation of treatment, at least 100 ml on day 14 after initiation of treatment, or at least 5 ml or at least 100 ml on day 28 after initiation of treatment. A method for improving symptoms of asthma or COPD in a subject in need thereof, comprising administering to the subject the pharmaceutical composition of any one of claims 1 to 32 or the dry powder formulation of claim 37. The pharmaceutical composition of any one of claims 1 to 32 or the dry powder formulation of claim 37, for use in a method of ameliorating the symptoms of asthma or COPD in a subject in need thereof. Use of the pharmaceutical composition of any one of claims 1 to 32 or the dry powder formulation of claim 37 for the manufacture of a medicament for ameliorating the symptoms of asthma or COPD in a subject in need thereof. The method, use or composition for use of any one of claims 53 to 55, wherein improvement in asthma symptoms refers to improvement compared to baseline in one or more of the following parameters: (i) mean Asthma Symptom Diary score, (ii) ACQ-6 score, (iii) AQLQ score and/or (iv) SGRQ score. The method, use or composition for use of claim 56, wherein improvement in asthma symptoms refers to improvement in ACQ-6 scores compared to baseline. The method, use or composition for use of any one of claims 53 to 57, wherein improvement means achieving a minimal clinically important difference (MCID) for each respective score. The method, use or composition for use of claim 56 or 57, wherein the baseline refers to the value of the respective score before starting the treatment with the pharmaceutical composition of any one of claims 1 to 32 or the dry powder formulation of claim 37. In the method, use or composition for use of claim 59, before means within one month of commencing the treatment. The method, use, or composition for use of claim 42 or any one of claims 44 to 60, wherein the asthma is moderate to severe asthma. The method, use or composition for use of claim 42 or any one of claims 44 to 61, wherein the pharmaceutical composition is administered by inhalation or intranasally. The method, use, or composition for use of claim 42 or any one of claims 44 to 62, wherein the pharmaceutical composition is delivered by a dry powder inhaler. The method, use or composition for use of claim 42 or any one of claims 44 to 63, wherein the pharmaceutical composition is administered or is to be administered in an amount comprising about 0.4 mg to about 8 mg of the antigen-binding fragment. The method, use or composition for use of claim 64, wherein the dose is administered or is to be administered daily. The method, use or composition for use of claim 65, wherein the dose is administered or is to be administered once a day (Q1D). The method, use or composition for use of any one of claims 64 to 66, wherein the pharmaceutical composition is administered or is to be administered in an amount comprising about 0.4 mg of the antigen-binding fragment. The method, use or composition for use of any one of claims 64 to 66, wherein the pharmaceutical composition is administered or is to be administered in an amount comprising about 2 mg of the antigen-binding fragment. The method, use or composition for use of any one of claims 64 to 66, wherein the pharmaceutical composition is administered or is to be administered in an amount comprising about 8 mg of the antigen-binding fragment. The method, use, or composition for use of claim 42 or any one of claims 44 to 69, wherein the asthma is uncontrolled moderate to severe asthma. The method, use, or composition for use of claim 42 or any one of claims 44 to 70, wherein the individual has a history of ≥ 1 severe exacerbation in the last 12 months prior to the treatment. The method, use, or composition for use of claim 42 or any one of claims 44 to 71, wherein the individual has a history of ≥ 2 severe exacerbations in the last 12 months prior to the treatment. The method, use, or composition for use of claim 42 or any one of claims 44 to 72, wherein the subject is co-administered with a background therapy. The method, use or composition for use of claim 73, wherein the individual has received the background therapy prior to the treatment. The method, use, or composition for use of claim 73 or 74, wherein the background therapy is selected from the group consisting of an inhaled corticosteroid; a leukotriene modulator; a long-acting beta agonist (LABA); a long-acting muscarinic antagonist (LAMA); combination therapy, such as fluticasone and salmeterol, adenine and formoterol, mometasone and formoterol, and fluticasone and vilanterol; theophylline; a short-acting beta agonist (SABA); ipratropium or a combination of ipratropium and albuterol; or oral corticosteroids. The method, use, or composition for use of any of claims 73 to 75, wherein the background therapy comprises a combination of a medium- or high-dose ICS (reported in accordance with GINA 2023) and a LABA (GINA step 4 or 5 therapy). The method, use or composition for use of any one of claims 38 to 76, wherein the subject has a baseline blood eosinophil count of ≥ 150 cells/μl. The method, use or composition for use of any one of claims 38 to 77, wherein the subject has a baseline blood eosinophil count of ≥ 300 cells/μl. The method, use, or composition for use of claim 77 or 78, wherein baseline is the blood eosinophil count before initiation of the treatment. In the method, use or composition for use of claim 79, before means within one month of commencing the treatment. The method, use or composition for use of any one of claims 38 to 80, wherein after treatment with the pharmaceutical composition of any one of claims 1 to 32 or the dry powder formulation of claim 37, the prevalence of ADA in the individual is less than 6%, less than 5% or less than 4%, and/or the incidence of ADA is less than 4%, less than 3% or less than 2%. A method of treating asthma in a subject in need thereof comprises administering to the subject a pharmaceutical composition comprising an antigen-binding fragment of an anti-TSLP antibody in an amount of about 0.4 mg to about 8 mg, wherein the pharmaceutical composition is administered once daily (Q1D) by inhalation. A pharmaceutical composition for use in a method of treating asthma in a subject in need thereof, wherein the pharmaceutical composition comprises an antigen-binding fragment of an anti-TSLP antibody in an amount of about 0.4 mg to about 8 mg, the antigen-binding fragment to be administered to the subject once daily (Q1D) by inhalation. Use of an antigen-binding fragment of an anti-TSLP antibody in the manufacture of a pharmaceutical composition for treating asthma by inhalation, wherein the pharmaceutical composition comprises a dose of about 0.4 mg to about 8 mg of the antigen-binding fragment. A method for treating asthma in a subject in need thereof comprises administering to the subject a pharmaceutical composition comprising an antigen-binding fragment of an anti-TSLP antibody in an amount of about 0.4 mg to about 8 mg, wherein the pharmaceutical composition is administered once daily (Q1D) by inhalation, and wherein the antigen-binding fragment is a Fab comprising: a. HCDR1 comprising the amino acid sequence of SEQ ID NO: 1; b. HCDR2 comprising the amino acid sequence of SEQ ID NO: 2; c. HCDR3 comprising the amino acid sequence of SEQ ID NO: 3; d. LCDR1 comprising the amino acid sequence of SEQ ID NO: 5; e. LCDR2 comprising the amino acid sequence of SEQ ID NO: 6; and f. LCDR3 comprising the amino acid sequence of SEQ ID NO: 7. A pharmaceutical composition for use in a method of treating asthma in a subject in need thereof, wherein the pharmaceutical composition comprises an antigen-binding fragment of an anti-TSLP antibody in an amount of about 0.4 mg to about 8 mg, the antigen-binding fragment to be administered to the subject once daily (Q1D) by inhalation, wherein the antigen-binding fragment is a Fab comprising: a. HCDR1 comprising the amino acid sequence of SEQ ID NO: 1; b. HCDR2 comprising the amino acid sequence of SEQ ID NO: 2; c. HCDR3 comprising the amino acid sequence of SEQ ID NO: 3; d. LCDR1 comprising the amino acid sequence of SEQ ID NO: 5; e. LCDR2 comprising the amino acid sequence of SEQ ID NO: 6; and f. LCDR3 comprising the amino acid sequence of SEQ ID NO: 7. Use of an antigen-binding fragment of an anti-TSLP antibody in the manufacture of a pharmaceutical composition for treating asthma, wherein the pharmaceutical composition comprises a dose of about 0.4 mg to about 8 mg of the antigen-binding fragment, wherein the antigen-binding fragment is a Fab comprising: a. HCDR1 comprising the amino acid sequence of SEQ ID NO: 1; b. HCDR2 comprising the amino acid sequence of SEQ ID NO: 2; c. HCDR3 comprising the amino acid sequence of SEQ ID NO: 3; d. LCDR1 comprising the amino acid sequence of SEQ ID NO: 5; e. LCDR2 comprising the amino acid sequence of SEQ ID NO: 6; and f. LCDR3 comprising the amino acid sequence of SEQ ID NO: 7, and the pharmaceutical composition is to be administered once daily (Q1D) by inhalation. The method, use or pharmaceutical composition for use of any one of claims 82 to 87, wherein the antigen-binding fragment comprises a VH domain having a sequence that is at least 95%, 90%, 85% or 80% identical to SEQ ID NO: 4, and a VL domain having a sequence that is at least 95%, 90%, 85% or 80% identical to SEQ ID NO: 8. The method, use or pharmaceutical composition for use of any one of claims 82 to 88, wherein the antigen-binding fragment comprises a VH domain comprising the sequence of SEQ ID NO: 4; and a VL domain comprising the sequence of SEQ ID NO: 8. The method, use or pharmaceutical composition for use of any one of claims 82 to 89, wherein the Fab is an IgG1 antibody. The method, use or pharmaceutical composition for use of claim 90, wherein the antigen-binding fragment comprises a first unit having the sequence shown in SEQ ID NO: 28 and a second unit having the sequence shown in SEQ ID NO: 29. The method, use, or pharmaceutical composition for use of any of claims 82 to 91, wherein the pharmaceutical composition comprises spray-dried particles comprising: g. about 5% (w/w) to about 15% (w/w) leucine; h. about 1% (w/w) to about 5% (w/w) trileucine; i. about 1% (w/w) to about 50% (w/w) an antigen-binding fragment of an anti-thymic stromal lymphopoietin (TSLP) antibody; j. a buffer; and k. a glass stabilizer. The method, use, or pharmaceutical composition for use of claim 92, comprising about 1% (w/w) to about 10% (w/w) histidine buffer and having a pH between about pH 5 and pH 6. The method, use, or pharmaceutical composition for use of claim 92 or 93, comprising about 1% (w/w) to about 3% (w/w) trileucine. The method, use or pharmaceutical composition for use of claim 94, comprising about 2% (w/w) trileucine. The method, use or pharmaceutical composition for use of any one of claims 92 to 95, comprising about 8% (w/w) to about 12% (w/w) leucine. The method, use, or pharmaceutical composition for use of claim 96, comprising approximately 10.5% (w/w) leucine. The method, use, or pharmaceutical composition for use of any one of claims 92 to 97, comprising about 1% (w/w) to about 5% (w/w) histidine buffer. The method, use, or pharmaceutical composition for use of claim 98, comprising about 2.5% (w/w) to about 3.5% (w/w) histidine buffer. The method, use, or pharmaceutical composition for use of claim 99, comprising approximately 3.14% (w/w) histidine buffer. The method, use or pharmaceutical composition for use of any one of claims 92 to 100, wherein the antigen-binding fragment is present at a concentration of about 2% (w/w), about 10% (w/w) or about 40% (w/w). The method, use or pharmaceutical composition for use of any one of claims 92 to 101, wherein the antigen-binding fragment is present at a concentration of about 2% (w/w). The method, use or pharmaceutical composition for use of any one of claims 92 to 102, wherein the antigen-binding fragment is present at a concentration of about 10% (w/w). The method, use or pharmaceutical composition for use of any one of claims 92 to 103, wherein the antigen-binding fragment is present at a concentration of about 40% (w/w). The method, use or pharmaceutical composition for use of any one of claims 92 to 104, wherein the glass stabilizer is selected from trehalose, sucrose, raffinose, inulin, dextran, mannitol and cyclodextrin. The method, use, or pharmaceutical composition for use of claim 105, wherein the glass stabilizer is trehalose. The method, use or pharmaceutical composition for use of claim 106, wherein the percentage (w/w) concentration of trehalose is replenished to about 100%. The method, use or pharmaceutical composition for use of any one of claims 92 to 107, wherein the composition does not contain a surfactant. The method, use, or pharmaceutical composition for use of any of claims 92 to 108, comprising: a. 2% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 84.2% (w/w) trehalose ±10%, and 1.3% (w/w) histidine ±10%, pH 5.5; b. 2% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 82.36% (w/w) trehalose ±10%, and 3.14% (w/w) histidine ±10%, pH 5.5; c. 2% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 80.5% (w/w) trehalose ±10%, and 5.0% (w/w) histidine ±10%, pH 5.5; d. 10% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 76.2% (w/w) trehalose ±10%, and 1.3% (w/w) histidine ±10%, pH 5.5; e. 10% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 74.36% (w/w) trehalose ±10%, and 3.14% (w/w) histidine ±10%, pH 5.5; f. 10% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 72.5% (w/w) trehalose ±10%, and 5.0% (w/w) histidine ±10%, pH 5.5; g. 40% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 44.36% (w/w) trehalose ±10%, and 3.14% (w/w) histidine ±10%, pH 5.5; h. 40% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 42.5% (w/w) trehalose ±10%, and 5.0% (w/w) histidine ±10%, pH 5.5; or i. 40% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 46.2% (w/w) trehalose ±10%, and 1.3% (w/w) histidine ±10%, pH 5.5. The method, use or pharmaceutical composition for use of any one of claims 92 to 109, wherein the asthma is moderate to severe asthma. The method, use or pharmaceutical composition for use of any one of claims 92 to 110, wherein the pharmaceutical composition is delivered by a dry powder inhaler. The method, use or composition for use of any one of claims 110 to 111, wherein the asthma is uncontrolled moderate to severe asthma. The method, use, or composition for use of any of claims 92 to 112, wherein the individual has a history of ≥ 1 severe exacerbation in the last 12 months prior to the treatment. The method, use, or composition for use of any of claims 92 to 113, wherein the individual has a history of ≥ 2 severe exacerbations in the last 12 months prior to the treatment. The method, use or composition for use of any one of claims 92 to 114, wherein the subject is co-administered with a background therapy. The method, use or composition for use of claim 115, wherein the individual has received the background therapy prior to the treatment. The method, use, or composition for use of claim 115 or 116, wherein the background therapy is selected from: an inhaled corticosteroid; a leukotriene modulator; a long-acting beta agonist (LABA); a long-acting muscarinic antagonist (LAMA); combination therapy, such as flutecortin and salmeterol, adenine and formoterol, mometasone and formoterol, and flutecortin and vilanterol; theophylline; a short-acting beta agonist (SABA); ipratropium or a combination of ipratropium and albuterol; or oral corticosteroids. The method, use, or composition for use of any of claims 115 to 117, wherein the background therapy comprises a combination of a medium- or high-dose ICS (reported in accordance with GINA 2023) and a LABA (GINA Step 4 or 5 therapy). The method, use, or composition for use of any one of claims 92 to 118, wherein the subject has a baseline blood eosinophil count of ≥ 150 cells/μl. The method, use, or composition for use of any one of claims 92 to 118, wherein the subject has a baseline blood eosinophil count of ≥ 300 cells/μl. The method, use, or composition for use of claim 119 or 120, wherein baseline is the blood eosinophil count before initiation of the treatment. In the method, use or composition for use of claim 121, before means within one month of commencing the treatment. A unit dose pharmaceutical composition comprising 0.2 mg to 16 mg of an antigen-binding fragment of an anti-TSLP antibody, wherein the pharmaceutical composition and the antigen-binding fragment are as described herein. A method of treating COPD in a subject in need thereof comprises administering to the subject a pharmaceutical composition comprising an antigen-binding fragment of an anti-TSLP antibody in an amount of about 0.4 mg to about 8 mg, wherein the pharmaceutical composition is administered once daily (Q1D) by inhalation. A pharmaceutical composition for use in a method of treating COPD in a subject in need thereof, wherein the pharmaceutical composition comprises an antigen-binding fragment of an anti-TSLP antibody in an amount of about 0.4 mg to about 8 mg, the antigen-binding fragment to be administered to the subject once daily (Q1D) by inhalation. Use of an antigen-binding fragment of an anti-TSLP antibody in the manufacture of a pharmaceutical composition for treating COPD by inhalation, wherein the pharmaceutical composition comprises a dose of about 0.4 mg to about 8 mg of the antigen-binding fragment. A method for treating COPD in a subject in need thereof comprises administering to the subject a pharmaceutical composition comprising an antigen-binding fragment of an anti-TSLP antibody in an amount of about 0.4 mg to about 8 mg, wherein the pharmaceutical composition is administered once daily (Q1D) by inhalation, and wherein the antigen-binding fragment is a Fab comprising: g. HCDR1 comprising the amino acid sequence of SEQ ID NO: 1; h. HCDR2 comprising the amino acid sequence of SEQ ID NO: 2; i. HCDR3 comprising the amino acid sequence of SEQ ID NO: 3; j. LCDR1 comprising the amino acid sequence of SEQ ID NO: 5; k. LCDR2 comprising the amino acid sequence of SEQ ID NO: 6; and l. LCDR3 comprising the amino acid sequence of SEQ ID NO: 7. A pharmaceutical composition for use in a method of treating COPD in a subject in need thereof, wherein the pharmaceutical composition comprises an antigen-binding fragment of an anti-TSLP antibody in an amount of about 0.4 mg to about 8 mg, the antigen-binding fragment to be administered to the subject once daily (Q1D) by inhalation, wherein the antigen-binding fragment is a Fab comprising: g. HCDR1 comprising the amino acid sequence of SEQ ID NO: 1; h. HCDR2 comprising the amino acid sequence of SEQ ID NO: 2; i. HCDR3 comprising the amino acid sequence of SEQ ID NO: 3; j. LCDR1 comprising the amino acid sequence of SEQ ID NO: 5; k. LCDR2 comprising the amino acid sequence of SEQ ID NO: 6; and l. LCDR3 comprising the amino acid sequence of SEQ ID NO: 7. Use of an antigen-binding fragment of an anti-TSLP antibody in the manufacture of a pharmaceutical composition for treating COPD, wherein the pharmaceutical composition comprises a dose of about 0.4 mg to about 8 mg of the antigen-binding fragment, wherein the antigen-binding fragment is a Fab comprising: l. HCDR1 comprising the amino acid sequence of SEQ ID NO: 1; m. HCDR2 comprising the amino acid sequence of SEQ ID NO: 2; n. HCDR3 comprising the amino acid sequence of SEQ ID NO: 3; o. LCDR1 comprising the amino acid sequence of SEQ ID NO: 5; p. LCDR2 comprising the amino acid sequence of SEQ ID NO: 6; and q. LCDR3 comprising the amino acid sequence of SEQ ID NO: 7, and the pharmaceutical composition is to be administered once daily (Q1D) by inhalation. The method, use or pharmaceutical composition for use of any one of claims 124 to 129, wherein the antigen-binding fragment comprises a VH domain having a sequence that is at least 95%, 90%, 85% or 80% identical to SEQ ID NO: 4, and a VL domain having a sequence that is at least 95%, 90%, 85% or 80% identical to SEQ ID NO: 8. The method, use or pharmaceutical composition for use of any one of claims 124 to 130, wherein the antigen-binding fragment comprises a VH domain comprising the sequence of SEQ ID NO: 4; and a VL domain comprising the sequence of SEQ ID NO: 8. The method, use or pharmaceutical composition for use of any one of claims 124 to 131, wherein the Fab is an IgG1 antibody. The method, use or pharmaceutical composition for use of claim 132, wherein the antigen-binding fragment comprises a first unit having the sequence shown in SEQ ID NO: 28 and a second unit having the sequence shown in SEQ ID NO: 29. The method, use, or pharmaceutical composition for use of any one of claims 124 to 133, wherein the pharmaceutical composition comprises spray-dried particles comprising: r. about 5% (w/w) to about 15% (w/w) leucine; s. about 1% (w/w) to about 5% (w/w) trileucine; t. about 1% (w/w) to about 50% (w/w) antigen-binding fragment of an anti-thymic stromal lymphopoietin (TSLP) antibody; u. a buffer; and v. a glass stabilizer. The method, use, or pharmaceutical composition for use of claim 134, comprising about 1% (w/w) to about 10% (w/w) histidine buffer and having a pH between about pH 5 and pH 6. The method, use, or pharmaceutical composition for use of claim 134 or 135, comprising about 1% (w/w) to about 3% (w/w) trileucine. The method, use, or pharmaceutical composition for use of claim 136, comprising about 2% (w/w) trileucine. The method, use or pharmaceutical composition for use of any one of claims 134 to 137, comprising about 8% (w/w) to about 12% (w/w) leucine. The method, use, or pharmaceutical composition for use of claim 138, comprising approximately 10.5% (w/w) leucine. The method, use, or pharmaceutical composition for use of any one of claims 134 to 139, comprising about 1% (w/w) to about 5% (w/w) histidine buffer. The method, use, or pharmaceutical composition for use of claim 140, comprising about 2.5% (w/w) to about 3.5% (w/w) histidine buffer. The method, use, or pharmaceutical composition for use of claim 141, comprising approximately 3.14% (w/w) histidine buffer. The method, use or pharmaceutical composition for use of any one of claims 134 to 142, wherein the antigen-binding fragment is present at a concentration of about 2% (w/w), about 10% (w/w) or about 40% (w/w). The method, use or pharmaceutical composition for use of any one of claims 134 to 143, wherein the antigen-binding fragment is present at a concentration of about 2% (w/w). The method, use or pharmaceutical composition for use of any one of claims 134 to 143, wherein the antigen-binding fragment is present at a concentration of about 10% (w/w). The method, use or pharmaceutical composition for use of any one of claims 134 to 143, wherein the antigen-binding fragment is present at a concentration of about 40% (w/w). The method, use or pharmaceutical composition for use of any one of claims 134 to 146, wherein the glass stabilizer is selected from trehalose, sucrose, raffinose, inulin, dextran, mannitol and cyclodextrin. The method, use, or pharmaceutical composition for use of claim 147, wherein the glass stabilizer is trehalose. The method, use or pharmaceutical composition for use of claim 148, wherein the percentage (w/w) concentration of trehalose is supplemented to about 100%. The method, use or pharmaceutical composition for use of any one of claims 134 to 149, wherein the composition does not contain a surfactant. The method, use, or pharmaceutical composition for use of any of claims 134 to 150, comprising: j. 2% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 84.2% (w/w) trehalose ±10%, and 1.3% (w/w) histidine ±10%, pH 5.5; k. 2% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 82.36% (w/w) trehalose ±10%, and 3.14% (w/w) histidine ±10%, pH 5.5; l. 2% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 80.5% (w/w) trehalose ±10%, and 5.0% (w/w) histidine ±10%, pH 5.5; m. 10% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 76.2% (w/w) trehalose ±10%, and 1.3% (w/w) histidine ±10%, pH 5.5; n. 10% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 74.36% (w/w) trehalose ±10%, and 3.14% (w/w) histidine ±10%, pH 5.5; o. 10% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 72.5% (w/w) trehalose ±10%, and 5.0% (w/w) histidine ±10%, pH 5.5; p. 40% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 44.36% (w/w) trehalose ±10%, and 3.14% (w/w) histidine ±10%, pH 5.5; q. 40% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 42.5% (w/w) trehalose ±10%, and 5.0% (w/w) histidine ±10%, pH 5.5; or r. 40% (w/w) antigen-binding fragment ±20%, 2% (w/w) trileucine ±10%, 10.5% (w/w) leucine ±10%, 46.2% (w/w) trehalose ±10%, and 1.3% (w/w) histidine ±10%, pH 5.5. The method, use or pharmaceutical composition for use of any one of claims 134 to 151, wherein the pharmaceutical composition is delivered by a dry powder inhaler. The method, use, or composition for use of any of claims 134 to 152, wherein the individual has a history of ≥ 1 severe exacerbation in the last 12 months prior to the treatment. The method, use, or composition for use of any of claims 134 to 153, wherein the individual has a history of ≥ 2 severe exacerbations in the last 12 months prior to the treatment. The method, use or composition for use of any one of claims 134 to 154, wherein the subject is co-administered with a background therapy. The method, use or composition for use of claim 155, wherein the individual has received the background therapy prior to the treatment. The method, use, or composition for use of claim 155 or 156, wherein the background therapy is selected from: an inhaled corticosteroid; a leukotriene modulator; a long-acting beta agonist (LABA); a long-acting muscarinic antagonist (LAMA); combination therapy, such as flutecortin and salmeterol, adenine and formoterol, mometasone and formoterol, and flutecortin and vilanterol; theophylline; a short-acting beta agonist (SABA); ipratropium or a combination of ipratropium and albuterol; or oral corticosteroids. The method, use, or composition for use of any of claims 155 to 157, wherein the background therapy comprises a combination of a medium- or high-dose ICS (reported in accordance with GINA 2023) and a LABA (GINA Step 4 or 5 therapy). The method, use, or composition for use of any one of claims 134 to 158, wherein the subject has a baseline blood eosinophil count of ≥ 150 cells/μl. The method, use, or composition for use of any one of claims 134 to 158, wherein the subject has a baseline blood eosinophil count of ≥ 300 cells/μl. The method, use, or composition for use of claim 159 or 160, wherein baseline is the blood eosinophil count before initiation of the treatment. In the method, use or composition for use of claim 161, before means within one month of commencing the treatment.