TW202140072A - Formulations of human anti-tslp antibodies and methods of treating inflammatory disease - Google Patents

Abstract

Provided herein are aqueous compositions comprising (a) an anti-TSLP antibody at a concentration greater than about 140 mg/mL, (b) a surfactant, and (c) at least one basic amino acid or a salt thereof. Also provided are aqueous compositions comprising aqueous compositions comprising (a) an anti-TSLP antibody at a concentration greater than about 140 mg/mL, (b) a surfactant, and (c) at least one calcium salt or magnesium salt. Related articles of manufacture, pre-filled syringes, and vials comprising the compositions of the present disclosure are also provided. Uses of the compositions for treating an inflammatory disease, e.g., atopic dermatitis, are provided herein. Also, methods of making a stable, liquid antibody comprising having a viscosity of less than about 100 cP is provided herein.

Description

Human anti-TSLP antibody formulations and methods for treating inflammatory diseases

The present disclosure relates to human anti-TSLP monoclonal antibodies, including high-concentration aqueous formulations of tezepelumab and its biosimilars. Incorporate electronically submitted materials by reference

Incorporated in its entirety by reference is the computer-readable nucleotide/amino acid sequence list submitted at the same time as this article, which is identified as follows: 9911-byte ASCII (text) file name "54250_Seqlisting.txt"; created in 2021 February 11th.

Related technology

In the most recent phase 2, randomized, double-blind, placebo-controlled clinical trial, teperumumab (also known as AMG 157 and MED9929) was administered to humans in doses ranging from 70 mg to 280 mg. Compared with subjects who received placebo, subjects who received tepelumab showed a lower rate of clinically significant asthma exacerbations.

The increase in protein concentration in pharmaceutical formulations can cause problems. For example, a formulation containing a high concentration of protein can cause aggregation, which can lead to the formation of high molecular weight species (HMWS). HMWS may be of particular interest in some protein formulations. Aggregation may also potentially affect the subcutaneous bioavailability and pharmacokinetics of therapeutic proteins, and may also lead to loss of protein biological activity and increased immunity. High-concentration protein formulations may lead to increased viscosity, which may adversely affect the filling and administration of pharmaceutical products.

Therefore, there is a need in the art for high-concentration formulations of teperumumab with reduced viscosity, high stability, and low aggregation levels.

This article provides for the first time data demonstrating the viscosity-lowering effect of certain excipients on high-concentration antibody formulations. The data supports the viscosity-reducing effect of basic amino acids or their salts and calcium or magnesium salts. The data also supports the stability of this high concentration antibody formulation.

Therefore, the present disclosure provides a composition, such as an aqueous composition, which comprises (a) an anti-TSLP antibody at a concentration greater than about 140 mg/mL, (b) a surfactant, and (c) at least one basic amino acid or Its salt. An aqueous composition is also considered, which comprises (a) an anti-TSLP antibody at a concentration greater than about 140 mg/mL, (b) a surfactant, and (c) at least one basic amino acid or salt thereof, wherein the composition Contains about 10 mM to about 200 mM basic amino acid or its salt. In an illustrative example, the basic amino acid is arginine. If necessary, the salt is an organic salt of arginine. In many aspects, the arginine is arginine acetate, arginine aspartate, arginine glutamate, arginine glycolate, arginine lactate, arginine Methanesulfonate, arginine propionate, or a combination thereof. In an illustrative example, the basic amino acid is histidine. If necessary, the salt is an organic salt of histidine. In an exemplary aspect, the histidine salt is histidine acetate, histidine aspartate, histidine glutamine, histidine glycolate, histidine lactate, histidine Methanesulfonate, histidine propionate, or a combination thereof. In different cases, the basic amino acid is lysine. If necessary, the salt is an organic salt of lysine acid. In many aspects, the lysine salt is lysine acetate, lysine aspartate, lysine glutamate, lysine glycolate, lysine lactate, lysine Methanesulfonate, lysine propionate, or a combination thereof.

The present disclosure also provides a composition, such as an aqueous composition, which comprises (a) an anti-TSLP antibody at a concentration greater than about 140 mg/mL, (b) a surfactant, and (c) at least one calcium salt or magnesium salt. An aqueous composition is also considered, which comprises (a) an anti-TSLP antibody at a concentration greater than about 140 mg/mL, (b) a surfactant, and (c) at least one calcium salt or magnesium salt, wherein the composition contains about 15 mM to about 150 mM calcium or magnesium salt. In an exemplary aspect, the calcium salt or magnesium salt comprises a relative ion lacking chloride. If necessary, the relative ion is acetate, aspartate, glutamate, glycolate, lactate, methanesulfonate, propionate, or a combination thereof. In various aspects, the calcium salt is calcium acetate, calcium aspartate, calcium glutamate, calcium glycolate, calcium lactate, calcium methanesulfonate, calcium propionate, or a combination thereof. In an illustrative example, the magnesium salt is magnesium acetate, magnesium aspartate, magnesium glutamine, magnesium glycolate, magnesium lactate, magnesium methanesulfonate, magnesium propionate, or a combination thereof. Under different circumstances, the composition of the present disclosure contains about 50 mM to about 150 mM basic amino acid or its salt or about 50 mM to about 150 mM calcium salt or magnesium salt.

In various aspects, the composition of the present disclosure further contains N-acetylarginine (NAR) and N-acetyl arginine (NAR) in an amount of about 50 mM to about 150 mM, or about 50 mM to about 250 mM, if necessary. Lysine, methionine, glycine, proline, sodium acetate, tris acetate, histamine, or calcium salt. In many aspects, the composition of the present disclosure includes (i) arginine and (ii) NAR and/or methionine. In various aspects, the arginine salt is arginine glutamate. Under different circumstances, the composition of the present disclosure includes (i) calcium salt and (ii) NAR and/or methionine. In an exemplary aspect, the calcium salt is calcium glutamate. In an exemplary aspect, the composition of the present disclosure uses about 160 mg/mL to about 250 mg/mL, as needed, about 160 mg/mL to about 225 mg/mL, for example, about 170 mg/mL to about 200 mg/mL, As needed, about 175 mg/mL to about 185 mg/mL, for example, a concentration of 180 mg/mL contains anti-TSLP antibodies. In an illustrative example, the composition of the present disclosure has a pH of about 4.5 to about 6.75, and optionally about 4.8 to about 6.0. In an exemplary aspect, the present disclosure of the composition at 23 ° C, viscosity 1000 s ^-1 is less than 100 cP, as required, at 23 ° C, 1000 s ^-1 is less than 75 cP, e.g., less than 60 cP, or less than 50 cP.

In an exemplary embodiment, the composition of the present disclosure includes an amphiphilic and/or nonionic surfactant. In various aspects, the surfactant is a polysorbate, such as polysorbate 20 or polysorbate 80 or mixtures thereof. If necessary, the surfactant is present in a concentration of less than or about 0.005% (w/v) to about 0.015% (w/v), and if necessary, about 0.010% (w/v) ± 0.0025% (w/v) Surfactant, for example, about 0.005% (w/v), 0.010% (w/v), or 0.015% (w/v) surfactant.

In various embodiments, the aqueous composition contains 25-190 mM arginine base and 25-200 mM glutamine. In various embodiments, the aqueous composition contains 140 mM arginine base and 150 mM glutamine. In various embodiments, the aqueous composition containing arginine and glutamate contains from 0 to 250 mM proline. In various embodiments, the aqueous composition comprises 80 mM arginine base, 85 mM glutamine and 100 mM L-proline. In various embodiments, the aqueous composition containing arginine and glutamate and optionally proline contains 0.01% (w/v) polysorbate 80. In various embodiments, the aqueous composition comprises 140 mM arginine base, 150 mM glutamine acid, and 0.01% (w/v) polysorbate 80. In various embodiments, the aqueous composition includes 80 mM arginine base, 85 mM glutamic acid, 100 mM L-proline, 0.01% (w/v) polysorbate 80.

In various embodiments, the aqueous composition contains 10-125 mM arginine base and 25-225 mM glutamine. In various embodiments, the aqueous composition contains 95 mM arginine base and 170 mM glutamine. In various embodiments, the aqueous composition containing arginine and glutamate contains from 0 to 220 mM proline. In various embodiments, the aqueous composition comprises 50 mM arginine base, 95 mM glutamine and 85 mM L-proline. In various embodiments, the aqueous composition containing arginine and glutamate contains 0.01% (w/v) polysorbate 80. In various embodiments, the aqueous composition comprises 95 mM arginine base, 170 mM glutamine acid, and 0.01% (w/v) polysorbate 80. In various embodiments, the aqueous composition comprises 50 mM arginine base, 95 mM glutamine, 85 mM L-proline, 0.01% (w/v) polysorbate 80.

In various embodiments, the aqueous composition contains 15-130 mM calcium and 30-300 mM glutamine. In various embodiments, the aqueous composition contains 100 mM calcium and 230 mM glutamine. In various embodiments, the aqueous composition containing calcium and glutamate contains from 0 to 250 mM proline. In various embodiments, the aqueous composition contains 60 mM calcium, 140 mM glutamine, and 70 mM L-proline. In various embodiments, the aqueous composition contains 15-195 mM calcium and 25-320 mM glutamine. In various embodiments, the aqueous composition contains 110 mM calcium and 240 mM glutamine. In various embodiments, the aqueous composition contains from 0 to 220 mM proline. In various embodiments, the aqueous composition contains 70 mM calcium, 145 mM glutamate, and 60 mM L-proline. In various embodiments, the aqueous composition containing calcium and glutamate contains 0.01% (w/v) polysorbate 80. In various embodiments, the aqueous composition contains 100 mM calcium, 230 mM glutamate, and 0.01% (w/v) polysorbate 80. In various embodiments, the aqueous composition contains 60 mM calcium, 140 mM glutamate, 70 mM L-proline, 0.01% (w/v) polysorbate 80. In various embodiments, the aqueous composition contains 110 mM calcium, 240 mM glutamate, and 0.01% (w/v) polysorbate 80. In various embodiments, the aqueous composition contains 70 mM calcium, 145 mM glutamate, 60 mM L-proline, 0.01% (w/v) polysorbate 80.

In various embodiments, the aqueous composition described herein has a pH of about 4.5 to about 6.75. In various embodiments, the aqueous composition has a pH of about 4.7 to about 6.0. In various embodiments, the aqueous composition has a pH of about 5.1 to about 5.7. In various embodiments, the aqueous composition has a pH of about 4.7 to about 5.3. In various embodiments, the aqueous composition described herein has a pH of about 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, or 5.7.

In different cases, the composition is isotonic or has a range of about 200 mOsm/kg to about 500 mOsm/kg, or about 225 mOsm/kg to about 400 mOsm/kg, or about 250 mOsm/kg to about 350 mOsm Osmotic pressure in the range of /kg. Optionally, the composition is isotonic or has an osmotic pressure greater than about 350 mOsm/kg.

In illustrative examples, the composition is suitable for short-term storage at 25°C, 30°C, or 40°C or long-term storage at about -30°C or about 2°C to about 8°C. For example, as determined by size exclusion chromatography (SEC), after 6 months of storage at 2°C to 8°C, less than 0.5% of the therapeutic protein is degraded, if necessary, where the therapeutic protein Contained in a glass vial or syringe. In different cases, as determined by size exclusion chromatography (SEC), less than about 5% of the antibody is degraded after storage at about 2°C to about 8°C for at least or about 12 months. In many aspects, as determined by size exclusion chromatography (SEC), after storage at about 2°C to about 8°C for about 20 months to about 26 months, less than about 5% of the antibody is degradation. In an exemplary example, as determined by size exclusion chromatography (SEC), less than about 5% of the antibody is degraded after storage at about 2°C to about 8°C for about 30 to about 40 months . In an exemplary example, as determined by size exclusion chromatography (SEC), less than about 5% of the antibody is degraded after storage at about 2°C to about 8°C for about 2 to about 3 years . For another example, as determined by size exclusion chromatography (SEC), less than 5% of the antibody is degraded after storage at 2°C to 8°C for about 24 months to about 36 months, if necessary, Among them, less than 2% of the antibodies are degraded after storage at 2°C to 8°C for 24 months or 36 months. In many respects, as determined by the SEC, after storage at about room temperature (eg 25°C) for at least 2 weeks (as needed, at least 1 month, at least 2 months, at least 3 months, After at least 4 months, at least 5 months, or at least 6 months), less than 5% of the antibody is degraded. In different cases, as determined by the SEC, storage at 2°C to 8°C for about 24 months to about 36 months, and then storage at about room temperature (for example, 25°C) for at least 2 weeks or at least After about 1 month or at least about 2 months, less than 5% of the antibodies are degraded. Optionally, as determined by size exclusion chromatography (SEC), less than about 5% of the antibody is degraded after storage at a temperature greater than about 20°C for at least or about 2 weeks, and storage for at least or about 4 weeks or about 8 weeks. In various aspects, the temperature is greater than or about 25°C or greater than or about 30°C or greater than or about 40°C.

This article further provides an article. In an exemplary embodiment, the product contains the composition of the present disclosure, and optionally contains about 1 mL to about 5 mL (for example, about 1 mL to about 3 mL) of the aqueous composition.

This document additionally provides a pre-filled syringe that contains the composition of the present disclosure, and optionally contains about 1 mL to about 5 mL (for example, about 1 mL to about 3 mL) of the composition.

There is further provided a vial, which contains the composition of the present disclosure, and optionally contains about 1 mL to about 5 mL (for example, about 1 mL to about 3 mL) of the aqueous composition.

An autoinjector containing the aqueous composition described herein is also provided. In various embodiments, the auto-injector is Ypsomed YpsoMate®. In various embodiments, the autoinjector is disclosed in WO 2018/226565, WO 2019/094138, WO 2019/178151, WO 20120/072577, WO 2020/081479, WO 2020/081480, PCT/US 20/70590, PCT/ US 20/70591, PCT/US 20/53180, PCT/US 20/53179, PCT/US 20/53178, or PCT/US 20/53176.

This article provides the use of the disclosed composition in the treatment of inflammatory diseases. In an exemplary aspect, the inflammatory disease is selected from the group consisting of asthma, atopic dermatitis, chronic obstructive pulmonary disease (COPD), eosinophilic esophagitis (EoE), nasal polyps, chronic spontaneous urticaria Measles, Ig-driven diseases (such as IgA nephropathy & lupus nephritis), eosinophilic gastritis, chronic sinusitis without nasal polyps, and idiopathic pulmonary fibrosis (IPF). If necessary, the inflammatory disease is atopic dermatitis. If necessary, the inflammatory disease is COPD.

The present disclosure provides methods for treating inflammatory diseases in subjects. In an exemplary embodiment, the method includes administering to the subject a therapeutically effective amount of a composition of the present disclosure. In many respects, the inflammatory disease is selected from the group consisting of asthma, atopic dermatitis, chronic obstructive pulmonary disease (COPD), eosinophilic esophagitis (EoE), nasal swelling, chronic spontaneous urticaria Measles, Ig-driven diseases (such as IgA nephropathy & lupus nephritis), eosinophilic gastritis, chronic sinusitis without nasal polyps, and idiopathic pulmonary fibrosis (IPF). If necessary, the inflammatory disease is atopic dermatitis. If necessary, the inflammatory disease is COPD. Under different circumstances, the composition of the present disclosure is administered to the subject by subcutaneous administration. In an illustrative example, about 1 mL to about 5 mL (eg, about 1 mL to about 3 mL) of the aqueous composition is administered to the subject.

A method for preparing a stable liquid antibody composition is further provided, the composition having a viscosity of less than about 100 cP and containing (A) an anti-TSLP antibody with a concentration greater than about 140 mg/mL, (B) a surfactant, and (C) a base Amino acid or its salt, calcium salt, magnesium salt, or a combination thereof. In an exemplary embodiment, the method includes: (i) combining the antibody with an aqueous solution containing about 50 mM to about 150 mM basic amino acid or a salt thereof, a calcium salt, a magnesium salt, or The combination thereof, and (ii) the addition of a surfactant to achieve a final concentration of about 0.01% (w/v) ± 0.005% (w/v) of the surfactant.

Based on the following detailed description of the summary, combined with the accompanying drawings, other aspects and advantages will be obvious to those familiar with the technology. Although these compositions, products, and methods are easily applied to various forms of implementation, the following description includes specific Implementation mode. Regarding the compositions, products, and methods described herein, expected optional features, including but not limited to components, their composition ranges, substituents, conditions, and steps, are selected from the various aspects, embodiments, and examples provided herein.

definition

The above description is provided only for a clear understanding, and should not be construed as unnecessary limitations, because the modifications within the scope of the present invention are obvious to those skilled in the art.

Throughout this specification and the scope of the following patent applications, unless the context requires otherwise, the word "comprise" and variations such as "comprises and comprising" should be understood as implicitly including the stated integers or steps or integers or The group of steps, rather than excluding any other integers or steps or groups of integers or steps.

Throughout this specification, when compositions are described as including components or materials, unless otherwise described, it is contemplated that such compositions may also consist essentially of or consist of any combination of the recited components or materials. Likewise, when methods are described as including specific steps, unless otherwise described, it is contemplated that such methods may also consist essentially of or consist of any combination of the recited steps. The present invention illustratively disclosed herein can be suitably practiced without any elements or steps not explicitly disclosed herein.

The method disclosed herein and the practice of its individual steps can be carried out manually and/or under the assistance of or automation provided by electronic equipment. Although many methods have been described with reference to specific embodiments, those skilled in the art should easily understand that other ways of performing actions related to these methods can be used. For example, unless otherwise described, the order of the steps may be changed without departing from the scope or spirit of the method. In addition, some individual steps can be combined, omitted or further subdivided into other steps.

Unless otherwise stated, the compositions and methods are expected to include embodiments containing any combination of one or more of other optional elements, features, and steps (including those shown in the drawings) described further below.

Regarding the jurisdiction that prohibits the granting of patents to human subjects, the meaning of “administering” the composition to human subjects should be limited to stipulating that human subjects will use any technique (such as oral, inhalation, local administration, Injection, insertion, etc.) self-administered controlled substances. The broadest reasonable interpretation is intended, which is consistent with the laws or regulations that define the subject matter that can be patented. In terms of the jurisdiction that does not prohibit patenting of methods performed on the human body, the "investment" composition includes both the methods performed on the human body and the above-mentioned activities.

It should be understood that each maximum numerical limit provided throughout this specification includes the range formed with each corresponding lower numerical limit as an alternative aspect, as if such a range was clearly written. Each minimum numerical limit provided throughout this specification will include the range formed with each higher numerical limit as an alternative, as if such a range was expressly written. Every numerical range provided throughout this specification will include every narrower numerical range that falls within such a wider numerical range, as if all such narrower numerical ranges are expressly written herein. The size and value disclosed herein should be understood to include the value stated in the disclosure and the corresponding accurate value. For example, a value described as "about 10 mM" should be understood to include "10 mM" as an alternative disclosure.

All patents, publications and references cited in this article are incorporated into this article in their entirety by reference. In the case of conflicts between this disclosure and the incorporated patents, publications and references, this disclosure shall prevail.

Unless otherwise stated, the following terms used in this application (including the specification and the scope of the patent application) have the definitions given below.

Unless the context clearly dictates otherwise, as used in the specification and the scope of the appended application, the indefinite article "a/an" and the definite article "the" include plural and singular referents.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which this disclosure belongs. The following references provide technicians with general definitions of many terms used in this disclosure, including but not limited to: Singleton et al., DICTIONARY OF MICROBIOLOGY AND MOLECULAR BIOLOGY [Dictionary of Microbiology and Molecular Biology] (2nd Edition 1994); THE CAMBRIDGE DICTIONARY OF SCIENCE AND TECHNOLOGY [Cambridge Science and Technology Dictionary] (Walker editor, 1988); THE GLOSSARY OF GENETICS [Genetics glossary], 5th edition, R. Rieger et al. (Editor), Springer Verlag [Springer Publishing Company] (1991); and Hale and Marham, THE HARPER COLLINS DICTIONARY OF BIOLOGY [Harper Collins Dictionary of Biology] (1991).

The term "about" or "approximately" means the acceptable error of a specific value as determined by those skilled in the art, which depends in part on how the value is measured or determined. In certain embodiments, the term "about" or "approximately" means within 1, 2, 3, or 4 standard deviations. In certain embodiments, the term "about" or "approximately" means 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, Within 5%, 4%, 3%, 2%, 1%, 0.5% or 0.05%. Whenever the term "about" or "approximately" precedes the first value in a series of two or more values, it should be understood that the term "about" or "approximately" applies to each value in the series.

The term “specifically binds” means “antigen-specific”, “specific to antigen”, “selective binding agent”, “specific binding agent”, “antigen target” or “immune reaction” with antigen. The antibody or polypeptide binds to the target antigen with greater affinity than other antigens of the related protein. It is expected herein that the agent specifically binds to target proteins, such as surface antigens (such as T cell receptors, CD3), cytokines (such as TSLP, IL-4, IL-5, IL-13, IL-17, IFN-g) , TNF-a) and its analogues.

The term "antibody" or "immunoglobulin" refers to a canonical tetrameric carbohydrate composed of two substantially full-length heavy chains and two substantially full-length light chains each comprising a variable region and a substantially full-length constant region protein. The antigen-binding portion can be produced by recombinant DNA technology or by enzymatic or chemical cleavage of intact antibodies. The term "antibody" includes monoclonal antibodies, multiple antibodies, chimeric antibodies, human antibodies, and humanized antibodies.

Antibody variants include antibody fragments and antibody-like proteins that become the structure of a standard tetrameric antibody. Generally, antibody variants include V regions that become constant regions, or alternatively, V regions are added to the constant regions in a non-standard manner as needed. Examples include multispecific antibodies (such as bispecific antibodies with additional V regions), antibody fragments that can bind antigen (such as Fab', F'(ab)2, Fv, single-chain antibodies, diabodies), including the foregoing The biparatopic peptides and recombinant peptides of the item (as long as they exhibit the desired biological activity).

Antibody fragments include the antigen-binding portion of antibodies, especially Fab, Fab', F(ab')2, Fv, domain antibodies (dAb), complementarity determining region (CDR) fragments, CDR grafted antibodies, single chain antibodies (scFv), Single-chain antibody fragments, chimeric antibodies, diabodies, triabodies, tetrabodies, minibodies, linear antibodies; chelating recombinant antibodies, triabodies or diabodies, intracellular antibodies, nanoantibodies, and small module immunity Drugs (SMIP), antigen-binding domain immunoglobulin fusion proteins, single domain antibodies (including camelized antibodies), VHH-containing antibodies or variants or derivatives thereof, and immunoglobulins containing enough to confer specific antigen binding to the polypeptide At least a part of the polypeptide (such as one, two, three, four, five or six CDR sequences), as long as the antibody retains the desired biological activity.

"Valency" refers to the number of antigen binding sites on each antibody or antibody fragment targeted to an epitope. A typical full-length IgG molecule or F(ab)2 is "bivalent" because it has two identical target binding sites. "Monovalent" antibody fragments, such as F(ab)' or scFc, have a single antigen binding site. Trivalent or tetravalent antigen binding proteins can also be engineered to be multivalent.

The term "monoclonal antibody" refers to an antibody obtained from a substantially homogeneous antibody population (that is, the individual antibodies that constitute the population are identical except for possible naturally occurring mutations that may exist in small amounts).

The term "inhibition of TSLP activity" includes inhibiting any one or more of the following:- -TSLP binds to its receptor; -Proliferation, activation or differentiation of cells expressing TSLPR in the presence of TSLP; -Inhibition of Th2 cytokine production in the presence of TSLP in the polarization assay; -Dendritic cells activate or mature in the presence of TSLP; -The release of cytokines from obese cells in the presence of TSLP. See, for example, column 6 and Example 8 of US Patent 7982016 B2, and Examples 7-10 of US 2012/0020988 A1.

The term "sample" or "biological sample" refers to a sample obtained from a subject for use in the method of the present invention, and includes urine, whole blood, plasma, serum, saliva, sputum, tissue sections, cerebrospinal fluid, Peripheral blood mononuclear cells with in vitro stimulation, peripheral blood mononuclear cells without in vitro stimulation, intestinal lymphoid tissue with in vitro stimulation, intestinal lymphoid tissue without in vitro stimulation, intestinal lavage fluid, bronchoalveolar lavage fluid , Nasal lavage fluid and induced sputum.

The term "treat (treat, treating and treatment)" refers to the temporary or permanent, partial or complete elimination, reduction, suppression or improvement of the clinical symptoms, manifestations, or progression of events, diseases, or conditions related to the inflammatory conditions described herein . As recognized in the related art, drugs used as therapeutic agents can reduce the severity of a given disease state, but it is not necessary to eliminate every manifestation of the disease to be considered a useful therapeutic agent. Similarly, a prophylactically administered treatment does not have to be fully effective in preventing the onset of the disorder in order to be a viable preventive agent. Only reduce the impact of the disease (for example, by reducing the number of symptoms or reducing the severity of symptoms, or by increasing the effectiveness of another treatment, or by producing another beneficial effect) or by reducing the disease in the subject The possibility of occurrence or deterioration is sufficient. One embodiment of the present disclosure relates to a method for determining the efficacy of a treatment, the method comprising administering to a patient a therapeutic agent in an amount and time sufficient to cause a sustained improvement beyond the indicator that reflects the severity of a particular condition Baseline.

The term "therapeutically effective amount" refers to the amount of the therapeutic agent that is effective to improve or alleviate the symptoms or signs associated with the disease or disorder.

The term "cytokines" as used herein refers to one or more small (5-20 kD) proteins released by cells, which are important for the interaction and communication between cells or for cell behaviors such as immune cell proliferation and differentiation. Specific role. The functions of cytokines in the immune system include promoting circulating white blood cells and lymphocytes to flow into immune collision sites; stimulating the development and proliferation of B cells, T cells, peripheral blood mononuclear cells (PBMC) and other immune cells; and providing antimicrobial active. Exemplary immune cytokines include, but are not limited to, IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-9, IL-10, IL-12, IL-13, IL-15, IL17A, IL-17F, IL-18, IL-21, IL-22, interferon (including IFNα, β and γ), tumor necrosis factor (including TNFα, β), transforming growth factor (Including TGFα, β), granulocyte colony stimulating factor (GCSF), granulocyte macrophage colony stimulating factor (GMCSF) and thymic stromal lymphopoietin (TSLP).

"Helper T cell (Th) 1 interleukin" or "Th1 specific interleukin" refers to the interleukin expressed (intracellular and/or secreted) by Th1 T cells, and includes IFN-g, TNF- a. IL-12. "Th2 cytokines" or "Th2 specific cytokines" refer to cytokines expressed (intracellular and/or secreted) by Th2 T cells, including IL-4, IL-5, IL-13 and IL- 10. "Th17 cytokines" or "Th17-specific cytokines" refer to cytokines expressed (intracellular and/or secreted) by Th17 T cells, including IL-17A, IL-17F, IL-22 and IL- twenty one. In addition to the Th17 cytokines listed herein, certain populations of Th17 cells also express IFN-g and/or IL-2. Multifunctional CTL cytokines include IFN-g, TNF-a, IL-2 and IL-17.

Low viscosity resistance TSLP Antibody composition

Teperumumab has shown efficacy in the strength range of 70 mg to 280 mg, and in some cases, the anti-TSLP antibody will be formulated at a dose of 110 mg/mL or 140 mg/mL. A formulation with a high protein concentration may exhibit increased viscosity, so that it may negatively affect the function of the device used to administer the antibody to the patient. Similarly, the ability of healthcare providers to manually inject drugs into patients may be affected. In addition, high viscosity may be prohibited during the manufacturing process. From the standpoint of protein stability, formulations with high protein concentrations are also challenging. For example, aggregation leading to the formation of high molecular weight species (HMWS) can occur in formulations containing high protein concentrations. Therefore, there is a need to provide low-viscosity, isotonic, liquid formulations of anti-TSLP antibodies (such as tepeluzumab) that are suitable for parenteral administration and can be used at low temperatures (e.g., 2°C-8°C and -30°C) long-term storage, or short-term storage at room temperature (for example, 20°C-25°C, for the convenience of the patient).

In order to overcome the high viscosity problem of formulations with high protein concentrations, the present disclosure provides a composition, such as an aqueous composition, which contains (a) an anti-TSLP antibody at a concentration greater than about 140 mg/mL, and (b) a surfactant , And (c) at least one basic amino acid or salt thereof. The present disclosure also provides a composition, such as an aqueous composition, which comprises (a) an anti-TSLP antibody at a concentration greater than about 140 mg/mL, (b) a surfactant, and (c) at least one calcium salt or magnesium salt. Based at least in part on the data provided herein, and not bound by any particular theory, the composition of the present disclosure represents a low-viscosity composition containing a high concentration of therapeutic protein, which can be administered to patients in need without being bound by high Viscosity causes any complications. Based at least in part on the data provided herein, and not bound by any particular theory, the composition of the present disclosure is highly stable because the composition of the present disclosure is at low temperatures (for example, 2°C-8°C and -30°C). ) Storage (short-term and/or long-term storage) or short-term storage at room temperature (e.g. about 20°C-25°C) shows minimal degradation.

Basic amino acid

In an exemplary embodiment, the composition of the present disclosure includes a basic amino acid or a salt thereof. As used herein, the term "basic amino acid" refers to an amino acid with a basic side chain at neutral pH. The pKa of basic amino acids is high enough that they tend to bind protons, thereby gaining a positive charge in the process. In an exemplary aspect, the basic amino acid includes a side chain that includes nitrogen that binds (and protonates) or releases a bond to protons (and deprotonates). In an exemplary aspect, the basic amino acid may be between the NH2 (deprotonated) and NH3+ (protonated) form or between the NH (deprotonated) and NH2+ (protonated) form or between the N (deprotonated) form or the N (deprotonated) form. The balance between protonation) and NH+ (protonation) forms. For basic amino acids at physiological pH, such as about pH 7.0, the protonated form predominates. In an exemplary aspect, the basic amino acid is arginine (Arg; R), lysine (Lys, K) or histidine (His, H). Although the basic amino acid may be a D-isomer or an L-isomer, in an illustrative example, the basic amino acid is an L-isomer of an amino acid, such as L-Arg, L-Lys, L-His. In an illustrative example, the basic amino acid is arginine. In an illustrative example, the basic amino acid is histidine. In different cases, the basic amino acid is lysine.

式IIn an exemplary aspect, the basic amino acid is a derivative of arginine, such as L-2-amino-3-guanidinopropionic acid, 4-guanidinobutyric acid. In an exemplary aspect, the basic amino acid comprises a structure having formula I:

Formula I

Wherein n is 1 to 16, or 1 to 10, or 1 to 7, or 1 to 6, or 2 to 6, or 2 or 3 or 4 or 5.

式IIIn an exemplary aspect, the basic amino acid is a derivative of lysine, such as 5-hydroxylysine, ornithine, N-acetyl-L-lysine, 2,4-diamino Butyric acid.

Formula II

Wherein n is 1 to 16, or 1 to 10, or 1 to 7, or 1 to 6, or 2 to 6, or 2 or 3 or 4 or 5. R ₁ and R ₂ are each independently selected from the group consisting of Group: H, C ₁ -C ₁₈ alkyl, (C ₁ -C ₁₈ alkyl) OH, (C ₁ -C ₁₈ alkyl) NH ₂ , (C ₁ -C ₁₈ alkyl) SH, (C _0- C ₄ alkyl) (C ₃ -C ₆ ) cycloalkyl, (C ₀ -C ₄ alkyl) (C ₂ -C ₅ heterocycle), (C ₀ -C ₄ alkyl) (C ₆ -C ₁₀ Aryl) R ₇ , and (C ₁ -C ₄ alkyl) (C ₃ -C ₉ heteroaryl), wherein R ₇ is H or OH.

In an exemplary aspect, the basic amino acid is a derivative of histidine, such as deaminated histidine, hydroxy-histidine, acetyl-histidine, homo-histidine , N-methyl histidine, α-methyl histidine, imidazole acetic acid, or α, α-dimethylimidazole acetic acid (DMIA).

In many aspects, the composition of the present disclosure includes a salt of a basic amino acid. In an illustrative example, the salt is a pharmaceutically acceptable salt. The term "pharmaceutically acceptable salt" as used herein refers to a salt of a compound that retains the biological activity of the parent compound and which is not biologically or otherwise undesirable. Such salts can be prepared on the spot during the final isolation and purification of the analog, or separately prepared by reacting the free base functional group with a suitable acid. Due to the presence of amine and/or carboxyl groups or similar groups, many of the compounds disclosed herein can form acid and/or basic salts.

Pharmaceutically acceptable acid addition salts can be prepared from inorganic acids and organic acids. Representative acid addition salts include (but are not limited to) acetate, adipate, alginate, citrate, aspartate, benzoate, besylate, bisulfate, butyric acid Salt, camphorate, camphorsulfonate, digluconate, glycerophosphate, hemisulfate, heptanoate, caproate, fumarate, hydrochloride, hydrobromide, hydroiodide, 2-Hydroxyethanesulfonate (isothiosulfate), lactate, maleate, methanesulfonate, nicotinate, 2-naphthalenesulfonate, oxalate, palmitate, fruit Glueate, persulfate, 3-phenylpropionate, picrate, trimethylacetate, propionate, succinate, tartrate, thiocyanate, phosphate, glutamate, Bicarbonate, p-toluenesulfonate and undecanoate. Salts derived from inorganic acids include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like. Salts derived from organic acids include acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, malic acid, malonic acid, succinic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid , Methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, etc. Examples of acids that can be used to form pharmaceutically acceptable acid addition salts include, for example, inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, and phosphoric acid; and organic acids such as oxalic acid, maleic acid, succinic acid, and citric acid.

The base addition salt can also be prepared on-site during the final separation and purification of the source of salicylic acid, or by combining the carboxylic acid-containing moiety with a suitable base, such as the hydroxide, carbonate, or carbonic acid of a pharmaceutically acceptable metal cation. Hydrogen salt, or prepared by reaction with amine or organic primary amine, secondary amine and tertiary amine. Pharmaceutically acceptable salts especially include (but are not limited to) alkali metal or alkaline earth metal-based cations, such as lithium, sodium, potassium, calcium, magnesium, and aluminum salts; and non-toxic quaternary amines and amine cations , Including ammonium, tetramethylammonium, tetraethylammonium, methylammonium, dimethylammonium, trimethylammonium, triethylammonium, diethylammonium and ethylammonium. Other representative organic amines that can be used to form base addition salts include, for example, ethylenediamine, ethanolamine, diethanolamine, piperidine, piperidine, and the like. Salts derived from organic bases include, but are not limited to, salts of primary amines, secondary amines, and tertiary amines.

In addition, basic nitrogen-containing groups can be quaternized with analogs of the present disclosure into lower alkyl halides, such as methyl, ethyl, propyl and butyl chlorides, bromides and iodides; long-chain halogenation Substances such as decyl, lauryl, myristyl, and stearyl chloride, bromide and iodide; aralkyl halides such as benzyl and phenethyl bromide. Thereby, water-soluble or oil-soluble or water-dispersible or oil-dispersible products are obtained.

In an exemplary aspect, the basic amino acid is arginine and the composition of the present disclosure includes arginine. In various aspects, the salt of arginine is an organic salt of arginine. In many aspects, the arginine is arginine acetate, arginine aspartate, arginine glutamate, arginine glycolate, arginine lactate, arginine Methanesulfonate, arginine propionate, or a combination thereof.

In an exemplary aspect, the basic amino acid is histidine and the composition of the present disclosure includes histidine. In various aspects, the histamine acid salt is an organic salt of histidine acid. In an exemplary aspect, the histidine salt is histidine acetate, histidine aspartate, histidine glutamine, histidine glycolate, histidine lactate, histidine Methanesulfonate, histidine propionate, or a combination thereof.

In an exemplary aspect, the basic amino acid is a lysine acid and the composition of the present disclosure includes a lysine salt. In various aspects, the lysine acid salt is an organic salt of lysine acid. In many aspects, the lysine salt is lysine acetate, lysine aspartate, lysine glutamate, lysine glycolate, lysine lactate, lysine Methanesulfonate, lysine propionate, or a combination thereof.

Under different circumstances, the composition of the present disclosure contains about 10 mM to about 300 mM, or about 50 mM to about 300 mM basic amino acid or its salt. In exemplary aspects, the composition of the present disclosure includes about 10 mM to about 200 mM, about 50 mM to about 250 mM, about 50 mM to about 200 mM, about 50 mM to about 150 mM, about 50 mM to about 100 mM , About 50 mM to about 90 mM, about 50 mM to about 80 mM, about 50 mM to about 70 mM, about 50 mM to about 60 mM, about 50 mM to about 55 mM, about 55 mM to about 200 mM, about 60 mM to about 200 mM, about 70 mM to about 200 mM, about 80 mM to about 200 mM, about 90 mM to about 200 mM, about 100 mM to about 200 mM, about 150 mM to about 200 mM, about 160 mM To about 200 mM, about 170 mM to about 200 mM, about 180 mM to about 200 mM, or about 190 mM to about 200 mM basic amino acid or salt thereof. Under different circumstances, the composition of the present disclosure contains about 50 mM to about 100 mM (for example, about 50 mM, about 60 mM, about 70 mM, about 80 mM, about 90 mM, about 100 mM) or about 100 mM to about 100 mM. 200 mM (for example, about 100 mM, about 110 mM, about 120 mM, about 130 mM, about 140 mM, about 150 mM, about 160 mM, about 170 mM, about 180 mM, about 190 mM, about 200 mM) alkaline Amino acid or its salt. Under different circumstances, the composition of the present disclosure contains about 50 mM to about 100 mM or about 50 mM to about 75 mM or about 75 mM to about 100 mM basic amino acid or its salt. Under different circumstances, the composition of the present disclosure contains about 100 mM to about 200 mM or about 100 mM to about 150 mM or about 150 mM to about 200 mM basic amino acid or its salt. In an exemplary aspect, the composition of the present disclosure includes about 10 mM to about 200 mM basic amino acid or a salt thereof.

In different cases, anti-TSLP antibodies (eg, teperumumab) are formulated with about 25 mM to about 190 mM arginine and about 25 mM to about 200 mM glutamine. In various aspects, an anti-TSLP antibody (e.g., Teperumumab) is combined with 100 mM to about 180 mM arginine (e.g., about 100 mM to about 170 mM, about 100 mM to about 160 mM, about 100 mM to about 150 mM, about 100 mM to about 140 mM, about 100 mM to about 130 mM, about 100 mM to about 120 mM, about 100 mM to about 110 mM, about 110 mM to about 180 mM, about 120 mM to about 180 mM , About 130 mM to about 180 mM, about 140 mM to about 180 mM, about 150 mM to about 180 mM, about 160 mM to about 180 mM, about 170 mM to about 180 mM, about 120 mM to about 170 mM, about 130 mM to about 160 mM, about 135 mM to about 155 mM) and about 110 mM to about 240 mM glutamate (for example, about 110 mM to about 180 mM, about 110 mM to about 170 mM, about 110 mM to about 160 mM, about 110 mM to about 150 mM, about 110 mM to about 140 mM, about 110 mM to about 130 mM, about 110 mM to about 120 mM, about 120 mM to about 180 mM, about 130 mM to about 180 mM , About 140 mM to about 180 mM, about 150 mM to about 180 mM, about 160 mM to about 180 mM, about 170 mM to about 180 mM, about 120 mM to about 170 mM, about 130 mM to about 160 mM, about 140 mM to about 160 mM, about 145 mM to about 155 mM glutamate). In various cases, anti-TSLP antibodies (for example, teperumumab) are formulated with about 135 mM to about 145 mM arginine and about 145 mM to about 155 mM glutamine. Under different circumstances, an anti-TSLP antibody (for example, teparumab) is formulated in about 10 mM to about 125 mM arginine and about 25 mM to about 225 mM glutamate. In various aspects, in about 55 mM to about 135 mM arginine (e.g., about 55 mM to about 125 mM, about 55 mM to about 115 mM, about 55 mM to about 105 mM, about 55 mM to about 95 mM, About 55 mM to about 85 mM, about 55 mM to about 75 mM, about 55 mM to about 65 mM, about 65 mM to about 135 mM, about 75 mM to about 135 mM, about 85 mM to about 135 mM, about 95 mM to about 135 mM, about 105 mM to about 135 mM, about 115 mM to about 145 mM, about 125 mM to about 135 mM, about 75 mM to about 115 mM, about 85 mM to about 105 mM arginine) and About 130 mM to about 210 mM glutamate (for example, about 130 mM to about 200 mM, about 130 mM to about 240 mM, about 130 mM to about 180 mM, about 130 mM to about 170 mM, about 130 mM to about 160 mM, about 130 mM to about 150 mM, about 130 mM to about 140 mM, about 140 mM to about 210 mM, about 150 mM to about 210 mM, about 160 mM to about 210 mM, about 170 mM to about 210 mM , About 180 mM to about 210 mM, about 190 mM to about 210 mM, about 200 mM to about 210 mM, about 150 mM to about 190 mM, about 160 mM to about 180 mM, about 165 mM to about 175 mM glutamine Acid salt), prepare an anti-TSLP antibody (for example, tepelumab). In an exemplary embodiment, the composition of the present disclosure contains more than 140 mg/mL teperumumab, about 85.5 mM to about 104.5 mM arginine, 153 mM to about 187 mM glutamine, and 0.01% ( w/v) Polysorbate 80. In an exemplary embodiment, the composition of the present disclosure comprises more than 140 mg/mL teperumumab, about 95 mM arginine, 170 mM glutamine, and 0.01% (w/v) polysorbate 80. If necessary, the pH is about 5.4 ± 0.2, or about 5.4 ± 0.1.

Calcium and magnesium salts

In an exemplary embodiment, the composition of the present disclosure includes calcium salt or magnesium salt. In various aspects, the calcium salt or the magnesium salt comprises any relative ions. In an exemplary aspect, the calcium salt or magnesium salt comprises a relative ion lacking chloride. In an illustrative example, the relative ion is acetate, aspartate, glutamate, glycolate, lactate, methanesulfonate, propionate, or a combination thereof. In various aspects, the calcium salt is calcium acetate, calcium aspartate, calcium glutamate, calcium glycolate, calcium lactate, calcium methanesulfonate, calcium propionate, or a combination thereof. In an illustrative example, the magnesium salt is magnesium acetate, magnesium aspartate, magnesium glutamine, magnesium glycolate, magnesium lactate, magnesium methanesulfonate, magnesium propionate, or a combination thereof.

Under different circumstances, the composition of the present disclosure contains about 15 mM to about 300 mM, about 15 mM to about 200 mM, or about 50 mM to about 150 mM calcium salt or magnesium salt. Under different circumstances, the composition of the present disclosure contains about 15 mM to about 300 mM, or about 50 mM to about 300 mM calcium salt or magnesium salt. In exemplary aspects, the composition of the present disclosure includes about 15 mM to about 200 mM, about 15 to about 150 mM, about 50 mM to about 250 mM, about 50 mM to about 200 mM, about 50 mM to about 150 mM, About 50 mM to about 100 mM, about 50 mM to about 90 mM, about 50 mM to about 80 mM, about 50 mM to about 70 mM, about 50 mM to about 60 mM, about 50 mM to about 55 mM, about 55 mM to about 200 mM, about 60 mM to about 200 mM, about 70 mM to about 200 mM, about 80 mM to about 200 mM, about 90 mM to about 200 mM, about 100 mM to about 200 mM, about 150 mM to About 200 mM, about 160 mM to about 200 mM, about 170 mM to about 200 mM, about 180 mM to about 200 mM, or about 190 mM to about 200 mM calcium salt or magnesium salt. Under different circumstances, the composition of the present disclosure contains about 50 mM to about 100 mM (for example, about 50 mM, about 60 mM, about 70 mM, about 80 mM, about 90 mM, about 100 mM) or about 100 mM to about 100 mM. 200 mM (for example, about 100 mM, about 110 mM, about 120 mM, about 130 mM, about 140 mM, about 150 mM, about 160 mM, about 170 mM, about 180 mM, about 190 mM, about 200 mM) calcium salt Or magnesium salt. Under different circumstances, the composition of the present disclosure contains about 15 mM to about 130 mM, about 50 mM to about 100 mM, or about 50 mM to about 75 mM or about 75 mM to about 100 mM calcium salt or magnesium salt. Under different circumstances, the composition of the present disclosure contains about 100 mM to about 200 mM or about 100 mM to about 150 mM or about 150 mM to about 200 mM calcium salt or magnesium salt.

In various cases, anti-TSLP antibodies (eg, teperumumab) are formulated with about 15 mM to about 130 mM calcium and about 30 mM to about 300 mM glutamine. In various aspects, an anti-TSLP antibody (e.g., teperumumab) is combined with about 70 mM to about 130 mM calcium (e.g., about 80 mM to about 130 mM, about 90 mM to about 130 mM, about 100 mM to about 130 mM, about 110 mM to about 130 mM, about 120 mM to about 130 mM, about 70 mM to about 120 mM, about 70 mM to about 110 mM, about 70 mM to about 100 mM, about 70 mM to about 90 mM, About 70 mM to about 80 mM, about 80 mM to about 110 mM, about 90 mM to about 110 mM, about 95 mM to about 105 mM calcium) and about 190 mM to about 270 mM glutamate (for example, about 190 mM To about 260 mM, about 190 mM to about 250 mM, about 190 mM to about 240 mM, about 190 mM to about 230 mM, about 190 mM to about 220 mM, about 190 mM to about 210 mM, about 190 mM to about 200 mM, about 200 mM to about 270 mM, about 210 mM to about 270 mM, about 220 mM to about 270 mM, about 230 mM to about 270 mM, about 240 mM to about 270 mM, about 250 mM to about 270 mM , About 260 mM to about 270 mM, about 210 mM to about 250 mM, about 220 mM to about 240 mM, about 230 mM to about 240 mM, about 225 mM to about 235 mM, or about 235 to 245 mM glutamine Salt) together. In an exemplary aspect, an anti-TSLP antibody (e.g., teperumumab) is combined with about 95 mM to about 105 mM calcium and about 225 mM to about 235 mM glutamate, or about 235 to 245 mM glutamate Preparation. Under different circumstances, an anti-TSLP antibody (for example, tepeluxumab) is formulated in about 15 mM to about 195 mM calcium and about 25 mM to about 320 mM glutamine. In various aspects, calcium is about 70 mM to about 150 mM (e.g., about 80 mM to about 150 mM, about 90 mM to about 150 mM, about 100 mM to about 150 mM, about 110 mM to about 150 mM, about 120 mM to about 150 mM, about 130 mM to about 150 mM, about 140 mM to about 150 mM, about 70 mM to about 140 mM, about 70 mM to about 130 mM, about 70 mM to about 120 mM, about 70 mM to About 110 mM, about 70 mM to about 100 mM, about 70 mM to about 90 mM, about 70 mM to about 80 mM, about 90 mM to about 130 mM, about 100 mM to about 120 mM, about 105 mM to about 115 mM calcium) and about 150 mM to about 230 mM glutamine (e.g., about 150 mM to about 220 mM, about 150 mM to about 210 mM, about 150 mM to about 200 mM, about 150 mM to about 190 mM, about 150 mM to about 180 mM, about 150 mM to about 170 mM, about 150 mM to about 160 mM, about 160 mM to about 230 mM, about 170 mM to about 230 mM, about 180 mM to about 230 mM, about 190 mM To about 230 mM, about 200 mM to about 230 mM, about 210 mM to about 230 mM, about 220 mM to about 230 mM, about 170 mM to about 210 mM, about 180 mM to about 200 mM, about 185 mM to about 195 mM glutamate) to prepare TSLP antibody (for example, teparumab). In an exemplary aspect, an anti-TSLP antibody (e.g., teparumab) is formulated in about 105 mM to about 115 mM calcium and about 225 mM to about 235 mM glutamate, or about 235 to 245 mM glutamate. anti). In an exemplary embodiment, the composition of the present disclosure contains more than 140 mg/mL teperumumab, about 99 mM to about 121 mM calcium, 171 mM to about 209 mM glutamate, and 0.01% (w/ v) Polysorbate 80. In an exemplary embodiment, the composition of the present disclosure includes more than 140 mg/mL teperumumab, about 110 mM calcium, 240 mM glutamate, and 0.01% (w/v) polysorbate 80. If necessary, the pH is about 5.0 ± 0.2, or about 5.0 ± 0.1.

Combination excipients

In various aspects, the compositions of the present disclosure include more than one excipient that reduces the viscosity of high protein concentration formulations. In many aspects, the composition of the present disclosure further includes one or more of the following: N-acetylarginine (NAR), N-acetyllysine, methionine, glycine, and proline , Sodium acetate, tris acetate, histamine, or calcium salt. In many aspects, the composition of the present disclosure includes (i) arginine and (ii) NAR and/or methionine. In various aspects, the arginine salt is arginine glutamate. Under different circumstances, the composition of the present disclosure includes (i) calcium salt and (ii) NAR and/or methionine. In an exemplary aspect, the calcium salt is calcium glutamate.

In many respects, N-Acetylarginine (NAR), N-Acetyllysine, Methionine, Glycine, Proline, Sodium Acetate, Tris Acetate, Histamine One or more of, and calcium salts are present in the composition in an amount of about 15 mM to about 300 mM, or about 50 mM to about 300 mM. In an exemplary aspect, the composition of the present disclosure includes one or more of the following: N-Acetylarginine (NAR), N-Acetyllysine, Methionine, Glycine, Proline , Sodium acetate, tris acetate, histamine, and calcium salt, they are about 15 mM to about 200 mM, about 15 to about 150 mM, about 50 mM to about 250 mM, about 50 mM to about 200 mM, About 50 mM to about 150 mM, about 50 mM to about 100 mM, about 50 mM to about 90 mM, about 50 mM to about 80 mM, about 50 mM to about 70 mM, about 50 mM to about 60 mM, about 50 mM to about 55 mM, about 55 mM to about 200 mM, about 60 mM to about 200 mM, about 70 mM to about 200 mM, about 80 mM to about 200 mM, about 90 mM to about 200 mM, about 100 mM to About 200 mM, about 150 mM to about 200 mM, about 160 mM to about 200 mM, about 170 mM to about 200 mM, about 180 mM to about 200 mM, or about 190 mM to about 200 mM is present in the composition In. Under different circumstances, the composition of the present disclosure includes one or more of the following: N-Acetylarginine (NAR), N-Acetyllysine, Methionine, Glycine, Proline , Sodium acetate, tris acetate, histamine, and calcium salt, they are about 50 mM to about 100 mM (for example, about 50 mM, about 60 mM, about 70 mM, about 80 mM, about 90 mM, about 100 mM) or about 100 mM to about 200 mM (e.g., about 100 mM, about 110 mM, about 120 mM, about 130 mM, about 140 mM, about 150 mM, about 160 mM, about 170 mM, about 180 mM, about 190 mM, about 200 mM) is present in the composition. Under different circumstances, the composition of the present disclosure includes one or more of the following: N-Acetylarginine (NAR), N-Acetyllysine, Methionine, Glycine, Proline , Sodium acetate, tris acetate, histamine, and calcium salt, they are about 15 mM to about 200 mM, about 50 mM to about 100 mM or about 50 mM to about 75 mM or about 75 mM to about 100 mM The amount is present in the composition. Under different circumstances, the composition of the present disclosure includes one or more of the following: N-Acetylarginine (NAR), N-Acetyllysine, Methionine, Glycine, Proline , Sodium acetate, tris acetate, histamine, and calcium salt, which are present in the composition in an amount of about 100 mM to about 200 mM or about 100 mM to about 150 mM or about 150 mM to about 200 mM .

In various aspects, when the basic amino acid or its salt or calcium salt or magnesium salt is combined with each other or with another viscosity-reducing excipient, the basic amino acid or its salt or calcium salt or magnesium can be reduced. The amount of salt. In many aspects, compared to a formulation that does not contain another viscosity-lowering excipient (such as proline), when a basic amino acid or its salt or a calcium salt or magnesium salt is combined with each other or another viscosity-reducing formulation When the excipients are combined, the amount of basic amino acid or its salt or calcium salt or magnesium salt is reduced by about 50%. In an illustrative example, the viscosity-reducing excipients present in the composition are in the range of about 0 mM to about 250 mM, about 0 mM to about 220 mM, about 50 mM to about 250 mM, about 50 mM to about 200 mM, About 50 mM to about 150 mM, about 50 mM to about 100 mM, about 50 mM to about 90 mM, about 50 mM to about 80 mM, about 50 mM to about 70 mM, about 50 mM to about 60 mM, about 50 mM to about 55 mM, about 55 mM to about 200 mM, about 60 mM to about 200 mM, about 70 mM to about 200 mM, about 80 mM to about 200 mM, about 90 mM to about 200 mM, about 100 mM to It is present in a total amount of about 200 mM, about 150 mM to about 200 mM, about 160 mM to about 200 mM, about 170 mM to about 200 mM, about 180 mM to about 200 mM, or about 190 mM to about 200 mM.

If necessary, the composition of the present disclosure contains one or more basic amino acids or their salts, and/or calcium salts and/or magnesium salts and/or one or more of the following: N-acetylarginine (NAR ), N-acetyl lysine, methionine, glycine, proline, sodium acetate, tris acetate, histidine, or calcium salt, and at about 50 mM to about 250 mM, About 50 mM to about 200 mM, about 50 mM to about 150 mM, about 50 mM to about 100 mM, about 50 mM to about 90 mM, about 50 mM to about 80 mM, about 50 mM to about 70 mM, about 50 mM to about 60 mM, about 50 mM to about 55 mM, about 55 mM to about 200 mM, about 60 mM to about 200 mM, about 70 mM to about 200 mM, about 80 mM to about 200 mM, about 90 mM to About 200 mM, about 100 mM to about 200 mM, about 150 mM to about 200 mM, about 160 mM to about 200 mM, about 170 mM to about 200 mM, about 180 mM to about 200 mM, or about 190 mM to about A total amount of 200 mM is present. In various embodiments, the composition described herein contains proline (such as L-proline), optionally from about 0 mM to about 250 mM, about 0 mM to about 220 mM, about 25 mM to about It is present in a total amount of 200 mM, about 50 mM to about 150 mM, or about 50 mM to about 100 mM. In various embodiments, proline is at about 40 mM, about 50 mM, about 60 mM, about 65 mM, about 70 mM, about 75 mM, about 80 mM, about 85 mM, about 90 mM, about 95 mM, About 100 mM, about 105 mM, about 110 mM, about 115 mM, about 120 mM, about 125 mM, about 130 mM, about 135 mM, about 140 mM, about 145 mM, about 150 mM, about 160 mM, about 170 It is present in an amount of mM, about 180 mM, about 190 mM, about 200 mM, about 210 mM, about 220 mM, about 230 mM, about 240 mM, or about 250 mM.

Under different circumstances, the composition of the present disclosure contains more than 140 mg/mL anti-TSLP antibody (for example, tepeluxumab), a surfactant, a basic amino acid or its salt, and proline. Under different circumstances, an anti-TSLP antibody (such as teperumumab) greater than 140 mg/mL is combined with about 10 mM to 200 mM, or about 50 mM to about 150 mM basic amino acid or its salt, and about 50 mM to about 250 mM proline, or about 50 mM to about 150 mM proline. If necessary, an anti-TSLP antibody greater than 140 mg/mL (for example, teperumumab) is formulated with about 50 mM to about 100 mM basic amino acid or its salt, and about 90 mM to about 150 mM proline . In many aspects, the salt of the basic amino acid is arginine glutamate, and a certain amount of arginine is added, and a certain amount of glutamate is added to obtain arginine glutamate The solution. In various aspects, an anti-TSLP antibody (e.g., teperumumab) greater than 140 mg/mL is combined with about 40 mM to about 120 mM arginine and about 45 mM to about 125 mM glutamate and about 60 mM to about 60 mM Prepared with approximately 140 mM proline. Optionally, an anti-TSLP antibody (for example, teperumumab) and about 50 mM to about 110 mM, about 60 mM to about 100 mM, about 70 mM to about 90 mM, or about 75 mM to about 85 mM spermine Acid, and about 55 mM to about 115 mM, about 65 mM to about 105 mM, about 75 mM to about 95 mM, about 80 mM to or about 90 mM glutamate, and about 70 mM to about 130 mM, about 80 mM to about 120 mM, about 90 mM to about 110 mM, or about 95 mM to about 105 mM proline is formulated together. In various aspects, anti-TSLP antibodies are formulated in about 10 mM to about 90 mM arginine and about 55 mM to about 135 mM glutamate and about 45 mM to about 125 mM proline. If necessary, in about 20 mM to about 80 mM, about 30 mM to about 70 mM, about 40 mM to about 60 mM, about 45 mM to about 55 mM arginine, and about 65 mM to about 125 mM, about 75 mM mM to about 115 mM, about 85 mM to about 105 mM, about 90 mM to about 100 mM glutamate, and about 55 mM to about 115 mM, about 65 mM to about 105 mM, about 75 mM to about 95 mM , About 80 mM to about 90 mM, or about 85 mM proline to prepare an anti-TSLP antibody (for example, tepelumab). In various aspects, the composition comprises greater than 140 mg/mL anti-TSLP antibody, about 10 mM to about 90 mM arginine, and about 55 mM to about 135 mM glutamine and about 45 mM to about 125 mM proline acid. If necessary, the composition contains an anti-TSLP antibody, such as teperumumab, about 20 mM to about 80 mM, about 30 mM to about 70 mM, about 40 mM to about 60 mM, about 45 mM to about 55 mM Amino acid, and about 65 mM to about 125 mM, about 75 mM to about 115 mM, about 85 mM to about 105 mM, about 90 mM to about 100 mM glutamate, and about 55 mM to about 115 mM, about 65 mM to about 105 mM, about 75 mM to about 95 mM, about 80 mM to about 90 mM, about 85 mM proline. Under different circumstances, the composition of the present disclosure contains more than 140 mg/mL teperumumab, about 45 mM to about 55 mM arginine, about 85.5 mM to about 104.5 mM glutamine and about 76.5 mM to about 93.5 mM proline.

Under different circumstances, the composition of the present disclosure contains more than 140 mg/mL anti-TSLP antibody (for example, tepeluxumab), surfactant, calcium or magnesium salt, and proline (for example, L-proline) . Under different circumstances, anti-TSLP antibodies greater than 140 mg/mL (such as teperumumab) are formulated with about 15 mM to about 150 mM calcium or magnesium salts, and about 50 mM to about 150 mM proline. If necessary, an anti-TSLP antibody greater than 140 mg/mL (for example, teperumumab) is formulated with about 50 mM to about 100 mM basic amino acid or its salt, and about 90 mM to about 150 mM proline . In many aspects, the calcium is calcium glutamate, and a certain amount of calcium is added and a certain amount of glutamate is added to obtain a solution containing calcium glutamate. Under different circumstances, the anti-TSLP antibody is formulated with 20 mM to about 100 mM calcium and about 100 mM to about 180 mM glutamate and about 30 mM to about 110 mM proline. If necessary, the anti-TSLP antibody (for example, teperumumab) is combined with about 30 mM to about 90 mM, about 40 mM to about 80 mM, about 50 mM to about 70 mM, about 55 mM to about 65 mM, or About 60 mM calcium, and about 110 mM to about 170 mM, about 120 mM to about 160 mM, about 130 mM to about 150 mM, about 135 mM to about 145 mM, or about 140 mM glutamate, and about 40 mM to about 100 mM, about 50 mM to about 90 mM, about 60 mM to about 80 mM, about 65 mM to about 75 mM, or about 70 mM proline. Under different circumstances, anti-TSLP antibodies are formulated in about 30 mM to about 110 mM calcium, about 105 mM to about 185 mM glutamate, and about 20 mM to about 100 mM proline. If necessary, in about 40 mM to about 100 mM, about 50 mM to about 90 mM, about 60 mM to about 80 mM, about 65 mM to about 75 mM, or about 75 mM calcium, and about 115 mM to about 175 mM , About 125 mM to about 165 mM, about 135 mM to about 155 mM, about 140 mM to about 150 mM, or about 145 mM glutamate, and about 30 mM to about 90 mM, about 40 mM to about 80 mM , About 50 mM to about 70 mM, about 55 mM to about 65 mM, or about 60 mM proline to prepare an anti-TSLP antibody (for example, tepeluxumab). In various cases, the composition contains greater than 140 mg/mL anti-TSLP antibody, about 30 mM to about 110 mM calcium, about 105 mM to about 185 mM glutamate, and about 20 mM to about 100 mM proline . If necessary, the composition contains an anti-TSLP antibody, such as teperumumab, about 40 mM to about 100 mM, about 50 mM to about 90 mM, about 60 mM to about 80 mM, about 65 mM to about 75 mM, Or about 75 mM calcium, and about 115 mM to about 175 mM, about 125 mM to about 165 mM, about 135 mM to about 155 mM, about 140 mM to about 150 mM, or about 145 mM glutamate, and about 30 mM to about 90 mM, about 40 mM to about 80 mM, about 50 mM to about 70 mM, about 55 mM to about 65 mM, or about 60 mM proline. Under different circumstances, the composition of the present disclosure contains more than 140 mg/mL teperumumab, about 63 mM to about 77 mM calcium, about 130.5 mM to about 159.5 mM glutamate, and about 54 mM to about 66 mM Proline.

Surfactant

In many aspects, the composition of the present disclosure includes a surfactant. Surfactants are amphiphilic (with polar head and hydrophobic tail) surfactants. Surfactant preferentially accumulates at the interface, thereby reducing interface tension. The use of surfactants can also help alleviate the formation of large protein particles. In some aspects, the surfactants present in the composition of the present disclosure are amphiphilic surfactants and/or nonionic surfactants. Exemplary surfactants include polyoxyethylene sorbitan fatty acid esters (e.g., polysorbate 20, polysorbate 80), alkyl aryl polyethers, such as oxyethylated alkyl phenols (e.g. Triton™ X -100), and poloxamers (such as Pluronics®, such as Pluronic® F68), and a combination of any of the above belonging to one surfactant class or belonging to multiple surfactant classes. In particular, polysorbate 20 and polysorbate 80 (and mixtures thereof as required) are contemplated. In an illustrative example, the surfactant is present in the composition at a concentration of less than or about 0.015% (w/v) ± 0.005% (w/v). For example, the formulation may contain about 0.005% (w/v) to about 0.015% (w/v) surfactant, such as about 0.005% (w/v), about 0.006% (w/v), about 0.007% ( w/v), about 0.008% (w/v), about 0.009% (w/v), about 0.010% (w/v), about 0.011% (w/v), about 0.012% (w/v), About 0.013% (w/v), about 0.014% (w/v), about 0.015% (w/v). In exemplary aspects, the formulation includes about 0.005% (w/v), 0.010% (w/v), or 0.015% (w/v) surfactant. In various aspects, the surfactant is a polysorbate, such as polysorbate 20 or polysorbate 80 or mixtures thereof. If necessary, the surfactant is present in a concentration of less than or about 0.005% (w/v) to about 0.015% (w/v), and if necessary, about 0.010% (w/v) ± 0.0025% (w/v) Surfactant, for example, about 0.005% (w/v), 0.010% (w/v), or 0.015% (w/v) surfactant.

Antibody concentration

In an exemplary aspect, the composition of the present disclosure is at a rate of greater than about 100 mg/mL and less than about 300 mg/mL or less than about 250 mg/mL, as needed, from about 160 mg/mL to about 250 mg/mL, such as about 180 mg/mL. The anti-TSLP antibody is included in a concentration of mg/mL to about 225 mg/mL, or about 180 mg/mL to about 200 mg/mL. In some aspects, the anti-TSLP antibody ranges from about 160 mg/mL to about 250 mg/mL, from about 160 mg/mL to about 240 mg/mL, from about 160 mg/mL to about 230 mg/mL, from about 160 mg/mL to About 220 mg/mL, about 160 mg/mL to about 210 mg/mL, about 160 mg/mL to about 200 mg/mL, about 160 mg/mL to about 190 mg/mL, about 160 mg/mL to about 180 mg/mL, about 160 mg/mL to about 170 mg/mL, about 160 mg/mL to about 165 mg/mL, about 165 mg/mL to about 250 mg/mL, about 170 mg/mL to about 250 mg/mL mL, about 180 mg/mL to about 250 mg/mL, about 190 mg/mL to about 200 mg/mL, about 210 mg/mL to about 250 mg/mL, about 220 mg/mL to about 250 mg/mL, Concentrations of about 230 mg/mL to about 250 mg/mL, and about 240 mg/mL to about 250 mg/mL are present in the composition. In some aspects, the anti-TSLP antibody is present in the composition at a concentration of about 180 mg/mL or about 190 mg/mL, about 200 mg/mL, or about 210 mg/mL.

In an exemplary aspect, the composition comprises about 160 mg/mL to about 250 mg/mL of anti-TSLP antibody, optionally about 160 mg/mL to about 225 mg/mL or about 160 mg/mL to about 200 mg/mL . In various aspects, the composition includes about 175 mg/mL to about 185 mg/mL of anti-TSLP antibody, as needed, about 175 mg/mL, about 176 mg/mL, about 177 mg/mL, about 178 mg/mL, About 179 mg/mL, about 180 mg/mL, about 181 mg/mL, about 182 mg/mL, about 183 mg/mL, about 184 mg/mL, about 185 mg/mL. In various aspects, the composition contains about 180 mg/mL of anti-TSLP antibody. Under different circumstances, the concentration of the anti-TSLP antibody ranges from about 189 mg/mL or about 190 mg/mL to about 230 mg/mL or about 231 mg/mL. If necessary, the concentration of anti-TSLP antibody is about 205 mg/mL to about 215 mg/mL, and about 210 mg/mL or about 205 mg/mL, about 206 mg/mL, about 207 mg/mL, about 208 mg as needed /mL, about 209 mg/mL, about 210 mg/mL, about 211 mg/mL, about 212 mg/mL, about 213 mg/mL, about 214 mg/mL, about 215 mg/mL.

In an exemplary aspect, the anti-TSLP antibody is at a concentration of about 140 mg/mL to about 210 mg/mL, for example, about 180 mg/mL ± 10%, about 200 mg/mL ± 10%, about 210 mg/mL ± 10% Exist in the composition.

Additional excipients

In an exemplary aspect, the composition of the present disclosure may include additional components. In various aspects, the composition contains any pharmaceutically acceptable ingredients, including, for example, acidifiers, additives, adsorbents, aerosol propellants, air displacement agents, alkalizers, anti-caking agents, anticoagulants, Antimicrobial preservatives, antioxidants, antibacterial agents, alkalis, binders, buffers, chelating agents, coating agents, colorants, desiccants, detergents, diluents, disinfectants, disintegrants, dispersants, dissolution enhancers Agents, dyes, lubricants, emulsifiers, emulsion stabilizers, fillers, film forming agents, flavor enhancers, flavoring agents, flow enhancers, gelling agents, granulating agents, humectants, lubricants, mucosal adhesives, Ointment base, ointment, oily vehicle, organic base, lozenge base, pigment, plasticizer, polishing agent, preservative, sequestrant, skin penetrant, solubilizer, solvent, stabilizer, suppository base, Surface active agent (surface active agent), surfactant (surfactant), suspending agent, sweetener, therapeutic agent, thickener, tonicity agent, toxic agent, viscosity increasing agent, water absorbing agent, water miscible cosolvent, water Softener or wetting agent. See, for example, Handbook of Pharmaceutical Excipients , Third Edition, AH Kibbe (Pharmaceutical Press [Medical Press], London, UK [London, UK], 2000), which is incorporated by reference in its entirety . Remington's Pharmaceutical Sciences , Sixteenth Edition, EW Martin (Mack Publishing Co., Easton, Pa. [Easton, Pennsylvania], 1980), which is cited as The full text is incorporated.

pH , Viscosity, and osmotic pressure

In an alternative aspect, the composition of the present disclosure is a liquid. In certain aspects, the liquid has a pH of less than about 6.0, if desired, less than about 5.7, or less than about 5.5. In some aspects, the pH is about 4.5 to about 5.7, about 4.5 to about 5.5, about 4.7 to about 5.3, about 4.8 to about 5.4, or about 5.0 to about 5.7, for example, about 4.7, about 4.8, about 4.9, about 5.0 , About 5.1, about 5.2, about 5.3, about 5.4, about 5.5, about 5.6, about 5.7. In some aspects, the pH is about 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, or 5.7. In an illustrative example, the composition of the present disclosure has a pH of about 4.5 to about 6.75, and optionally about 4.8 to about 6.0.

In some aspects, the composition is characterized by a reduced viscosity relative to a liquid composition that does not contain proline. In an exemplary embodiment, the composition is characterized in that when the concentration of the anti-TSLP antibody is less than 155 mg/mL, the viscosity at 23°C is less than about 24 centipoise (cP), and when the concentration of the anti-TSLP antibody is When the concentration is about 110 mg/mL, the viscosity is about 6 cP, or when the concentration of anti-TSLP antibody is about 140 mg/mL, the viscosity is about 15 cP. In certain aspects, when the concentration of the anti-TSLP antibody is less than 155 mg/mL (for example, about 110 mg/mL, about 140 mg/mL), the composition is characterized by a viscosity ranging from about 5 cP to about 20 cP, for example about 5 cP to about 15 cP, about 5 cP to about 10 cP, about 10 cP to about 20 cP, about 15 cP to about 20 cP, or about 5 cP, about 6 cP, about 7 cP, about 8 cP, about 9 cP, about 10 cP, about 11 cP, about 12 cP, about 13 cP, about 14 cP, about 15 cP, about 16 cP, about 17 cP, about 18 cP, about 19 cP, about 20 cP, about 21 cP, About 22 cP.

In some aspects, when the concentration of the anti-TSLP antibody is 180 mg/mL or greater (e.g., about 180 mg/ml, about 210 mg/mL, about 240 mg/mL), the composition is characterized by a viscosity of about 5 cP to about 25 cP, for example, about 5 cP to about 20 cP, about 5 cP to about 15 cP, about 5 cP to about 10 cP, about 10 cP to about 25 cP, about 15 cP to about 20 cP, or about 5 cP, about 6 cP, about 7 cP, about 8 cP, about 9 cP, about 10 cP, about 11 cP, about 12 cP, about 13 cP, about 14 cP, about 15 cP, about 16 cP, about 17 cP , About 18 cP, about 19 cP, about 20 cP, about 21 cP, about 22 cP, about 23 cP, about 24 cP, about 25 cP. In an exemplary aspect, when the concentration of the antibody is about 100 mg/mL to about 180 mg/mL, the composition has a viscosity of about 15 cP ± 5 cP or about 20 cP ± 5 cP. Unless otherwise specified, all viscosities disclosed herein refer to the viscosity measured using a rotary viscometer at 23°C and a shear rate of about 1000 l/s.

In an exemplary aspect, the present disclosure of the composition at 23 ° C, viscosity 1000 s ^-1 is less than 100 cP, as required, at 23 ° C, 1000 s ^-1 is less than 75 cP, e.g., less than 60 cP, or less than 50 cP.

In an exemplary aspect, the composition is intended for subcutaneous administration to a subject, so the composition is isotonic with the intended site of administration. For example, in some aspects, the osmotic pressure of the composition is in the range of about 270 to about 350 mOsm/kG, or about 285 to about 345 mOsm/kG, or about 300 to about 315 mOsm/kG. For example, if the solution is in a form intended for parenteral administration, it can be made isotonic with blood (about 300 mOsm/kG osmotic pressure). In an exemplary aspect, the aqueous pharmaceutical formulation has a range of about 200 mOsm/kg to about 500 mOsm/kg, or about 225 mOsm/kg to about 400 mOsm/kg, or about 250 mOsm/kg to about 350 mOsm/kg. Osmotic pressure within the range. Optionally, the composition is isotonic or has an osmotic pressure greater than about 350 mOsm/kg.

stability

In different cases, as determined by size exclusion chromatography (SEC), less than about 5% of the antibody is degraded after storage at about 2°C to about 8°C for at least or about 12 months. In many aspects, as determined by size exclusion chromatography (SEC), after storage at about 2°C to about 8°C for about 20 months to about 26 months, less than about 5% of the antibody is degradation. In an exemplary example, as determined by size exclusion chromatography (SEC), less than about 5% of the antibody is degraded after storage at about 2°C to about 8°C for about 30 to about 40 months . In an exemplary example, as determined by size exclusion chromatography (SEC), less than about 5% of the antibody is degraded after storage at about 2°C to about 8°C for about 2 to about 3 years . For another example, as determined by size exclusion chromatography (SEC), less than 5% of the antibody is degraded after storage at 2°C to 8°C for about 24 months to about 36 months, if necessary, Among them, less than 2% of the antibodies are degraded after storage at 2°C to 8°C for 24 months or 36 months. In many respects, as determined by the SEC, after storage at about room temperature (eg 25°C) for at least 2 weeks (as needed, at least 1 month, at least 2 months, at least 3 months, After at least 4 months, at least 5 months, or at least 6 months), less than 5% of the antibody is degraded. In different cases, as determined by the SEC, storage at 2°C to 8°C for about 24 months to about 36 months, and then storage at about room temperature (for example, 25°C) for at least 2 weeks or at least After about 1 month or at least about 2 months, less than 5% of the antibodies are degraded. Optionally, as determined by size exclusion chromatography (SEC), less than about 5% of the antibody is degraded after storage at a temperature greater than about 20°C for at least or about 2 weeks, and storage for at least or about 4 weeks or about 8 weeks. In various aspects, the temperature is greater than or about 25°C or greater than or about 30°C or greater than or about 40°C.

Products, syringes, and vials

This article further provides an article. In an exemplary embodiment, the product contains the composition of the present disclosure, and optionally contains about 1 mL to about 5 mL (for example, about 1 mL to about 3 mL) of the aqueous composition. In an exemplary aspect of the present disclosure, the composition is provided in a form for storage or use, for example, in a single-use vial, a single-use syringe, or glass, glass-lined, glass-coated main container, or auto-injector. In an exemplary aspect, the composition is provided in a single-use system bag or polycarbonate carafe for refrigerated storage. In an alternative aspect, the composition is contained in a glass vial or syringe for storage at 2°C to 8°C. This document additionally provides a pre-filled syringe that contains the composition of the present disclosure, and optionally contains about 1 mL to about 5 mL (for example, about 1 mL to about 3 mL) of the composition. A vial is further provided, which contains the composition of the present disclosure, and optionally contains about 1 mL to about 5 mL (for example, about 1 mL to about 3 mL) of the aqueous composition. In various aspects, the articles, pre-filled syringes, and vials contain about 2 mL to about 3 mL (e.g., about 2.1 mL, about 2.2 mL, about 2.3 mL, about 2.4 mL, about 2.5 mL, about 2.6 mL, about 2.7 mL). mL, about 2.8 mL, about 2.9 mL) of the composition of the present disclosure, and in many aspects, the composition contains tepelumumab at a concentration of about 180 mg/mL to provide about 420 mg of tepelumumab anti.

In an illustrative example, the composition is provided for use in an off-the-shelf and/or designed for self-administration delivery system. In an exemplary aspect, the composition is provided in a pre-filled syringe, or an auto-injector, a pen-type syringe, a dual-chamber pen, and the like. Such products are known in the art and commercially available. See, for example, Shire, Steven, Monoclonal Antibodies: Meeting the Challenges in Manufacturing, Formulation, Delivery and Stability of Final Drug Product, Chapter 8: Development of delivery device technology to deal with the challenges of highly viscous mAb formulations at high concentration [Chapter 8: Development of delivery device technology to deal with the challenges of highly viscous mAb formulations at high concentration], Woodhead Publishing Society], Cambridge, UK, pp. 153-162 (2015). In an exemplary aspect, the composition is provided for YpsoMate™ automatic injection, YpsoMate™ 2.25 autoinjector, or VarioJect™ (YpsoMed, Burgdorf, Switzerland). Other auto-injectors include, for example, SelfDose™ patient-controlled syringes, BD Physioject™ disposable auto-injectors, and Autoject® II syringes (Owen Mumford, Oxfordshire, UK). In various embodiments, the auto-injector is a Ypsomed YpsoMate® auto-injector. Additional auto-injectors anticipated in this method are disclosed in International Patent Publications WO 2018/226565, WO 2019/094138, WO 2019/178151, WO 20120/072577, WO 2020/081479, WO 2020/081480, and International Patent Application Numbers In PCT/US 20/70590, PCT/US 20/70591, PCT/US 20/53180, PCT/US 20/53179, PCT/US 20/53178, and PCT/US 20/53176, these documents are incorporated by reference Incorporated into this article.

The composition of the present disclosure may be suitable for administration by any acceptable route, including parenteral, and especially subcutaneous administration. For example, subcutaneous administration can be administered to the upper arm, thigh, or abdomen. Other routes include, for example, intravenous, intradermal, intramuscular, intraperitoneal, intranodular, and intrasplenic. The subcutaneous route is better.

If the composition is in a form intended for administration to a subject, it can be made isotonic with the intended administration site. For example, if the solution is in a form intended for parenteral administration, it can be made isotonic with blood. The composition is usually sterile. In some embodiments, this can be achieved by filtration with a sterile filter membrane. In certain embodiments, the parenteral composition is generally placed in a container with a sterile access port, such as an intravenous solution bag, or a vial with a stopper pierceable by a hypodermic injection needle, or a pre-filled syringe. In certain embodiments, the composition can be stored in a ready-to-use form.

anti- TSLP antibody

The composition of the present disclosure includes anti-TSLP antibodies. In an exemplary embodiment, the anti-TSLP antibody specifically binds to the TSLP polypeptide shown in amino acids 29-159 of SEQ ID NO: 2. Thymic stromal lymphopoietin (TSLP) is a cytokine derived from epithelial cells, which is produced in response to pro-inflammatory stimuli, and is mainly produced in dendritic cells (Gilliet, J Exp Med. [Journal of Experimental Medicine ] 197: 1059-1067, 2003; Soumelis, Nat Immunol. [Natural Immunology] 3: 673-680, 2002; Reche, J Immunol. [Journal of Immunology] 167: 336-343, 2001), obese cells (Allakhverdi , J Exp Med. [Journal of Experimental Medicine] 204: 253-258, 2007), and the activity on CD34+ precursor cells to drive allergic inflammatory responses. Swedin et al., Pharmacol Ther [Pharmacology and Therapeutics] 169: 13-34 (2017). TSLP sends messages through a heterodimeric receptor composed of the interleukin (IL)-7 receptor alpha (IL-7Rα) chain and the common gamma chain-like receptor (TSLPR) (Pandey, Nat Immunol. [Natural Immunology ] 1: 59-64, 2000; Park, J Exp Med. [Journal of Experimental Medicine] 192: 659-669, 2000).

Compared with controls, human TSLP mRNA in the airways of individuals with asthma (Brightling et al., J Allergy Clin Immunol [Journal of Allergy and Clinical Immunity] 121: 5-10 quiz 1-2 (2008); Ortega et al., NEJM [New England Medicine Journal] 371: 1198-1207 (2014)) and protein levels (Ortega et al., (2014), ibid.) have increased, and the degree of this manifestation is related to the severity of the disease. Brightling et al., (2008), ibid. Recent studies have demonstrated that single nucleotide polymorphisms in the human TSLP locus are related to the prevention of asthma, atopic asthma, and airway overreaction, which indicates that differential regulation of TSLP gene expression can affect disease susceptibility. (To et al., BMC Public Health [BMC Public Health] 12: 204 (2012); XOLAIR® (omalizumab): Highlights of Prescribing Information 2016. (At https://www .gene.com/download/pdf/xolair_prescribing.pdf.); Bleecker et al., The Lancet [The Lancet] 388: 2115-2127 (2016). These data indicate that targeting TSLP can inhibit multiple organisms related to asthma way.

Early non-clinical studies of TSLP have shown that after TSLP is released from airway epithelial cells or stromal cells, it activates obese cells, dendritic cells and T cells to release Th2 cytokines (such as IL-4/13/5). Recently published human data prove that there is a good correlation between tissue TSLP gene and protein expression, Th2 gene signature score and tissue eosinophils in severe asthma. Therefore, anti-TSLP target therapy may be effective in asthma patients with Th2-type inflammation (Shikotra et al., J Allergy Clin Immunol. [Journal of Allergy and Clinical Immunity] 129 (1): 104-11, 2012).

Data from other studies indicate that TSLP can cause airway inflammation through pathways unrelated to Th2, such as crosstalk between airway smooth muscle and obese cells (Allakhverdi et al., J Allergy Clin Immunol. [Journal of Allergy and Clinical Immunity] 123 (4) : 958-60, 2009; Shikotra et al., ibid). TSLP can also promote the differentiation of T cells into Th-17-producing cells, so that increased neutrophil inflammation is common in more severe asthma (Tanaka et al., Clin Exp Allergy. [Clinical and Experimental Allergy] 39 (1): 89-100, 2009). These data and other emerging evidence indicate that blocking TSLP can be used to suppress multiple biological pathways, including but not limited to pathways involving Th2 cytokines (IL-4/13/5).

Antibodies specific to TSLP are expected to be useful in the treatment of asthma, including severe asthma, eosinophilic asthma, non-eosinophilic/low eosinophilic asthma and other forms of asthma described herein.

Specific binding agents (such as antibodies and antibody variants or fragments) that bind to target antigens (such as TSLP) can be used in the methods of the present disclosure. In one embodiment, the specific binding agent is an antibody. The antibodies can be single cloned (MAb); recombinant; chimeric; humanized, such as complementarity determining region (CDR) grafted; human; antibody variants, including single chains; and/or double Specific; and fragments thereof; variants; or derivatives thereof. Antibody fragments include those parts of the antibody that bind to the epitope on the polypeptide of interest. Examples of such fragments include Fab and F(ab') fragments produced by enzymatic cleavage of full-length antibodies. Other binding fragments include fragments produced by recombinant DNA technology, such as the expression of recombinant plastids containing nucleic acid sequences encoding antibody variable regions.

Monoclonal antibodies can be modified for use as therapeutic or diagnostic agents. One embodiment is a "chimeric" antibody, in which a part of the heavy chain (H) and/or light chain (L) is the same or homologous to the corresponding sequence in an antibody derived from a specific species or belonging to a specific antibody class or subclass, and The remainder of the chain is the same or homologous to the corresponding sequence in an antibody derived from another species or belonging to another antibody class or subclass. Fragments of such antibodies are also included, as long as they exhibit the desired biological activity. See U.S. Patent No. 4,816,567; Morrison et al., 1985, Proc. Natl. Acad. Sci. [Proceedings of the National Academy of Sciences] 81: 6851-55.

In another embodiment, the monoclonal antibody system is "humanized" antibodies. Methods for humanizing non-human antibodies are well known in the art. See U.S. Patent Nos. 5,585,089 and 5,693,762. Generally, a humanized antibody has one or more amino acid residues introduced into it from a source that is non-human. For example, methods described in the art can be used (Jones et al., 1986, Nature [Nature] 321: 522-25; Riechmann et al., 1998, Nature [Nature] 332: 323-27; Verhoeyen et al., 1988, Science [Science] 239: 1534-36), by substituting at least a part of the rodent complementarity determining region for the corresponding region of the human antibody to carry out humanization.

This disclosure also covers human antibodies and antibody variants (including antibody fragments) that bind TSLP. Use transgenic animals (such as mice) that can produce human antibody repertoire in the absence of endogenous immunoglobulin production, by immunizing with polypeptide antigens (that is, having at least 6 adjacent amino acids), combined with carriers as needed , To produce such antibodies. See, for example, Jakobovits et al., 1993, Proc. Natl. Acad. Sci. [Proceedings of the National Academy of Sciences] 90: 2551-55; Jakobovits et al., 1993, Nature [Nature] 362: 255-58; Bruggermann et al., 1993, Year in Immuno. [Annual Immunology] 7: 33. See also PCT application number PCT/US 96/05928 and PCT/US 93/06926. Other methods are described in U.S. Patent No. 5,545,807, PCT Application No. PCT/US 91/245 and PCT/GB 89/01207, and European Patent No. 546073 B1 and 546073 A1. Human antibodies can also be produced by expression of recombinant DNA in host cells or by expression in fusion tumor cells as described herein.

Chimeric, CDR grafted, and humanized antibodies and/or antibody variants are usually produced by recombinant methods. The nucleic acid encoding the antibody is introduced into the host cell and performed using the materials and procedures described herein. In a preferred embodiment, antibodies are produced in mammalian host cells such as CHO cells. Monoclonal (eg, human) antibodies can be produced by expression of recombinant DNA in host cells or by expression in fusion tumor cells as described herein.

Antibodies and antibody variants (including antibody fragments) that can be used in the methods of the present invention comprise anti-TSLP antibodies comprising: (A) a light chain variable domain comprising: (i) comprising SEQ The light chain CDR1 sequence of the amino acid sequence shown in ID NO: 3; (ii) the light chain CDR2 sequence containing the amino acid sequence shown in SEQ ID NO: 4; and (iii) the light chain CDR2 sequence containing the amino acid sequence shown in SEQ ID NO: The light chain CDR3 sequence of the amino acid sequence shown in 5; and (B) the heavy chain variable domain comprising: (i) containing the amino group shown in SEQ ID NO: 6 The heavy chain CDR1 sequence of the acid sequence; (ii) the heavy chain CDR2 sequence containing the amino acid sequence shown in SEQ ID NO: 7, and (iii) the heavy chain CDR2 sequence containing the amino acid sequence shown in SEQ ID NO: 8 Heavy chain CDR3 sequence.

It is also expected to comprise the following antibodies or antibody variants: (A) a light chain variable domain selected from the group consisting of: (i) having at least 80% with SEQ ID NO: 12 ( For example, about 85%, about 90%, about 95%, greater than 95%) identical amino acid sequences; (ii) having at least 80% (for example, about 85%, about 90%) with SEQ ID NO: 11 , About 95%, greater than 95%) an amino acid sequence encoded by a polynucleotide sequence of identity; (iii) a polynucleotide that hybridizes to the complementary sequence of the polynucleotide composed of SEQ ID NO: 11 under moderately stringent conditions Acid-encoded amino acid sequence; and (B) a heavy chain variable domain, the heavy chain variable domain being selected from the group consisting of: (i) having at least 80% with SEQ ID NO: 10 (for example, About 85%, about 90%, about 95%, greater than 95%) identical amino acid sequences; (ii) having at least 80% (for example, about 85%, about 90%, about 95%, greater than 95%) an amino acid sequence encoded by a polynucleotide sequence with identity; (iii) a polynucleotide encoding a polynucleotide that hybridizes to the complementary sequence of the polynucleotide composed of SEQ ID NO: 9 under moderately stringent conditions Or (C) the light chain variable domain of (A) and the heavy chain variable domain of (A), wherein the antibody or antibody variant is the same as the amino acid 29 of SEQ ID NO: 2 The TSLP polypeptide shown in -159 specifically binds.

In an exemplary example, the anti-TSLP antibody comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 13, a light chain containing the amino acid sequence of SEQ ID NO: 14, or an amine containing SEQ ID NO: 13 The heavy chain containing the amino acid sequence of SEQ ID NO: 14 and the light chain containing the amino acid sequence of SEQ ID NO: 14.

Teperumumab has the following exemplary anti-TSLP antibodies: (A) a light chain variable domain, the light chain variable domain comprising: (i) containing the amino acid shown in SEQ ID NO: 3 The light chain CDR1 sequence of the sequence; (ii) the light chain CDR2 sequence containing the amino acid sequence shown in SEQ ID NO: 4; and (iii) the light chain CDR2 sequence containing the amino acid sequence shown in SEQ ID NO: 5 Chain CDR3 sequence; and (B) a heavy chain variable domain, the heavy chain variable domain comprising: (i) a heavy chain CDR1 sequence containing the amino acid sequence shown in SEQ ID NO: 6; (ii) The heavy chain CDR2 sequence containing the amino acid sequence shown in SEQ ID NO: 7, and (iii) the heavy chain CDR3 sequence containing the amino acid sequence shown in SEQ ID NO: 8.

Teperumumab also contains:

(A) The light chain variable domain, the light chain variable domain is selected from the group consisting of: (i) An amino acid sequence with at least 80% identity with SEQ ID NO: 12; (ii) An amino acid sequence encoded by a polynucleotide sequence having at least 80% identity with SEQ ID NO: 11; (iii) The amino acid sequence encoded by the polynucleotide that hybridizes with the complementary sequence of the polynucleotide composed of SEQ ID NO: 11 under moderately stringent conditions; and

(B) Heavy chain variable domain, the heavy chain variable domain is selected from the group consisting of: (i) An amino acid sequence with at least 80% identity with SEQ ID NO: 10; (ii) An amino acid sequence encoded by a polynucleotide sequence having at least 80% identity with SEQ ID NO: 9; (iii) The amino acid sequence encoded by the polynucleotide that hybridizes with the complementary sequence of the polynucleotide composed of SEQ ID NO: 9 under moderately stringent conditions; or

(C) (A) light chain variable domain and (B) heavy chain variable domain.

Other exemplary anti-TSLP resistance systems are known in the art. See, for example, International Patent Application Publication No. WO 2017/042701, WO 2016/142426, WO 2010/017468, U.S. Patent Application Publication No. US 2012/0020988, and U.S. Patent No. 8,637,019. In an exemplary aspect, the anti-TSLP system is an antibody disclosed in one of these publications.

In various embodiments, the anti-TSLP antibody or its antibody variant system is bivalent and selected from the group consisting of: human antibodies, humanized antibodies, chimeric antibodies, monoclonal antibodies, recombinant antibodies, antigen-binding antibodies Fragments, single-chain antibodies, monomeric antibodies, diabodies, triabodies, tetrabodies, Fab fragments, IgG1 antibodies, IgG2 antibodies, IgG3 antibodies, and IgG4 antibodies. In an exemplary aspect, the anti-TSLP anti-system IgG2 antibody.

In various embodiments, the anti-TSLP antibody variant is selected from the group consisting of diabodies, tribodies, tetrabodies, Fab fragments, single domain antibodies, scFv, wherein the dosage is adjusted so that the binding sites The point is equimolar relative to the binding site of the administered bivalent antibody. In an exemplary aspect, the two binding sites of the antibody have the same binding as TSLP.

The antibody or antibody variant is expected to be an IgG2 antibody. An exemplary sequence of the human IgG2 constant region can be obtained from the Uniprot database under Uniprot number P01859, which is incorporated herein by reference. Information including sequence information about other antibody heavy and light chain constant regions can also be publicly obtained through the Uniprot database and other databases well-known in the field of antibody engineering and production.

In certain embodiments, antibody derivatives include tetrameric glycosylated antibodies in which the number and/or type of glycosylation sites are changed compared to the amino acid sequence of the parent polypeptide. In certain embodiments, the variant contains more or fewer N-linked glycosylation sites than the native protein. Alternatively, a substitution that eliminates this sequence will remove the existing N-linked carbohydrate chain. The rearrangement of N-linked carbohydrate chains is also provided, in which one or more N-linked glycosylation sites (usually naturally occurring N-linked glycosylation sites) are eliminated and one or more new N-linked sites are created Of the site. Other preferred antibody variants include cysteine variants, in which one or more cysteine residues are deleted or substituted for another amino acid (such as serine ). Cysteine variants can be useful when the antibody must be refolded into a biologically active conformation, such as after isolation of insoluble inclusion bodies. Cysteine variants generally have fewer cysteine residues than natural proteins, and usually have an even number in order to minimize interactions caused by unpaired cysteine.

The required amino acid substitutions (whether conservative or non-conservative) can be determined by those skilled in the art when such substitutions are needed. In some embodiments, amino acid substitutions can be used to identify important residues of antibodies to human TSLP, or to increase or decrease the affinity of antibodies to human TSLP as described herein.

According to certain embodiments, the preferred amino acid substitutions are the following: (1) reduce the sensitivity to protein hydrolysis; (2) reduce the sensitivity to oxidation; (3) change the binding used to form protein complexes Affinity; (4) change the binding affinity and/or (4) confer or change other physiochemical or functional properties on such polypeptides. According to certain embodiments, single or multiple amino acid substitutions (in certain embodiments It is a conservative amino acid substitution). In certain embodiments, conservative amino acid substitutions generally do not substantially change the structural characteristics of the parent sequence (for example, replacing the amino acid should not tend to break the helix present in the parent sequence, or destroy the characteristics of the parent sequence. Other types of secondary structure). Examples of technically recognized polypeptide secondary and tertiary structures are described in Proteins, Structures and Molecular Principles (Edited by Creighton, WH Freeman and Company [WH Freeman and Company], New York (1984)); Introduction to Protein Structure [edited by C. Branden and J. Tooze, Garland Publishing [Garland Publishing], New York, New York State (1991)); and Thornton et al. Nature [Nature] 354: 105 (1991) ), and each of these documents is incorporated herein by reference.

Consistent with the foregoing, in some aspects, the composition of the present disclosure contains about 110 mg/mL to about 140 mg/mL anti-TSLP antibody, about 0.01% (w/v) ± 0.005% (w/v) polysorbate 80 , Greater than about 2.5% (w/v) and less than about 3.0% (w/v) L-proline, and about 20 mM to about 30 mM acetate, wherein the viscosity of the composition is less than about 20 cP (for example 15 cP), and the pH is less than about 5.5, if necessary, about 5.2. Optionally, the anti-TSLP antibody comprises (A) a light chain variable domain comprising: (i) a light chain CDR1 sequence containing the amino acid sequence shown in SEQ ID NO: 3; ii) a light chain CDR2 sequence containing the amino acid sequence shown in SEQ ID NO: 4; and (iii) a light chain CDR3 sequence containing the amino acid sequence shown in SEQ ID NO: 5; and (B) A heavy chain variable domain comprising: (i) a heavy chain CDR1 sequence containing the amino acid sequence shown in SEQ ID NO: 6; (ii) a heavy chain CDR1 sequence containing the amino acid sequence shown in SEQ ID NO: 7 The heavy chain CDR2 sequence of the amino acid sequence shown, and (iii) the heavy chain CDR3 sequence of the amino acid sequence shown in SEQ ID NO: 8. In an illustrative example, the composition contains about 110 mg/mL of anti-TSLP antibody (for example, tepelumab), 0.01% (w/v) polysorbate 80, about 2.5% (w/v) to About 3.0% (w/v) L-proline, and about 20 mM to about 22 mM acetate, wherein the pH of the composition is about 5.2, wherein the anti-TSLP antibody optionally contains: (A) light chain may A variable domain comprising: (i) a light chain CDR1 sequence containing the amino acid sequence shown in SEQ ID NO: 3; (ii) a light chain CDR1 sequence containing the amine shown in SEQ ID NO: 4 And (iii) the light chain CDR3 sequence containing the amino acid sequence shown in SEQ ID NO: 5; and (B) the heavy chain variable domain, the heavy chain variable structure The domain comprises: (i) the heavy chain CDR1 sequence containing the amino acid sequence shown in SEQ ID NO: 6; (ii) the heavy chain CDR2 sequence containing the amino acid sequence shown in SEQ ID NO: 7, and (iii) The heavy chain CDR3 sequence containing the amino acid sequence shown in SEQ ID NO: 8. In an alternative example, the composition contains about 140 mg/mL of anti-TSLP antibody (for example, teperumumab), 0.01% (w/v) polysorbate 80, and about 2.6% (w/v) To about 2.7% (w/v) L-proline, and about 23 mM to about 25 mM acetate, wherein the pH of the composition is about 5.2, wherein the anti-TSLP antibody optionally contains: (A) light chain A variable domain comprising: (i) a light chain CDR1 sequence containing the amino acid sequence shown in SEQ ID NO: 3; (ii) a light chain CDR1 sequence containing the amino acid sequence shown in SEQ ID NO: 4 The light chain CDR2 sequence of the amino acid sequence; and (iii) the light chain CDR3 sequence containing the amino acid sequence shown in SEQ ID NO: 5; and (B) the heavy chain variable domain, the heavy chain variable The domain comprises: (i) the heavy chain CDR1 sequence containing the amino acid sequence shown in SEQ ID NO: 6; (ii) the heavy chain CDR2 sequence containing the amino acid sequence shown in SEQ ID NO: 7, And (iii) the heavy chain CDR3 sequence containing the amino acid sequence shown in SEQ ID NO: 8.

Consistent with the foregoing, in some aspects, the composition of the present disclosure contains about 180 mg/mL to about 210 mg/mL anti-TSLP antibody, about 0.01% (w/v) ± 0.005% (w/v) polysorbate 80 , 15-190 mM arginine base, 25-200 mM glutamate and 0-250 mM proline acid, where the viscosity of the composition is less than about 22 cP (for example, 15, 17 or 20 cP) at 23°C And the pH is less than about 5.7, if necessary, about 5.4. Optionally, the anti-TSLP antibody comprises (A) a light chain variable domain comprising: (i) a light chain CDR1 sequence containing the amino acid sequence shown in SEQ ID NO: 3; ii) a light chain CDR2 sequence containing the amino acid sequence shown in SEQ ID NO: 4; and (iii) a light chain CDR3 sequence containing the amino acid sequence shown in SEQ ID NO: 5; and (B) A heavy chain variable domain comprising: (i) a heavy chain CDR1 sequence containing the amino acid sequence shown in SEQ ID NO: 6; (ii) a heavy chain CDR1 sequence containing the amino acid sequence shown in SEQ ID NO: 7 The heavy chain CDR2 sequence of the amino acid sequence shown, and (iii) the heavy chain CDR3 sequence of the amino acid sequence shown in SEQ ID NO: 8. In various embodiments, the aqueous composition contains 140 mM arginine base and 150 mM glutamine. In various embodiments, the aqueous composition comprises 140 mM arginine base, 150 mM glutamate, 0.01% (w/v) polysorbate 80, pH 5.4. In various embodiments, the aqueous composition comprises 80 mM arginine base, 85 mM glutamine and 100 mM L-proline. In various embodiments, the aqueous composition comprises 80 mM arginine, 85 mM glutamine, 100 mM L-proline, 0.01% (w/v) polysorbate 80, pH 5.4.

Consistent with the foregoing, in some aspects, the composition of the present disclosure contains about 180 mg/mL to about 210 mg/mL anti-TSLP antibody, about 0.01% (w/v) ± 0.005% (w/v) polysorbate 80 , 10-125 mM arginine, 25-225 mM glutamine and 0-250 mM proline, where the viscosity of the composition is less than about 22 cP at 23°C (for example, 15, 17 or 20 cP) And the pH is less than about 5.7, if necessary, about 5.4. Optionally, the anti-TSLP antibody comprises (A) a light chain variable domain comprising: (i) a light chain CDR1 sequence containing the amino acid sequence shown in SEQ ID NO: 3; ii) a light chain CDR2 sequence containing the amino acid sequence shown in SEQ ID NO: 4; and (iii) a light chain CDR3 sequence containing the amino acid sequence shown in SEQ ID NO: 5; and (B) A heavy chain variable domain comprising: (i) a heavy chain CDR1 sequence containing the amino acid sequence shown in SEQ ID NO: 6; (ii) a heavy chain CDR1 sequence containing the amino acid sequence shown in SEQ ID NO: 7 The heavy chain CDR2 sequence of the amino acid sequence shown, and (iii) the heavy chain CDR3 sequence of the amino acid sequence shown in SEQ ID NO: 8. In various embodiments, the aqueous composition contains 95 mM arginine base and 170 mM glutamine. In various embodiments, the aqueous composition comprises 50 mM arginine base, 95 mM glutamine and 85 mM L-proline. In various embodiments, the aqueous composition contains 95 mM arginine base, 170 mM glutamine, 0.01% (w/v) polysorbate 80, pH 5.4. In various embodiments, the aqueous composition comprises 50 mM arginine base, 95 mM glutamine and 85 mM L-proline, 0.01% (w/v) polysorbate 80, pH 5.4.

Consistent with the foregoing, in some aspects, the composition of the present disclosure contains about 180 mg/mL to about 210 mg/mL anti-TSLP antibody, about 0.01% (w/v) ± 0.005% (w/v) polysorbate 80 , Contains 15-130 mM calcium, 30-300 mM glutamate and 0-250 mM proline, wherein the viscosity of the composition is less than about 20 cP (such as 15 or 17 cP) at 23°C, and the pH Less than about 5.5, if necessary, about 5.0. Optionally, the anti-TSLP antibody comprises (A) a light chain variable domain comprising: (i) a light chain CDR1 sequence containing the amino acid sequence shown in SEQ ID NO: 3; ii) a light chain CDR2 sequence containing the amino acid sequence shown in SEQ ID NO: 4; and (iii) a light chain CDR3 sequence containing the amino acid sequence shown in SEQ ID NO: 5; and (B) A heavy chain variable domain comprising: (i) a heavy chain CDR1 sequence containing the amino acid sequence shown in SEQ ID NO: 6; (ii) a heavy chain CDR1 sequence containing the amino acid sequence shown in SEQ ID NO: 7 The heavy chain CDR2 sequence of the amino acid sequence shown, and (iii) the heavy chain CDR3 sequence of the amino acid sequence shown in SEQ ID NO: 8. In various embodiments, the aqueous composition contains 100 mM calcium and 230 mM glutamine. In various embodiments, the aqueous composition contains 60 mM calcium, 140 mM glutamine, and 70 mM L-proline. In various embodiments, the aqueous composition contains 100 mM calcium, 230 mM glutamate, 0.01% (w/v) polysorbate 80, and pH 5.0. In various embodiments, the aqueous composition contains 60 mM calcium, 140 mM glutamate, 70 mM L-proline, 0.01% (w/v) polysorbate 80, pH 5.0.

Consistent with the foregoing, in some aspects, the composition of the present disclosure contains about 180 mg/mL to about 210 mg/mL anti-TSLP antibody, about 0.01% (w/v) ± 0.005% (w/v) polysorbate 80 , 15-195 mM calcium and 25-320 mM and 0-220 mM proline, wherein the viscosity of the composition is less than about 20 cP (such as 15 or 17 cP) at 23°C, and the pH is less than about 5.5, depending on Need, about 5.0. Optionally, the anti-TSLP antibody comprises (A) a light chain variable domain comprising: (i) a light chain CDR1 sequence containing the amino acid sequence shown in SEQ ID NO: 3; ii) a light chain CDR2 sequence containing the amino acid sequence shown in SEQ ID NO: 4; and (iii) a light chain CDR3 sequence containing the amino acid sequence shown in SEQ ID NO: 5; and (B) A heavy chain variable domain comprising: (i) a heavy chain CDR1 sequence containing the amino acid sequence shown in SEQ ID NO: 6; (ii) a heavy chain CDR1 sequence containing the amino acid sequence shown in SEQ ID NO: 7 The heavy chain CDR2 sequence of the amino acid sequence shown, and (iii) the heavy chain CDR3 sequence of the amino acid sequence shown in SEQ ID NO: 8. In various embodiments, the aqueous composition contains 110 mM calcium and 240 mM glutamine. In various embodiments, the aqueous composition contains 70 mM calcium, 145 mM glutamate, and 60 mM L-proline. In various embodiments, the aqueous composition contains 110 mM calcium, 240 mM glutamate, 0.01% (w/v) polysorbate 80, and pH 5.0. In various embodiments, the aqueous composition contains 70 mM calcium, 145 mM glutamate, 60 mM L-proline, 0.01% (w/v) polysorbate 80, pH 5.0.

Instructions

Without being bound by a particular theory and based at least in part on the data provided herein, the composition of the present disclosure is particularly suitable for treating patients with inflammatory diseases. As used herein, "inflammatory disease" refers to a medical condition involving abnormal inflammation caused by the immune system attacking the body's own cells or tissues, which may lead to chronic pain, redness, swelling, stiffness, and damage to normal tissues . Inflammatory diseases include, for example, asthma, chronic peptic ulcer, tuberculosis, periodontitis, sinusitis, active hepatitis, joint adhesive spondylitis, rheumatoid arthritis, chronic obstructive pulmonary disease (COPD), Crohn Disease, ulcerative colitis, osteoarthritis, atherosclerosis, systemic lupus erythematosus, atopic dermatitis, eosinophilic esophagitis (EoE), nasal polyps, chronic spontaneous urticaria, Ig-driven Diseases (such as IgA nephropathy & lupus nephritis), eosinophilic gastritis, chronic sinusitis without nasal polyps, and idiopathic pulmonary fibrosis (IPF), etc. In an exemplary aspect, the inflammatory disease is asthma, atopic dermatitis, COPD, eosinophilic esophagitis (EoE), nasal polyps and chronic spontaneous urticaria, Ig-driven diseases (such as IgA nephropathy & lupus Nephritis), eosinophilic gastritis, chronic sinusitis without nasal polyps, and idiopathic pulmonary fibrosis (IPF). In an exemplary aspect, the inflammatory disease is atopic dermatitis (AD). In many respects, this inflammatory disease is asthma. In many respects, the inflammatory disease is COPD.

Therefore, this article provides the use of the composition of the present disclosure for the treatment of inflammatory diseases. In an exemplary aspect, the inflammatory disease is selected from the group consisting of asthma, atopic dermatitis, chronic obstructive pulmonary disease (COPD), eosinophilic esophagitis (EoE), nasal polyps, chronic spontaneous urticaria Measles, Ig-driven diseases (such as IgA nephropathy & lupus nephritis), eosinophilic gastritis, chronic sinusitis without nasal polyps, and idiopathic pulmonary fibrosis (IPF). If necessary, the inflammatory disease is atopic dermatitis. In many respects, this inflammatory disease is asthma. In many respects, the inflammatory disease is COPD. The present disclosure also provides methods for treating inflammatory diseases in subjects. In an exemplary embodiment, the method includes administering to the subject a therapeutically effective amount of a composition of the present disclosure. In many respects, the inflammatory disease is selected from the group consisting of asthma, atopic dermatitis, chronic obstructive pulmonary disease (COPD), eosinophilic esophagitis (EoE), nasal swelling, chronic spontaneous urticaria Measles, Ig-driven diseases (such as IgA nephropathy & lupus nephritis), eosinophilic gastritis, chronic sinusitis without nasal polyps, and idiopathic pulmonary fibrosis (IPF). If necessary, the inflammatory disease is atopic dermatitis. Under different circumstances, the composition of the present disclosure is administered to the subject by subcutaneous administration. In an illustrative example, about 1 mL to about 5 mL (eg, about 1 mL to about 3 mL) of the aqueous composition is administered to the subject.

asthma

The term "asthma" as used herein refers to allergic asthma, non-allergic asthma, eosinophilic asthma, and non-eosinophilic asthma.

The term "allergic asthma" as used herein refers to asthma caused by one or more inhaled allergens. Such patients have positive IgE Fluorescence Enzyme Immunoassay (FEIA) levels for one or more allergens that cause asthma.

Generally, most allergic asthma is related to Th2 type inflammation.

The term "non-allergic asthma" refers to patients with low eosinophils, low Th2, or low IgE at the time of diagnosis. Generally, in the IgE Fluorescence Enzyme Immunoassay (FEIA), patients with "non-allergic asthma" have a negative reaction to a group of allergens including region-specific allergens. Except for those with low IgE, those patients often have a low eosinophil count or no eosinophil count and a low Th2 count at the time of diagnosis.

As used herein, the term "severe asthma" refers to asthma that requires high-intensity treatment (eg GINA steps 4 and 5) to maintain good control or despite high-intensity treatment but still not well controlled (GINA, Global Strategy for Asthma Management and Prevention [Global Asthma Management and Prevention Strategy]. Global Initiative for Asthma [Global Initiative for Asthma] (GINA, December 2012).

The term "eosinophilic asthma" as used herein refers to patients with asthma who have a screening blood eosinophil count of ≥ 250 cells/μL. "Low eosinophilic" asthma refers to patients with asthma with less than 250 cells/uL of blood or serum.

The term "Th2-type inflammation" as used herein refers to subjects with a screening blood eosinophil count of ≥ 140 cells/μL and a screening total serum IgE level of> 100 IU/mL (Corren et al., N Engl J Med. [New England Journal of Medicine] 22; 365 (12): 1088-98, 2011). "Th2 high" asthma population or profile refers to subjects with IgE> 100 IU/mL and blood eosinophil count ≥ 140 cells/μL. "Th2 low" asthma cluster system refers to subjects with IgE <100 IU/mL and blood eosinophil count ≤ 140 cells/μL

Atopic dermatitis

Under different circumstances, the inflammatory disease is atopic dermatitis (AD), which is a common allergic inflammatory skin disease, also known as eczema. AD is the most common skin disease in children and is characterized by severe itching and inflammation of the skin, as well as chronic mossy, multi-scaly plaques. Although the cause of AD is unclear, the current theory believes that AD is a major skill disorder that causes defects in other atopic diseases. AD is reviewed in Kapur et al., Atopic dermatitis. [Atopic Dermatitis] Allergy Asthma Clin Immunol [Allergy Asthma and Clinical Immunology] 14 , 52 (2018) doi:10.1186/s13223-018-0281-6. Like asthma and allergic rhinitis, AD also involves T helper type 2 (Th2) cell-mediated allergic inflammation, which is caused by the secretion of IL-4, IL-5, IL-13, and TNFα by CD4+ T cells. These cytokines cause B cells to produce IgE antibodies, and IgG binds to obese cells to promote the initiation of allergic reactions and drive white blood cells into the skin dermis. Indra, Exper Rev Proteomics [Proteomics expert review] 10 (4): 309-311 (2013). In an exemplary aspect, AD is characterized by higher performance of TSLP. In various aspects, AD is characterized by TSLP secretion by epidermal keratinocytes, which triggers TH2 cytokine-related inflammation.

In an exemplary aspect, AD is chronic and can occur periodically. In various aspects, the patient experiences one or more of the following AD symptoms: • Dry skin, itching (may be severe, especially at night), • Red to brown or gray spots (for example, on hands, feet, ankles, wrists, neck, upper chest, eyelids, inner elbows or knees, face or scalp), • Small, raised bumps, • Thickened and cracked scaly skin, and • Scratched, sensitive, swollen skin due to scratching. (From mayoclinic.org/diseases-conditions/atopic-dermatitis-eczema/symptoms-causes/syc-20353273)

In many aspects, AD coexists with one or more of asthma, hay fever, chronic itching, scaly skin, skin infections, irritant hand dermatitis, allergic rhinitis, allergic contact dermatitis, or sleep problems.

In various embodiments, the use of the disclosed composition eliminates the need for corticosteroid therapy or other drugs for the treatment of AD.

In an alternative embodiment, the method includes administering the composition of the present disclosure in combination with another anti-inflammatory or anti-AD treatment. For example, in various aspects, the method further comprises administering corticosteroids (such as prednisone) or calcineurin inhibitors (such as tacrolimus, pimecrolimus), Antibiotics or biological products (such as dupilumab). In an exemplary aspect, the method further comprises phototherapy treatment, such as phototherapy with sunlight, UVA or UVB.

Subject

The subject is expected to be human. The subject can be an adult, adolescent, or a child.

In various aspects, the subject exhibits signs or symptoms of an inflammatory disease such as AD or COPD, such as any one or more of the above. In various aspects, the subject also suffers from one or more of asthma, hay fever, chronic itching, scaly skin, skin infection, irritant hand dermatitis, allergic contact dermatitis, or sleep problems. In various aspects, the subject has previously been or is on anti-AD or anti-inflammatory therapy (eg, corticosteroids (eg Praisu) or calcineurin inhibitors (eg tacrolimus, pimecrolimus) , Antibiotics or biological products (such as Dupi Luzumab)) for treatment. In an exemplary aspect, the method further comprises phototherapy treatment, such as phototherapy with sunlight, UVA or UVB. If necessary, subjects have never received any of the following treatments: corticosteroids (e.g. Praisu) or calcineurin inhibitors (e.g. tacrolimus, pimecrolimus), antibiotics or biological products (e.g. Dupiluzumab). In an exemplary aspect, the method further comprises phototherapy treatment, such as phototherapy with sunlight, UVA or UVB.

Treatment plan, dosage and route of administration

The therapeutic antibody (or antibody variant) composition can be delivered to the patient at multiple locations. Multiple administrations can be carried out at the same time or can be administered over a period of time. In some cases, it may be beneficial to provide a continuous flow of the therapeutic composition. Other therapies can be administered periodically, such as hourly, daily, weekly, every 2 weeks, every 3 weeks, monthly, or at longer intervals.

In various embodiments, the amount of a therapeutic agent (such as a bivalent antibody with two TSLP binding sites) at a given dose may vary according to the size of the individual to be administered the therapy and the characteristics of the condition being treated.

In an exemplary treatment, the dosage of the anti-TSLP antibody or antibody variant provided by the composition is in the range of about 210 mg to about 420 mg per day. For example, the dose can be provided in about 210 mg, 280 mg, or 420 mg. In various embodiments, the composition comprising an anti-TSLP antibody or antibody variant has an amount of about 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360 , 370, 380, 390, 400, 410, or 420 mg per dose. These concentrations can be administered in a single dosage form or in multiple doses. The above dose is given every two weeks or every four weeks. In various embodiments, the anti-TSLP antibody or antibody variant is administered in a single dose of 280 mg or 420 mg every two weeks or every four weeks. In various embodiments, the anti-TSLP antibody or antibody variant is administered in a single dose of 210 mg every two weeks or every four weeks. In various embodiments, a composition containing greater than about 100 mg/mL, or greater than about 140 mg/ml of an anti-TSLP antibody as described herein is administered to the subject every two weeks or every four weeks.

For antibody variants, the amount of antibody variant should be such that the number of TSLP binding sites in the dose is equal to the number of TSLP binding sites of the above-mentioned standard bivalent antibody.

It is expected that the composition of the present disclosure including the anti-TSLP antibody or antibody variant will be administered every 2 weeks or every 4 weeks in a period of less than 4 months, 6 months, 9 months, 1 year or longer. In various embodiments, the administration is subcutaneous or intravenous.

Production method

This article further provides methods for preparing the composition of the present disclosure. Therefore, a method for preparing a stable liquid antibody composition with a viscosity less than about 100 cP is further provided, the composition comprising (A) an anti-TSLP antibody at a concentration greater than about 140 mg/mL, (B) a surfactant, and (C ) Basic amino acids or their salts, calcium salts, magnesium salts, or combinations thereof. In an exemplary embodiment, the method includes: (i) combining the antibody with a basic amino acid or its salt, calcium salt, magnesium salt, or The combined aqueous solution combination and (ii) addition of surfactants to achieve a final concentration of about 0.01% (w/v) ± 0.005% (w/v) surfactants.

In an exemplary aspect, the composition comprises about 160 mg/mL to about 250 mg/mL of anti-TSLP antibody, optionally about 180 mg/mL to about 225 mg/mL or about 180 mg/mL to about 200 mg/mL . Optionally, the composition contains about 160 mg/mL to about 250 mg/mL of anti-TSLP antibody, and about 165 mg/mL to about 225 mg/mL or about 165 mg/mL to about 200 mg/mL as needed. In various aspects, the composition includes about 175 mg/mL to about 185 mg/mL of anti-TSLP antibody, as needed, about 175 mg/mL, about 176 mg/mL, about 177 mg/mL, about 178 mg/mL, About 179 mg/mL, about 180 mg/mL, about 181 mg/mL, about 182 mg/mL, about 183 mg/mL, about 184 mg/mL, about 185 mg/mL. In various aspects, the composition contains about 180 mg/mL of anti-TSLP antibody. Under different circumstances, the concentration of the anti-TSLP antibody ranges from about 189 mg/mL or about 190 mg/mL to about 230 mg/mL or about 231 mg/mL. If necessary, the concentration of anti-TSLP antibody is about 205 mg/mL to about 215 mg/mL, and about 210 mg/mL or about 205 mg/mL, about 206 mg/mL, about 207 mg/mL, about 208 mg as needed /mL, about 209 mg/mL, about 210 mg/mL, about 211 mg/mL, about 212 mg/mL, about 213 mg/mL, about 214 mg/mL, about 215 mg/mL. In an exemplary aspect, the aqueous solution contains an organic salt of arginine, lysine, or histidine. In an exemplary aspect, the aqueous solution comprises arginine acetate, arginine aspartate, arginine glutamate, arginine glycolate, arginine lactate, arginine methanesulfonate Salt, arginine propionate, histidine acetate, histidine aspartate, histidine glutamate, histidine glycolate, histidine lactate, histidine methyl Sulfonate, histidine propionate, lysine acetate, lysine aspartate, lysine glutamine, lysine glycolate, lysine lactate, lysine Methanesulfonate, lysine propionate, calcium acetate, calcium aspartate, calcium glutamine, calcium glycolate, calcium lactate, calcium methanesulfonate, calcium propionate, magnesium acetate, magnesium aspartate , Magnesium glutamate, magnesium glycolate, magnesium lactate, magnesium methanesulfonate, magnesium propionate, or a combination thereof. In various aspects, the aqueous solution contains about 15 mM to about 200 mM or about 50 mM to about 150 mM salt (alkaline amino acid salt, calcium salt, magnesium salt). In some aspects, the surfactant is polysorbate 80 or polysorbate 20. In an illustrative example, the surfactant is polysorbate 80, and the final concentration of PS80 is about 0.01% (w/v). In an exemplary embodiment, the anti-TSLP antibody system is Teperumumab.

Pharmacy box

The present disclosure also provides a kit, which includes the composition described herein, as well as package inserts, package labels, instructions, or other markings that guide or expose any method or implementation disclosed herein. In certain embodiments, the present disclosure provides kits for generating single-dose administration units. In certain embodiments of the present disclosure, a kit including single-chamber and multi-chamber pre-filled syringes (such as liquid syringes) is included.

The following examples are given only to illustrate the present invention, but not to limit its scope in any way. Instance

Throughout the examples provided herein, the following abbreviations are used: DF, diafiltration; PS80, polysorbate 80; SEC, size exclusion chromatography, F#, formulation number. In addition, throughout the examples, the composition of the DF buffer used to make the final formulation containing tepeluxumab and the estimated concentration of the components of the final formulation are provided.

The final concentration of some components of the final formulation analyzed (for example, after storage, as needed, for stability, viscosity) is different from the concentration of DF or dialysis buffer, depending on the presence or absence of relative ions. In the absence of counter ions, the formulation has low ionic strength. In this case, acetate is co-concentrated with tepelumab so that the final formulation contains a higher acetate concentration relative to the concentration of DF or dialysis buffer. For example, when neither the DF buffer nor the final formulation contains a salt (such as arginine HCl) and therefore has a low ionic strength, the use of a DF buffer containing 10 mM acetate results in the inclusion of 110 mg/mL teparuzumab About 20 mM to about 22 mM acetate is present in the formulation (pH 5.2). Similarly, in the absence of salts (such as arginine HCl), a DF buffer containing 10 mM acetate results in a formulation containing 140 mg/mL tepeluzumab (pH 5.2) in the presence of about 23 mM to about 25 mM acetate. When a salt (such as arginine HCl) is present, acetate is not co-concentrated with teperumumab, and therefore the acetate concentration of the DF buffer and the acetate concentration of the final composition are generally equal. In addition, excipients may be subject to size exclusion, or may be affected by non-specific interactions. For example, in a 110 mg/mL tepilumab formulation, the proline concentration can be as much as about 16.67% lower than the concentration indicated in the DF buffer, and in a 140 mg/mL tepilumab formulation, The proline concentration can be as much as about 10% to about 13.3% lower than the concentration indicated in the DF buffer. Based on the above, and throughout the following examples, taking into account the excipient exclusion and acetate co-concentration effects described above, the concentrations of the components of the final formulation are provided. Example 1

This example demonstrates an exemplary method of producing a high-concentration tepelumab formulation.

A series of studies have been conducted to develop formulations containing high concentrations (for example,> 70 mg/mL) of Teperumumab. Because the formulation is intended for subcutaneous administration, the formulation needs to be isotonic and exhibit a viscosity suitable for the route of administration. For example, the formulation of tepelumab (110 mg/mL) needs to have a viscosity of about 15 cP at 23°C. It is also expected that the formulation will be stable after storage for more than 1 year (eg, at least 2 or 3 years) at 2°C to 8°C.

In order to prepare a high-concentration formulation of tepelumab, the initial solution containing tepelumab (70 mg/mL) in acetate (pH 5.2) was dialyzed against diafiltration (DF) buffer. A total of 10 buffer changes were used to achieve a complete buffer exchange. Using a centrifuge-concentrator, the buffer-exchanged tepeluzumab solution was over-concentrated to the concentration of tepeluzumab, which was about 110% of the target tepeluzumab concentration. For example, the buffer-exchanged tepeluzumab solution is over-concentrated to about 200 mg/mL to bring the target tepeluzumab concentration to 180 mg/mL. Using the same DF buffer used in the buffer exchange step, this over-concentrated solution was diluted to the target teperumumab concentration.

Following the above procedure, a series of tepelumumab formulations with different concentrations of tepelumumab (ranging from about 150 mg/mL to about 250 mg/mL) were prepared for viscosity studies. Two DF buffers are used in the following formulations: a first DF buffer containing arginine (arginine glutamate) and a second DF buffer containing proline. Arginine is present in DF buffer at a concentration of 150 mM, and proline is present in DF buffer at a concentration of 3% (w/v). A rotary viscometer was used to test the viscosity of samples of each formulation at 23°C. The reported viscosity value is at a shear rate of ^{1000 s -1.}

Figure 1 provides the graphical results of this determination. Figure 1 plots the viscosity of each formulation as a function of the concentration of Teperumumab. As shown in the figure, no matter what DF buffer is used, as the concentration of Teperumumab increases, the viscosity will also increase. For the formulations prepared with the first DF buffer, the viscosity was generally lower, which supports the use of spermine in the low viscosity formulation of tepeluxumab.

Since lysine is similar in structure to arginine (because both are basic amino acids), lysine glutamate was tested as an excipient for viscosity reduction and compared with arginine. The formulations of glutamine salt were compared. Each formulation contained about 195 mg/mL of teperumumab, and the DF buffer used to prepare the final formulation contained 100 mM arginine glutamine or 100 mM lysine glutamine. A rotary viscometer was used to test the viscosity of samples of each formulation at 23°C. The reported viscosity value is at a shear rate of ^{1000 s -1.} The viscosity of the formulation containing lysine glutamate was 48.9 cP, while the viscosity of the formulation containing arginine glutamate was 35.3 cP. These data indicate that the two basic amino acids or their salts perform well in reducing the viscosity of formulations containing high concentrations of teperumumab. Example 2

This example demonstrates the viscosity-reducing effect of different excipients on the formulation of Teperumumab.

Since arginine and lysine can well reduce the viscosity of formulations containing high concentrations of teperumumab, various arginine and lysine acid analogues are used to incorporate the samples of teperumumab. Then test the viscosity of these samples. The arginine salts used in this study are arginine hydrochloride, arginine acetate, and arginine glutamine. The lysine analogue (N-acetyl lysine (NAK)) was tested together with samples containing lysine. The effects of calcium and sodium salts on viscosity were also tested. In particular, calcium chloride, calcium acetate, and sodium chloride are used.

By accurately mixing a concentrated (5x or 10x) stock solution containing one excipient into an aliquot of the concentrated tepeluxumab solution, a sample containing one of the above-mentioned excipients is generated. By using a dialysis tube with a molecular weight cut-off of 10,000, the aqueous solution containing 110 mg/mL teperumumab was dialyzed against 10 mM sodium acetate (pH 4.4), and then the dialysate was concentrated using an Amicon Ultra centrifugal concentrator to provide An aqueous composition containing ≥ 230 mg/mL Tepe Luzumab was used to prepare a concentrated Tepe Luzumab solution. This method creates a sample set with matching concentrations.

Before the viscosity test, after 5-fold dilution by weight, the concentration of each sample was confirmed by UV absorption slope spectroscopy using Solo-VPE (C-Technologies). Make sure that the concentration of tepelumab in each sample is approximately 210 mg/mL. The concentration of each excipient in the sample is about 100 mM or about 150 mM. Figure 2 shows the concentration of each excipient. A Seven Easy pH meter (Mettler Toledo) was used to determine the final pH of the sample.

The viscosity of each sample was basically tested as described in Example 1 and compared with a control sample containing no excipients or containing proline or sucrose.

The results are shown in Figure 2. As shown in Figure 2, all samples containing arginine, lysine or NAK showed a lower viscosity than the control. Although the calcium acetate-containing sample showed a lower viscosity than the control, the calcium chloride-containing sample showed the same viscosity as one of the controls (proline control). Sodium chloride and arginine HCl can only slightly reduce the viscosity performance. Although the viscosity of the samples with chloride-containing excipients was lower than the control, the performance of these samples was not as good as the chloride-free excipients. Example 3

This example demonstrates the viscosity-reducing effect of different excipients on high-concentration teperumumab formulations.

Another study was conducted to test the viscosity-lowering effect of another arginine salt. In this study, it was determined that the concentration of teparumab in each sample was approximately 190 mg/mL, and the samples contained 60 mM of one of the following excipients: arginine hydrochloride, arginine Acetate, arginine glutamate, arginine propionate, arginine aspartate, arginine methanesulfonate, arginine glycolate, or arginine phosphate. In this study, a control sample without excipients was prepared and tested. The samples were prepared and tested for viscosity essentially as described in Examples 1 and 2.

The results are provided in Figure 3. As shown in Figure 3, all samples containing arginine showed a lower viscosity than the control. Among the samples containing spermine, the samples containing arginine hydrochloride showed the highest viscosity, which is consistent with the data in Figure 2, which indicates that samples containing chloride-containing excipients are usually not well used. Used as an excipient to reduce viscosity. Example 4

This example demonstrates the effect of combined excipients on viscosity.

Since arginine acetate and calcium acetate are very effective in reducing the viscosity of high-concentration teperumumab samples (Figure 2), the results of combining these excipients with each other or with another excipient were measured. Effect. NAK, sodium acetate, and proline were combined with arginine acetate in three different samples of teparumab, and in another sample, calcium acetate was combined with proline. As a control, a sample containing no excipients or a sample containing sodium acetate or proline was used. In addition, in this study, the test polyvinylpyrrolidone (PVP) content ranged from 0.5% (w/v) to 3% (w/v). Each sample contained approximately 210 mg/mL tepelumab and was prepared according to the procedure described in Example 2. A summary of the samples is described in Table 1. [Table 1] sample# Excipients in the sample 1 No excipient-control 2 Proline (150 mM) 3 Acetic acid Na (150mM) 4 Arg acetate (75 mM) + proline (75 mM) 5 Arg acetate (75 mM) + NAK (75 mM) 6 Arg acetate (75 mM) + acetic acid Ca (75 mM) 7 Arg acetate (75 mM) + Na acetate (100 mM) 8 PVP (0.5% (w/v)) 9 PVP (1.0% (w/v)) 10 PVP (2.0% (w/v)) 11 PVP (3.0% (w/v))

The viscosity was determined essentially as described in Example 1.

The results are shown in Figure 4. As shown in Figure 4, the combination of proline or sodium acetate and arginine acetate resulted in a lower viscosity relative to the samples containing proline as the sole excipient or sodium acetate as the sole excipient. The combination of proline and calcium acetate also resulted in a decrease in viscosity compared to samples containing only proline. Sodium acetate only appears to reduce viscosity when combined with arginine acetate. The samples containing both arginine acetate and calcium acetate showed the lowest viscosity of all the samples tested in this study. PVP at any test concentration could not reduce the viscosity of the Teperumumab sample at all.

Additional studies were conducted to test the combination of spermine salt with a second excipient. In this study, glycine and tris acetate were tested with or without arginine acetate. As a control, samples containing no excipients or samples containing proline or tris hydrochloride were used in this study. In addition, samples containing other arginine, arginine propionate and arginine mesylate were tested. In addition, samples containing tris hydrochloride were prepared and tested. Each sample contained approximately 210 mg/mL tepelumab and was prepared according to the procedure described in Example 2. Table 2 provides a summary of the excipients and their concentrations in each sample. [Table 2] sample# Excipients in the sample 12 No excipient-control 13 Proline (150 mM) 14 Glycine (150 mM) 15 Tris HCl (150 mM) 16 Tris acetate (150 mM) 17 Arginine propionate (150 mM) 18 Arginine aspartate (150 mM) 19 Arginine mesylate (150 mM) 20 Arginine aspartate (75 mM) + Tris acetate (75 mM) twenty one Arginine aspartate (75 mM) + glycine (75 mM)

The viscosity of the formulation was tested and compared to a control formulation that contained no excipients or contained 150 mM proline. The viscosity was determined essentially as described in Example 1.

Figure 5 provides the viscosity results for each formulation. As shown in the figure, the combination of arginine acetate and tris acetate or glycine resulted in a lower viscosity compared to samples containing only tris acetate or only glycine. The poor performance of Tris hydrochloride as a viscosity-reducing agent is consistent with previous studies, which showed that chloride-containing excipients cannot reduce the viscosity of high-concentration tepirumumab samples very well. Example 5

This example demonstrates the effect of several excipients in reducing the viscosity of samples containing high concentrations of teperumumab.

Basic amino acids, arginine, and lysine performed well in reducing the viscosity of formulations containing high concentrations of teperumumab. In one study, a sample containing histidine was used and compared with a sample containing arginine. In particular, a sample containing about 190 mg/mL tepeluzumab was prepared according to the procedure described in Example 2. The sample contains 60 mM histidine, or 60 mM arginine (arginine hydrochloride, arginine glutamate, arginine propionate, arginine aspartate, arginine Methanesulfonate, arginine glycolate, arginine phosphate). Samples containing 60 mM N-acetylarginine (NAR) or 60 mM methionine were also prepared. The viscosity of the formulation was tested essentially as described in Example 1 and compared with a control formulation without excipients.

The results are shown in Figure 6. As shown in Figure 6, all arginine salts, except spermine hydrochloride, were able to reduce the viscosity of high-concentration teparuzumab samples very well. The histidine sample is as effective as most spermine. Samples containing NAR and Met are as effective as arginine hydrochloride in reducing viscosity. Example 6

This example demonstrates the effect of several combinations of excipients containing arginine or calcium salt on the viscosity-reducing effect of samples containing high concentrations of tepeluxumab.

In the preliminary study, samples containing approximately 190 mg/mL tepirumumab and 60 mM NAR and samples containing approximately 190 mg/mL tepirumumab and 60 mM methionine were prepared. The viscosity of each sample was measured basically as described in Example 1. As shown in Figure 7, NAR and methionine reduced the viscosity to below 60 cP.

Since arginine acetate and calcium acetate are very effective in reducing the viscosity of high-concentration teperumumab formulations (Figure 2), the effect of combining these excipients with NAR or methionine was tested . To this end, a series containing about 180 mg/mL or about 210 mg/mL tepirumumab and arginine glutamine or calcium glutamine ( Alone or in combination with NAR or methionine, and in some cases with both NAR and methionine) samples. The pH of each sample is in the range of 5.1 to 5.3. Figure 8 provides the concentration of each of the above excipients in each sample and the pH and viscosity of each sample.

As shown in Fig. 8, the viscosity of the sample containing a lower concentration of tepelumab was lower than the viscosity of the sample containing a higher concentration of tepelumab. The viscosity range of samples with a lower concentration of tepilumab is about 22 cP to about 30 cP, while the viscosity of samples with a higher concentration of tepilumab is in the range of about 48 cP to about 64 cP. Generally, the viscosity of a sample containing calcium glutamate is lower than that of a sample containing arginine glutamate. For lower concentrations of teparumab, the addition of NAR, methionine, or both will further reduce the viscosity of the sample. Example 7

This example demonstrates the viscosity-reducing effect of several excipients on formulations containing high concentrations of Teperumumab.

Prepared according to the procedure described in Example 2 containing about 210 mg/mL tepirumumab and arginine or other excipients known to reduce viscosity (i.e. β-alanine, creatine and L -Serine) samples. Each sample contains 150 mM of one of the following: β-alanine, creatine, L-serine, proline, or arginine (arginine propionate, arginine aspartate) , Arginine methanesulfonate, arginine glycolate, phosphate). Test the viscosity of the sample and compare it with a control sample without excipients or with 150 mM proline. The viscosity was determined essentially as described in Example 1.

The results are shown in Figure 9A. Consistent with previous studies, arginine can reduce the viscosity of samples containing high concentrations of tepeluxumab (Figure 9A). The viscosity ranges from about 45 cP to about 65 cP. β-alanine, creatine, and L-serine were not able to reduce the viscosity of the Teperumumab formulation very well (Figure 9A).

In another study, a formulation containing about 195 mg/mL teperumumab and betaine, taurine, or proline was prepared following the procedure described in Example 2. Test the viscosity of the sample and compare it with a control sample without excipients. The viscosity was determined essentially as described in Example 1.

The results are shown in Figure 9B. Consistent with previous studies, samples containing arginine had the greatest reduction in viscosity. The sample containing taurine had a moderate effect on the viscosity; the viscosity was slightly lower than that of the sample containing proline (Figure 9B). The samples containing betaine had no effect on viscosity (Figure 9B). Example 8

This example demonstrates the effect of magnesium salt on the viscosity of a sample with a high concentration of Taipa Luzumab.

A sample containing approximately 210 mg/mL tepelumab and 150 mM magnesium acetate was prepared according to the procedure described in Example 2. Test the viscosity of this sample and compare it with a control sample that contains no excipients or contains 150 mM proline. In addition, as a positive control, a sample containing 150 mM arginine acetate was prepared and tested for viscosity. In order to determine whether a higher amount of proline can well reduce the viscosity of samples with high teliperuzumab concentration, a formulation containing 300 mM proline was prepared and tested. The viscosity was determined essentially as described in Example 1.

The results are shown in Figure 10. As shown in the figure, magnesium acetate is as effective as arginine acetate, and can reduce the viscosity of the Teperumumab sample to just below 60 cP. Proline of either concentration (150 mM or 300 mM) will not reduce the viscosity of the sample (as well as arginine acetate and magnesium acetate), but the viscosity of the two samples containing proline is lower than that without excipients. The viscosity of the control of the form. These data indicate that the magnesium salt has a viscosity-lowering effect on the Tepa Luzumab formulation. Example 9

This example demonstrates the effect of histamine on the viscosity of the concentration of Gaotaipe Luzumab.

According to the procedure described in Example 2, a sample containing about 210 mg/mL teperumumab and arginine, histamine, calcium salt, or a combination thereof was prepared. The arginine salts used in this study are arginine glycolate and arginine glutamate. The histamine salts used in this study are histidine glycolate and histidine glutamate. The calcium salt used in this study is calcium acetate. A summary of excipients for each sample is described in Table 3. [table 3] sample# Excipients in the sample twenty two Control-no excipients twenty three Proline (150 mM) twenty four Arginine Glutamine (100 mM) 25 Arginine glycolate (100 mM) 26 Arginine glutamate (50 mM) and arginine glycolate (50 mM) 27 Histidine glutamate (100 mM) 28 Histidine glycolate (100 mM) 29 Histidine glutamate (50 mM) and histidine glycolate (50 mM) 30 Calcium acetate (100 mM) 31 Arginine glutamate (33 mM) and histidine glutamate (33 mM) and calcium acetate (33 mM) 32 Arginine glycolate (33 mM) and histidine glycolate (33 mM) and calcium acetate (33 mM)

The viscosity of the sample was tested and compared with a control formulation without excipients or with 150 mM proline. The viscosity was determined essentially as described in Example 1.

The results are shown in Figure 11. As shown in this figure, all samples containing arginine, histamine, calcium salt, or a combination thereof reduced the viscosity to less than 75 cP, while the control showed a viscosity higher than 100 cP. Example 10

This example demonstrates the effect of arginine glycolate and arginine aspartate on the viscosity of a high taipelumab concentration formulation.

Prepared according to the procedure described in Example 2 containing about 210 mg/mL teperumumab and arginine glycolate (at a concentration of 50 mM, 75 mM, 125 mM, or 150 mM) or arginine aspartate Amino acid salt (concentration of 50 mM, 75 mM, 125 mM, or 150 mM) samples. Test the viscosity of the sample and compare it with a control without excipients or with 150 mM proline. The viscosity was determined essentially as described in Example 1.

The results are shown in Figure 12. As shown in the figure, all samples containing arginine at any concentration reduced the viscosity to less than 100 cP, while the control showed a viscosity higher than 110 cP. Although arginine glycolate has a greater effect, increasing the amount of arginine will reduce viscosity.

This example demonstrates the dose-viscosity-lowering effect of arginine. Example 11

This example demonstrates the effect of pH on the viscosity of formulations with high telumumab concentration.

A sample containing approximately 210 mg/mL teperumumab and 150 mM arginine aspartate or 150 mM histidine acetate was prepared according to the procedure described in Example 2. For the arginine aspartate sample, the pH varied from 4.75 to 5.7, and for the histidine acetate sample, the pH varied from 5.5 to 6.5. Test the viscosity of the sample and compare it with a control without excipients or with 150 mM proline. The viscosity was determined essentially as described in Example 1.

The results are shown in Figures 13A and 13B. As shown in the figure, the salts reduce the viscosity to less than 50 cP (for histamine) or less than 60 cP (for arginine), and the changing pH has no negative effect on the viscosity reduction. .

These data support that formulations containing high concentrations of teperumumab and arginine or histamine and having a pH of about 4.75 to about 6.5 exhibit reduced viscosity. Example 12

This example demonstrates the stability of a Teperumumab formulation containing arginine after storage for 1 week at 40°C.

A series of samples were prepared according to the procedure described in Example 2. Each sample contained approximately 195 mg/mL teperumumab and 150 mM arginine. Test the stability of the sample and compare it with a control that does not contain excipients or contains 150 mM proline. For stability testing, the samples were filled into containers and then stored at 40°C for up to 1 week. The sample is tested by size exclusion chromatography (SEC) to determine the stability of the sample at multiple storage time points. The percentage of high molecular weight (HMW) substances in the sample is reported and reflects the amount of HMW substances formed after the storage period. The results of the stability measurement are shown in Table 4 below. Shows the percentage of high molecular weight (HMW) substances for each sample. The lower the HMW%, the higher the stability of the formulation. [Table 4] excipient % HMW Control (no excipients) 3.42 Proline 3.25 Arg HCl 10.61 Arg acetate 7.93 Arg Glutamate 4.37 Arg glycolate 6.39 Arg propionate 16.55 Arg mesylate 8.33 Arg aspartate 4.47 Arg Phosphate 3.14

These data indicate that the identity of the relative ion affects the formation of HMW, thereby affecting the stability of teparumab. Example 13

This example demonstrates the viscosity and stability of an exemplary formulation of Teperumumab of the present disclosure after storage at -30°C, 5°C, and 25°C for up to 6 months.

In order to study the effect of high pacilinumab concentration formulations on viscosity and stability after storage at different temperatures and different times, and to evaluate the robustness of the formulations when subjected to the stress of the drug manufacturing process and related processes, four types of formulations were prepared. A formulation containing a basic amino acid or its salt, calcium salt or magnesium salt and about 180 mg/mL teperumumab, and then stored and/or subjected to one or more drug manufacturing processes and related process stresses, such Stress includes stress from one or more kinds of freezing/thawing, formulation merging/mixing, bioburden reduction filtration, drug storage, sterile filtration, filling, inspection, simulated labeling/packaging, and simulated transportation.

For each formulation, a unique diafiltration (DF) buffer was used to UF/DF treatment of the initial solution containing teperumumab. Add surfactant after UF/DF. Table 5 lists the components of each DF buffer used to obtain the four formulations and the amount of surfactant added after UF/DF. As a control, the fifth formulation (A5) was prepared using DF buffer containing 10 mM acetate, 261 mM (3.0% (w/v)) L-proline (pH 5.2), and added after UF/DF Surfactant 0.010% (w/v) polysorbate 80 (PS 80). [table 5] Formulation Teperumumab ( mg/mL ) Basic amino acid or its salt, calcium salt or magnesium salt Additional excipients Surfactant Final pH A1 180 120 mM calcium glutamate none 0.01% (w/v) PS80 5.0 A2 180 155 mM arginine glutamate none 0.01% (w/v) PS80 5.4 A3 180 60 mM calcium glutamate 140 mM proline 0.01% (w/v) PS80 5.0 A4 180 80 mM arginine glutamate 100 mM proline 0.01% (w/v) PS80 5.4 A5 180 n/a 10 mM acetate 261 mM proline 0.01% (w/v) PS80 5.2

Calcium glutamate is prepared by combining calcium hydroxide and glutamate, and arginine glutamate is prepared by combining arginine base and glutamine (glutamate). The final pH is reached by titration with glutamic acid and measured with a Seven Easy pH meter (Mettler Toledo). After 5-fold dilution by weight, Solo-VPE (C Technology Company) was used to confirm the concentration of teparumab in each formulation by UV absorption slope spectroscopy.

The final concentration of certain components of the final formulation analyzed (for example, after storage, as needed, for stability, viscosity) is different from the concentration of DF buffer. In this case, the components (such as acetate, glutamate, and calcium) are co-concentrated with Teperumumab so that the final formulation contains a higher concentration of components relative to the concentration of the components in the DF buffer. Points (e.g. acetate, glutamate, calcium). For example, in a final formulation containing 180 mg/mL tepirumumab, the glutamate concentration in the DF buffer was increased to 15%, and in the final formulation containing 180 mg/mL tepirumumab , The calcium concentration in DF buffer increased to 50%.

In addition, excipients may be subject to size exclusion, or may be affected by non-specific interactions. For example, in a 110 mg/mL tepilumab formulation, the proline concentration can be as much as about 16.67% lower than the concentration indicated in the DF buffer, and in a 140 mg/mL tepilumab formulation, The proline concentration can be as much as about 10% to about 13.3% lower than the concentration indicated in the DF buffer. For another example, in a 180 mg/mL tepirumumab formulation, the proline concentration may be about 15% lower than the concentration indicated in the DF buffer, and the arginine concentration may be about 40% lower than the concentration indicated in the DF buffer. %-45%.

The viscosity and stability after storage of the formulations in Table 5 were tested by a series of measurements used to evaluate product quality. The AR-G2 cone-plate rheometer (TA instruments) was used to measure the viscosity at 23°C at a shear rate of 1000 1/sec. Unless otherwise stated, viscosity is measured in the absence of surfactants. To measure the stability, a sample of each formulation was filled into a container, and then placed at a temperature of about -30°C to about 40°C (e.g., -30°C, 5°C, 25°C, 40°C ) Store up to 6 months (e.g. 1 week, 2 weeks, 4 weeks, 3 months, 6 months). The samples are tested by size exclusion chromatography (SEC) to determine the stability of the formulation at multiple storage time points. The percentage of high molecular weight (HMW) material and the percentage of main peak for each formulation are reported. The percentage of the main peak reflects the amount of teparumab (in monomeric form) remaining after the specified storage time.

As shown in Figures 15A-15D and Figure 16, arginine glutamate (ArgGlu) and arginine glutamate proline (ArgGluPro) formulations provide the best stability while maintaining reduced viscosity sex. A six-month study conducted at temperatures of -30°C, 5°C, and 25°C confirmed that the ArgGlu and ArgGluPro formulations showed minimal protein aggregation. ArgGlu and ArgGluPro formulations are also more stable than calcium glutamate (CaGlu) or calcium glutamate proline (CaGluPro) formulations under high stress at 40°C after 4 weeks.

Capillary electrophoresis-sodium dodecyl sulfate (CE-SDS) is used to separate denatured protein size variants under non-reducing (nr) or reducing conditions (RCE-SDS). Under RCE-SDS, the release and stability of heavy and light chains were measured. Under the test conditions, the heavy chain + light chain release ≥ 98%, and the heavy chain + light chain stability ≥ 96% (Figure 17).

Viscosity analysis showed that the viscosity of the ArgGlu and ArgGluPro formulations remained below 25 cP, and remained at about 20-22 cP or less (Figure 18).

Stability analysis at temperatures of -30°C, 5°C, and 25°C at six-month time points confirmed that the ArgGlu and ArgGluPro formulations showed the least protein aggregation. Overall, these conditions indicate that both arginine glutamate and arginine glutamate proline formulations are effective in reducing viscosity and protein aggregation in anti-TSLP antibody formulations.

Based on the results, an exemplary expected excipient range is listed in Table 6. The amounts of surfactants expected are those in Table 5. "Formulated with" refers to the amount of excipients in the diafiltration (DF) buffer. "In ... formulation" refers to the final estimated concentration of excipients after mixing, taking into account any repulsive characteristics or co-concentration effects produced by the mixture. [Table 6] Effective range of DF buffer concentration (mM ) Exemplary DF buffer concentration target ( mM ) (without proline) Exemplary DF buffer concentration target ( mM ) (including proline) Effective range of drug product concentration ( mM ) Exemplary drug product concentration target ( mM ) (without proline) Exemplary drug product concentration target ( mM ) (including proline) Prepared with In ...... in preparation Arginine glutamate formulation Arginine 25-190 140 80 10-125 95 50 Glutamate 25-200 150 85 25-225 170 95 Proline 0-250 0 100 0-220 0 85 Calcium glutamate formulation calcium 15-130 100 60 15-195 110 70 Glutamate 30-300 230 140 25-320 240 145 Proline 0-250 0 70 0-220 0 60

All references (including publications, patent applications, and patents) cited herein are incorporated by reference to the extent that each reference is individually and clearly indicated to be incorporated by reference and to the extent that The whole content is explained here.

Unless otherwise indicated herein or the context is clearly contradictory, the terms "a/an" and "the" are used in the context of describing the content of this disclosure (especially in the context of the following claims) And similar indicators will be regarded as covering both the singular and the plural. Unless otherwise described, the terms "include", "have", "include" and "contain" will be regarded as open terms (also meaning "including (but not limited to)").

The range of values quoted herein is only intended to be used as a shorthand method of individually referring to each individual value falling within the range and each end point, unless otherwise stated herein, and each individual value and end point is Incorporate into this specification as if it were individually cited in this document.

Unless otherwise indicated herein or otherwise clearly contradictory to the context, all methods described herein can be performed in any suitable order. Unless otherwise claimed, the use of any and all examples or exemplary language (such as "such as") provided herein is only intended to better describe the content of this disclosure, and does not impose restrictions on the scope of the content of this disclosure. The language in the specification should not be interpreted as indicating that any unclaimed element is necessary to practice this disclosure.

This article describes the preferred embodiments of the present disclosure, including the best way known to the inventors for implementing the present disclosure. After reading the above description, changes to those preferred embodiments will be obvious to those of ordinary skill in the art. The inventors expect those skilled in the art to adopt such changes as appropriate, and the inventors intend to implement the present disclosure in ways other than those specifically described herein. Therefore, this disclosure includes all modifications and equivalents permitted by applicable laws on the subject matter described in the appended claims. In addition, unless otherwise indicated herein or the context is otherwise clearly contradictory, this disclosure covers any combination of the above elements with all possible changes.

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[Figure 1] A graph plotting the viscosity (cP) of two different formulations containing therapeutic protein as a function of protein concentration (mg/mL).

[Figure 2] A graph of the viscosity (cP) of several different tepelumab formulations containing the indicated excipients (in the indicated amounts of 100 mM or 150 mM). The viscosity of the control without excipients is also provided. Each formulation contained teperumumab at a concentration of approximately 210 mg/mL.

[Figure 3] is a graph of the viscosity (cP) of several different Teperumumab formulations containing the indicated excipients (in the indicated amount, 60 mM). The viscosity of the control without excipients is also provided. Each formulation contains tepelumab at a concentration of approximately 190 mg/mL.

[Figure 4] is a graph of the viscosity (cP) of several different tepelumab formulations containing one or more indicated excipients (in the indicated amounts). The viscosity of the control without excipients is also provided. Each formulation contained teperumumab at a concentration of approximately 210 mg/mL. The column next to 0.5% PVP represents a formulation containing 100 mM sodium acetate and 75 mM arginine acetate. The stated% is% (w/v).

[Figure 5] is a graph of the viscosity (cP) of several different formulations of tepelumab containing one or more indicated excipients (in the indicated amount (mM)). The viscosity of the control without excipients is also provided. Each formulation contained teperumumab at a concentration of approximately 210 mg/mL.

[Figure 6] is a graph of the viscosity (cP) of several different formulations of Teperumumab containing the indicated excipients (in the indicated amount (60 mM)). The viscosity of the control without excipients is also provided. Each formulation contains tepelumab at a concentration of approximately 190 mg/mL.

[Figure 7] is a graph of the viscosity (cP) of two different Teperumumab formulations containing the indicated excipients (in the indicated amount (60 mM)). The viscosity of the control without excipients is also provided. Each formulation contains tepelumab at a concentration of approximately 190 mg/mL.

[Figure 8] is a table of the protein concentration, viscosity, and pH of several different tepelumab formulations containing one or more indicated excipients (in indicated amounts (mM)).

[Fig. 9A] is a graph of the viscosity (cP) of several different tepelumab formulations containing the indicated excipients (in the indicated amount (150 mM)). The viscosity of the control without excipients is also provided. Each formulation contained teperumumab at a concentration of approximately 210 mg/mL.

[Figure 9B] is a graph of the viscosity (cP) of several different tepelumab formulations containing the indicated excipients (in the indicated amount (90 mM)). The viscosity of the control without excipients is also provided. Each formulation contains tepelumab at a concentration of approximately 195 mg/mL.

[Figure 10] is a graph of the viscosity (cP) of several different tepelumab formulations containing the indicated excipients (in the indicated amount (mM)). The viscosity of the control without excipients is also provided. Each formulation contained teperumumab at a concentration of approximately 210 mg/mL.

[Figure 11] is a graph of the viscosity (cP) of several different formulations of Teperumumab containing the indicated excipients (in the indicated amount (150 mM)). The viscosity of the control without excipients is also provided. Each formulation contained teperumumab at a concentration of approximately 210 mg/mL.

[Figure 12] is a graph of the viscosity (cP) of several different Teperumumab formulations containing the indicated excipients (in the indicated amounts (50 mM-150 mM)). The viscosity of the control without excipients is also provided. Each formulation contained teperumumab at a concentration of approximately 210 mg/mL.

[Figure 13A] is a graph of the viscosity (cP) of several different formulations of teparumab containing 150 mM arginine acetate (at the indicated pH (4.75-5.7)). The viscosity of a control without excipients and a formulation containing 150 mM proline are also provided. Each formulation contained teperumumab at a concentration of approximately 210 mg/mL.

[Figure 13B] is a graph of the viscosity (cP) of several different formulations of Teperumumab containing 60 mM histidine acetate (at the indicated pH (5.5-6.5)). Each formulation contained teperumumab at a concentration of approximately 210 mg/mL.

[Figure 14] A graph of the viscosity (cP) of several different formulations of Teperumumab containing one or more indicated excipients (in the indicated amounts (33 mM-150 mM)). The viscosity of the control without excipients is also provided. Each formulation contained teperumumab at a concentration of approximately 210 mg/mL.

[Figure 15A-15D] shows size exclusion chromatography (SEC) analysis of different anti-TSLP formulations under stress conditions: Figure 15A, -30°C; Figure 15B, 5°C, Figure 15C, 25°C , Figure 15D, 40°C.

[Figure 16] shows the cation exchange chromatography (CEX) analysis (main peak %) of different anti-TSLP formulations under stress conditions.

[Figure 17] RCE-SDS analysis showing the release and stability of heavy and light chains under various storage conditions within 6 months.

[Figure 18] shows the viscosity of different anti-TSLP formulations under stress conditions.

none

Claims (122)

An aqueous composition comprising (a) an anti-TSLP antibody at a concentration greater than about 140 mg/mL, (b) a surfactant, and (c) at least one basic amino acid or salt thereof, wherein the composition contains about 10 mM to about 200 mM basic amino acid or its salt. The aqueous composition according to claim 1, wherein the basic amino acid is arginine. The aqueous composition according to claim 2, which comprises an organic salt of arginine. The aqueous composition according to claim 3, wherein the arginine acid salt is arginine acetate, arginine aspartate, arginine glutamine, arginine glycolate, spermine Acid lactate, arginine mesylate, arginine propionate, or a combination thereof. The aqueous composition according to claim 1, wherein the basic amino acid is histidine. The aqueous composition according to claim 5, which comprises an organic salt of histidine. The aqueous composition according to claim 6, wherein the histidine salt is histidine acetate, histidine aspartate, histidine glutamine, histidine glycolate, histamine Acid lactate, histidine methanesulfonate, histidine propionate, or a combination thereof. The aqueous composition according to claim 1, wherein the basic amino acid is lysine. The aqueous composition according to claim 8, which comprises an organic salt of lysine acid. The aqueous composition according to claim 9, wherein the lysine salt is lysine acetate, lysine aspartate, lysine glutamate, lysine glycolate, and lysine Acid lactate, lysine mesylate, lysine propionate, or combinations thereof. An aqueous composition comprising (a) an anti-TSLP antibody at a concentration greater than about 140 mg/mL, (b) a surfactant, and (c) at least one calcium salt or magnesium salt, wherein the composition contains about 15 mM to Approximately 150 mM calcium or magnesium salt. The aqueous composition according to claim 12, wherein the calcium salt or magnesium salt contains a relative ion lacking chloride. The aqueous composition according to claim 12, wherein the relative ion is acetate, aspartate, glutamate, glycolate, lactate, methanesulfonate, propionate, or a combination thereof. The aqueous composition according to claim 13, wherein the calcium salt is calcium acetate, calcium aspartate, calcium glutamate, calcium glycolate, calcium lactate, calcium methanesulfonate, calcium propionate, or a combination thereof. The aqueous composition according to claim 13, wherein the magnesium salt is magnesium acetate, magnesium aspartate, magnesium glutamate, magnesium glycolate, magnesium lactate, magnesium methanesulfonate, magnesium propionate, or a combination thereof. The aqueous composition according to any one of the preceding claims, which further comprises N-acetylarginine (NAR), N-acetyllysine, methionine, glycine, and proline Acid, sodium acetate, tris acetate, histamine, or calcium salt, as needed, in an amount of about 50 mM to about 250 mM. The aqueous composition according to claim 16, which comprises arginine and NAR or methionine or a combination thereof. The aqueous composition according to claim 17, wherein the arginine salt is arginine glutamate. The aqueous composition according to claim 18, which comprises 25-190 mM arginine base and 25-200 mM glutamine. The aqueous composition according to claim 18 or 19, which comprises 140 mM arginine base and 150 mM glutamine. The aqueous composition according to any one of claims 18 to 20, which contains from 0 to 250 mM proline. The aqueous composition according to claim 21, which comprises 80 mM arginine, 85 mM glutamine and 100 mM L-proline. The aqueous composition according to any one of claims 18 to 22, which contains 0.01% (w/v) polysorbate 80. The composition according to claim 23, which comprises 140 mM arginine base, 150 mM glutamine acid, and 0.01% (w/v) polysorbate 80. The composition according to claim 23, which comprises 80 mM arginine, 85 mM glutamine, 100 mM L-proline, and 0.01% (w/v) polysorbate 80. The aqueous composition according to claim 18, which contains 10-125 mM arginine base and 25-225 mM glutamine. The aqueous composition according to claim 18 or 26, which comprises 95 mM arginine base and 170 mM glutamine. The aqueous composition according to any one of claims 26 to 27, which contains from 0 to 220 mM proline. The aqueous composition according to claim 28, which comprises 50 mM arginine base, 95 mM glutamine and 85 mM L-proline. The aqueous composition according to any one of claims 26 to 29, which contains 0.01% (w/v) polysorbate 80. The aqueous composition according to claim 30, which comprises 95 mM arginine base, 170 mM glutamic acid, and 0.01% (w/v) polysorbate 80. The aqueous composition according to claim 30, which comprises 50 mM arginine, 95 mM glutamic acid and 85 mM L-proline, 0.01% (w/v) polysorbate 80. The aqueous composition according to any one of the preceding claims, wherein the anti-TSLP antibody comprises: (A) A light chain variable domain, the light chain variable domain comprising: (i) a light chain CDR1 sequence containing the amino acid sequence shown in SEQ ID NO: 3; (ii) a light chain CDR1 sequence containing SEQ ID NO: The light chain CDR2 sequence of the amino acid sequence shown in 4; and (iii) the light chain CDR3 sequence of the amino acid sequence shown in SEQ ID NO: 5; and (B) A heavy chain variable domain, the heavy chain variable domain comprising: (i) a heavy chain CDR1 sequence containing the amino acid sequence shown in SEQ ID NO: 6; (ii) a heavy chain CDR1 sequence containing SEQ ID NO: The heavy chain CDR2 sequence of the amino acid sequence shown in 7 and (iii) the heavy chain CDR3 sequence of the amino acid sequence shown in SEQ ID NO: 8. The aqueous composition of claim 33, wherein the anti-TSLP antibody comprises: (A) The light chain variable domain, the light chain variable domain is selected from the group consisting of: i. An amino acid sequence with at least 80% identity with SEQ ID NO: 12; ii. An amino acid sequence encoded by a polynucleotide sequence having at least 80% identity with SEQ ID NO: 11; or iii. The amino acid sequence encoded by the polynucleotide that hybridizes to the complementary sequence of the polynucleotide composed of SEQ ID NO: 11 under moderately stringent conditions; or (B) Heavy chain variable domain, the heavy chain variable domain is selected from the group consisting of: i. An amino acid sequence with at least 80% identity with SEQ ID NO: 10; ii. An amino acid sequence encoded by a polynucleotide sequence having at least 80% identity with SEQ ID NO: 9; or iii. The amino acid sequence encoded by the polynucleotide that hybridizes to the complementary sequence of the polynucleotide composed of SEQ ID NO: 9 under moderately stringent conditions; or (C) The light chain variable domain of (A) and the heavy chain variable domain of (B). The aqueous composition according to claim 17, which comprises a calcium salt and NAR or methionine or a combination thereof. The aqueous composition according to claim 35, wherein the calcium salt is calcium glutamate. The aqueous composition according to claim 36, which contains 15-130 mM calcium and 30-300 mM glutamine. The aqueous composition according to claim 36 or 37, which contains 100 mM calcium and 230 mM glutamine. The aqueous composition according to any one of claims 35 to 38, which contains from 0 to 250 mM proline. The aqueous composition according to claim 39, which contains 60 mM calcium, 140 mM glutamate and 70 mM L-proline. The aqueous composition according to claim 35, which contains 15-195 mM calcium and 25-320 mM glutamine. The aqueous composition according to claim 35 or 41, which contains 110 mM calcium and 240 mM glutamine. The aqueous composition according to any one of claims 35 or 41 to 42, which contains from 0 to 220 mM proline. The aqueous composition according to claim 43, which contains 70 mM calcium, 145 mM glutamate and 60 mM L-proline. The aqueous composition according to any one of claims 35 to 44, which contains 0.01% (w/v) polysorbate 80. The aqueous composition according to claim 45, which contains 100 mM calcium, 230 mM glutamate, and 0.01% (w/v) polysorbate 80. The aqueous composition according to claim 45, which comprises 60 mM calcium, 140 mM glutamate, 70 mM L-proline, and 0.01% (w/v) polysorbate 80. The aqueous composition according to claim 45, which contains 110 mM calcium, 240 mM glutamate, and 0.01% (w/v) polysorbate 80. The aqueous composition according to claim 45, which contains 70 mM calcium, 145 mM glutamate, 60 mM L-proline, and 0.01% (w/v) polysorbate 80. The aqueous composition according to any one of claims 35 to 49, wherein the anti-TSLP antibody comprises: (A) A light chain variable domain, the light chain variable domain comprising: (i) a light chain CDR1 sequence containing the amino acid sequence shown in SEQ ID NO: 3; (ii) a light chain CDR1 sequence containing SEQ ID NO: The light chain CDR2 sequence of the amino acid sequence shown in 4; and (iii) the light chain CDR3 sequence of the amino acid sequence shown in SEQ ID NO: 5; and (B) A heavy chain variable domain comprising: (i) a heavy chain CDR1 sequence containing the amino acid sequence shown in SEQ ID NO: 6; (ii) a heavy chain CDR1 sequence containing SEQ ID NO: The heavy chain CDR2 sequence of the amino acid sequence shown in 7 and (iii) the heavy chain CDR3 sequence of the amino acid sequence shown in SEQ ID NO: 8. The aqueous composition of claim 50, wherein the anti-TSLP antibody comprises: (A) The light chain variable domain, the light chain variable domain is selected from the group consisting of: iv. An amino acid sequence with at least 80% identity with SEQ ID NO: 12; v. An amino acid sequence encoded by a polynucleotide sequence having at least 80% identity with SEQ ID NO: 11; or vi. The amino acid sequence encoded by the polynucleotide that hybridizes to the complementary sequence of the polynucleotide composed of SEQ ID NO: 11 under moderately stringent conditions; or (B) Heavy chain variable domain, which is selected from the group consisting of: iv. An amino acid sequence with at least 80% identity with SEQ ID NO: 10; v. An amino acid sequence encoded by a polynucleotide sequence having at least 80% identity with SEQ ID NO: 9; or vi. The amino acid sequence encoded by the polynucleotide that hybridizes with the complementary sequence of the polynucleotide composed of SEQ ID NO: 9 under moderately stringent conditions; or (C) The light chain variable domain of (A) and the heavy chain variable domain of (B). The aqueous composition according to any one of the preceding claims, wherein the concentration of the anti-TSLP antibody is about 160 mg/mL to about 250 mg/mL. The aqueous composition of claim 52, wherein the concentration of the anti-TSLP antibody is about 165 mg/mL to about 225 mg/mL. The aqueous composition according to claim 53, wherein the concentration of the anti-TSLP antibody is about 165 mg/mL to about 200 mg/mL. The aqueous composition according to claim 54, wherein the concentration of the anti-TSLP antibody is about 175 mg/mL to about 185 mg/mL, if necessary, about 180 mg/mL. The aqueous composition of claim 52, wherein the concentration of the anti-TSLP antibody is about 189 mg/mL to about 231 mg/mL. The aqueous composition according to claim 52, wherein the concentration of the anti-TSLP antibody is about 205 mg/mL to about 215 mg/mL, and if necessary, about 210 mg/mL. The aqueous composition according to any one of the preceding claims, which has a pH of about 4.5 to about 6.75. The aqueous composition according to claim 58, which has a pH of about 4.8 to about 6.0. The aqueous composition according to claim 59, which contains 25-190 mM arginine base and 25-200 mM glutamine. The aqueous composition according to claim 59 or 60, which contains 140 mM arginine base and 150 mM glutamine. The aqueous composition according to any one of claims 60 to 61, which contains from 0 to 250 mM proline. The aqueous composition according to claim 62, which comprises 80 mM arginine base, 85 mM glutamine and 100 mM L-proline. The aqueous composition according to any one of claims 60 to 63, which contains 0.01% (w/v) polysorbate 80 and has a pH of 5.4. The aqueous composition according to claim 60, which comprises 140 mM arginine base, 150 mM glutamine acid, 0.01% (w/v) polysorbate 80, pH 5.4. The aqueous composition according to claim 60, which comprises 80 mM arginine, 85 mM glutamic acid, 100 mM L-proline, 0.01% (w/v) polysorbate 80, pH 5.4. The aqueous composition according to claim 59, which contains 10-125 mM arginine base and 25-225 mM glutamine. The aqueous composition according to claim 67, which comprises 95 mM arginine base and 170 mM glutamine. The aqueous composition according to any one of claim 67 or 68, which contains from 0 to 220 mM proline. The aqueous composition according to claim 69, which comprises 50 mM arginine base, 95 mM glutamine and 85 mM L-proline. The aqueous composition according to any one of claims 67 to 70, which contains 0.01% (w/v) polysorbate 80 and has a pH of 5.4. The aqueous composition according to claim 71, which comprises 95 mM arginine base, 170 mM glutamic acid, 0.01% (w/v) polysorbate 80, pH 5.4. The aqueous composition according to claim 71, which comprises 50 mM arginine, 95 mM glutamic acid, 85 mM L-proline, 0.01% (w/v) polysorbate 80, pH 5.4. The aqueous composition according to claim 59, which contains 15-130 mM calcium and 30-300 mM glutamine. The aqueous composition according to claim 74, which contains 100 mM calcium and 230 mM glutamine. The aqueous composition according to any one of claim 74 or 75, which contains from 0 to 250 mM proline. The aqueous composition according to claim 76, which comprises 60 mM calcium, 140 mM glutamate and 70 mM L-proline. The aqueous composition according to claim 59, which contains 15-195 mM calcium and 25-320 mM glutamate. The aqueous composition according to claim 78, which contains 110 mM calcium and 240 mM glutamine. The aqueous composition according to any one of claim 78 or 79, which contains from 0 to 220 mM proline. The aqueous composition according to claim 80, which contains 70 mM calcium, 145 mM glutamate and 60 mM L-proline. The aqueous composition according to any one of claims 78 to 81, which contains 0.01% (w/v) polysorbate 80 and has a pH of 5.0. The aqueous composition according to claim 82, which comprises 100 mM calcium, 230 mM glutamate, 0.01% (w/v) polysorbate 80, and pH 5.0. The aqueous composition according to claim 82, which comprises 60 mM calcium, 140 mM glutamate, 70 mM L-proline, 0.01% (w/v) polysorbate 80, pH 5.0. The aqueous composition according to claim 82, which contains 110 mM calcium, 240 mM glutamate, 0.01% (w/v) polysorbate 80, and pH 5.0. The aqueous composition according to claim 82, which contains 70 mM calcium, 145 mM glutamate, 60 mM L-proline, 0.01% (w/v) polysorbate 80, pH 5.0. The aqueous composition according to any one of the preceding claims, wherein the ^{viscosity of the aqueous composition at 23°C and 1000s -1} is less than 100 cP. The aqueous composition according to claim 87, wherein the ^{viscosity of the aqueous composition at 23°C and 1000s -1} is less than 75 cP. The aqueous composition according to claim 88, wherein the ^{viscosity of the aqueous composition at 23°C and 1000s -1} is less than 60 cP. The aqueous composition according to claim 89, wherein the ^{viscosity of the aqueous composition at 23°C and 1000s -1} is less than 50 cP. The aqueous composition according to any one of the preceding claims, wherein the surfactant is amphiphilic and/or nonionic. The composition according to claim 91, wherein the surfactant is polysorbate. The composition according to claim 92, wherein the surfactant is polysorbate 20 or polysorbate 80, or a mixture thereof. The composition according to any one of the preceding claims, which comprises a surfactant at a concentration of less than or about 0.005% (w/v) to about 0.015% (w/v). The composition according to claim 94, which contains about 0.010% (w/v) ± 0.0025% (w/v) surfactant. The composition according to claim 95, which contains about 0.005% (w/v), 0.010% (w/v), or 0.015% (w/v) surfactant. The composition according to any one of the preceding claims, wherein the composition is isotonic or has a range from about 200 mOsm/kg to about 500 mOsm/kg, or from about 225 mOsm/kg to about 400 mOsm/kg, Or an osmotic pressure in the range of about 250 mOsm/kg to about 350 mOsm/kg. The composition according to any one of the preceding claims, wherein the composition is isotonic or has an osmotic pressure greater than about 350 mOsm/kg. The aqueous composition according to any one of the preceding claims, wherein as determined by size exclusion chromatography (SEC), after storage at 2°C to 8°C for at least or about 12 months, less than 5% of antibodies are degraded. The aqueous composition according to claim 99, wherein as determined by size exclusion chromatography (SEC), after storage at 2°C to 8°C for about 20 months to about 26 months, less than 5 % Of antibodies are degraded. The aqueous composition according to claim 100, wherein as determined by size exclusion chromatography (SEC), after storage at 2°C to 8°C for about 30 to about 40 months, less than 5% The antibody is degraded. The aqueous composition according to any one of the preceding claims, wherein, as determined by size exclusion chromatography (SEC), less than 5% after storage at a temperature greater than 20°C for at least or about 2 weeks The antibody is degraded and stored for at least or about 4 or 8 weeks as needed. The aqueous composition of claim 102, wherein the temperature is greater than or about 25°C or greater than or about 30°C or greater than or about 40°C. The aqueous composition according to any one of the preceding claims, wherein the anti-TSLP anti-system IgG2 antibody. The aqueous composition according to any one of claims 33 to 104, wherein the anti-TSLP antibody comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 13 and one of the heavy chain containing the amino acid sequence of SEQ ID NO: 14 A light chain, or a heavy chain containing the amino acid sequence of SEQ ID NO: 13, and a light chain containing the amino acid sequence of SEQ ID NO: 14. The aqueous composition according to any one of claims 33 to 105, wherein the anti-TSLP antibody specifically binds to the TSLP polypeptide shown in amino acids 29-159 of SEQ ID NO: 2. The aqueous composition according to any one of claims 1 to 106, wherein the two binding sites of the anti-TSLP antibody have the same binding as TSLP. A product comprising the aqueous composition according to any one of the preceding claims, the product comprising about 1 mL to about 5 mL of the aqueous composition as needed. A pre-filled syringe containing the aqueous composition according to any one of the preceding claims, and the syringe containing about 1 mL to about 5 mL of the aqueous composition as needed. A vial containing the aqueous composition according to any one of the preceding claims, the vial containing about 1 mL to about 5 mL of the aqueous composition as needed. An auto-injector containing the aqueous composition described in claim 107. The auto-injector according to claim 111, wherein the auto-injector is Ypsomed YpsoMate®. The auto-injector according to claim 111, wherein the auto-injector is disclosed in WO 2018/226565, WO 2019/094138, WO 2019/178151, WO 20120/072577, WO2020/081479, WO 2020/081480, PCT/US 20/ 70590, PCT/US 20/70591, PCT/US 20/53180, PCT/US 20/53179, PCT/US 20/53178, or PCT/US 20/53176. The use of the aqueous composition according to any one of the preceding claims for the treatment of inflammatory diseases. The use according to claim 114, wherein the inflammatory disease is selected from the group consisting of asthma, atopic dermatitis, chronic obstructive pulmonary disease (COPD), nasal polyps, chronic spontaneous urticaria, eosinophils Esophagitis (EoE), Ig-driven diseases (such as IgA nephropathy & lupus nephritis), eosinophilic gastritis, chronic sinusitis without nasal polyps, and idiopathic pulmonary fibrosis (IPF). The use according to claim 115, wherein the inflammatory disease is atopic dermatitis. A method for treating an inflammatory disease in a subject, the method comprising administering to the subject a therapeutically effective amount of the aqueous composition according to any one of the preceding claims. The method according to claim 117, wherein the inflammatory disease is selected from the group consisting of asthma, atopic dermatitis, chronic obstructive pulmonary disease (COPD), nasal polyps, chronic spontaneous urticaria, eosinophils Esophagitis (EoE), Ig-driven diseases (such as IgA nephropathy & lupus nephritis), eosinophilic gastritis, chronic sinusitis without nasal polyps, and idiopathic pulmonary fibrosis (IPF). The method according to claim 118, wherein the inflammatory disease is atopic dermatitis. The method according to any one of claims 117 to 119, wherein the aqueous composition is administered to the subject by subcutaneous administration. The method according to any one of claims 117 to 120, wherein about 10 mL to about 20 mL of the aqueous composition is administered to the subject. A method for preparing a stable liquid antibody composition having a viscosity of less than about 100 cP and containing (A) anti-TSLP antibody at a concentration greater than about 140 mg/mL, (B) surfactant, and (C) alkaline An amino acid or a salt thereof, a calcium salt, a magnesium salt, or a combination thereof, the method comprising: (i) combining the antibody with an aqueous solution, the aqueous solution comprising about 50 mM to about 150 mM basic amino acid Or its salt, calcium salt, magnesium salt, or a combination thereof, and (ii) adding a surfactant to achieve a final concentration of about 0.01% (w/v) ± 0.005% (w/v) of the surfactant.

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