Stable liquid formulation of Anti-il-4rα antibody

Abstract

The present invention relates to: a liquid formulation including an anti-IL-4Rα antibody, a buffering agent, saccharides and methionine, and having a pH of 4.9 to 5.5; a device comprising same; and a use thereof for treating IL-4Rα-related conditions. The liquid formulation can improve the viscosity while ensuring the stability of the antibody, and thus can be effectively used as a medicine for treating IL-4Rα-related conditions.

Description

Stable liquid formulation of anti-IL-4Rα antibody

[0001] A stable liquid formulation of an anti-IL-4Rα antibody, a device comprising the same, and their use for treating an IL-4Rα-associated condition are disclosed.

[0002] Antibody drugs have larger molecular weights and complex secondary and higher-order structures compared to typical protein drugs, which can lead to physicochemical instability. For this reason, the development of optimal formulations that ensure quality and stability throughout the entire process of antibody drugs, from manufacturing and storage to patient administration, is essential. Protein instability can be caused by various external factors, such as temperature, light, and chemical factors, and can lead to decreased activity, reduced efficacy, or immunogenicity when administered to the human body. Therefore, improving the instability of antibody drugs and maintaining optimal quality until administration to patients is crucial. To achieve optimal quality, methods such as changing buffer solutions, assessing optimal pH, and adding stabilizers are used. Because the material properties of each antibody protein vary, and the optimal combination of buffer solution, pH, and stabilizer can vary, developing a formulation appropriate for the target substance is essential to address these issues.

[0003] For example, dupilumab, an anti-IL-4Rα antibody, is available from Dupixent. ®) is manufactured and sold under the trade name Dupixent. Dupixent 200 mg is a 1.14 mL liquid formulation consisting of 200 mg anti-IL4Rα IgG, 1.2 mg sodium acetate, 3.5 mg histidine, 12 mg arginine, 57 mg sucrose, and 2.3 mg polysorbate 80, and Dupixent 300 mg is a 2.00 mL liquid formulation consisting of 300 mg anti-IL4Rα IgG, 2.0 mg sodium acetate, 6.2 mg histidine, 10.5 mg arginine, 100 mg sucrose, and 4 mg polysorbate 80. To stabilize liquid formulations containing various antibody proteins, such as dupilumab, various conditions, such as the type and concentration of each component, are being reviewed and applied, such as adding sugars, amino acids, buffers, and surfactants, or applying a specific pH, as in the Dupixent product composition.

[0004] In addition to ensuring the stability described above, there is a need to develop a high-concentration formulation that increases therapeutic activity while reducing the number of administrations through high-dose injections in small volumes to improve patient convenience during injection. When developing a formulation for a high-concentration formulation, there are limitations such as protein instability due to protein aggregation and decreased injectability due to increased viscosity as the protein concentration increases. Therefore, it is important to develop a formulation that can improve viscosity while ensuring protein stability. Typically, sugars are used as stabilizers for protein stabilization, but in the case of formulations containing such stabilizers, viscosity increases accelerate when the protein concentration increases, limiting the ability to improve viscosity while ensuring protein stability. To solve this problem, it is possible to develop a formulation that can simultaneously improve viscosity by including a viscosity-reducing agent to prevent protein aggregation and thereby ensure protein stability.

[0005] However, to determine whether viscosity has improved and whether protein stability has improved until administration to patients, key quality assessments must be conducted under storage temperature conditions and up to the expiration date. This requires a long-term validation of the developed formulation. To effectively assess protein stability and optimal formulation in a short period of time, exposure to heat (temperature) and chemicals, factors that cause structural and physicochemical denaturation of proteins, can be analyzed for new quality indicators to select the optimal formulation combination.

[0006] The present disclosure relates to a stable liquid formulation of an anti-IL-4Rα antibody, such as dupilumab. Specifically, the present disclosure provides a liquid formulation that can improve the viscosity of an anti-IL-4Rα antibody, such as dupilumab, while ensuring the stability of the anti-IL-4Rα antibody throughout the entire process from manufacturing, storage, and administration to a patient. The liquid formulation has improved stability and viscosity compared to commercially available formulations of anti-IL-4Rα antibodies, such as dupilumab (e.g., Dupixent), making it applicable to high-concentration formulations, thereby increasing patient convenience.

[0007] One aspect is to provide a liquid formulation comprising an anti-IL-4Rα antibody; a buffer; a sugar; and methionine, wherein the pH is about 4.9 to about 5.5.

[0008] Another aspect is to provide a device comprising the liquid formulation.

[0009] Another aspect provides a method of treating an IL-4Rα associated condition, comprising administering the liquid formulation to a subject in need thereof.

[0010] Another aspect provides the use of the liquid formulation in the manufacture of a medicament for treating an IL-4Rα associated condition.

[0011] Unless otherwise defined, all technical terms used herein have the same meaning as commonly understood by those skilled in the art in the relevant field of the present invention. Furthermore, while preferred methods and samples are described herein, similar or equivalent methods are also included within the scope of the present invention. Furthermore, numerical values ​​described herein are considered to include the meaning of "about," even if not explicitly stated. The contents of all publications cited as references herein are incorporated herein by reference in their entirety.

[0012] The terms "about" or "approximately" in this specification can generally be interpreted to mean a value or range that is within 10%, 5%, 4%, 3%, 2%, or 1% above or below a given value or range.

[0013] One aspect provides a stable liquid formulation of an anti-IL-4Rα antibody, specifically,

[0014] anti-IL-4Rα antibody;

[0015] buffer;

[0016] sugars; and

[0017] Contains methionine,

[0018] The pH is about 4.9 to about 5.5,

[0019] Provides liquid formulation.

[0020] (1) Antibodies

[0021] The term "antibody" as used herein may be interpreted to mean a full-length antibody or an antigen-binding fragment thereof. The antibody includes a monoclonal antibody, a polyclonal antibody, a humanized antibody, a human antibody, and a chimeric antibody.

[0022] The term "antigen-binding fragment" refers to a fragment comprising the antigen-binding site of an antibody. For example, an antigen-binding fragment includes, but is not limited to, a Fab fragment, an F(ab')2 fragment, an Fc fragment, or an scFv fragment.

[0023] In the present specification, the antibody may be an anti-IL-4Rα antibody. The anti-IL-4Rα antibody may refer to any antibody that binds to the interleukin-4 receptor alpha chain (IL-4Rα). The anti-IL-4Rα antibody may inhibit IL-4 and IL-13 signaling.

[0024] The above anti-IL-4Rα antibody may be dupilumab (CAS No. 1190264-60-8). Therefore, the above liquid formulation may be a stable liquid formulation of dupilumab. Dupilumab is a fully human monoclonal antibody that binds to IL-4Rα and belongs to IgG4. Dupilumab is Dupixent. ® ) is sold under the trade name Dupixent. After receiving approval from the U.S. Food and Drug Administration (FDA) for the treatment of atopic dermatitis, Dupixent is expanding its indications to various type 2 inflammatory diseases such as asthma, chronic rhinosinusitis with nasal polyposis (CRSwNP), and eosinophilic esophagitis (EoE). The sequence of dupilumab is known, and it can be produced by general methods known in the art. More detailed information on dupilumab can be easily obtained by those skilled in the art from known databases.

[0025] The term "dupilumab" herein may also be interpreted to mean dupilumab having a modified amino acid sequence (deletion, insertion, and / or substitution) and / or a modified glycosylation characteristic, as long as it does not affect the polypeptide function.

[0026] The above anti-IL-4Rα antibody may be included in a therapeutically effective amount in a liquid formulation.

[0027] The concentration of the anti-IL-4Rα antibody is about 1 mg / mL to about 300 mg / mL, about 1 mg / mL to about 250 mg / mL, about 1 mg / mL to about 200 mg / mL, about 1 mg / mL to about 175 mg / mL, about 5 mg / mL to about 300 mg / mL, about 5 mg / mL to about 250 mg / mL, about 5 mg / mL to about 200 mg / mL, about 5 mg / mL to about 175 mg / mL, about 10 mg / mL to about 300 mg / mL, about 10 mg / mL to about 250 mg / mL, about 10 mg / mL to about 200 mg / mL, about 10 mg / mL to about 175 mg / mL, about 25 mg / mL to about 300 mg / mL, about 25 mg / mL to about 250 mg / mL, about 25 mg / mL to about 200 mg / mL, about 25 mg / mL to about 175 mg / mL, about 50 mg / mL to about 300 mg / mL, about 50 mg / mL to about 250 mg / mL, about 50 mg / mL to about 200 mg / mL, about 50 mg / mL to about 175 mg / mL, about 75 mg / mL to about 300 mg / mL, about 75 mg / mL to about 250 mg / mL, about 75 mg / mL to about 200 mg / mL, about 75 mg / mL to about 175 mg / mL, about 100 mg / mL to about 300 mg / mL, about 100 mg / mL to about 250 mg / mL, about 100 mg / mL to about 200 mg / mL, about 100 mg / mL to about 175 mg / mL, about 125 mg / mL to about 300 mg / mL, about 125 mg / mL to about 250 mg / mL, about 125 mg / mL to about 200 mg / mL, about 125 mg / mL to about 175 mg / mL, about 145 mg / mL to about 300 mg / mL, about 145 mg / mL to about 250 mg / mL, about 145 mg / mL to about 200 mg / mL, about 145 mg / mL to about 180 mg / mL,About 145 mg / mL to about 175 mg / mL, about 149 mg / mL to about 151 mg / mL, about 150 mg / mL to about 300 mg / mL, about 150 mg / mL to about 250 mg / mL, about 150 mg / mL to about 225 mg / mL, about 150 mg / mL to about 200 mg / mL, about 150 mg / mL to about 180 mg / mL, about 150 mg / mL to about 175 mg / mL, about 170 mg / mL to about 300 mg / mL, about 170 mg / mL to about 250 mg / mL, about 170 mg / mL to about 225 mg / mL, about 170 mg / mL to about 200 mg / mL, about 170 mg / mL to about 180 mg / mL, or about 174 mg / mL may be about 176 mg / mL.

[0028] In one specific example, the concentration of the anti-IL-4Rα antibody may be about 100 mg / mL to about 200 mg / mL.

[0029] In one specific example, the concentration of the anti-IL-4Rα antibody may be about 125 mg / mL to about 200 mg / mL.

[0030] In one specific example, the concentration of the anti-IL-4Rα antibody may be about 150 mg / mL to about 200 mg / mL.

[0031] In one specific example, the concentration of the anti-IL-4Rα antibody may be about 150 mg / mL to about 175 mg / mL.

[0032] In one specific example, the concentration of the anti-IL-4Rα antibody may be 135 mg / mL to 165 mg / mL.

[0033] In one specific example, the concentration of the anti-IL-4Rα antibody may be 158 mg / mL to 192 mg / mL.

[0034] In certain embodiments, the concentration of the anti-IL-4Rα antibody may be about 150 mg / mL.

[0035] In certain embodiments, the concentration of the anti-IL-4Rα antibody may be about 175 mg / mL.

[0036] The concentration of the anti-IL-4Rα antibody may be high. Accordingly, the liquid formulation may be a high-concentration liquid formulation of the anti-IL-4Rα antibody. The high concentration may mean, but is not limited to, 100 mg / mL or more or 150 mg / mL or more, for example, 100 mg / mL to 300 mg / mL or 150 mg / mL to 300 mg / mL.

[0037] (2) Buffer and pH

[0038] Liquid formulations according to one aspect include a buffer. The buffer can function to regulate the pH of the formulation. The buffer can maintain the pH of the liquid formulation at a certain value or within a certain range. Therefore, the buffer can serve to provide a pH of a certain value or within a certain range to the liquid formulation.

[0039] The above buffer may be used without limitation in its type as long as it is applicable to biopharmaceuticals.

[0040] The buffer may include at least one selected from acetate, histidine, phosphate, citrate, succinate, malate, tartarate, carbonate, salts thereof, and hydrates thereof.

[0041] The term "salt" may be a pharmaceutically acceptable salt. The salt may include an inorganic acid salt, an organic acid salt, a metal salt, etc. of the compound. The inorganic acid salt may be a hydrochloride, a bromate, a phosphate, a sulfate, or a disulfate. The organic acid salt may be a formate, an acetate, a propionate, a lactate, an oxalate, a tartrate, a malate, a maleate, a citrate, a fumarate, a besylate, a camsylate, an edisyl salt, a trichloroacetic acid, a trifluoroacetate, a benzoate, a gluconate, a methanesulfonate, a glycolate, a succinate, a 4-toluenesulfonate, a galacturonate, an embonate, a glutamate, an ethanesulfonate, a benzenesulfonate, a p-toluenesulfonate, or an aspartate. The metal salt may be a calcium salt, a sodium salt, a magnesium salt, a strontium salt, or a potassium salt.

[0042] The term "hydrate" refers to a substance containing water molecules within its molecules. The hydrate may be a monohydrate, a dihydrate, or a trihydrate.

[0043] The pH of the liquid formulation according to one aspect may be from about 4.9 to about 5.5. The pH of the liquid formulation may be provided by a buffer. Alternatively, the pH of the liquid formulation may be adjusted by the addition of any acid (e.g., HCl) or base (e.g., NaOH).

[0044] Specifically, the pH of the formulation can be any range or any value selected from about 4.9 to about 5.5. For example, the pH can be about 4.9 to about 5.5, about 4.9 to about 5.4, about 4.9 to about 5.3, about 4.9 to about 5.2, about 5.0 to about 5.5, about 5.0 to about 5.4, about 5.0 to about 5.3, about 5.0 to about 5.2, about 5.1 to about 5.5, about 5.1 to about 5.4, or about 5.1 to about 5.3.

[0045] In one embodiment, the pH of the liquid formulation can be about 4.9, about 5.0, about 5.1, about 5.2, about 5.3, about 5.4, or about 5.5.

[0046] In one embodiment, the buffer may include one or more selected from acetate, histidine, salts thereof, and hydrates thereof.

[0047] In one embodiment, the buffer may include one or more selected from acetate, a salt thereof, and a hydrate thereof. In one embodiment, the buffer may include acetate. In one embodiment, the buffer may be acetate. In one embodiment, the buffer may include sodium acetate. In one embodiment, the buffer may be sodium acetate.

[0048] The concentration of the buffer may be any range or any value selected from about 0.1 mM to about 50 mM. For example, the concentration of the buffer may be about 0.1 mM to about 50 mM, about 0.1 mM to about 40 mM, about 0.1 mM to about 30 mM, about 1 mM to about 50 mM, about 1 mM to about 40 mM, about 1 mM to about 30 mM, about 5 mM to about 50 mM, about 5 mM to about 40 mM, about 5 mM to about 35 mM, about 5 mM to about 30 mM, about 10 mM to about 50 mM, about 10 mM to about 40 mM, about 10 mM to about 35 mM, about 10 mM to about 30 mM, about 15 mM to about 50 mM, about 15 mM to about 40 mM, about 15 mM to about 35 mM, about 15 mM to about 30 mM, about 20 mM to about 50 mM, about 20 mM to about 40 mM, about 20 mM to about 35 mM, about 20 mM to about 30 mM, about 25 mM to about 50 mM, about 25 mM to about 40 mM, about 25 mM to about 35 mM, about 27 mM to about 50 mM, about 27 mM to about 40 mM, about 27 mM to about 35 mM, or about 27 mM to about 33 mM.

[0049] In certain embodiments, the concentration of the buffer may be about 30 mM.

[0050] The concentration of the acetate may be any range or any value selected from about 0.1 mM to about 50 mM. For example, the concentration of the acetate may be about 0.1 mM to about 50 mM, about 0.1 mM to about 40 mM, about 0.1 mM to about 30 mM, about 1 mM to about 50 mM, about 1 mM to about 40 mM, about 1 mM to about 30 mM, about 5 mM to about 50 mM, about 5 mM to about 40 mM, about 5 mM to about 35 mM, about 5 mM to about 30 mM, about 10 mM to about 50 mM, about 10 mM to about 40 mM, about 10 mM to about 35 mM, about 10 mM to about 30 mM, about 15 mM to about 50 mM, about 15 mM to about 40 mM, about 15 mM to about 35 mM, about 15 mM to about 30 mM, about 20 mM to about 50 mM, about 20 mM to about 40 mM, about 20 mM to about 35 mM, about 20 mM to about 30 mM, about 25 mM to about 50 mM, about 25 mM to about 40 mM, about 25 mM to about 35 mM, about 27 mM to about 50 mM, about 27 mM to about 40 mM, about 27 mM to about 35 mM, or about 27 mM to about 33 mM.

[0051] In certain embodiments, the concentration of the acetate may be about 30 mM.

[0052] (3) Sugars

[0053] Liquid formulations according to the aspect of the invention contain saccharides.

[0054] The above sugar may include at least one selected from sugar and sugar alcohol.

[0055] The sugar may be a monosaccharide, a disaccharide, an oligosaccharide, or a polysaccharide. The sugar may include one or more selected from sucrose, trehalose, galactose, mannose, maltose, lactose, fructose, and glucose.

[0056] The sugar alcohol described above is a general term for polyols having two or more hydroxyl groups, which are formed by reducing an aldehyde group or a ketone group of a sugar to an alcohol group. The sugar alcohol may include at least one selected from mannitol, sorbitol, xylitol, arabitol, erythritol, lactitol, maltitol, and inositol. The sugar alcohol includes an anhydride or hydrate of the sugar alcohol. For example, trehalose may include not only trehalose but also trehalose dihydrate.

[0057] The concentration of the sugar can be freely adjusted within a range that maintains the stability of the antibody, and can vary individually depending on each specific sugar type. The concentration of the sugar can be any range or value selected from about 0.1% (w / v) to about 20% (w / v). For example, the concentration of the sugar may be from about 0.1% (w / v) to about 20% (w / v), from about 0.1% (w / v) to about 15% (w / v), from about 0.1% (w / v) to about 12% (w / v), from about 0.1% (w / v) to about 10% (w / v), from about 0.1% (w / v) to about 8% (w / v), from about 0.1% (w / v) to about 6% (w / v), from about 0.1% (w / v) to about 5.5% (w / v), from about 0.1% (w / v) to about 5% (w / v), from about 0.1% (w / v) to about 4.5% (w / v), from about 0.1% (w / v) to about 4% (w / v), from about 0.1% (w / v) to about 3.5% (w / v), about 0.1% (w / v) to about 3% (w / v), about 0.1% (w / v) to about 2.5% (w / v), about 1% (w / v) to about 20% (w / v), about 1% (w / v) to about 15% (w / v), about 1% (w / v) to about 12% (w / v), about 1% (w / v) to about 10% (w / v), about 1% (w / v) to about 8% (w / v), about 1% (w / v) to about 6% (w / v), about 1% (w / v) to about 5.5% (w / v), about 1% (w / v) to about 5% (w / v), about 1% (w / v) to about 4.5% (w / v), about 1% (w / v) to about 4% (w / v), about 1% (w / v) to about 3.5% (w / v), about 1% (w / v) to about 3% (w / v), about 1% (w / v) to about 2.5% (w / v), about 2% (w / v) to about 20% (w / v), about 2% (w / v) to about 15% (w / v), about 2% (w / v) to about 12% (w / v), about 2% (w / v) to about 10% (w / v), about 2% (w / v) to about 8% (w / v), about 2% (w / v) to about 6% (w / v), about 2% (w / v) to about 5.5% (w / v), about 2% (w / v) to about 5% (w / v), about 2% (w / v) to about 4.5% (w / v), about 2% (w / v) to about 4% (w / v), about 2% (w / v) to about 3.5% (w / v), about 2% (w / v) to about 3% (w / v), about 2% (w / v) to about 2.5% (w / v), about 3% (w / v) to about 20% (w / v), about 3% (w / v) to about 15% (w / v), about 3% (w / v) to about 12% (w / v), about 3% (w / v) to about 10% (w / v), about 3% (w / v) to about 8% (w / v), about 3% (w / v) to about 6% (w / v), about 3% (w / v) to about 5.5% (w / v), about 3% (w / v) to about 5% (w / v), about 3% (w / v) to about 4.5% (w / v), about 3% (w / v) to about 4% (w / v), about 3% (w / v) to about 3.5% (w / v), about 3.5% (w / v) to about 20% (w / v), about 3.5% (w / v) to about 15% (w / v), about 3.5% (w / v) to about 12% (w / v), about 3.5% (w / v) to about 10% (w / v), about 3.5% (w / v) to about 8% (w / v), about 3.5% (w / v) to about 6% (w / v), about 3.5% (w / v) to about 5.5% (w / v), about 3.5% (w / v) to about 5% (w / v), about 3.5% (w / v) to about 4.5% (w / v), about 3.5% (w / v) to about 4.2% (w / v), about 3.5% (w / v) to about 4% (w / v), about 3.8% (w / v) to about 12% (w / v), about 3.8% (w / v) to about 10% (w / v), about 3.8% (w / v) to about 8% (w / v), about 3.8% (w / v) to about 5% (w / v), about 3.8% (w / v) to about 4.5% (w / v), about 3.8% (w / v) to about 4.2% (w / v), about 4% (w / v) to about 20% (w / v), about 4% (w / v) to about 15% (w / v), about 4% (w / v) to about 12% (w / v), about 4% (w / v) to about 10% (w / v), about 4% (w / v) to about 8% (w / v), about 4% (w / v) to about 6% (w / v), about 4% (w / v) to about 5.5% (w / v), about 4% (w / v) to about 5% (w / v), about 4% (w / v) to about 4.5% (w / v), about 5% (w / v) to about 20% (w / v), about 5% (w / v) to about 15% (w / v), about 5% (w / v) to about 12% (w / v), about 5% (w / v) to about 10% (w / v), about 5% (w / v) to about 8% (w / v), about 5% (w / v) to about 6% (w / v), about 5% (w / v) to about 5.7% (w / v), about 5% (w / v) to about 5.5% (w / v), about 5.3% (w / v) to about 20% (w / v), about 5.3% (w / v) to about 15% (w / v), about 5.3% (w / v) to about 12% (w / v), about 5.3% (w / v) to about 10% (w / v), about 5.3% (w / v) to about 8% (w / v), or about 5.3% (w / v) to about 6% (w / v), about 5.3% (w / v) to about 5.7% (w / v), about 5.5% (w / v) to about 20% (w / v), about 5.5% (w / v) to about 15% (w / v), about 5.5% (w / v) to about 12% (w / v), about 5.5% (w / v) to about 10% (w / v), about 5.5% (w / v) to about 8% (w / v), Or about 5.It can be from 5% (w / v) to about 6% (w / v). The concentration of the sugar alcohol can be freely adjusted within a range that maintains the stability of the antibody, and can vary individually depending on each specific sugar alcohol type. The concentration of the sugar alcohol can be any range or any value selected from about 0.1% (w / v) to about 20% (w / v). For example, the concentration of the sugar alcohol may be from about 0.1% (w / v) to about 20% (w / v), from about 0.1% (w / v) to about 15% (w / v), from about 0.1% (w / v) to about 12% (w / v), from about 0.1% (w / v) to about 10% (w / v), from about 0.1% (w / v) to about 8% (w / v), from about 0.1% (w / v) to about 5% (w / v), from about 0.1% (w / v) to about 4% (w / v), from about 1% (w / v) to about 20% (w / v), from about 1% (w / v) to about 15% (w / v), from about 1% (w / v) to about 12% (w / v), from about 1% (w / v) to about 10% (w / v), from about It can be from 1% (w / v) to about 8% (w / v), from about 1% (w / v) to about 7% (w / v), from about 1% (w / v) to about 6% (w / v), from about 1% (w / v) to about 5% (w / v), from about 1% (w / v) to about 4% (w / v), from about 1% (w / v) to about 3% (w / v), or from about 1% (w / v) to about 2% (w / v).

[0058] In one specific embodiment, the sugar may include one or more selected from sucrose, trehalose, and mannitol.

[0059] In certain embodiments, the sugar may comprise sucrose. In certain embodiments, the sugar may be sucrose.

[0060] The concentration of the sucrose may be any range or any value selected from about 0.1% (w / v) to about 20% (w / v). For example, the concentration of the sucrose may be from about 0.1% (w / v) to about 20% (w / v), from about 0.1% (w / v) to about 15% (w / v), from about 0.1% (w / v) to about 12% (w / v), from about 0.1% (w / v) to about 10% (w / v), from about 0.1% (w / v) to about 8% (w / v), from about 0.1% (w / v) to about 6% (w / v), from about 0.1% (w / v) to about 5.5% (w / v), from about 0.1% (w / v) to about 5% (w / v), from about 0.1% (w / v) to about 4.5% (w / v), from about 0.1% (w / v) to about 4% (w / v), from about 0.1% (w / v) to about 3.5% (w / v), about 0.1% (w / v) to about 3% (w / v), about 0.1% (w / v) to about 2.5% (w / v), about 1% (w / v) to about 20% (w / v), about 1% (w / v) to about 15% (w / v), about 1% (w / v) to about 12% (w / v), about 1% (w / v) to about 10% (w / v), about 1% (w / v) to about 8% (w / v), about 1% (w / v) to about 6% (w / v), about 1% (w / v) to about 5.5% (w / v), about 1% (w / v) to about 5% (w / v), about 1% (w / v) to about 4.5% (w / v), about 1% (w / v) to about 4% (w / v), about 1% (w / v) to about 3.5% (w / v), about 1% (w / v) to about 3% (w / v), about 1% (w / v) to about 2.5% (w / v), about 2% (w / v) to about 20% (w / v), about 2% (w / v) to about 15% (w / v), about 2% (w / v) to about 12% (w / v), about 2% (w / v) to about 10% (w / v), about 2% (w / v) to about 8% (w / v), about 2% (w / v) to about 6% (w / v), about 2% (w / v) to about 5.5% (w / v), about 2% (w / v) to about 5% (w / v), about 2% (w / v) to about 4.5% (w / v), about 2% (w / v) to about 4% (w / v), about 2% (w / v) to about 3.5% (w / v), about 2% (w / v) to about 3% (w / v), about 2% (w / v) to about 2.5% (w / v), about 3% (w / v) to about 20% (w / v), about 3% (w / v) to about 15% (w / v), about 3% (w / v) to about 12% (w / v), about 3% (w / v) to about 10% (w / v), about 3% (w / v) to about 8% (w / v), about 3% (w / v) to about 6% (w / v), about 3% (w / v) to about 5.5% (w / v), about 3% (w / v) to about 5% (w / v), about 3% (w / v) to about 4.5% (w / v), about 3% (w / v) to about 4% (w / v), about 3% (w / v) to about 3.5% (w / v), about 3.5% (w / v) to about 20% (w / v), about 3.5% (w / v) to about 15% (w / v), about 3.5% (w / v) to about 12% (w / v), about 3.5% (w / v) to about 10% (w / v), about 3.5% (w / v) to about 8% (w / v), about 3.5% (w / v) to about 6% (w / v), about 3.5% (w / v) to about 5.5% (w / v), about 3.5% (w / v) to about 5% (w / v), about 3.5% (w / v) to about 4.5% (w / v), about 3.5% (w / v) to about 4.2% (w / v), about 3.5% (w / v) to about 4% (w / v), about 3.8% (w / v) to about 12% (w / v), about 3.8% (w / v) to about 10% (w / v), about 3.8% (w / v) to about 8% (w / v), about 3.8% (w / v) to about 5% (w / v), about 3.8% (w / v) to about 4.5% (w / v), about 3.8% (w / v) to about 4.2% (w / v), about 4% (w / v) to about 20% (w / v), about 4% (w / v) to about 15% (w / v), about 4% (w / v) to about 12% (w / v), about 4% (w / v) to about 10% (w / v), about 4% (w / v) to about 8% (w / v), about 4% (w / v) to about 6% (w / v), about 4% (w / v) to about 5.5% (w / v), about 4% (w / v) to about 5% (w / v), about 4% (w / v) to about 4.5% (w / v), about 5% (w / v) to about 20% (w / v), about 5% (w / v) to about 15% (w / v), about 5% (w / v) to about 12% (w / v), about 5% (w / v) to about 10% (w / v), about 5% (w / v) to about 8% (w / v), about 5% (w / v) to about 6% (w / v), about 5% (w / v) to about 5.7% (w / v), about 5% (w / v) to about 5.5% (w / v), about 5.3% (w / v) to about 20% (w / v), about 5.3% (w / v) to about 15% (w / v), about 5.3% (w / v) to about 12% (w / v), about 5.3% (w / v) to about 10% (w / v), about 5.3% (w / v) to about 8% (w / v), or about 5.3% (w / v) to about 6% (w / v), about 5.3% (w / v) to about 5.7% (w / v), about 5.5% (w / v) to about 20% (w / v), about 5.5% (w / v) to about 15% (w / v), about 5.5% (w / v) to about 12% (w / v), about 5.5% (w / v) to about 10% (w / v), about 5.5% (w / v) to about 8% (w / v), or about 5.5% (w / v) to about 6% (w / v). In one embodiment, the concentration of sucrose may be about 2% (w / v) to about 6% (w / v).

[0061] In certain embodiments, the concentration of sucrose may be about 4% (w / v).

[0062] In certain embodiments, the concentration of sucrose may be about 5.5% (w / v).

[0063] (4) Amino acids

[0064] Liquid formulations according to the aspect of the invention contain amino acids.

[0065] Liquid formulations according to the aspect of the invention contain methionine.

[0066] The concentration of the methionine is about 1 mM to about 400 mM, about 2.7 mM to about 400 mM, about 2.7 mM to about 300 mM, about 2.7 mM to about 200 mM, about 2.7 mM to about 100 mM, about 2.7 mM to about 90 mM, about 2.7 mM to about 50 mM, about 2.7 mM to about 40 mM, about 2.7 mM to about 35 mM, about 2.7 mM to about 30 mM, about 10 mM to about 400 mM, about 10 mM to about 300 mM, about 10 mM to about 200 mM, about 10 mM to about 100 mM, about 10 mM to about 90 mM, about 10 mM to about 50 mM, about 10 mM to about 40 mM, about 10 mM to about 35 mM, about 10 mM to about 30 mM, about 15 mM to about 100 mM, about 15 mM to about 50 mM, about 15 mM to about 40 mM, about 15 mM to about 35 mM, about 15 mM to about 30 mM, about 20 mM to about 100 mM, about 20 mM to about 90 mM, about 20 mM to about 80 mM, about 20 mM to about 70 mM, about 20 mM to about 60 mM, about 20 mM to about 50 mM, about 20 mM to about 40 mM, about 20 mM to about 35 mM, about 20 mM to about 30 mM, about 20 mM to about 28 mM, about 22 mM to about 100 mM, about 22 mM to about 90 mM, about 22 mM to about 50 mM, about 22 mM to about 40 mM, about 22 It can be from about 25 mM to about 35 mM, from about 22 mM to about 30 mM, from about 22 mM to about 28 mM, from about 25 mM to about 100 mM, from about 25 mM to about 90 mM, from about 25 mM to about 50 mM, from about 25 mM to about 40 mM, from about 25 mM to about 35 mM, or from about 25 mM to about 30 mM.

[0067] In one specific embodiment, the concentration of methionine can be any range or any value selected from about 2.7 mM to about 90 mM.

[0068] In one specific example, the concentration of methionine may be from about 20 mM to about 90 mM.

[0069] In certain embodiments, the concentration of methionine may be about 20 mM to about 30 mM.

[0070] In certain embodiments, the concentration of methionine may be about 25 mM.

[0071] The above methionine may have one or more functions of a viscosity reducing agent, a stabilizer, and an antioxidant. The above methionine may have all of the functions of a viscosity reducing agent, a stabilizer, and an antioxidant.

[0072] The term "viscosity reducing agent" or "viscosity reducer" refers to a substance that lowers the viscosity of a liquid substance.

[0073] The term "stabilizer" refers to a substance added to a substance during preservation to prevent changes in state or chemical change. The stabilizer may be a thermal stabilizer. The stabilizer may inhibit antibody aggregation or denaturation.

[0074] The term "antioxidant" is a general term for substances that prevent oxidation, and specifically, in this specification, may mean a substance that prevents oxidation of a protein (e.g., an antibody).

[0075] The above methionine increases the aggregation temperature (T) of the antibody in the liquid formulation. agg) can increase T. When methionine is included in the anti-IL-4Rα antibody-containing liquid formulation, the liquid formulation has a higher T than when arginine or histidine is included. agg Therefore, the methionine can increase the thermal stability of the anti-IL-4Rα antibody-containing liquid formulation.

[0076] The above methionine has a diffusion interaction parameter (K) of the liquid formulation D ) can increase. When methionine is included in the anti-IL-4Rα antibody-containing liquid formulation, the liquid formulation has a higher K than when it includes sucrose, trehalose, sorbitol, mannitol, arginine, lysine, histidine, or NaCl. D Therefore, the methionine can increase the chemical stability of the anti-IL-4Rα antibody-containing liquid formulation by inhibiting the aggregation of the anti-IL-4Rα antibody in the liquid formulation.

[0077] The above methionine can reduce the viscosity (cP) of the liquid formulation. When methionine is included in the anti-IL-4Rα antibody-containing liquid formulation, the liquid formulation can have a lower viscosity than when methionine is not included. When methionine is added to the anti-IL-4Rα antibody-containing liquid formulation containing a saccharide, the liquid formulation can have a lower viscosity than when methionine is not added. When lysine is replaced with methionine in the anti-IL-4Rα antibody-containing liquid formulation containing a saccharide and lysine, the liquid formulation can have a lower viscosity than when lysine is not replaced with methionine. Therefore, since the methionine reduces the viscosity of the anti-IL-4Rα antibody-containing liquid formulation, it can be used as a viscosity-reducing agent.

[0078] The above methionine may reduce the high molecular weight species content ratio (%HMW) of the antibody in the liquid formulation. When methionine is included in the anti-IL-4Rα antibody-containing liquid formulation, the %HMW of the antibody in the liquid formulation may be reduced when stored under the same conditions (e.g., 4 weeks under accelerated stability conditions of 25±2°C and 60±5% RH, or 4 weeks under harsh stability conditions of 40±2°C and 75+5% RH) compared to when methionine is not included. When methionine is added to a liquid formulation containing anti-IL-4Rα antibody with a saccharide, the %HMW of the antibody in the liquid formulation may decrease when stored under the same conditions (e.g., 4 weeks under accelerated stability conditions of 25±2°C and 60±5% RH, or 4 weeks under harsh stability conditions of 40±2°C and 75+5% RH) compared to when methionine is not added. When lysine is replaced with methionine in a liquid formulation containing anti-IL-4Rα antibody with a saccharide and lysine, the %HMW of the antibody in the liquid formulation may decrease when stored under the same conditions (e.g., 4 weeks under accelerated stability conditions of 25±2°C and 60±5% RH, or 4 weeks under harsh stability conditions of 40±2°C and 75+5% RH) compared to when lysine is not replaced with methionine. Therefore, the methionine can increase the stability of the anti-IL-4Rα antibody-containing liquid formulation by inhibiting the aggregation of anti-IL-4Rα antibodies in the liquid formulation.

[0079] The above methionine can reduce the acidic species content ratio (%Acidic) of the antibody in the liquid formulation. When methionine is included in the anti-IL-4Rα antibody-containing liquid formulation, the %Acidic of the antibody in the liquid formulation can be reduced when stored under the same conditions (e.g., 4 weeks under accelerated stability conditions of 25±2°C and 60±5% RH) compared to when methionine is not included. Therefore, the methionine can reduce the production of acidic species of the anti-IL-4Rα antibody in the liquid formulation, thereby increasing the stability of the anti-IL-4Rα antibody-containing liquid formulation.

[0080] The above methionine is the oxidation rate (%Met) of the methionine residue in the antibody in the liquid formulation 257 ) can be reduced. When methionine is included in the anti-IL-4Rα antibody-containing liquid formulation, the %Met of the antibody in the liquid formulation is reduced when stored under the same conditions (e.g., 4 weeks under accelerated stability conditions of 25±2°C and 60±5% RH) compared to when methionine is not included. 257 This can be reduced. Therefore, the methionine can increase the stability of the anti-IL-4Rα antibody-containing liquid formulation by inhibiting oxidation of the anti-IL-4Rα antibody in the liquid formulation.

[0081] The liquid formulation according to one aspect may additionally include histidine. Accordingly, the liquid formulation may be a liquid formulation comprising an anti-IL-4Rα antibody; a buffer; a saccharide; methionine; and histidine, and having a pH of about 4.9 to about 5.5.

[0082] The concentration of the histidine may be about 0.1 mM to about 400 mM, about 0.1 mM to about 92.4 mM, about 0.5 mM to about 400 mM, about 0.5 mM to about 92.4 mM, about 1 mM to about 400 mM, about 10 mM to about 400 mM, about 20 mM to about 400 mM, about 20 mM to about 300 mM, about 20 mM to about 200 mM, about 20 mM to about 100 mM, or about 20 mM to about 92.4 mM.

[0083] In one specific embodiment, the concentration of histidine can be any range or any value selected from about 20 mM to about 92.4 mM. For example, the concentration of histidine may be about 20 mM to about 92.4 mM, about 20 mM to about 90 mM, about 20 mM to about 80 mM, about 20 mM to about 60 mM, about 30 mM to about 92.4 mM, about 30 mM to about 90 mM, about 30 mM to about 80 mM, about 30 mM to about 60 mM, about 40 mM to about 92.4 mM, about 40 mM to about 90 mM, about 40 mM to about 80 mM, about 40 mM to about 60 mM, about 45 mM to about 92.4 mM, about 45 mM to about 90 mM, about 45 mM to about 80 mM, about 45 mM to about 60 mM, about 45 mM to about 55 mM, about 50 mM to about 92.4 mM, about 50 mM to about It may be 90 mM, about 50 mM to about 80 mM, or about 50 mM to about 60 mM.

[0084] In one specific example, the histidine may have the function of a buffer.

[0085] In one specific example, the histidine may have the function of a stabilizer.

[0086] The histidine can reduce the acidic species content ratio (%Acidic) of the antibody in the liquid formulation. When histidine is included in the anti-IL-4Rα antibody-containing liquid formulation, the %Acidic of the antibody in the liquid formulation can be reduced when stored under the same conditions (e.g., 4 weeks under accelerated stability conditions of 25±2°C and 60±5% RH) compared to when histidine is not included. Therefore, the histidine can reduce the production of acidic species of the anti-IL-4Rα antibody in the liquid formulation, thereby increasing the stability of the anti-IL-4Rα antibody-containing liquid formulation.

[0087] The above histidine is the oxidation rate (%Met) of methionine residues in the antibody in the liquid formulation 257 ) can be reduced. When histidine is included in the anti-IL-4Rα antibody-containing liquid formulation, the %Met of the antibody in the liquid formulation is reduced when stored under the same conditions (e.g., 4 weeks under accelerated stability conditions of 25±2°C and 60±5% RH) compared to when histidine is not included. 257 This can be reduced. Therefore, the histidine can increase the stability of the anti-IL-4Rα antibody-containing liquid formulation by inhibiting the oxidation of the anti-IL-4Rα antibody in the liquid formulation.

[0088] According to one aspect, the liquid formulation may be arginine-free. The liquid formulation may not contain arginine as a viscosity-reducing agent. The arginine may include arginine or a salt thereof, such as arginine hydrochloride.

[0089] The liquid formulation according to the aspect may not contain any amino acids other than methionine and histidine.

[0090] The term 'free of component A' or 'substantially free of A' may be interpreted to include cases where component A is not present at all, or where component A is present in trace amounts that do not substantially affect the properties of the formulation, or where it is present in undetectable amounts.

[0091] In this specification, the phrase "does not contain arginine" may be interpreted to mean that the arginine component is not present in the formulation, or is contained in an amount that is insufficient to function as the intended viscosity reducing agent in the formulation.

[0092] Since the above liquid formulation contains methionine instead of arginine, both viscosity and %HMW are reduced compared to Dupixent containing arginine, so viscosity and stability may be improved simultaneously.

[0093] (5) Surfactant

[0094] Liquid formulations according to the aspect may additionally contain a surfactant.

[0095] Accordingly, the liquid formulation may be a liquid formulation comprising an anti-IL-4Rα antibody; a buffer; a sugar; methionine; and a surfactant, and having a pH of about 4.9 to about 5.5.

[0096] The above liquid formulation may be a liquid formulation comprising an anti-IL-4Rα antibody; a buffer; a sugar; methionine; histidine; and a surfactant, and having a pH of about 4.9 to about 5.5.

[0097] The surfactant may be selected from any pharmaceutically acceptable surfactants capable of evenly dispersing a protein (e.g., antibody) in a liquid formulation medium.

[0098] The above surfactant may be a nonionic surfactant.

[0099] Specifically, the surfactant may be at least one selected from the group consisting of polysorbate, poloxamer, sorbitan ester of other fatty acids, polyethylene-polypropylene glycol, polyoxyethylene compound, and sodium dodecyl sulfate (SDS).

[0100] The above polysorbate may include polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 65, polysorbate 80, and polysorbate 85.

[0101] The above poloxamer may include a PEO-PPO-PEO copolymer (PEO is poly(ethylene oxide) and PPO is poly(propylene oxide)).

[0102] The above sorbitan esters of other fatty acids may mean sorbitan esters of other fatty acids than polysorbates, and may include, for example, sorbitan polyethoxylates.

[0103] The above polyoxyethylene compound may include polyoxyethylene-stearate, polyoxyethylene alkyl ether (alkyl: C1-C30), polyoxyethylene monolyl ether, alkylphenyl polyoxyethylene copolymer (alkyl: C1-C30), etc.

[0104] In one specific embodiment, the surfactant may be a polysorbate.

[0105] In one embodiment, the surfactant may include one or more selected from polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 65, polysorbate 80, and polysorbate 85.

[0106] In one embodiment, the surfactant may include polysorbate 20, polysorbate 80, or a combination thereof.

[0107] In certain embodiments, the surfactant may comprise polysorbate 80. In certain embodiments, the surfactant may be polysorbate 80.

[0108] The concentration of the surfactant may be any range or any value selected from about 0.01% (w / v) to about 0.9% (w / v). For example, the concentration of the surfactant may be from about 0.01% (w / v) to about 0.9% (w / v), from about 0.01% (w / v) to about 0.5% (w / v), from about 0.1% (w / v) to about 0.9% (w / v), from about 0.1% (w / v) to about 0.5% (w / v), from about 0.1% (w / v) to about 0.4% (w / v), from about 0.1% (w / v) to about 0.3% (w / v), from about 0.15% (w / v) to about 0.9% (w / v), from about 0.15% (w / v) to about 0.5% (w / v), from about 0.15% (w / v) to about 0.4% (w / v), from about 0.15% (w / v) to about 0.3% (w / v), or about 0.15% (w / v) to about 0.25% (w / v).

[0109] In certain embodiments, the concentration of the surfactant may be about 0.2% (w / v).

[0110] (6) Diluent

[0111] Liquid formulations according to the aspect may additionally contain a diluent.

[0112] The diluent may be an aqueous carrier. The aqueous carrier may be a pharmaceutically acceptable carrier that is safe and non-toxic when administered to humans, such as water, saline solution, Ringer's solution, dextrose, or a mixture thereof.

[0113] In one specific example, the diluent may be water. Accordingly, the liquid formulation may be an aqueous liquid formulation.

[0114] (7) Formulation

[0115] The term "liquid formulation" means a formulation in liquid form.

[0116] The liquid formulation according to the aspect is a stable liquid formulation of anti-IL-4Rα antibody.

[0117] The liquid formulation according to the aspect may be a pharmaceutical formulation of an anti-IL-4Rα antibody.

[0118] The term "pharmaceutical composition" or "pharmaceutical preparation" means a preparation that allows the biological activity of the active ingredient to be effectively carried out and does not contain additional components that are seriously toxic to the subject to which the preparation is administered.

[0119] The term "pharmaceutical formulation" refers to the product of a process that combines an active drug with chemicals to produce the final drug product.

[0120] The term "pharmaceutically acceptable" may refer to excipients, carriers, vehicles, diluents, additives, salts, etc. that are suitable for administration to a subject.

[0121] Liquid formulations according to the daily pattern are Dupixent ® ) may be a biosimilar. The liquid formulation according to the aspect has improved stability and improved viscosity compared to Dupixent.

[0122] The term "biosimilar," also known as "biogeneric," refers to a copy of an original biopharmaceutical. Because biopharmaceuticals are produced through cells rather than synthetic chemicals, they cannot be perfectly identical to the original drug. Therefore, a biopharmaceutical copy is called a biosimilar, meaning it is similar, though not identical, to the original drug.

[0123] Liquid formulations according to the aspect of the work may be selected from the following items:

[0124] 1) A liquid formulation comprising an anti-IL-4Rα antibody; a buffer; a sugar; and methionine, and having a pH of about 4.9 to about 5.5;

[0125] 2) A liquid formulation comprising an anti-IL-4Rα antibody; a buffer; a sugar; methionine; and histidine, and having a pH of about 4.9 to about 5.5;

[0126] 3) A liquid formulation comprising an anti-IL-4Rα antibody; a buffer; a sugar; methionine; and a surfactant, and having a pH of about 4.9 to about 5.5;

[0127] 4) A liquid formulation comprising an anti-IL-4Rα antibody; a buffer; a sugar; methionine; histidine; and a surfactant, and having a pH of about 4.9 to about 5.5;

[0128] 5) A liquid formulation comprising about 5 mg / mL to about 300 mg / mL of dupilumab; about 10 mM to about 40 mM of acetate; about 2% (w / v) to about 10% (w / v) of sucrose; about 10 mM to about 100 mM of methionine; about 20 mM to about 92.4 mM of histidine; and about 0.1% (w / v) to about 0.9% (w / v) of polysorbate 80, and having a pH of about 4.9 to about 5.5;

[0129] 6) A liquid formulation comprising 100 mg / mL to 200 mg / mL of dupilumab; 30 mM±5 mM of acetate; 4%±1.0% (w / v) of sucrose; 25 mM±5 mM of methionine; 20 mM to 92.4 mM of histidine; and 0.2%±0.1% (w / v) of polysorbate 80, and having a pH of about 4.9 to about 5.5;

[0130] 7) A liquid formulation comprising 175 mg / mL of dupilumab; 30 mM±5 mM of acetate; 4%±1.0% (w / v) of sucrose; 25 mM±5 mM of methionine; 20 mM to 92.4 mM of histidine; and 0.2%±0.1% (w / v) of polysorbate 80, and having a pH of about 4.9 to about 5.5;

[0131] 8) A liquid formulation comprising 175 mg / mL of dupilumab; 30 mM±5 mM of acetate; 4%±1.0% (w / v) of sucrose; 25 mM±5 mM of methionine; 20 mM±5 mM of histidine; and 0.2%±0.1% (w / v) of polysorbate 80, having a pH of about 4.9;

[0132] 9) A liquid formulation comprising 175 mg / mL of dupilumab; 30 mM±5 mM of acetate; 4%±1.0% (w / v) of sucrose; 25 mM±5 mM of methionine; 50 mM±5 mM of histidine; and 0.2%±0.1% (w / v) of polysorbate 80, having a pH of about 4.8;

[0133] 10) A liquid formulation comprising 175 mg / mL of dupilumab; 30 mM±5 mM of acetate; 4%±1.0% (w / v) of sucrose; 25 mM±5 mM of methionine; 50 mM±5 mM of histidine; and 0.2%±0.1% (w / v) of polysorbate 80, having a pH of about 5.2;

[0134] 11) A liquid formulation comprising 175 mg / mL of dupilumab; 30 mM±5 mM of acetate; 4%±1.0% (w / v) of sucrose; 25 mM±5 mM of methionine; 80 mM±5 mM of histidine; and 0.2%±0.1% (w / v) of polysorbate 80, having a pH of about 4.9;

[0135] 12) A liquid formulation comprising 175 mg / mL of dupilumab; 30 mM±5 mM of acetate; 4%±1.0% (w / v) of sucrose; 25 mM±5 mM of methionine; 80 mM±5 mM of histidine; and 0.2%±0.1% (w / v) of polysorbate 80, having a pH of about 5.5;

[0136] 13) A liquid formulation comprising 175 mg / mL of dupilumab; 30 mM±5 mM of acetate; 4%±1.0% (w / v) of sucrose; 25 mM±5 mM of methionine; 92.4 mM±5 mM of histidine; and 0.2%±0.1% (w / v) of polysorbate 80, having a pH of about 5.2;

[0137] 14) A liquid formulation comprising 100 mg / mL to 200 mg / mL of dupilumab; 30 mM±5 mM of acetate; 5.5%±1.0% (w / v) of sucrose; 25 mM±5 mM of methionine; 20 mM to 92.4 mM of histidine; and 0.2%±0.1% (w / v) of polysorbate 80, and having a pH of about 4.9 to about 5.5;

[0138] 15) A liquid formulation comprising 175 mg / mL of dupilumab; 30 mM±5 mM of acetate; 5.5%±1.0% (w / v) of sucrose; 25 mM±5 mM of methionine; 20 mM to 92.4 mM of histidine; and 0.2%±0.1% (w / v) of polysorbate 80, and having a pH of about 4.9 to about 5.5;

[0139] 16) A liquid formulation comprising 175 mg / mL of dupilumab; 30 mM±5 mM of acetate; 5.5%±1.0% (w / v) of sucrose; 25 mM±5 mM of methionine; 20 mM±5 mM of histidine; and 0.2%±0.1% (w / v) of polysorbate 80, having a pH of about 4.9;

[0140] 17) A liquid formulation comprising 175 mg / mL of dupilumab; 30 mM±5 mM of acetate; 5.5%±1.0% (w / v) of sucrose; 25 mM±5 mM of methionine; 50 mM±5 mM of histidine; and 0.2%±0.1% (w / v) of polysorbate 80, having a pH of about 4.8;

[0141] 18) A liquid formulation comprising 175 mg / mL of dupilumab; 30 mM±5 mM of acetate; 5.5%±1.0% (w / v) of sucrose; 25 mM±5 mM of methionine; 50 mM±5 mM of histidine; and 0.2%±0.1% (w / v) of polysorbate 80, having a pH of about 5.2;

[0142] 19) A liquid formulation comprising 175 mg / mL of dupilumab; 30 mM±5 mM of acetate; 5.5%±1.0% (w / v) of sucrose; 25 mM±5 mM of methionine; 80 mM±5 mM of histidine; and 0.2%±0.1% (w / v) of polysorbate 80, having a pH of about 4.9;

[0143] 20) A liquid formulation comprising 175 mg / mL of dupilumab; 30 mM±5 mM of acetate; 5.5%±1.0% (w / v) of sucrose; 25 mM±5 mM of methionine; 80 mM±5 mM of histidine; and 0.2%±0.1% (w / v) of polysorbate 80, having a pH of about 5.5; or

[0144] 21) A liquid formulation comprising 175 mg / mL of dupilumab; 30 mM±5 mM of acetate; 5.5%±1.0% (w / v) of sucrose; 25 mM±5 mM of methionine; 92.4 mM±5 mM of histidine; and 0.2%±0.1% (w / v) of polysorbate 80, having a pH of about 5.2.

[0145] (8) Stability and viscosity

[0146] Liquid formulations according to the aspect can have low viscosity while ensuring the stability of the antibody.

[0147] In the examples of this specification, three experiments were performed. First, T agg and K D Through measurements, we compared and evaluated the thermal and chemical stability of different excipients, confirming the superiority of methionine among excipients. Second, we evaluated viscosity, %HMW, %Acidic, and %Met257 to confirm the functions of methionine as a viscosity reducer, stabilizer, and antioxidant. Third, through a comparative evaluation of viscosity and %HMW with the Dupixent formulation, we confirmed that the methionine-containing anti-IL-4Rα antibody liquid formulation can simultaneously improve stability and viscosity compared to commercialized formulations.

[0148] The term "stability" means that an antibody (e.g., dupilumab) contained in a formulation substantially retains its physical stability, chemical stability, and / or biological activity before and after administration, during further manufacturing processes, storage, or preservation. For example, it can be understood to mean that the degree of loss of stability, such as aggregation, degradation, denaturation (acidic or basic), oxidation, etc. of the antibody contained in the formulation is 20% or less, 15% or less, 10% or less, or 5% or less compared to the initial storage. Accordingly, "excellent stability" or "improved stability" can mean a low protein aggregation rate, a low protein degradation rate, a low protein denaturation rate, a low amino acid oxidation rate, etc. during storage. Physical stability, chemical stability, and / or biological activity can be evaluated by conventionally known methods.

[0149] A method for evaluating the stability of antibodies contained in a formulation in a short period of time is to expose the formulation to heat (temperature), which is a factor that causes structural and physicochemical denaturation of proteins, and then measure the aggregation temperature (T agg ), diffusion interaction parameter (K D ) etc. For example, "improved stability" is measured T for the formulation aggor K D The value may mean an increase of 0.01% or more, 0.1% or more, 0.5% or more, 1% or more, 2% or more, 3% or more, 4% or more, 5% or more, 10% or more, 15% or more, or 20% or more compared to the existing formulation. T agg or K D An increase in the value may indicate improved stability by reducing the extent to which antibodies within the formulation aggregation due to external factors (e.g., heat, temperature).

[0150] The term "aggregate" may refer to high molecular weight (HMW) species formed by aggregation of antibody proteins. The term "protein aggregation rate" may be expressed as the percentage of high molecular weight species content (%HMW) of the antibody in the formulation at a given point in time. %HMW may be measured by, but is not limited to, size exclusion chromatography (SEC). For example, "improved stability" may mean that the %HMW of the antibody measured for the formulation is reduced by at least 0.01%, at least 0.1%, at least 0.5%, at least 1%, at least 2%, at least 3%, at least 4%, at least 5%, at least 10%, at least 15%, or at least 20% compared to the original formulation. A decrease in %HMW may mean that the degree to which the antibody in the formulation undergoes aggregation is reduced, thereby improving stability.

[0151] The term "protein denaturation" may be expressed as the percentage acidic species content (%Acidic) of the antibody within the formulation at a given point in time. %Acidic may be measured by, but is not limited to, imaged capillary isoelectric focusing (icIEF). For example, "improved stability" may mean that the %Acidic of the antibody measured for the formulation is reduced by at least 0.01%, at least 0.1%, at least 0.5%, at least 1%, at least 2%, at least 3%, at least 4%, at least 5%, at least 10%, at least 15%, or at least 20% compared to the original formulation. A decrease in %Acidic may mean that the extent to which the antibody within the formulation undergoes denaturation is reduced, thereby improving stability.

[0152] The term "amino acid oxidation rate" may refer to the percentage of oxidized amino acids among the amino acids of an antibody protein (e.g., the percentage of oxidized methionine). In one embodiment, the amino acid oxidation rate is methionine (Met), which is the 257th amino acid residue of dupilumab, an anti-IL-4Rα antibody. 257 ) Oxidation ratio (%Met) 257 ) can mean %Met 257 can be measured by, but is not limited to, liquid chromatography-mass spectrometry (LC-MS). For example, "improved stability" means the %Met of dupilumab measured for the formulation. 257 This may mean a decrease of 0.01% or more, 0.1% or more, 0.5% or more, 1% or more, 2% or more, 3% or more, 4% or more, 5% or more, 10% or more, 15% or more, or 20% or more compared to the existing formulation. %Met 257 A decrease in may indicate improved stability due to a decrease in the extent to which the antibody within the formulation undergoes oxidation.

[0153] The stability assessment may be performed immediately after manufacturing the formulation; or after a certain period of storage under accelerated stability conditions or stress stability conditions. The accelerated stability conditions may include conditions used in accelerated tests for pharmaceuticals, such as a temperature of 25±2°C and a relative humidity (RH) of 60±5%. The stress stability conditions may include conditions used in stress tests for pharmaceuticals, such as a temperature of 40±2°C and a relative humidity (RH) of 75+5%.

[0154] The above liquid formulation has an increased coagulation temperature (T agg ) may have.

[0155] The above liquid formulation contains methionine, thereby increasing T compared to the anti-IL-4Rα antibody liquid formulation containing arginine. agg It may have .

[0156] The above liquid formulation has an increased diffusion interaction parameter (K D ) may have.

[0157] The above liquid formulation contains methionine, thereby increasing K compared to the anti-IL-4Rα antibody liquid formulation containing arginine. D The liquid formulation may have increased K by including methionine compared to anti-IL-4Rα antibody liquid formulations containing sucrose, trehalose, sorbitol, mannitol, arginine, lysine, histidine, or NaCl. D The liquid formulation may have an increased K compared to the anti-IL-4Rα antibody liquid formulation comprising a dual buffer of 12.5 mM acetate and 20 mM histidine, pH 5.9, 5% (w / v) sucrose, and 50 mM arginine by including methionine. D It may have .

[0158] The above liquid formulation may have improved viscosity. "Improving" the viscosity may mean "reducing" the viscosity. Accordingly, the above liquid formulation may have reduced viscosity.

[0159] The above liquid formulation may have improved viscosity by including methionine compared to an anti-IL-4Rα antibody liquid formulation that does not include methionine.

[0160] In one embodiment, it was confirmed that viscosity was improved when methionine was added to a liquid formulation of an anti-IL-4Rα antibody containing a saccharide. Furthermore, it was confirmed that methionine functioned better as a viscosity-reducing agent than lysine in a liquid formulation of an anti-IL-4Rα antibody containing a saccharide.

[0161] The liquid formulation according to one aspect may have a reduced viscosity compared to Dupixent. In one embodiment, the liquid formulation according to one aspect may have a reduced viscosity compared to the viscosity of 11.1 cP (centiPoise) of Dupixent 200 mg. In another embodiment, the liquid formulation according to one aspect may have a reduced viscosity compared to the viscosity of 300 mg of Dupixent.

[0162] The viscosity of the liquid formulation may be less than 11.1 cP (centiPoise). The viscosity of the liquid formulation may be less than 11.1 cP, 11.0 cP or less, 10.5 cP or less, 10.0 cP or less, 9.5 cP or less, 9.0 cP or less, 8.5 cP or less, 8.0 cP or less, 7.5 cP or less, 7.0 cP or less, or 6.5 cP or less. The viscosity of the above liquid formulation is 5.0 cP or more and less than 11.1 cP, 5.0 cP to 11.0 cP, 5.0 cP to 10.5 cP, 5.0 cP to 10.0 cP, 5.0 cP to 9.5 cP, 5.0 cP to 9.0 cP, 5.0 cP to 8.5 cP, 5.0 cP to 8.0 cP, 5.0 cP to 7.5 cP, 5.0 cP to 7.0 cP, 5.0 cP to 6.5 cP, 5.5 cP or more and less than 11.1 cP, 5.5 cP to 11.0 cP, 5.5 cP to 10.5 cP, 5.5 cP to 10.0 cP, 5.5 cP to 9.5 cP, 5.5 cP to 9.0 cP, 5.5 cP to 8.5 cP, 5.5 cP to 8.0 cP, 5.5 cP to 7.5 cP, 5.5 cP to 7.0 cP, 5.5 cP to 6.5 cP, 6.0 cP or more to less than 11.1 cP, 6.0 cP to 11.0 cP, 6.0 cP to 10.5 cP, 6.0 cP to 10.0 cP, 6.0 cP to 9.5 cP, 6.0 cP to 9.0 cP, 6.0 cP to 8.5 cP, 6.0 cP to 8.0 cP, 6.0 cP to 7.5 cP, 6.0 cP to 7.0 cP, 6.0 cP to 6.5 cP, 6.5 cP or more but less than 11.1 cP, 6.5 cP to 11.0 cP, 6.5 cP to 10.5 cP, 6.5 cP to 10.0 cP, 6.5 cP to 9.5 cP, 6.5 cP to 9.0 cP, 6.5 cP to 8.5 cP, 6.5 cP to 8.0 cP, 6.5 cP to 7.5 cP, 6.5 cP to 7.0 cP, 7.0 cP or more to less than 11.1 cP, 7.0 cP to 11.0 cP, 7.0 cP to 10.5 cP, 7.0 cP to 10.0 cP, 7.0 cP to 9.5 cP, 7.0 cP to 9.0 cP, 7.0 cP to 8.5 cP, 7.0 cP to 8.0 cP, 7.5 cP or more to less than 11.1 cP, 7.5 cP to 11.0 cP, 7.5 cP to 10.5 cP, 7.5 cP to 10.0 cP, 7.5 cP to 9.5 cP, 7.5 cP to 9.0 cP, 7.5 cP to 8.5 cP, It may be 7.5 cP to 8.0 cP, 8.0 cP or more to less than 11.1 cP, 8.0 cP to 11.0 cP, 8.0 cP to 10.5 cP, 8.0 cP to 10.0 cP, 8.0 cP to 9.5 cP, 8.0 cP to 9.0 cP, or 8.0 cP to 8.5 cP.

[0163] In one specific embodiment, the viscosity of the liquid formulation is about 6.0 cP, about 6.1 cP, about 6.2 cP, about 6.3 cP, about 6.4 cP, about 6.5 cP, about 6.6 cP, about 6.7 cP, about 6.8 cP, about 6.9 cP, about 7.0 cP, about 7.1 cP, about 7.2 cP, about 7.3 cP, about 7.4 cP, about 7.5 cP, about 7.6 cP, about 7.7 cP, about 7.8 cP, about 7.9 cP, about 8.0 cP, about 8.1 cP, about 8.2 cP, about 8.3 cP, about 8.4 cP, about 8.5 cP, about 8.6 cP, about 8.7 cP, about 8.8 cP, about 8.9 cP, about 9.0 cP, about 9.1 cP, about 9.2 cP, about 9.3 cP, about 9.4 cP, or about 9.5 cP.

[0164] The above liquid formulation may have a viscosity that is reduced by at least 10%, at least 15%, at least 20%, at least 25%, or at least 30% relative to the viscosity of the Dupixent formulation.

[0165] The above liquid formulation may have a reduced high molecular weight species content (%HMW).

[0166] The above liquid formulation may have a reduced %HMW by including methionine compared to an anti-IL-4Rα antibody liquid formulation that does not include methionine.

[0167] The above liquid formulation may have a %HMW of the antibody measured after 4 weeks of storage at 40°C reduced by at least 5%, at least 10%, at least 15%, or at least 20% compared to a formulation that does not contain methionine.

[0168] The above liquid formulation may have a %HMW of antibody measured after 4 weeks of storage at 25°C that is reduced by at least 5%, at least 10%, or at least 15% compared to a formulation that does not contain methionine.

[0169] The above liquid formulation may have a %HMW of antibody measured after storage at 25°C for 4 weeks of 3.1% or less, 3.0% or less, 2.9% or less, 2.8% or less, 2.7% or less, 2.6% or less, 2.5% or less, 2.4% or less, or 2.3% or less.

[0170] The liquid formulation may have a %HMW of antibody measured immediately after manufacture of less than 2.03%, 2.02% or less, 2.01% or less, 2.00% or less, 1.99% or less, 1.98% or less, 1.97% or less, 1.96% or less, 1.95% or less, or 1.94% or less.

[0171] The above liquid formulation may have a reduced acid species content (%Acidic).

[0172] The above liquid formulation may have a reduced %Acidic by including methionine compared to an anti-IL-4Rα antibody liquid formulation that does not include methionine.

[0173] The above liquid formulation may have a %Acidic of antibody measured after 4 weeks of storage at 25°C reduced by at least 1%, at least 2%, at least 3%, at least 4%, at least 5%, or at least 7% compared to a formulation that does not contain methionine.

[0174] The above liquid formulation may have a %Acidic of antibody measured after 4 weeks of storage at 25°C of less than 41%, less than 40%, or less than 39%.

[0175] The above liquid formulation has a reduced oxidation rate of methionine residues (%Met 257 ) can have.

[0176] The above liquid formulation contains methionine, thereby reducing %Met compared to anti-IL-4Rα antibody liquid formulations that do not contain methionine. 257 can have

[0177] The above liquid formulation was stored for 4 weeks at 25°C and the %Met of the antibody was measured. 257 This may be reduced by 50% or more, 55% or more, or 60% or more compared to a formulation that does not contain methionine.

[0178] The above liquid formulation was stored for 4 weeks at 25°C and the %Met of the antibody was measured. 257 This can be less than 12%, less than 10%, less than 8%, less than 6%, or less than 5%.

[0179] (9) Device

[0180] Another aspect provides a device comprising a liquid formulation according to the above aspect.

[0181] The device is primarily used for parenteral administration. Parenteral administration may include, for example, subcutaneous, intramuscular, intravenous, intraperitoneal, intracerebrospinal, intraarticular, intrasynovial, or intrathecal administration. The device may be accompanied by instructions for administration.

[0182] The device may contain the liquid formulation in a container selected from, but not limited to, a syringe, a pre-filled syringe, a pre-filled pen, a microinfusor, an autoinjector, a bottle, a vial, and a tube.

[0183] The above device may be in a single-dose form or a multiple-dose form.

[0184] In one embodiment, the device may comprise the liquid formulation in a prefilled syringe. The prefilled syringe may be a single-dose prefilled syringe.

[0185] In one specific embodiment, the device may comprise the liquid formulation in a prefilled pen. The prefilled pen may be a single-dose prefilled pen.

[0186] (10) Treatment of diseases

[0187] Another aspect provides a method for treating an IL-4Rα-associated condition, comprising administering to a subject in need thereof a liquid formulation according to the above aspect. The liquid formulation may be in a form contained in a device.

[0188] Another aspect provides the use of a liquid formulation according to the above aspect in the manufacture of a medicament for treating an IL-4Rα associated condition.

[0189] The method of treating the IL-4Rα associated condition may further comprise, prior to the administering step, a step of identifying a subject in need of administration of an anti-IL-4Rα antibody (e.g., dupilumab).

[0190] The subject may be a subject requiring administration of a liquid formulation containing the anti-IL-4Rα antibody (e.g., dupilumab). The subject requiring administration of the liquid formulation containing the anti-IL-4Rα antibody (e.g., dupilumab) may be a subject having a disease or disorder that can be significantly treated (e.g., symptoms can be eliminated, alleviated, relieved, or improved, etc.) by administration of the anti-IL-4Rα antibody. The subject may be selected from mammals, including humans.

[0191] The above liquid formulation can be administered in a pharmaceutically effective amount.

[0192] The IL-4Rα associated condition may include any condition, disease, or disorder that can be treated by administration of an anti-IL-4Rα antibody. The IL-4Rα associated condition that can be treated by administration of an anti-IL-4Rα antibody (e.g., dupilumab) may include any indication currently approved or that may be approved in the future for an anti-IL-4Rα antibody (e.g., dupilumab).

[0193] The IL-4Rα-associated condition may be an IL-4-mediated disease and / or an IL-13-mediated disease. The IL-4-mediated disease may include any disease that can be treated by inhibition of IL-4 signaling. The IL-13-mediated disease may include any disease that can be treated by inhibition of IL-13 signaling. The IL-4Rα-associated condition may include any disease that can be treated by dual blockade of IL-4 and IL-13.

[0194] The IL-4Rα-related condition may include an inflammatory disease, an allergic disease, or an autoimmune disease. The inflammatory disease may be a Type 2 inflammatory disease.

[0195] The IL-4Rα-related condition may be any one selected from, but is not limited to, atopic dermatitis, asthma, chronic rhinosinusitis (CRS), chronic rhinosinusitis with nasal polyposis (CRSwNP), allergic fungal rhinosinusitis (AFRS), eosinophilic esophagitis (EoE), prurigo nodularis (PN), bullous pemhigoid (BP), chronic spontaneous urticarial (CSU), chronic inducible urticarial (CIndU), and chronic obstructive pulmonary disease (COPD).

[0196] The above atopic dermatitis may be moderate-to-severe atopic dermatitis.

[0197] The above asthma may be moderate-to-severe asthma.

[0198] (11) Route of administration and dosage

[0199] Liquid formulations, depending on the mode of administration, may be administered parenterally. Such parenteral routes may include subcutaneous administration, intravenous administration, etc. Parenteral administration may be by bolus injection or continuous infusion.

[0200] In certain embodiments, the liquid formulation may be for subcutaneous injection.

[0201] The above liquid formulation may be formulated into a formulation suitable for the above administration route. For example, the above liquid formulation may be formulated into an injection, a ready-to-use injectable, etc., but is not limited thereto.

[0202] The liquid formulation may be formulated to contain the entire amount or a pharmaceutically effective amount of the anti-IL-4Rα antibody (e.g., dupilumab) in one formulation, or may be formulated to contain two or more (e.g., two, three, four, five, six, seven, eight, nine, or ten) formulations. The liquid formulation may be included in a single-dose or multi-dose form of the device.

[0203] The above liquid formulation may be administered to the body in one dose (e.g., within 1 minute, within 30 seconds, within 20 seconds, or within 10 seconds); or may be administered to the body gradually over a period of, but not limited to, 5 minutes, 10 minutes, 30 minutes, 60 minutes, 90 minutes, 120 minutes, 150 minutes, 180 minutes, 210 minutes, or 240 minutes.

[0204] The target of administration of the above liquid formulation may be selected from mammals including primates (e.g., humans, etc.), rodents (e.g., mice, rats, guinea pigs, hamsters, rabbits, etc.), cats, dogs, pigs, cows, horses, etc.

[0205] The pharmaceutically effective amount of the above liquid formulation or the anti-IL-4Rα antibody (e.g., dupilumab) contained therein may refer to an amount or dosage that can exhibit a desired pharmacological effect, such as elimination, reduction, alleviation, or improvement of symptoms. The pharmaceutically effective amount may be determined in various ways depending on factors such as the formulation method, administration method, patient's age, weight, sex, pathological condition (severity of condition), food, administration time, administration interval, administration route, excretion rate, response sensitivity, previous therapy, clinical history, etc. The dosage may be adjusted according to the judgment of the physician in charge. The pharmaceutically effective amount may be administered at once or administered in two or more divided doses.

[0206] For example, the liquid formulation may be administered once every two to four weeks over a period of two weeks or more at a dose such that the anti-IL-4Rα antibody (e.g., dupilumab) is 300 mg, 250 mg, 200 mg, 150 mg, or 100 mg.

[0207] The above liquid formulation can be prepared as a general bulk formulation, and the components of the liquid formulation can be adjusted to a higher concentration than that required for administration and used after being suitably diluted before administration.

[0208] Liquid formulations of anti-IL-4Rα antibodies according to one aspect may exhibit improved stability and viscosity compared to existing commercially available formulations. Therefore, the liquid formulations may exhibit high stability and low viscosity even when containing high antibody concentrations, thereby increasing patient convenience during injection. Accordingly, the liquid formulations may be useful as pharmaceuticals for treating IL-4Rα-related conditions.

[0209] Figure 1 shows the coagulation temperature (T) for 10 formulations with different types of excipients and formulations with Dupixent composition. agg ) is a graph showing the results of measurement.

[0210] Figure 2 shows the diffusion interaction parameter (K) for 10 formulations with different types of excipients and formulations with Dupixent composition. D ) is a graph showing the results of measurement.

[0211] Figure 3 is a graph showing the results of viscosity measurements for formulations containing methionine and formulations not containing methionine.

[0212] Figure 4 is a graph showing the results of analyzing the high molecular weight species content ratio (%HMW) for formulations containing methionine and formulations not containing methionine.

[0213] Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the following examples are intended only to illustrate the present invention and are not intended to limit the scope of the present invention.

[0214]

[0215] [Experimental Method]

[0216] 1. Coagulation temperature (T agg ) measurement

[0217] The aggregation temperature was measured using dynamic light scattering (DLS) measurement. Specifically, each sample was diluted with each buffer and loaded into a 96-well plate. The plate was loaded into a DynoPro® Plate ReaderTM II (Wyatt Technology). The sample was heated at a rate of 0.25°C / min in the range of 25 to 80°C, and the size of the aggregates in the formulation was measured using DLS. The stability was evaluated by measuring the temperature at which the size of the aggregates changed, and the aggregation temperature (T agg ) The higher the value, the more stable the formulation.

[0218] 2. Diffusion interaction parameter K D analyze ​

[0219] To evaluate whether the binding force in the fluid phase is attractive or repulsive as the protein concentration increases when the sample is exposed at room temperature, the diffusion interaction parameter, K D was analyzed. This is a parameter that enables evaluation or ranking of excellent formulations according to aggregation tendency by evaluating the binding force between substances that causes irreversible aggregation. Specifically, each sample was diluted using a buffer (protein concentration: 2, 4, 6, 8, 10 mg / mL) and loaded into a 96-well plate. The plate was loaded into a DynaPro® Plate ReaderTM II (Wayatt Technology). The diffusion coefficient value according to the protein concentration of the prepared sample was measured, and then the slope and y-intercept of the graph were used to calculate K D If the value is positive, the repulsion between substances is dominant, so the cohesion tendency is weak, and if it is negative, the attraction is dominant, so the cohesion tendency is strong. K D The larger the positive number, the more stable the formulation.

[0220] 3. Viscosity measurement

[0221] The viscosity of the sample was measured using a viscometer (manufacturer: RheoSense, model number: VROC initium one plus). The analysis was conducted at a temperature of 25°C during the viscosity measurement. The same sample was divided into 11 segments and the slope fit R was calculated. 2 The viscosity value of each sample was calculated by taking the average value for values ​​greater than 0.9995.

[0222] 4. Analysis of high molecular weight species content (% high molecular weight species, %HMW)

[0223] The percentage of high molecular weight species (%HMW) was determined using Waters size exclusion chromatography (SEC). The protein was separated into three peaks based on its molecular weight. These three peaks correspond to the HMW peak (protein aggregation), the Monomer peak, and the LMW peak (protein degradation), in descending order of retention time (i.e., increasing protein molecular weight).

[0224] - %HMW = {area of ​​HMW / area of ​​(HMW + monomer + LMW)}*100

[0225] Each formulation sample for stability testing was stored in a chamber maintaining accelerated stability conditions of 25±2℃ and 60±5% relative humidity (RH), or in a chamber maintaining harsh stability conditions of 40±2℃ and 75±5% RH.

[0226] 5. Analysis of acid species content ratio (%Acidic)

[0227] The % acidity of the sample was measured using Imaging Capillary Isoelectric Focusing (icIEF). The sample was injected into the iCE3 system (Protein Simple, USA), and the isoelectric point (pI) and relative area of ​​protein charge variants were measured using the iCE CFR software. % acidity represents the relative area of ​​protein charge variants detected before the main peak. Measuring these indicates the identification of protein charge variants. When proteins undergo stress, they may aggregate or change their charge. Therefore, the charge variant of a protein can be a factor in determining its stability.

[0228] - %Acidic = {area of ​​Acidic / area of ​​(Acidic + Main + Basic)}*100

[0229] Each formulation sample for stability testing was stored in a chamber maintaining accelerated stability conditions of 25±2℃ and 60±5% RH.

[0230] 6. Oxidation level analysis

[0231] Using LC-MS (liquid-chromatography-mass spectrometry) from Waters, the 257th amino acid residue of the protein, methionine (Met) was identified. 257 ) was measured. After performing the LC-MS experiment, MassLynx and BioPharmaLynx TM Using the program, the oxidation rate of methionine, the 257th amino acid residue (%Met) 257 ) was analyzed.

[0232] %Met 257= {intensity of oxidized Met 257 / (intensity of oxidized Met 257 + intensity of non-oxidized Met 257 )}*100

[0233] Each formulation sample for stability testing was stored in a chamber maintaining accelerated stability conditions of 25±2℃ and 60±5% RH.

[0234]

[0235] [Example]

[0236] Example 1. T with or without methionine agg evaluation

[0237] Using dupilumab (CAS No. 1190264-60-8) as an anti-IL-4Rα antibody, aqueous liquid formulations having the compositions shown in Table 1 below were prepared. The aggregation temperature (T) of each formulation agg ) was measured.

[0238] T agg An increase in T means that the temperature at which aggregation is formed has increased. Therefore, the increased T agg means improved thermal stability.

[0239] No. a Buffer pH Excipient T agg (℃) 120 mM Histidine 6.08% (w / v) Sucrose 68.5 129% (w / v) Trehalose 69.4 134.7% (w / v) Sorbitol 69.5 443.13% (w / v) Mannitol 68.6 25140 mM Arginine 66.7 66135 mM Lysine 67.6 1770 mM Histidine 67.3 6890 mM Methionine 67.4 49250 mM Glycine 68.3 2100.8% (w / v) NaCl 67.4 411 b 12.5 mM acetate + 20 mM histidine 5.95% (w / v) sucrose + 50 mM arginine 67.89

[0240] a In all formulations The concentration of antibodies is T agg Set to 5 mg / mL according to the required concentration of the analytical method.

[0241] b Formulation 11 has the same composition as Dupixent in terms of buffer, pH, sugar, and amino acid conditions.

[0242] Figure 1 shows the coagulation temperature (T) for 10 formulations with different types of excipients and formulations with Dupixent composition. agg ) is a graph showing the results of measurement.

[0243] As a result, as shown in Table 1 and Figure 1, Formulation 8 containing methionine had a higher aggregation temperature (T) than Formulations 5 and 7 containing other types of amino acids, such as arginine or histidine. agg ) was shown.

[0244] Therefore, Formulation 8 containing methionine is T agg It was confirmed that this increased thermal stability was improved.

[0245] Example 2. K with or without methionine D evaluation

[0246] Table 2 below shows K for the same formulation as Example 1. D This is the result of measuring. Attractive interaction between proteins is the main cause of protein aggregation. K D An increase in K indicates a decrease in the inter-protein attraction. Therefore, increased K D means improved stability.

[0247] No. a Buffer pH Excipient K D(mL / g) 120 mM Histidine 6.08% (w / v) Sucrose - 2029% (w / v) Trehalose - 1434.7% (w / v) Sorbitol - 643.13% (w / v) Mannitol - 55140 mM Arginine - 36135 mM Lysine - 6770 mM Histidine - 7890 mM Methionine - 19250 mM Glycine 0100.8% (w / v) NaCl - 611 b 12.5 mM acetate + 20 mM histidine 5.95% (w / v) sucrose + 50 mM arginine-9

[0248] a In all formulations, the concentration of antibody was K D Set to 2, 4, 6, 8, or 10 mg / mL depending on the required concentration of the analytical method.

[0249] b Formulation 11 has the same composition as Dupixent in terms of buffer, pH, sugar, and amino acid conditions.

[0250] Figure 2 shows the diffusion interaction parameter (K) for 10 formulations with different types of excipients and formulations with Dupixent composition. D ) is a graph showing the results of measurement.

[0251] As a result, as shown in Table 2 and Figure 2, Formulation 8 containing methionine has a higher K than Formulations 1 to 7 and 10 containing sucrose, trehalose, sorbitol, mannitol, arginine, lysine, histidine, or NaCl. D It was confirmed that the K of formulation 8 containing methionine was increased compared to formulation 11 having the Dupixent composition. D It was confirmed that there was a significant increase.

[0252] Therefore, it was confirmed that Formulation 8 containing methionine had excellent stability by reducing the inter-protein attraction, which is the main cause of protein aggregation.

[0253] Example 3. Evaluation of viscosity and %HMW according to the presence or absence of methionine

[0254] Aqueous liquid formulations having the compositions shown in Table 3 below were prepared. For each formulation, the viscosity at 25°C was measured, and the %HMW under accelerated stability conditions (25°C, 4 weeks) or harsh stability conditions (40°C, 4 weeks) was measured.

[0255] Formulations 9', 10', and 11' are formulations that additionally contain methionine in formulations 9, 10, and 11 that contain sugar.

[0256] Formulations 12' and 13' are formulations in which lysine is replaced with methionine in formulations 12 and 13 containing sugar and lysine.

[0257] No. a ClassificationExcipient 1Excipient 2Excipient 3Viscosity (cP)%HMW(25℃, 4 weeks)%HMW(40℃, 4 weeks)9Added methionine8% (w / v) Trehalose33.94.277.189'8% (w / v) Trehalose30 mM Methionine23.73.366.01104.3% (w / v) Trehalose80 mM Histidine16.82.594.4210'4.3% (w / v) Trehalose80 mM Histidine30 mM Methionine12.42.334.01114.3% (w / v) Trehalose70 mM Arginine12.73.145.2011'4.3% (w / v) Trehalose70 mM Arginine30 mM Methionine 11.5 2.7 14.6 3 12 Lysine replaced with methionine 4.3% (w / v) Trehalose 120 mM Glycine 30 mM Lysine 31.4 3.6 8 6.8 8 12' 4.3% (w / v) Trehalose 120 mM Glycine 30 mM Methionine 27.6 3.2 5 5.8 6 13 1.8% (w / v) Mannitol 80 mM Histidine 30 mM Lysine 15.3 2.4 9 4.3 2 13' 1.8% (w / v) Mannitol 80 mM Histidine 30 mM Methionine 13.4 2.3 5 4.0 5

[0258] aAll formulations contained 175 mg / mL dupilumab, 10 mM sodium acetate, 0.2% (w / v) polysorbate 80, and had a pH of 6.0.

[0259] As a result, as shown in Table 3, when methionine was added to the formulation containing sugar, viscosity, %HMW 25℃ / 4주 , %HMW 40℃ / 4주 It was confirmed from the results of Formulations 9', 10', and 11' that simultaneous improvements were observed. Through these results, it was found that methionine has excellent functions as a viscosity reducing agent and stabilizer.

[0260] Also, when lysine was replaced with methionine in a formulation containing sugar and lysine, viscosity, %HMW 25℃ / 4주 , %HMW 40℃ / 4주 It was confirmed from the results of Formulations 12' and 13' that simultaneous improvements were observed. Through these results, it was found that methionine has superior functions as a viscosity reducer and stabilizer compared to lysine.

[0261] Example 4. %HMW, %Acidic, %Met according to the presence or absence of methionine 257 evaluation

[0262] Aqueous liquid formulations having the compositions shown in Table 4 below were prepared. %HMW, %Acidic, and %Met under accelerated stability conditions (25°C, 4 weeks) of each formulation 257 was measured.

[0263] Formulation 14' is Formulation 14 containing sugars and amino acids (i.e., histidine and methionine) but excluding histidine.

[0264] Formulation 14'' is Formulation 14 containing sugars and amino acids (i.e., histidine and methionine) but excluding methionine.

[0265] No. a Excipient 1 Excipient 2 Excipient 3%HMW(25℃ / 4 weeks)%Acidic(25℃ / 4 weeks)%Met 257(25℃ / 4 weeks) 144% (w / v) sucrose 70 mM histidine 25 mM methionine 2.26 38.5 4.6 14'4% (w / v) sucrose histidine-free 25 mM methionine 3.10 39.7 5.4 14'4% (w / v) sucrose 70 mM histidine methionine-free 2.5 6 4 1.6 12.5

[0266] a All formulations contain 175 mg / mL dupilumab, 15 mM sodium acetate, 0.2% (w / v) polysorbate 80, and have a pH of 5.5.

[0267] As a result, as shown in Table 4, when methionine was excluded from the formulation containing sugars and amino acids, %HMW, %Acidic, and %Met under accelerated stability conditions at 25℃ / 4 weeks 257 It was confirmed from the results of Formulation 14'' that this increased simultaneously. In particular, the %Acidic and %Met of the formulation excluding methionine (Formulation 14') compared to the formulation excluding histidine (Formulation 14') 257 This was further increased. Through these results, we were able to confirm the function of methionine as a stabilizer and antioxidant.

[0268] Example 5. Evaluation of viscosity and %HMW of formulations containing and not containing methionine

[0269] Aqueous liquid formulations having the compositions shown in Table 5 below were prepared. The viscosity of each formulation at 25°C was measured, and %HMW was measured immediately after preparation (0 day, Initial).

[0270] Formulations 15 to 20 are methionine-containing formulations with different histidine concentrations and pHs.

[0271] Formulation 21 is a methionine-free formulation and has the same composition as Dupixent.

[0272] No. aBuffer pH Part 1 Part 2 Part 3 Viscosity (cP) %HMW (Initial) 1530 mM Sodium Acetate 4.94% (w / v) Sucrose 20 mM Histidine 25 mM Methionine 8.7 1.93 16 4.84% (w / v) Sucrose 50 mM Histidine 25 mM Methionine 8.3 1.94 17 5.24% (w / v) Sucrose 50 mM Histidine 25 mM Methionine 9.0 1.97 18 4.94% (w / v) Sucrose 80 mM Histidine 25 mM Methionine 7.6 1.95 19 5.54% (w / v) Sucrose 80 mM Histidine 25 mM Methionine 8.7 2.0 12 0 5.24% (w / v) Sucrose 92.4 mM Histidine 25 mM Methionine 8.81.9321 b 12.5 mM sodium acetate + 20 mM histidine 5.95% (w / v) sucrose 50 mM arginine - 11.12.03

[0273] a All formulations contained 175 mg / mL dupilumab and 0.2% (w / v) polysorbate 80.

[0274] b Formulation 21 is a Dupixent formulation.

[0275] Figure 3 is a graph showing the results of viscosity measurements for formulations containing methionine and formulations not containing methionine.

[0276] Figure 4 is a graph showing the results of analyzing the high molecular weight species content ratio (%HMW) for formulations containing methionine and formulations not containing methionine.

[0277] As a result, as shown in Table 5, FIG. 3, and FIG. 4, the formulations containing 20 to 92.4 mM histidine and 25 mM methionine having a pH range of 4.9 to 5.5 (Formulations 15 to 20) were confirmed to simultaneously improve viscosity and %HMW compared to the Dupixent formulation not containing methionine (Formulation 21).

[0278]

[0279] The foregoing description of the present invention is provided for illustrative purposes only. Those skilled in the art will readily appreciate that the present invention can be readily modified into other specific forms without altering the technical spirit or essential characteristics of the present invention. Therefore, the embodiments described above should be understood as illustrative in all respects and not restrictive.

Claims

1. Anti-IL-4Rα antibody; buffer; sugars; and Contains methionine, The pH is about 4.9 to about 5.5, Liquid formulation.

2. A liquid formulation according to claim 1, wherein the anti-IL-4Rα antibody is dupilumab.

3. A liquid formulation according to claim 1, wherein the concentration of the anti-IL-4Rα antibody is from about 5 mg / mL to about 300 mg / mL.

4. A liquid formulation according to claim 1, wherein the concentration of the anti-IL-4Rα antibody is about 100 mg / mL to about 200 mg / mL.

5. A liquid formulation according to claim 1, wherein the concentration of the anti-IL-4Rα antibody is from about 125 mg / mL to about 200 mg / mL.

6. A liquid formulation according to claim 1, wherein the buffer comprises acetate.

7. A liquid formulation according to claim 1, wherein the buffer is sodium acetate.

8. A liquid formulation according to any one of claims 1 to 7, wherein the concentration of the buffer is from about 10 mM to about 50 mM.

9. A liquid formulation according to claim 6, wherein the concentration of the acetate is from about 10 mM to about 40 mM.

10. A liquid formulation according to claim 6, wherein the concentration of the acetate is from about 15 mM to about 35 mM.

11. A liquid formulation according to claim 1, wherein the sugar comprises at least one selected from sugar and sugar alcohol.

12. A liquid formulation according to claim 11, wherein the sugar comprises at least one selected from sucrose, trehalose, galactose, mannose, maltose, lactose, fructose, and glucose.

13. A liquid formulation according to claim 11, wherein the sugar alcohol comprises at least one selected from mannitol, sorbitol, xylitol, arabitol, erythritol, lactitol, maltitol, and inositol.

14. A liquid formulation according to claim 11, wherein the sugar comprises at least one selected from sucrose, trehalose, and mannitol.

15. A liquid formulation according to claim 11, wherein the sugar is sucrose.

16. A liquid formulation according to claim 11, wherein the concentration of the sugar is from about 1% (w / v) to about 20% (w / v).

17. A liquid formulation according to claim 11, wherein the concentration of the sugar alcohol is from about 1% (w / v) to about 20% (w / v).

18. A liquid formulation according to claim 15, wherein the concentration of sucrose is from about 1% (w / v) to about 20% (w / v).

19. A liquid formulation according to claim 1, wherein the concentration of methionine is from about 2.7 mM to about 90 mM.

20. A liquid formulation according to claim 1, wherein the concentration of methionine is about 20 mM to about 30 mM.

21. A liquid formulation according to claim 1, wherein the methionine has at least one function of a viscosity reducing agent, a stabilizer, and an antioxidant.

22. A liquid formulation according to any one of claims 1 to 21, further comprising histidine.

23. A liquid formulation according to claim 12, wherein the concentration of histidine is from about 20 mM to about 92.4 mM.

24. A liquid formulation according to claim 12, wherein the concentration of histidine is from about 30 mM to about 92.4 mM.

25. A liquid formulation according to any one of claims 1 to 24, which does not contain arginine.

26. A liquid formulation according to any one of claims 1 to 25, further comprising a surfactant.

27. A liquid formulation according to claim 26, wherein the surfactant comprises a nonionic surfactant.

28. A liquid formulation according to claim 26, wherein the surfactant comprises polysorbate 20, polysorbate 80, or a combination thereof.

29. A liquid formulation according to any one of claims 26 to 28, wherein the concentration of the surfactant is 0.01% (w / v) to 0.9% (w / v).

30. A liquid formulation according to any one of claims 26 to 28, wherein the concentration of the surfactant is from about 0.1% (w / v) to about 0.3% (w / v).

31. A liquid formulation according to any one of claims 1 to 30, wherein the viscosity of the liquid formulation is 5.0 cP to 11.0 cP.

32. In any one of claims 1 to 30, The high molecular weight species content (%HMW) of the antibody measured after 4 weeks of storage at 25°C is 3.1% or less; The acidic species content (%Acidic) of the antibody measured after 4 weeks of storage at 25°C is less than 41%; or Oxidation rate of methionine residues in the above antibodies (%Met) measured after 4 weeks of storage at 25°C 257 ) is less than 12%, liquid formulation.

33. A liquid formulation according to any one of claims 1 to 30, wherein the high molecular weight species content ratio (%HMW) of the antibody measured immediately after manufacture is less than 2.03%.

34. Dupilumab at about 5 mg / mL to about 300 mg / mL; Acetate from about 10 mM to about 40 mM; About 10 mM to about 100 mM methionine; Histidine from about 20 mM to about 92.4 mM; About 2% (w / v) to about 10% (w / v) sucrose; and Containing about 0.1% (w / v) to about 0.9% (w / v) of polysorbate 80, The pH is about 4.9 to about 5.5, Liquid formulation. Dupilumab at 35.100 mg / mL to 200 mg / mL; 30 mM±5 mM acetate; 25 mM±5 mM methionine; 20 mM to 92.4 mM histidine; 4%±1.0% (w / v) sucrose; and Contains 0.2%±0.1% (w / v) of polysorbate 80, The pH is about 4.9 to about 5.5, Liquid formulation. Dupilumab at 36.100 mg / mL to 200 mg / mL; 30 mM±5 mM acetate; 25 mM±5 mM methionine; 20 mM to 92.4 mM histidine; 5.5%±1.0% (w / v) sucrose; and Contains 0.2%±0.1% (w / v) of polysorbate 80, The pH is about 4.9 to about 5.5, Liquid formulation.

37. A liquid formulation for subcutaneous injection according to any one of claims 1 to 36.

38. A device comprising a liquid formulation of any one of claims 1 to 37.

39. A device according to claim 38, comprising the liquid formulation in a container selected from a syringe, a pre-filled syringe, a pre-filled pen, a microinfusor, an autoinjector, a bottle, a vial, and a tube.

40. A method of treating an IL-4Rα associated condition, comprising administering to a subject in need thereof a liquid formulation of any one of claims 1 to 37.

41. A method for treating an IL-4Rα-associated condition according to claim 40, wherein the IL-4Rα-associated condition is an IL-4-mediated disease or an IL-13-mediated disease.

42. In claim 40, the IL-4Rα-related condition is any one selected from atopic dermatitis, asthma, chronic rhinosinusitis (CRS), chronic rhinosinusitis with nasal polyposis (CRSwNP), allergic fungal rhinosinusitis (AFRS), eosinophilic esophagitis (EoE), prurigo nodularis (PN), bullous pemhigoid (BP), chronic spontaneous urticarial (CSU), chronic inducible urticarial (CIndU), and chronic obstructive pulmonary disease (COPD). A method for treating a condition associated with IL-4Rα.

43. Use of a liquid formulation of any one of claims 1 to 37 in the manufacture of a medicament for treating an IL-4Rα associated condition.

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