WO2025116637A1 - STABLE LIQUID FORMULATION OF ANTI-IL-4Rα ANTIBODY - Google Patents

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

Stable liquid formulations of anti-IL-4Rα antibodies, devices comprising same, and uses thereof for treating IL-4Rα associated conditions.

Antibody drugs have a large molecular weight compared to general protein drugs, and their secondary/higher order structures are complex, which can cause physicochemical instability. For this reason, antibody drugs require the development of an optimal formulation that guarantees quality and stability throughout the entire process from manufacturing, storage, and administration to patients. Protein instability can be caused by various external factors such as temperature, light, and chemical factors, which can lead to decreased activity, decreased efficacy, or immunogenicity when administered to the human body. Therefore, it is important to improve the instability of such antibody drugs and maintain optimal quality until they are administered to patients. To achieve optimal quality, methods such as changing the buffer solution, evaluating the optimal pH, and adding stabilizers are used. Since the material properties of each antibody protein are different, and the optimal combination of buffer solution, pH, and stabilizer may be different, a formulation suitable for the target substance is required to solve this problem.

For example, dupilumab, an anti-IL-4Rα antibody, is manufactured and sold under the trade name Dupixent ^® . Dupixent 200 mg is a 1.14 mL liquid formulation containing 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 containing 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, such as adding sugars, amino acids, buffers, and surfactants, or applying a specific pH, as in the composition of Dupixent products, are being reviewed and applied.

In addition to ensuring the stability described above, in consideration of patient convenience during injection, there is a demand for the development of a high-concentration formulation that reduces the number of administrations through high-dose injections in a small volume while increasing therapeutic activity. When developing a formulation for a high-concentration formulation, as the protein concentration increases, there are limitations such as protein instability due to protein aggregation and inhibition of injectability due to increased viscosity, so it is important to develop a formulation that can improve viscosity while ensuring protein stability. Normally, sugars are used as stabilizers for protein stabilization, but in the case of formulations containing such stabilizers, there is a phenomenon in which the viscosity increase is accelerated when the protein concentration increases, so there is a limit to improving viscosity while ensuring protein stability. To solve this problem, it is possible to develop a formulation that can simultaneously improve viscosity and ensure protein stability by inhibiting protein aggregation by including a viscosity-reducing agent.

However, in order to confirm whether the viscosity is improved and whether the protein stability is improved until it is administered to the patient, it is necessary to evaluate the main quality under the storage temperature condition until the expiration date, so a long period of time is required for the verification of the developed formulation. In order to effectively maintain protein stability and evaluate the optimal formulation in a short period of time, the optimal formulation combination can be selected by analyzing the new quality indicators after exposure to heat (temperature) and chemicals, which are factors that cause structural and physicochemical denaturation of proteins.

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 secure the stability of an anti-IL-4Rα antibody such as dupilumab throughout the entire process of manufacturing, storage, and administration to a patient, while at the same time improving the viscosity of the formulation. The liquid formulation has improved stability and viscosity compared to commercialized formulations of anti-IL-4Rα antibodies such as dupilumab (e.g., Dupixent), and thus can be applied to high-concentration preparations, thereby increasing patient convenience.

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

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

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

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

All technical terms used in this specification, unless otherwise defined, are used in the same meaning as commonly understood by those skilled in the art in the relevant field of the present invention. In addition, although preferred methods or samples are described in this specification, similar or equivalent ones are also included in the scope of the present invention. In addition, the numerical values described in this specification are considered to include the meaning of "about" even if not specified. The contents of all publications mentioned as references in this specification are incorporated herein by reference in their entirety.

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

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

anti-IL-4Rα antibody;

buffer;

sugars; and

Contains methionine,

The pH is about 4.9 to about 5.5,

Provides a liquid formulation.

(1) Antibody

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.

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

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.

The above anti-IL-4Rα antibody may be dupilumab (CAS No. 1190264-60-8). Therefore, the 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 sold under the trade name Dupixent ^® . After receiving approval from the U.S. Food and Drug Administration (FDA) as a treatment for 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 can be produced by general methods known in the art. More detailed information about dupilumab is readily available to those skilled in the art from publicly known databases.

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.

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

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 to about 176 mg/mL.

In one specific embodiment, the concentration of the anti-IL-4Rα antibody can be about 100 mg/mL to about 200 mg/mL.

In one specific embodiment, the concentration of the anti-IL-4Rα antibody can be about 125 mg/mL to about 200 mg/mL.

In one specific embodiment, the concentration of the anti-IL-4Rα antibody can be about 150 mg/mL to about 200 mg/mL.

In one specific embodiment, the concentration of the anti-IL-4Rα antibody can be about 150 mg/mL to about 175 mg/mL.

In one specific example, the concentration of the anti-IL-4Rα antibody can be from 135 mg/mL to 165 mg/mL.

In one specific example, the concentration of the anti-IL-4Rα antibody can be from 158 mg/mL to 192 mg/mL.

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

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

The concentration of the anti-IL-4Rα antibody may be a high concentration. 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.

(2) Buffer and pH

The liquid formulation according to the aspect includes a buffer. The buffer can have the function of controlling the pH of the formulation. The pH of the liquid formulation can be maintained at a certain value or a certain range by the buffer. Therefore, the buffer can play a role of providing a certain value or a certain range of pH to the liquid formulation.

The above buffering agent can be used without limitation in its type as long as it is applicable to biopharmaceuticals.

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

The term "salt" may be a pharmaceutically acceptable salt. The salt may include an inorganic acid salt, an organic acid salt, a metal salt, and the like 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 emboxide, 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.

The term "hydrate" means a substance containing water molecules within its molecules. The hydrate may be a monohydrate, a dihydrate, or a trihydrate.

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).

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.

In one specific 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.

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

In one embodiment, the buffering agent can include one or more selected from acetate, a salt thereof, and a hydrate thereof. In one embodiment, the buffering agent can include acetate. In one embodiment, the buffering agent can be acetate. In one embodiment, the buffering agent can include sodium acetate. In one embodiment, the buffering agent can be sodium acetate.

The concentration of the buffer can 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.

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

The concentration of the acetate can 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.

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

(3) Sugar

Liquid formulations according to the aspect of the invention contain saccharides.

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

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

The above sugar alcohol is a general term for polyols having two or more hydroxy groups, which are made into alcohol groups by reducing an aldehyde group or a ketone group of a sugar. 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.

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 any 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 5.5% (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 may vary individually depending on the type of each specific sugar alcohol. 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 1% (w/v) to about 8% (w/v), about 1% (w/v) to about 7% (w/v), about 1% (w/v) to about 6% (w/v), about 1% (w/v) to about 5% (w/v), about 1% (w/v) to about 4% (w/v), about 1% (w/v) to about 3% (w/v), or about 1% (w/v) to about 2% (w/v).

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

In certain embodiments, the sugar may comprise sucrose. In certain embodiments, the sugar may be sucrose.

The concentration of the above sucrose can be any range or any value selected from about 0.1% (w/v) to about 20% (w/v). For example, the concentration of 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 specific embodiment, the concentration of sucrose can be about 2% (w/v) to about 6% (w/v).

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

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

(4) Amino acids

The liquid formulation according to the aspect contains amino acids.

Liquid formulations according to the invention contain methionine.

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 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.

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.

In one specific embodiment, the concentration of methionine can be from about 20 mM to about 90 mM.

In certain embodiments, the concentration of methionine can be from about 20 mM to about 30 mM.

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

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.

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

The term "stabilizer" refers to a substance added to prevent a change in state or chemical change when preserving a substance. The stabilizer may be a thermal stabilizer. The stabilizer may inhibit aggregation or denaturation of antibodies.

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).

The above methionine can increase the aggregation temperature (T _agg ) of the antibody in the liquid formulation. When methionine is included in the anti-IL-4Rα antibody-containing liquid formulation, the liquid formulation can exhibit a higher T _agg compared to when arginine or histidine is included. Therefore, the methionine can increase the thermal stability of the anti-IL-4Rα antibody-containing liquid formulation.

The above methionine can increase the diffusion interaction parameter (K _D ) of the liquid formulation. When methionine is included in the anti-IL-4Rα antibody-containing liquid formulation, the liquid formulation can exhibit a higher K _D compared to when sucrose, trehalose, sorbitol, mannitol, arginine, lysine, histidine, or NaCl is included. Therefore, the methionine can inhibit aggregation of the anti-IL-4Rα antibody in the liquid formulation, thereby increasing the chemical stability of the anti-IL-4Rα antibody-containing liquid formulation.

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 sugar, 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 sugar 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.

The above methionine can decrease 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 can 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 included. When methionine is added to a liquid formulation containing an 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 an 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 within the liquid formulation.

The above methionine can reduce the acid 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 acid species of the anti-IL-4Rα antibody in the liquid formulation, thereby increasing the stability of the anti-IL-4Rα antibody-containing liquid formulation.

The above methionine can reduce the oxidation rate (% Met ₂₅₇ ) of the methionine residue in the antibody in the liquid formulation. When methionine is included in the anti-IL-4Rα antibody-containing liquid formulation, the % Met 257 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 inhibit oxidation of the anti-IL-4Rα antibody in the liquid formulation, thereby increasing the stability of the anti-IL-4Rα antibody-containing liquid formulation.

The liquid formulation according to one aspect may additionally comprise histidine. Accordingly, the liquid formulation may be 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.

The concentration of said histidine can be from about 0.1 mM to about 400 mM, from about 0.1 mM to about 92.4 mM, from about 0.5 mM to about 400 mM, from about 0.5 mM to about 92.4 mM, from about 1 mM to about 400 mM, from about 10 mM to about 400 mM, from about 20 mM to about 400 mM, from about 20 mM to about 300 mM, from about 20 mM to about 200 mM, from about 20 mM to about 100 mM, or from about 20 mM to about 92.4 mM.

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 90 mM, about 50 mM to about 80 mM, or about 50 mM to about 60 mM.

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

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

The histidine above can reduce the acid 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 acid species of the anti-IL-4Rα antibody in the liquid formulation, thereby increasing the stability of the anti-IL-4Rα antibody-containing liquid formulation.

The histidine above can reduce the oxidation rate (% Met ₂₅₇ ) of the methionine residue in the antibody in the liquid formulation. When histidine is included in the anti-IL-4Rα antibody-containing liquid formulation, the % Met 257 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 inhibit oxidation of the anti-IL-4Rα antibody in the liquid formulation, thereby increasing the stability of the anti-IL-4Rα antibody-containing liquid formulation.

The liquid formulation according to one aspect may be free of arginine. The liquid formulation may not contain arginine as a viscosity reducing agent. The arginine may contain arginine or a salt thereof, for example, arginine hydrochloride.

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

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.

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

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.

(5) Surfactant

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

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.

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.

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

The above surfactant may be a nonionic surfactant.

Specifically, the surfactant may be at least one selected from the group consisting of polysorbate, poloxamer, sorbitan esters of other fatty acids, polyethylene-polypropylene glycol, polyoxyethylene compounds, and sodium dodecyl sulphate (SDS).

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

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

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.

The above polyoxyethylene compound may include polyoxyethylene-stearate, polyoxyethylene alkyl ether (alkyl: C1-C30), polyoxyethylene monoryl ether, alkylphenyl polyoxyethylene copolymer (alkyl: C1-C30), and the like.

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

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

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

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

The concentration of the surfactant can be any range or 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).

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

(6) Diluent

Liquid formulations, depending on the aspect, may additionally contain a diluent.

The above 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.

In one specific embodiment, the diluent may be water. Accordingly, the liquid formulation may be an aqueous liquid formulation.

(7) Formulation

The term "liquid formulation" means a formulation in liquid form.

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

The liquid formulation according to one aspect may be a pharmaceutical formulation of an anti-IL-4Rα antibody.

The term "pharmaceutical composition" or "pharmaceutical preparation" means a preparation which allows the biological activity of an active ingredient to be effectively carried out and which does not contain additional components which have significant toxicity to the subject to which the preparation is administered.

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

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

The liquid formulation according to one aspect may be a Dupixent ^® biosimilar. The liquid formulation according to one aspect has improved stability and improved viscosity compared to Dupixent.

The term "biosimilar" is also called "biogeneric" and refers to a copy of an original biopharmaceutical. Since biopharmaceuticals are not synthesized chemical products but are produced through cells, they cannot be exactly the same as the original drug. Therefore, a copy of a biopharmaceutical is called a biosimilar because it is similar, but not identical, to the original drug.

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

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;

2) 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;

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;

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;

5) A liquid formulation comprising about 5 mg/mL to about 300 mg/mL of dupilumab; about 10 mM to about 40 mM 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, wherein the pH is about 4.9 to about 5.5;

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, wherein the pH is about 4.9 to about 5.5;

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, wherein the pH is about 4.9 to about 5.5;

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, and having a pH of about 4.9;

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, and having a pH of about 4.8;

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, and having a pH of about 5.2;

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, and having a pH of about 4.9;

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, and having a pH of about 5.5;

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, and having a pH of about 5.2;

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, wherein the pH is about 4.9 to about 5.5;

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, wherein the pH is about 4.9 to about 5.5;

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, and having a pH of about 4.9;

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, and having a pH of about 4.8;

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, and having a pH of about 5.2;

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, and having a pH of about 4.9;

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, and having a pH of about 5.5; or

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, and having a pH of about 5.2.

(8) Stability and viscosity

Liquid formulations according to the aspect can have low viscosity while ensuring the stability of antibodies.

In the examples of this specification, three experiments were performed. First, the thermal stability and chemical stability according to the type of excipient were compared and evaluated through T _agg and K _D measurements, thereby confirming the superiority of methionine among the excipients. Second, the functions of methionine as a viscosity reducer, stabilizer, and antioxidant were confirmed through evaluation of viscosity, %HMW, %Acidic, and %Met257. Third, through comparative evaluation of viscosity and %HMW with the Dupixent formulation, it was confirmed that the liquid formulation of anti-IL-4Rα antibody containing methionine can simultaneously improve stability and viscosity compared to the commercialized formulation.

The term "stability" means that the antibody (e.g., dupilumab) contained in the 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, decomposition, denaturation (acidic or alkaline), 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", "improved stability" can mean low protein aggregation rate, low protein degradation rate, low protein denaturation rate, low amino acid oxidation rate, etc. during storage. Physical stability, chemical stability and/or biological activity can be evaluated by methods commonly known in the art.

One 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" may mean that the T _agg or K _D value measured for the formulation increases by 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. An increase in the T _agg or K _D value may mean that the degree to which antibodies in the formulation aggregation occurs due to external factors (e.g., heat, temperature) is reduced, thereby improving stability.

The term "aggregate" may refer to high molecular weight (HMW) species formed by the aggregation of antibody proteins. The term "protein aggregation percentage" may be expressed as the percentage of high molecular weight species content (%HMW) of antibody in the formulation at a given point in time. The %HMW may be measured by, but is not limited to, size exclusion chromatography (SEC). For example, "improved stability" may mean that the %HMW of antibody measured for the formulation decreases 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 prior formulation. A decrease in %HMW may mean that the degree to which antibodies in the formulation aggregate is reduced, thereby improving stability.

The term "protein denaturation" can be expressed as the percentage acidic species content (%Acidic) of the antibody within the formulation at a point in time. The %Acidic can be measured by, but is not limited to, imaged capillary isoelectric focusing (icIEF). For example, "improved stability" can mean a decrease in the %Acidic of the antibody measured for the formulation of 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 prior formulation. A decrease in the %Acidic can mean that the extent to which the antibody within the formulation undergoes denaturation is reduced, thereby improving stability.

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

The above stability evaluation can 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 can include conditions used in an accelerated test for a drug product, and can be, for example, a temperature of 25±2°C and a relative humidity (RH) of 60±5%. The stress stability conditions can include conditions used in a stress test for a drug product, and can be, for example, a temperature of 40±2°C and a relative humidity (RH) of 75+5%.

The above liquid formulation may have an increased agglomeration temperature (T _agg ).

The above liquid formulation may have an increased T _agg by including methionine compared to the anti-IL-4Rα antibody liquid formulation containing arginine.

The above liquid formulation may have an increased diffusion interaction parameter (K _D ).

The liquid formulation may have an increased K _D by including methionine, compared to an anti-IL-4Rα antibody liquid formulation comprising arginine. The liquid formulation may have an increased K _{D by including methionine, compared to an anti-IL-4Rα antibody liquid formulation comprising sucrose, trehalose, sorbitol, mannitol, arginine, lysine, histidine, or NaCl. The liquid formulation may have an increased K D} by including methionine, compared to an 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.

The above liquid formulation may have improved viscosity. The 'improvement' of the viscosity may mean a 'reduction' of the viscosity. Accordingly, the above liquid formulation may have reduced viscosity.

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

In one embodiment, it was confirmed that the viscosity was improved when methionine was added to the anti-IL-4Rα antibody liquid formulation containing sugar. In addition, it was confirmed that methionine functioned better as a viscosity reducing agent than lysine in the anti-IL-4Rα antibody liquid formulation containing sugar.

The liquid formulation according to one aspect can have a reduced viscosity relative to Dupixent. In one embodiment, the liquid formulation according to one aspect can have a reduced viscosity relative to the viscosity of 11.1 cP (centiPoise) of Dupixent 200 mg. In another embodiment, the liquid formulation according to one aspect can have a reduced viscosity relative to the viscosity of 300 mg of Dupixent.

The viscosity of the liquid formulation can be less than 11.1 cP (centiPoise). The viscosity of the liquid formulation can be less than 11.1 cP, less than or equal to 11.0 cP, less than or equal to 10.5 cP, less than or equal to 10.0 cP, less than or equal to 9.5 cP, less than or equal to 9.0 cP, less than or equal to 8.5 cP, less than or equal to 8.0 cP, less than or equal to 7.5 cP, less than or equal to 7.0 cP, or less than or equal to 6.5 cP. 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 but 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, 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 there is.

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, can be 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.

The liquid formulation may have a viscosity 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.

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

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

The liquid formulation may exhibit a decrease in %HMW of antibodies of at least 5%, at least 10%, at least 15%, or at least 20% after storage for 4 weeks at 40°C, compared to a formulation that does not contain methionine.

The liquid formulation may exhibit a decrease in %HMW of antibodies by at least 5%, at least 10%, or at least 15% after 4 weeks of storage at 25°C, compared to a formulation that does not contain methionine.

The liquid formulation may have a %HMW of antibody 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 after storage for 4 weeks at 25°C.

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

The above liquid formulation may have a reduced acidic species content (%Acidic).

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

The liquid formulation may have a decrease in the %Acidic of antibodies measured after 4 weeks of storage at 25°C of 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.

The liquid formulation may have a %Acidic of antibody of less than 41%, less than or equal to 40%, or less than or equal to 39% after storage for 4 weeks at 25°C.

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

The above liquid formulation may have a reduced %Met ₂₅₇ by including methionine compared to a liquid formulation of anti-IL-4Rα antibody that does not include methionine.

The liquid formulation can exhibit a decrease in the %Met ₂₅₇ of antibodies by at least 50%, at least 55%, or at least 60% after 4 weeks of storage at 25°C, compared to a formulation that does not contain methionine.

The liquid formulation may have a %Met ₂₅₇ of antibody of less than 12%, or less than 10%, or less than 8%, or less than 6%, or less than 5%, measured after storage for 4 weeks at 25°C.

(9) Device

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

The device is primarily intended for parenteral administration. The 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.

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.

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

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

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.

(10) Treatment of disease

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

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.

The method of treating the above 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).

The subject may be a subject in need of administration of a liquid formulation containing the anti-IL-4Rα antibody (e.g., dupilumab). The subject in need of 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., elimination, reduction, alleviation, or improvement of symptoms, etc.) by administration of the anti-IL-4Rα antibody. The subject may be selected from mammals, including humans.

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

The IL-4Rα associated condition can 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) can include any indication currently approved or that may be approved in the future for an anti-IL-4Rα antibody (e.g., dupilumab).

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.

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.

The above 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).

The above atopic dermatitis may be moderate-to-severe atopic dermatitis.

The above asthma may be moderate-to-severe asthma.

(11) Route of administration and dosage

Liquid formulations according to the aspect may be administered by parenteral route. The parenteral route may include subcutaneous administration, intravenous administration, etc. Parenteral administration may be by bolus injection or continuous infusion.

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

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, an injectable ready-to-use form, etc., but is not limited thereto.

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 a single formulation or divided into two or more (e.g., two, three, four, five, six, seven, eight, nine, or ten) formulations. The liquid formulation may be contained in a single-dose or multi-dose form of the device.

The above liquid formulation may be administered to the body in one dose, such that the entire amount of the anti-IL-4Rα antibody (e.g., dupilumab) contained in the formulation is administered to the body at one time (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 or more, 10 minutes or more, 30 minutes or more, 60 minutes or more, 90 minutes or more, 120 minutes or more, 150 minutes or more, 180 minutes or more, 210 minutes or more, or 240 minutes or more.

The administration target of the above liquid formulation can 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.

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 variously determined by 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, and the like. 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.

For example, the liquid formulation can 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.

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.

The liquid formulation of anti-IL-4Rα antibody according to one aspect may have improved stability and improved viscosity compared to existing commercialized formulations. Therefore, the liquid formulation may have high stability and low viscosity even when containing a high concentration of antibody, thereby increasing patient convenience upon injection. Therefore, the liquid formulation may be usefully used as a medicine for treating IL-4Rα-related conditions.

Figure 1 is a graph showing the results of measuring the coagulation temperature (T _agg ) for 10 formulations with different types of excipients and a formulation having a Dupixent composition.

Figure 2 is a graph showing the results of measuring the diffusion interaction parameter (K _D ) for 10 formulations with different types of excipients and a formulation having a Dupixent composition.

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

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.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the following examples are provided only to illustrate the present invention and the scope of the present invention is not limited thereby.

　

[Experimental Method]

1. Measurement of coagulation temperature (T _agg )

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 is evaluated by measuring the temperature at which the size of the aggregates changes, and the higher the aggregation temperature (T _agg ) value, the more stable the formulation.

2. Analysis of diffusion interaction parameter K _D

In order to evaluate whether the binding force in the fluid phase is attractive or repulsive as the protein concentration increases when the sample is exposed to room temperature, the diffusion interaction parameter, K _D , was analyzed. This is a parameter that evaluates the binding force between substances that causes irreversible aggregation, enabling evaluation or ranking of excellent formulations according to aggregation tendency. 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 K _D was calculated using the slope and y-intercept of the graph. If the value is positive, the repulsion between substances is dominant, indicating a weak aggregation tendency, and if it is negative, the attractive force is dominant, indicating a strong aggregation tendency. The larger and more positive K _D is, the more stable the formulation is.

3. Viscosity measurement

The viscosity of the sample was measured using a viscometer (manufacturer: RheoSense, model name: VROC initium one plus). The analysis was conducted under a temperature condition of 25°C when measuring the viscosity. After measuring 11 segments for the same sample, the average value was calculated for the values with a slope fit R ² value of 0.9995 or higher, and the viscosity value of each sample was calculated.

4. Analysis of high molecular weight species content (%HMW)

The percentage of high molecular weight species (%HMW) was determined using size exclusion chromatography (SEC) from Waters. It is separated into three peaks according to the molecular weight of the protein, and these three peaks correspond to the HMW peak (protein aggregation), Monomer peak, and LMW peak (protein degradation) in order of decreasing retention time, i.e. increasing protein molecular weight.

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

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

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

The % Acidic value 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 the relative area of the protein charge variant were measured using the iCE CFR software. The % Acidic represents the relative area of the protein charge variant detected in front of the main peak. Measuring these is to identify the protein charge variant. When a protein is stressed and changes, aggregation or charge changes occur. Therefore, the charge variant of a protein can be a factor to confirm its stability.

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

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

6. Analysis of oxidation level

The mass of methionine (Met ₂₅₇ ), the 257th amino acid residue of the protein, was measured using liquid-chromatography-mass spectrometry (LC-MS) from Waters. After performing the LC-MS experiment, the oxidation rate of methionine, the 257th amino acid residue, (%Met ₂₅₇ ) was analyzed using MassLynx and BioPharmaLynx ^TM programs.

%Met ₂₅₇ = {intensity of oxidated Met ₂₅₇ /(intensity of oxidated Met ₂₅₇ + intensity of non-oxidated Met ₂₅₇ )}*100

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

[Example]

Example 1. Evaluation of T _agg according to the presence or absence of methionine

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 _agg ) of each formulation was measured.

An increase in T _agg indicates an increase in the temperature at which aggregation occurs. Therefore, increased T _agg indicates improved thermal stability.

No. ^a Buffer pH Excipient T _agg (℃) 1 20 mM histidine 6.0 8% (w/v) sucrose 68.51 2 9% (w/v) trehalose 69.41 3 4.7% (w/v) sorbitol 69.54 4 3.13% (w/v) mannitol 68.62 5 140 mM arginine 66.76 6 135 mM lysine 67.61 7 70 mM histidine 67.36 8 90 mM methionine 67.44 9 250 mM glycine 68.32 10 0.8% (w/v) NaCl 67.44 11 ^b 12.5 mM acetate + 20 mM histidine 5.9 5% (w/v) sucrose + 50 mM arginine 67.89

^a in all formulations The concentration of antibody was set to 5 mg/mL according to the required concentration of the T _agg assay.

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

Figure 1 is a graph showing the results of measuring the coagulation temperature (T _agg ) for 10 formulations with different types of excipients and a formulation having a Dupixent composition.

As a result, as shown in Table 1 and Figure 1, Formulation 8 containing methionine exhibited a higher aggregation temperature (T _agg ) compared to Formulations 5 and 7 containing other types of amino acids, such as arginine or histidine.

Therefore, it was confirmed that Formulation 8 containing methionine had improved thermal stability with an increase in T _agg .

Example 2. Evaluation of K _D according to the presence or absence of methionine

Table 2 below shows the results of measuring K _D for the same formulation as Example 1. Attractive interaction between proteins is the main cause of protein aggregation. An increase in K _D means a decrease in attractive interaction between proteins. Therefore, an increased K _D means improved stability.

No. ^a Buffer pH Excipient K _D (mL/g) 1 20 mM histidine 6.0 8% (w/v) sucrose -20 2 9% (w/v) trehalose -14 3 4.7% (w/v) sorbitol -6 4 3.13% (w/v) mannitol -5 5 140 mM arginine -3 6 135 mM lysine -6 7 70 mM histidine -7 8 90 mM methionine -1 9 250 mM glycine 0 10 0.8% (w/v) NaCl -6 11 ^b 12.5 mM acetate + 20 mM histidine 5.9 5% (w/v) sucrose + 50 mM arginine -9

^a In all formulations, the antibody concentration was set to 2, 4, 6, 8, or 10 mg/mL depending on the required concentration of the K _D assay.

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

Figure 2 is a graph showing the results of measuring the diffusion interaction parameter (K _D ) for 10 formulations with different types of excipients and a formulation having a Dupixent composition.

As a result, as shown in Table 2 and Figure 2, it was confirmed that Formulation 8 including methionine had an increased K _D compared to Formulations 1 to 7 and 10 including sucrose, trehalose, sorbitol, mannitol, arginine, lysine, histidine, or NaCl. In addition, it was confirmed that Formulation 8 including methionine had a significantly increased K _D compared to Formulation 11 having a Dupixent composition.

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.

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

Aqueous liquid formulations having the compositions shown in Table 3 below were prepared. For each formulation, the viscosity at a temperature of 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.

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

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

No. ^a classification Sibling 1 Brother 2 Brother 3 Viscosity (cP) %HMW
(25℃, 4 weeks) %HMW
(40℃, 4 weeks) 9 Add methionine 8% (w/v) trehalose 33.9 4.27 7.18 9' 8% (w/v) trehalose 30 mM methionine 23.7 3.36 6.01 10 4.3% (w/v) trehalose 80 mM histidine 16.8 2.59 4.42 10' 4.3% (w/v) trehalose 80 mM histidine 30 mM methionine 12.4 2.33 4.01 11 4.3% (w/v) trehalose 70 mM arginine 12.7 3.14 5.20 11' 4.3% (w/v) trehalose 70 mM arginine 30 mM methionine 11.5 2.71 4.63 12 Replace lysine with methionine 4.3% (w/v) trehalose 120 mM glycine 30 mM lysine 31.4 3.68 6.88 12' 4.3% (w/v) trehalose 120 mM glycine 30 mM methionine 27.6 3.25 5.86 13 1.8% (w/v) mannitol 80 mM histidine 30 mM lysine 15.3 2.49 4.32 13' 1.8% (w/v) mannitol 80 mM histidine 30 mM methionine 13.4 2.35 4.05

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

As a result, as shown in Table 3, it was confirmed from the results of Formulations 9', 10', and 11' that when methionine was added to the formulation containing sugar, viscosity, %HMW _{25℃/4 weeks} , and %HMW _{40℃/4 weeks} were simultaneously improved. Through these results, it was found that methionine has excellent functions as a viscosity reducing agent and stabilizer.

In addition, it was confirmed from the results of Formulations 12' and 13' that when lysine was replaced with methionine in the formulation containing sugar and lysine, viscosity, %HMW _{25℃/4 weeks} , and %HMW _{40℃/4 weeks} were improved simultaneously. Through these results, it was found that methionine has a superior function as a viscosity reducing agent and stabilizer compared to lysine.

Example 4. Evaluation of %HMW, %Acidic, %Met ₂₅₇ according to the presence or absence of methionine

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

Formulation 14' excludes histidine from Formulation 14, which contains sugars and amino acids (i.e., histidine and methionine).

Formulation 14'' excludes methionine from Formulation 14, which contains sugars and amino acids (i.e., histidine and methionine).

No. ^a Sibling 1 Brother 2 Brother 3 %HMW
(25℃/4 weeks) %Acidic
(25℃/4 weeks) %Met ₂₅₇
(25℃/4 weeks) 14 4% (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 2.56 41.6 12.5

^a All formulations contained 175 mg/mL dupilumab, 15 mM sodium acetate, 0.2% (w/v) polysorbate 80, and had a pH of 5.5.

As a result, as shown in Table 4, it was confirmed from the results of Formulation 14'' that when methionine was excluded from the formulation containing sugars and amino acids, %HMW, %Acidic, and %Met ₂₅₇ simultaneously increased under accelerated stability conditions at 25℃/4 weeks. In particular, the %Acidic and %Met ₂₅₇ of the formulation excluding methionine (Formulation 14') increased more than those of the formulation excluding histidine (Formulation 14'). Through these results, the function of methionine as a stabilizer and antioxidant could be confirmed.

Example 5. Evaluation of viscosity and %HMW of formulations containing and without methionine

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

Formulations 15 to 20 are methionine-containing formulations having different histidine concentrations and pHs.

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

No. ^a Buffer pH Sibling 1 Brother 2 Brother 3 Viscosity (cP) %HMW
(Initial) 15 30 mM sodium acetate 4.9 4% (w/v) sucrose 20 mM histidine 25 mM methionine 8.7 1.93 16 4.8 4% (w/v) sucrose 50 mM histidine 25 mM methionine 8.3 1.94 17 5.2 4% (w/v) sucrose 50 mM histidine 25 mM methionine 9.0 1.97 18 4.9 4% (w/v) sucrose 80 mM histidine 25 mM methionine 7.6 1.95 19 5.5 4% (w/v) sucrose 80 mM histidine 25 mM methionine 8.7 2.01 20 5.2 4% (w/v) sucrose 92.4 mM histidine 25 mM methionine 8.8 1.93 21 ^b 12.5 mM sodium acetate + 20 mM histidine 5.9 5% (w/v) sucrose 50 mM arginine - 11.1 2.03

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

^b Formulation 21 is a Dupixent formulation.

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

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.

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).

The above description of the present invention is for illustrative purposes only, and those skilled in the art will understand that the present invention can be easily modified into other specific forms without changing the technical idea or essential characteristics of the present invention. Therefore, it should be understood that the embodiments described above are exemplary in all respects and not restrictive.

Claims (43)

anti-IL-4Rα antibody; buffer; sugars; and Contains methionine, The pH is about 4.9 to about 5.5, Liquid formulation. A liquid formulation according to claim 1, wherein the anti-IL-4Rα antibody is dupilumab. 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. 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. 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. A liquid formulation according to claim 1, wherein the buffer comprises acetate. A liquid formulation according to claim 1, wherein the buffer is sodium acetate. 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. A liquid formulation according to claim 6, wherein the concentration of the acetate is from about 10 mM to about 40 mM. A liquid formulation according to claim 6, wherein the concentration of the acetate is from about 15 mM to about 35 mM. A liquid formulation according to claim 1, wherein the sugar comprises at least one selected from sugar and sugar alcohol. 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. 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. A liquid formulation according to claim 11, wherein the sugar comprises at least one selected from sucrose, trehalose, and mannitol. A liquid formulation according to claim 11, wherein the sugar is sucrose. A liquid formulation according to claim 11, wherein the concentration of the sugar is from about 1% (w/v) to about 20% (w/v). 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). A liquid formulation according to claim 15, wherein the concentration of sucrose is from about 1% (w/v) to about 20% (w/v). A liquid formulation according to claim 1, wherein the concentration of methionine is from about 2.7 mM to about 90 mM. A liquid formulation according to claim 1, wherein the concentration of methionine is about 20 mM to about 30 mM. 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. A liquid formulation according to any one of claims 1 to 21, further comprising histidine. A liquid formulation according to claim 12, wherein the concentration of histidine is from about 20 mM to about 92.4 mM. A liquid formulation according to claim 12, wherein the concentration of histidine is from about 30 mM to about 92.4 mM. A liquid formulation according to any one of claims 1 to 24, which does not contain arginine. A liquid formulation according to any one of claims 1 to 25, further comprising a surfactant. A liquid formulation according to claim 26, wherein the surfactant comprises a nonionic surfactant. A liquid formulation according to claim 26, wherein the surfactant comprises polysorbate 20, polysorbate 80, or a combination thereof. 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). 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). 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. 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 A liquid formulation, wherein the oxidation rate of methionine residues (%Met ₂₅₇ ) in the antibody measured after 4 weeks of storage at 25°C is less than 12%. 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%. 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 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 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. A liquid formulation for subcutaneous injection according to any one of claims 1 to 36. A device comprising a liquid formulation of any one of claims 1 to 37. A device according to claim 38, wherein the liquid formulation is contained 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. 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. 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. 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). Methods for treating IL-4Rα-associated conditions. 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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