TW202102262A - Stabilized formulations containing anti-interleukin-4 receptor (il-4r) antibodies - Google Patents

Abstract

The present invention provides pharmaceutical formulations comprising a human antibody that specifically binds to human interleukin-4 receptor (hIL-4R). The formulations may contain, in addition to an anti-hIL-4R antibody, at least one amino acid, at least one sugar, or at least one non-ionic surfactant. The pharmaceutical formulations of the present invention exhibit a substantial degree of antibody stability after storage for several months.

Description

Stabilizing formulation containing anti-interleukin-4 receptor (IL-4R) antibody

The present invention relates to the field of therapeutic antibody formulations. More specifically, the present invention relates to the field of pharmaceutical formulations comprising human antibodies that specifically bind to the human interleukin-4 receptor.

Sequence Listing

At the same time, this patent specification is accompanied by a ST.25 matching text file of the sequence table. The content of the text file is incorporated here by quotation. Incorporate the paper sequence list with the same content as the text file in accordance with ST.25 as a part of this patent specification.

Therapeutic macromolecules (such as antibodies) must be formulated in a way that not only makes the molecules suitable for patient administration, but also maintains stability during storage and subsequent use. For example, unless the solution is properly prepared, the therapeutic antibody in the liquid solution is prone to degradation, aggregation, or undesirable chemical modification. The stability of the antibody in the liquid formulation depends not only on the type of excipients used in the formulation, but also on the relative amount and ratio of the excipients. In addition, other factors must be considered in addition to stability when preparing a liquid antibody formulation. Examples of such additional considerations include the viscosity of the solution and the antibody concentration that can be adapted to a known formulation, as well as the visual quality or appeal of the formulation. Therefore, when formulating a therapeutic antibody, care must be taken to maintain a formulation that is stable, contains the appropriate antibody, and has the appropriate viscosity and other properties to facilitate the administration of the formulation to the patient.

Human interleukin-4 receptor alpha (hIL-4Rα) antibodies are an example of therapeutically related macromolecules that require proper formulation. Anti-hIL-4Rα antibodies are clinically used to treat or prevent diseases such as atopic dermatitis and allergic asthma, and other conditions. Range of anti-hIL-4Rα antibodies Examples have been specifically mentioned in US Patent Nos. 7,605,237, 7,608,693, 7,465,450 and 7,186,809; and US Patent Application Nos. 2010-0047254 and 2010-0021476.

Although anti-hIL-4Rα antibodies are well-known, there is still an urgent need in the art for a novel pharmaceutical formulation containing anti-hIL-4Rα antibodies that is stable enough and suitable for administration to patients.

Summary of Invention

The present invention satisfies the above-mentioned needs by providing a pharmaceutical formulation containing a human antibody capable of specifically binding to human interleukin-4 receptor α (hIL-4Rα).

In one aspect, a liquid pharmaceutical formulation is provided, comprising: (i) a human antibody capable of specifically binding to human interleukin-4 receptor α (hIL-4Rα); (ii) a buffer; ( iii) An organic cosolvent; (iv) a heat stabilizer; and (v) a viscosity reducer.

In a specific embodiment, the concentration of the antibody is about 150 mg/ml ± 50 mg/ml. In another specific embodiment, the concentration of the antibody is about 150 mg/ml ± 15 mg/ml. In a specific embodiment, the concentration of the antibody is about 150 mg/ml.

In a specific embodiment, the antibody comprises any one or more amino acid sequences of SEQ ID NO: 1-8. In a specific embodiment, the antibody comprises (a) heavy chain complementarity determining regions 1, 2 and 3 (HCDR1-HCDR2-HCDR3) containing SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, respectively One heavy chain variable region (HCVR); and (b) contains the light chain complementarity determining regions 1, 2 and 3 of SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, respectively (LCDR1-LCDR2-LCDR3 ) A light chain variable region (LCVR). In a specific embodiment, the antibody comprises an HCVR and an LCVR containing the amino acid sequences of SEQ ID NO:1 and SEQ ID NO:5, respectively.

In a specific embodiment, the antibody comprises any one or more amino acid sequences of SEQ ID NO: 9-16. In a specific embodiment, the antibody comprises (a) heavy chain complementarity determining regions 1, 2 and 3 (HCDR1-HCDR2-HCDR3) containing SEQ ID NO: 10, SEQ ID NO: 11 and SEQ ID NO: 12, respectively One heavy chain variable region (HCVR); and (b) contains the light chain complementarity determining regions 1, 2 and 3 of SEQ ID NO: 14, SEQ ID NO: 15 and SEQ ID NO: 16, respectively (LCDR1-LCDR2-LCDR3 ) A light chain variable region (LCVR). In a specific embodiment, The antibody contains an HCVR and an LCVR containing the amino acid sequences of SEQ ID NO: 9 and SEQ ID NO: 13, respectively.

In a specific embodiment, the antibody comprises any one or more amino acid sequences of SEQ ID NO: 17-24. In a specific embodiment, the antibody comprises (a) heavy chain complementarity determining regions 1, 2 and 3 (HCDR1-HCDR2-HCDR3) containing SEQ ID NO: 18, SEQ ID NO: 19 and SEQ ID NO: 20, respectively One heavy chain variable region (HCVR); and (b) contains the light chain complementarity determining regions 1, 2 and 3 of SEQ ID NO: 22, SEQ ID NO: 23 and SEQ ID NO: 24 respectively (LCDR1-LCDR2-LCDR3 ) A light chain variable region (LCVR). In a specific embodiment, the antibody comprises an HCVR and an LCVR containing the amino acid sequences of SEQ ID NO: 17 and SEQ ID NO: 21, respectively.

In a specific embodiment, the pH of the liquid formulation is about 5.9±0.5. In a specific embodiment, the pH of the liquid formulation is about 5.9±0.1. In a specific embodiment, the liquid pharmaceutical buffer contains one or more buffers that can buffer from about pH 5.6 to about pH 6.2.

In a specific embodiment, the liquid pharmaceutical formulation includes a buffer system containing at least two buffers. In a specific embodiment, the buffer system includes a first buffer solution having an effective pH range of 3.6 to 5.6 and a second buffer solution having an effective pH range of 5.5 to 7.4. In a specific embodiment, the first buffer has a pKa of about 4.8±0.3 and the second buffer has a pKa of about 6.0±0.3. In a specific embodiment, the first buffer is an acetate buffer and the second buffer is a histidine buffer. In a specific embodiment, the concentration of the acetate is 12.5±1.9mM and the concentration of histidine is 20±3mM.

In a specific embodiment, the organic co-solvent is a non-ionic polymer containing polyoxyethylene groups. In some specific embodiments, the organic cosolvent is any one or more of polysorbate 20, poloxamer 181, and polyethylene glycol 3350. In a specific embodiment, the organic co-solvent is polysorbate 20.

In a specific embodiment, the concentration of the organic co-solvent is from about 0.2%±0.03% to about 1%±0.15% "weight by volume" or "w/v", where, for example, 0.1 g/ml=10% and 0.01 g/ml=1%). In a specific embodiment, the concentration of the organic co-solvent is about 0.2%±0.03% w/v polysorbate 20.

In a specific embodiment, the heat stabilizer is a sugar. In a specific embodiment Among them, the sugar is selected from the group consisting of sucrose, mannose and trehalose. In a specific embodiment, the heat stabilizer is sucrose.

In a specific embodiment, the concentration of the heat stabilizer is from about 0.9%±0.135% w/v to about 10%±1.5% w/v. In a specific embodiment, the heat stabilizer is sucrose at a concentration of about 5%±0.75% w/v.

In a specific embodiment, the viscosity reducing agent is selected from the group consisting of arginine hydrochloride, sodium thiocyanate, ammonium thiocyanate, ammonium sulfate, ammonium chloride, calcium chloride, zinc chloride and sodium acetate Group of salt. In a specific embodiment, the viscosity reducing agent is L-arginine hydrochloride.

In a specific embodiment, the concentration of the viscosity reducing agent is less than about 100 mM. In a specific embodiment, the concentration of the viscosity reducing agent is 50 mM ± 7.5 mM. In another specific embodiment, the concentration of the viscosity reducing agent is 25 mM ± 3.75 mM. In a specific embodiment, the viscosity reducing agent is L-arginine hydrochloride at a concentration of 25 mM ± 3.75 mM.

In a specific embodiment, the viscosity of the liquid pharmaceutical formulation is less than or equal to about 35±3.5 centipoise (cPoise). In a specific embodiment, the viscosity is about 21.5±13.5 centipoise, about 11±1.1 centipoise, or about 8.5±0.85 centipoise. In a specific embodiment, the viscosity of the liquid pharmaceutical formulation is about 8.5±0.85 centipoise.

In a specific embodiment, the molar osmolality of the liquid pharmaceutical formulation is less than about 450 mOsm/kg. In a specific embodiment, the molar osmolality of the liquid pharmaceutical formulation is about 290±20 mOsm/kg.

In a specific embodiment, at least 90% or at least 95% of the anti-hIL in its natural form is recovered from the liquid pharmaceutical formulation after the liquid pharmaceutical formulation is stored at 5°C for six months when measured by size exclusion chromatography. -4Rα antibody. In a specific embodiment, at least 98% of the antibody in its natural form can be recovered from the liquid pharmaceutical formulation after storage at 5°C for six months when measured by size exclusion chromatography.

In a specific embodiment, at least 90% of the antibody in its natural form can be recovered from the liquid pharmaceutical formulation after storage at 45°C for eight weeks when measured by size exclusion chromatography.

In a specific embodiment, less than 45% of the antibody in acid form can be recovered from the liquid pharmaceutical formulation after storage at 45°C for eight weeks when measured by cation exchange chromatography.

In a specific embodiment, when measured by size exclusion exchange chromatography The antibody recovered from the liquid pharmaceutical formulation after storage at 25°C for six months was less than about 4% aggregated.

In one aspect, a liquid pharmaceutical formulation is provided, comprising: (i) about 150 mg/ml ± 50 mg/ml of a human antibody capable of specifically binding hIL-4Rα, wherein the antibody comprises SEQ ID NO: 1 and The heavy chain variable region (HCVR) and the light chain variable region (LCVR) of the amino acid sequence of SEQ ID NO: 5; (ii) about 12.5mg/ml±2mM acetate; (iii) about 20mM±3mM (Iv) about 5%±0.75%(w/v) of sucrose; (v) about 0.2%±0.03%(w/v) of polysorbate 20; and (vi) about Approximately 25mM±3.75mM arginine at 5.9±0.5 pH.

In a specific embodiment, the liquid pharmaceutical formulation has a viscosity of from about 8.5±0.85 centipoise to about 11±1.1 centipoise. In a specific embodiment, the viscosity of the liquid pharmaceutical formulation is about 8.5±0.85 centipoise.

In a specific embodiment, the liquid pharmaceutical formulation is physiologically isotonic. In a specific embodiment, the molar osmolality of the liquid pharmaceutical formulation is about 290±20 mOsm/kg.

In a specific embodiment, at least 98% of the anti-hIL-4Rα antibody in its natural form is recovered from the liquid pharmaceutical formulation after storage at 5°C for six months when measured by size exclusion chromatography.

In a specific embodiment, at least 90% of the anti-hIL-4Rα antibody in its natural form is recovered from the liquid pharmaceutical formulation after storage at 45°C for eight weeks when measured by size exclusion chromatography.

In a specific embodiment, less than 45% of the antibody in acid form can be recovered from the liquid pharmaceutical formulation after storage at 45°C for eight weeks when measured by cation exchange chromatography.

In a specific embodiment, the antibody recovered from the liquid pharmaceutical formulation after storage at 25°C for six months when measured by size exclusion exchange chromatography is less than about 4% aggregated.

In one aspect, a diazepam low-viscosity isotonic liquid pharmaceutical formulation containing at least 100 mg/ml diazepam anti-hIL-4Rα antibody is provided. In a specific embodiment, the concentration of the antibody is about 150 mg/ml ± 50 mg/ml. In a specific embodiment, the concentration of the antibody is about 150 mg/ml ± 15 mg/ml.

In a specific embodiment, the antibody comprises any one or more amino acid sequences of SEQ ID NO: 1-8. In a specific embodiment, the antibody comprises a heavy chain variable region (HCVR) and A light chain variable region (LCVR), wherein the HCVR/LCVR combination includes heavy chain and light chain complements containing amino acid sequences of SEQ ID NO:2-3-4 and SEQ ID NO:6-7-8, respectively The determining region (HCDR1-HCDR2-HCDR3/LCDR1-LCDR2-LCDR3). In a specific embodiment, one HCVR and one LCVR of the anti-hIL-4Rα antibody.

In some embodiments, the viscosity of the formulation is less than 35±3.5 centipoise, less than 20±2 centipoise, less than 15±1.5 centipoise, or less than 10±1 centipoise. In a specific embodiment, the liquid formulation has a viscosity of about 8.5 ± 2.5 centipoise.

In a specific embodiment, the formulation has a physiologically compatible molar osmolality. In a specific embodiment, the formulation contains a molar osmolality of 290±20 mOsm/kg.

In a specific embodiment, the antibody has stability at about 5°C for at least about six months. In a specific embodiment, at least about 98% of the antibodies retain their natural configuration when stored at 5°C for about six months when measured by size exclusion chromatography.

In a specific embodiment, the antibody has storage stability at about 45°C for at least about eight weeks. In a specific embodiment, at least about 90% of the antibodies retain their natural configuration when stored at 45°C for about eight weeks when measured by size exclusion chromatography. In a specific embodiment, less than about 45% of the antibody is in acidic form when stored at 45°C for about eight weeks when measured by cation exchange chromatography.

In a specific embodiment, the antibody has a storage stability of at least about six months at about 25°C. In a specific embodiment, less than about 4% of the antibody contains aggregated form when stored at 25°C for about six months when measured by size exclusion chromatography.

In a specific embodiment, the formulation includes a buffer having a pH of about 5.9±0.5. In a specific embodiment, the buffer includes an acetate buffer and a set of amino acid buffers. In a specific embodiment, the concentration of the acetate is 12.5mM±1.9mM and the concentration of the histidine is 20mM±3mM.

In a specific embodiment, the formulation includes an organic co-solvent at a concentration of from about 0.2%±0.03% to about 1%±0.15% w/v. In a specific embodiment, the organic co-solvent is a non-ionic polymer containing polyoxyethylene groups. In some specific embodiments, the organic cosolvent is any one or more of polysorbate 20, poloxamer 181, and polyethylene glycol 3350. In a particular In a specific embodiment, the organic cosolvent is polysorbate 20 with a concentration of about 0.2%±0.03% w/v.

In a specific embodiment, the formulation includes a heat stabilizer at a concentration of from about 0.9% ± 0.135% w/v to about 10% ± 1.5% w/v. In a specific embodiment, the heat stabilizer is sucrose. In a specific embodiment, the sugar is selected from the group consisting of sucrose, mannose and trehalose. In a specific embodiment, the heat stabilizer is sucrose with a concentration of about 5%±0.75% w/v.

In a specific embodiment, the formulation includes a viscosity reducing agent at a concentration of no more than about 100 mM. In a specific embodiment, the viscosity reducing agent is arginine. In a specific embodiment, the viscosity reducing agent is L-arginine hydrochloride with a concentration of 25 mM ± 3.75 mM.

In a specific embodiment, the diazepam low-viscosity isotonic liquid pharmaceutical formulation has a viscosity of about 8.5±2.5 centipoise and a molar osmolality of about 290±20 mOsm/kg, and contains: (i) 150 mg/ml ±15mg/ml anti-hIL-4Rα antibody, wherein the antibody contains one HCVR and one LCVR containing the amino acid sequences of SEQ ID NO:1 and SEQ ID NO:5, respectively; (ii) 12.5mM±1.9mM acetic acid Salt; (iii) 20mg/ml±3mM histidine; (iv) 0.2%±0.03% w/v polysorbate 20; (v) 5%±0.75% w/v sucrose; and ( vi) 25mM ± 3.75mM L-arginine hydrochloride. According to this specific example, (i) when measured by size exclusion chromatography, at least about 98% of the antibodies retain their natural configuration when stored at 5°C for at least about six months; (ii) when measured by size exclusion chromatography When stored at 45°C for at least about eight weeks, at least about 90% of the antibody retains its natural configuration; (iii) When stored at 45°C for eight weeks when measured by cation exchange chromatography, less than about 45% is in acid form Antibody; and (iv) less than about 4% of the antibody contains aggregated form when stored at 25°C for about six months when measured by size exclusion chromatography.

In one aspect, a liquid pharmaceutical formulation in any of the above aspects is provided in the container. In a specific embodiment, the container is a glass bottle. In another embodiment, the container is a microsyringe. In another embodiment, the container is a syringe. In a specific embodiment, the syringe includes a fluorocarbon coated piston. In a specific embodiment, the syringe is a low-tungsten syringe.

Other specific embodiments of the present invention can be better understood from the summary described in detail later.

Detailed description of the invention

Before describing the present invention, it should be understood that the present invention is not limited to the specific specific embodiments and experimental conditions, so different methods and conditions are possible.

Unless otherwise specified, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art. The word "approximately" here when used in relation to a specific stated value or range of values means that the value does not exceed 1% of the stated value. For example, the expression of "about 100" described here includes 99 and 101 and all values in between (such as 99.1, 99.2, 99.3, 99.4, etc.).

Although any similar or identical methods and materials described herein can also be applied in the practice or testing of the present invention, the preferred methods and materials are described in detail below. Herein, all the publications mentioned in this text are incorporated by reference to fully describe the present invention.

Pharmaceutical formulations

The statement of "pharmaceutical formulation" here means a combination of at least one active ingredient (for example, small molecules, macromolecules, compounds, etc. that can exhibit biological effects in humans or non-human animals) and at least one inactive ingredient, which is when When the active ingredient is mixed with one or more additional inactive ingredients, it can be therapeutically administered to humans or non-human animals. Unless expressly stated otherwise, the term "formulation" herein means "pharmaceutical formulation". The present invention provides pharmaceutical formulations comprising at least one therapeutic polypeptide. According to certain embodiments of the present invention, the therapeutic polypeptide can specifically bind to an antibody of human interleukin-4 antibody α (hIL-4Rα) or an antigen-binding fragment thereof. More specifically, the present invention includes (i) a human antibody that specifically binds to hIL-4Rα; (ii) an acetate/histidine buffer system; (iii) an organic aid of a nonionic surfactant Solvent; (iv) a carbohydrate heat stabilizer; and (v) a pharmaceutical formulation of a viscosity reducer. Specific exemplary ingredients and formulations contained in the present invention are described in detail below.

Antibodies that specifically bind to hIL-4R

The pharmaceutical formulation of the present invention comprises a human antibody or antigen-binding fragment thereof that specifically binds hIL-4Rα. The term "hIL-4Rα" as used herein means a human cytokine receptor that specifically binds to interleukin-4 (IL-4). In certain specific embodiments, the pharmaceutical preparations contained in the present invention The antibody in the substance specifically binds to the extracellular region of hIL-4Rα. An exemplary human IL-4 receptor alpha (hIL-4Rα) amino acid sequence is shown in SEQ ID NO:25. Antibodies against hIL-4Rα are described in U.S. Patent Nos. 7,605,237 and 7,608,693. The extracellular region of hIL-4Rα is shown in the amino acid sequence of SEQ ID NO:26.

The term "antibody" as used herein generally refers to immunoglobulin molecules and their multimers comprising four polypeptide chains, two heavy chains (H) and two light chains (L) connected to each other by disulfide chains. (Such as IgM); however, immunoglobulin molecules composed only of heavy chains (ie no light chains) also fall within the definition of "antibody". Each heavy chain includes a heavy chain variable region (herein referred to as HCVR or V _H for short) and a heavy chain constant region. The heavy chain constant region contains three domains, CH1, CH2 and CH3. Each light chain comprises a light chain variable region (abbreviated herein as LCVR or V _L) and a light chain constant region. The constant region of the light chain contains a domain (CL1). The V _H and V _L, may then be further subdivided into more interspersed conserved region called the framework region (FR) of the hypervariable regions called complementarity determining regions (CDRs). FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4 configuration consisting of: each of V _H and V _L, is composed of three CDRs and four FRs in the following order from the amino terminal to carboxy-terminus.

Unless expressly stated otherwise, the term "antibody" as described herein shall be deemed to include complete antibody molecules and antigen-binding fragments thereof. The "antigen-binding region" or "antigen-binding fragment" (or simply referred to as "antibody region" or "antibody fragment") of an antibody as described herein means the one that retains the ability to specifically bind hIL-4Rα or its epitope (epitope) One or more fragments of an antibody.

The "isolated antibody" as used herein means an antibody that is substantially free of other antibodies with different antigen specificities (for example, an isolated antibody that can specifically bind to hIL-4Rα, which does not substantially bind to hIL-4Rα except for hIL-4Rα). Antibodies to the antigen).

The term "specifically binds" and the like means that an antibody or antigen-binding fragment thereof forms a complex with an antigen that is relatively stable under physiological conditions. Specific binding is characterized by a ^{dissociation constant of at least about 1×10 −6} M or higher. The methods for determining whether two molecules specifically bind are already known in the art and include, for example, the dialysis equilibrium method, the surface plasma resonance method, and the like. However, an isolated antibody that specifically binds hIL-4Rα has cross-reactivity with other antigens, such as IL-4R (homologous gene) from other strains. In the context of the present invention, a multispecific (bispecific) antibody that binds hIL-4Rα and one or more additional antigens is considered to "specifically bind" hIL-4Rα. In addition, an isolated antibody can be substantially free of other cellular substances or chemicals.

Exemplary anti-hIL-4Rα antibodies incorporated in the pharmaceutical formulations of the present invention are described in US 7,605,237 and US 7,608,693, which are incorporated herein by reference.

According to some specific embodiments of the present invention, the anti-hIL-4Rα antibody system comprises a heavy chain variable region of a subtype of IGHV 3-9 and a human IgG1 (please See Barbie and Lefranc, Human Immunoglobulin Kappa Variable Region (IGKV) Gene and Linkage (IGKJ) Fragment, Exp. Clin. Immunogenet. 1998, 15: 171~183; and Scavener, D. et al., Human Immunoglobulin The protein display of heavy chain, kappa and lambda variable regions and junction regions, Exp. Clin. Immunogenet. 1999, 16: 234~240).

In some embodiments, the anti-hIL-4Rα includes at least one amino acid substitution, which results in a change in the charge of the exposed surface of the antibody relative to the germline IGHV 3-9 sequence or the germline IGKV 2-28 sequence. The germline IGHV 3~9 and germline IGKV 2~28 sequences shown here, as well as the amino acid position designation numbers and the International Immunogenetics (IMGT) Information System have been described in Lefranc, M.-P. et al. , IMGT ^® , International Immunogenetics Information System ^® , Nucl. Acids Res. , 37, D1006~D1012 (2009). In some embodiments, the exposed surface includes a complementarity determining region (CDR). In some embodiments, the one or more substitutions of the amino acid are selected from (a) a basic amino acid is substituted for a natural amino acid in the CDR2 of IGHV 3~9 (such as at position 58); b) A natural amino acid is substituted for an amino acid in CDR3 of IGHV 3~9 (as in position 107); and (c) a natural amino acid is substituted for an amino acid in CDR1 of IGKV 2~28 (as in position 33). A combination of basic amino acids. The special arrangement of an antibody, especially in the charge distribution of the environmental interface (for example, in the CDR), will make the stability of the antibody in the solution unpredictable.

In some embodiments, the anti-hIL-4Rα antibody comprises at least one amino acid substitution, which is produced in the framework region of the antibody variable region relative to the germline IGHV 3-9 sequence or the germline IGKV 2-28 sequence Changes in torsional strain. In some embodiments, the one or more substitutions of the amino acid are selected from (a) a proline substitution in the framework region 3 (FR3) of IGHV 3~9 (such as at position 96) in a non-proline Amino acid; and (b) a non-proline amino acid substituted with a combination of proline acid in the framework region 2 (FR2) (such as at position 46) of IGKV 2-28. Changing the peptide chain specifically refers to the ability of a framework region to affect the twisting ability of the CDR and the solvent interface, which will make the stability of the antibody in the solution unpredictable.

According to some specific embodiments of the present invention, the anti-hIL-4Rα antibody or its antigen binding The combined fragment includes a heavy chain complementarity determining region (HCDR) of SEQ ID NO: 2, an HCDR2 of SEQ ID NO: 3, and an HCDR3 of SEQ ID NO: 4. In certain embodiments, the anti-hIL-4Rα antibody or antigen-binding fragment thereof comprises an HCVD of SEQ ID NO:1.

According to some specific embodiments of the present invention, the anti-hIL-4Rα antibody or antigen-binding fragment thereof comprises a light chain (kappa) complementarity determining region (LCDR)1 of SEQ ID NO: 6, an LCDR2 of SEQ ID NO: 7, And an LCDR3 of SEQ ID NO: 8. In certain embodiments, the anti-hIL-4Rα antibody or antigen-binding fragment thereof comprises an LCVD of SEQ ID NO:5.

According to some other specific embodiments of the present invention, the anti-hIL-4Rα antibody or antigen-binding fragment thereof comprises an HCDR1 of SEQ ID NO: 10, an HCDR2 of SEQ ID NO: 11, an HCDR3 of SEQ ID NO: 12, and SEQ ID NO: 12 An LCDR1 of ID NO:14, an LCDR2 of SEQ ID NO:15, and an LCDR3 of SEQ ID NO:16. In some specific embodiments, the anti-hIL-4Rα antibody or antigen-binding fragment thereof comprises an HCVD of SEQ ID NO: 9 and an LCVD of SEQ ID NO: 13.

According to some other specific embodiments of the present invention, the anti-hIL-4Rα antibody or antigen-binding fragment thereof comprises an HCDR1 of SEQ ID NO: 18, an HCDR2 of SEQ ID NO: 19, an HCDR3 of SEQ ID NO: 20, and SEQ ID NO: 20. An LCDR1 of ID NO: 22, an LCDR2 of SEQ ID NO: 23, and an LCDR3 of SEQ ID NO: 24. In certain embodiments, the anti-hIL-4Rα antibody or antigen-binding fragment thereof comprises an HCVD of SEQ ID NO: 17 and an LCVD of SEQ ID NO: 21.

The non-limiting exemplary antibody in the examples here is called "mAb1". This antibody is also known as H4H098P in US 7,608,693. mAb1 (H4H098 P) contains an HCVR/LCVR amino acid sequence pair with SEQ ID NO:1/5, and HCDR1-HCDR2 representing SEQ ID NO:2-3-4/SEQ ID NO:6-7-8 -HCDR3/LCDR1-LCDR2-LCDR3 domain.

Another non-limiting exemplary antibody used in the practice of the present invention is called "mAb2". This antibody is also known as H4H083P in US 7,608,693. mAb2 (H4H083P) contains an HCVR/LCVR amino acid sequence pair with SEQ ID NO: 9/13, and HCDR1-HCDR2- representing SEQ ID NO: 10-11-12/SEQ ID NO: 14-15-16 HCDR3/LCDR1-LCDR2-LCDR3 domain.

Yet another non-limiting exemplary antibody used in the practice of the present invention is called "mAb3". This antibody is also known as H4H095P in US 7,608,693. mAb3 (H4H095P) contains an HCVR/LCVR amino acid sequence pair with SEQ ID NO: 17/21, and HCDR1-HCDR2- representing SEQ ID NO: 18-19-20/SEQ ID NO: 22-23-24 HCDR3/LCDR1-LCDR2-LCDR3 domain.

The content of the antibody or the antigen-binding fragment thereof in the pharmaceutical formulation of the present invention depends on the specific properties of the formulation and the specific conditions and applications in which the formulation is intended to be used. In certain embodiments, the pharmaceutical formulation contains about 100±10 to about 200±20 mg/ml, about 110±11 to about 190±19 mg/ml, about 120±12 to about 180±18 mg/ml, A liquid formulation of antibody of about 130±13 to about 170±17 mg/ml, about 140±14 to about 160±16 mg/ml, or about 150±15 mg/ml. For example, the formulation of the present invention contains about 90mg/ml, about 95mg/ml, about 100mg/ml, about 105mg/ml, about 110mg/ml, about 115mg/ml, about 120mg/ml, about 125mg/ml, about 130mg/ ml, about 131mg/ml, about 132mg/ml, about 133mg/ml, about 134mg/ml, about 135mg/ml, about 140mg/ml, about 145mg/ml, about 150mg/ml, about 155mg/ml, about 160mg/ ml, about 165mg/ml, about 170mg/ml, about 175mg/ml, about 180mg/ml, about 185mg/ml, about 190mg/ml, about 195mg/ml, or about 200mg/ml specifically binds hIL-4Rα The antibody or its antigen-binding fragment.

Excipients and pH

The pharmaceutical formulation of the present invention contains one or more excipients. The term "excipient" as used herein means any non-therapeutic substance added to the formulation to provide the desired consistency, viscosity, or stabilizing effect.

In certain embodiments, the pharmaceutical formulation of the present invention contains at least one organic co-solvent capable of stabilizing the type and amount of the hIL-4Rα antibody under vigorous operations such as shaking. In some specific embodiments, the term "stable" means that the formation of aggregated antibodies of the total amount of antibodies (on a molar basis) can be prevented by more than 2% during vigorous operation. In some embodiments, the vigorous operation involves shaking the solution containing the antibody and the organic co-solvent for about 120 minutes.

In some embodiments, the organic co-solvent is a nonionic surfactant, such as alkyl poly(oxyethylene). Specific nonionic surfactants that can be incorporated into the formulations of the present invention include, for example, polysorbate esters such as polysorbate 20, polysorbate 28, polysorbate 40, polysorbate 60, polysorbate 65, polysorbate 80, polysorbate 81, and poly-sorbate 85; poloxamers such as poloxamer 181, poloxamer 188, polo-xamer 407; or polyethylene glycol (PEG). Polysorbate 20 is also known as TWEEN 20, sorbitol monolaurate and polyoxyethylene sorbitol monolaurate. Poloxamer 181 is also known as PLURONIC F68.

The content of the organic co-solvent in the pharmaceutical formulation of the present invention depends on the special properties of the formulation and the specific conditions and applications in which the formulation is intended to be used. In certain embodiments, the surfactant content in the formulation is about 0.1±0.01% to about 2±0.2%. For example, the formulation of the present invention may contain about 0.09%, about 0.10%, about 0.11%, about 0.12%, about 0.13%, about 0.14%, about 0.15%, about 0.16%, about 0.17%, about 0.18%, about 0.19%, about 0.20%, about 0.21%, about 0.22%, about 0.23%, about 0.24%, about 0.25%, about 0.26%, about 0.27%, about 0.28%, about 0.29%, or about 0.30% of polysorbate 20 Or poloxamer 181. For example, the formulation of the present invention may contain about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, about 1%, about 1.1%, about 1.2%, about 1.3%, about 1.4%, about 1.5%, about 1.6%, about 1.7%, about 1.8%, about 1.9%, or about 2.0% PEG 3350.

Exemplary organic cosolvents that can stabilize the hIL-4Rα antibody contain 0.2%±0.02% polysorbate 20, 0.2%±0.02% polyxamer 181, or 1%±0.1% PEG 3350.

The pharmaceutical formulations of the present invention also contain one or more heat stabilizers that can stabilize the type and amount of the hIL-4Rα antibody under heat stress. In some embodiments, the term "stable" means that when a solution containing the antibody and heat stabilizer is stored at about 45° C. for up to about 28 days, it can maintain greater than about 92% of the antibody in its natural configuration. In some specific embodiments, the term "stable" means that when a solution containing the antibody and heat stabilizer is stored at about 45°C for up to about 28 days, it can maintain less than about 5% of the aggregated antibody.

In some embodiments, the heat stabilizer is a sugar or sugar alcohol selected from sucrose, trehalose and mannose or any combination thereof, and its content in the formulation depends on the specific situation and the formulation to be used. It depends on the purpose. In certain embodiments, the formulation contains about 2.5% to about 10%, about 3% to about 9.5%, about 3.5% to about 9%, about 4% to about 8.5%, about 4.5% to about 8. %, about 5% to about 7.5%, about 5.5% to about 7%, or about 6.0% to about 6.5% sugar or sugar alcohol. For example, the pharmaceutical formulation of the present invention may contain about 2.5%±0.375%, about 3%±0.45%, About 3.5%±0.825%, about 4.0%±0.6%, about 4.5%±0.675%, about 5.0%±0.75%, about 5.5%±0.825%, about 6.0%±0.9%, about 6.5%±0.975%, about 7.0%±1.05%, about 7.5%±1.125%, about 8.0%±1.2%, about 8.5%±1.275%, about 9.0%±1.35%, or about 10.0%±1.5% sugar or sugar alcohol (such as sucrose, Trehalose or mannose).

The pharmaceutical formulations of the present invention may also contain a buffer or buffer system to maintain a stable pH and help stabilize the hIL-4Rα antibody. In some specific embodiments, the term "stable" means that when a solution containing the antibody and buffer is stored at about 45°C for up to about 14 days, it can maintain less than 3.0%±0.5% of aggregated antibody. In some embodiments, the term "stable" means that when the solution containing the antibody and buffer is stored at about 25°C for up to about 6 months, it can maintain less than 3.7% ± 0.5% of aggregated antibody . In some specific embodiments, the term "stable" means that when a solution containing the antibody and buffer is stored at about 45°C for up to about 14 days, it can maintain at least 95% ± as determined by size exclusion chromatography. 0.5% natural conformation antibody. In some specific embodiments, the term "stable" means that when the solution containing the antibody and buffer is stored at about 25° C. for up to about 6 months, it can maintain at least 96% as determined by size exclusion chromatography. ±0.5% of natural conformation antibody. In some specific embodiments, the term "stable" means that when the solution containing the antibody and buffer is stored at about 45°C for up to about 14 days, it can maintain at least 62% ± as determined by cation exchange chromatography. 0.5% neutral conformation antibody. In some specific embodiments, the term "stable" means that when the solution containing the antibody and buffer is stored at about 25°C for up to about 6 months, it can maintain at least 54% as determined by cation exchange chromatography. ±0.5% neutral conformation antibody. "Neutral configuration" refers to the part of the antibody dialyzed from the main peak in the ion exchange resin, which usually has a "alkaline" peak on one side and a more "acidic" peak on the other side.

The pharmaceutical formulation of the present invention has a pH of from about 5.2 to about 6.4. For example, the pharmaceutical formulation of the present invention may have a range from about 5.2, about 5.3, about 5.4, about 5.5, about 5.6, about 5.7, about 5.8, about 5.9, about 6.0, about 6.1, about 6.2, about 6.3, or about 6.4.的pH. In some embodiments, the pH is about 5.3±0.2, about 5.9±0.2, or about 6.0±0.2.

In some embodiments, the buffer or buffer system includes at least one buffer that completely overlaps or is within a partial range of pH 5.2 to 6.4. In a specific embodiment, the buffer or buffer system includes two buffers, the first of which is in the effective pH range of 3.6 to 5.6 and the second of which is in the effective pH range of 5.5 to 7.4. In a specific embodiment, the first buffer has about The pKa of 4.8±0.3 and the second buffer have a pKa of about 6.0±0.3. In some embodiments, the buffer system includes acetate buffer and histidine buffer. In some specific embodiments, each 1 part molar ratio of acetate contains about 1.3 to 1.9 parts of histidine. In some specific embodiments, each 1 molar ratio of acetate contains about 1.6±0.25 parts of histidine. In certain embodiments, the concentration of the acetate is about 2.5 mM to about 22.5 mM, about 3.0 mM to about 22 mM, about 3.5 mM to about 21.5 mM, about 4.0 mM to about 21.0 mM, about 4.5 mM to about 20.5mM, about 5.0mM to about 20mM, about 5.5mM to about 19.5mM, about 6.0mM to about 19.0mM, about 6.5mM to about 18.5mM, about 7.0mM to about 18.0mM, about 7.5mM to about 17.5mM, About 8.0 mM to about 17.0 mM, about 8.5 mM to about 16.5 mM, about 9.0 mM to about 16.0 mM, about 9.5 mM to about 15.5 mM, about 10.0 mM to about 15.0 mM, about 10.5 mM to about 14.5 mM, about 12.5 mM to about 1.875 mM, about 11.0 mM to about 14.0 mM, about 11.5 mM to about 13.5 mM, or about 12.0 mM to about 13.0 mM. In certain embodiments, the concentration of the histidine is about 10 mM to about 30 mM, about 11 mM to about 29 mM, about 12 mM to about 28 mM, about 13 mM to about 27 mM, about 14 mM to about 26 mM, about 15 mM to about 25 mM. , About 16mM to about 24mM, about 17mM to about 23mM, about 18mM to about 22mM, or about 19mM to about 21mM. In some embodiments, the slow system includes about 12.5 mM acetate and about 20 mM histidine at about pH 5.9.

The pharmaceutical formulations of the present invention also contain one or more excipients for maintaining low viscosity or reducing the viscosity of formulations containing high concentrations of protein (for example, usually >100 mg/ml protein). In some embodiments, the arginine content of the formulation is sufficient to maintain the viscosity of the liquid formulation at less than about 35cPoise, less than about 30cPoise, less than about 25cPoise, less than about 20cPoise, less than about 15cPoise, low Below about 14cPoise, below about 13cPoise, below about 12cPoise, below about 10cPoise, or below about 9cPoise.

In certain specific embodiments, the pharmaceutical formulation of the present invention preferably contains L-arginine hydrochloride at an arginine concentration of about 25mM±3.75mM, about 50mM±7.5mM, or about 100mM±15mM . In certain embodiments, the concentration of arginine is about 20 mM to about 30 mM, about 21 mM to about 29 mM, about 21.25 mM to about 28.75 mM, about 22 mM to about 28 mM, about 23 mM to about 27 mM, or about 24 mM To about 26mM.

Representative formulation

According to one aspect of the present invention, the pharmaceutical formulation is a low-viscosity, usually physiologically isotonic liquid formulation, which contains: (i) specifically binds hIL-4Rα (such as mAb1, mAb2 or mAb3 [as described above] ]) a human antibody with a concentration of about 100mg/ml or higher; (ii) a buffer system sufficient to buffer at a pH of about 5.9±0.6; (iii) a sugar specially used as a thermal stabilizer; (iv) protect the internal structure of the antibody Complete organic co-solvent; and (v) monoamino acid that maintains viscosity for easy subcutaneous injection.

According to a specific embodiment, the pharmaceutical formulation includes: (i) specifically binds hIL-4Rα and includes a substituted IGHV type 3-9 heavy chain variable region and a substituted IGLV type 2-28 light chain variable region ( Such as mAb1) human IgG1 antibody with a concentration of about 100mg/ml to about 200mg/ml; (ii) a buffer system containing acetate and histidine sufficient to buffer at a pH of about 5.9±0.6; (iii) as a thermal stabilizer Sucrose; (iv) polysorbate as an organic co-solvent; and (v) arginine as a viscosity reducer.

According to a specific embodiment, the pharmaceutical formulation comprises: (i) specifically binds hIL-4Rα and comprises an HCDR1 of SEQ ID NO: 2 and an HCDR2 of SEQ ID NO: 3 and an HCDR3 of SEQ ID NO: 4 , An LCDR1 of SEQ ID NO:6, an LCDR2 of SEQ ID NO:7, and an LCDR3 of SEQ ID NO:8 human IgG1 antibody with a concentration of about 150mg/ml±25mg/ml; (ii) enough to buffer at about About 12.5mM±1.9mM acetate and about 20mM±3mM histidine with pH 5.9±0.3; (iii) about 5%±0.75% w/v sucrose; (iv) about 0.2%±0.03% w/v polysorbate 20; and (v) arginine as L-arginine hydrochloride at about 25mM±3.75mM.

Other non-limiting examples of the pharmaceutical formulations of the present invention have been described elsewhere herein, including the examples described below.

Stability and viscosity of pharmaceutical formulations

The pharmaceutical formulations of the present invention generally exhibit a high degree of stability. The term "safety" of the pharmaceutical formulation here means that the antibody in the pharmaceutical formulation maintains an acceptable degree of chemical structure or biological function under defined storage conditions. Even though the chemical structure or biological function of the antibody contained in a formulation cannot be maintained at 100% after storage for a period of time, it is still in a stable state. In some cases, after storage for a period of time, the structure or function of the antibody is maintained at about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% can be considered "stable".

The stability can be determined by particularly measuring the percentage of natural antibodies retained in the formulation after storage at a defined temperature for a period of time. In particular, the percentage of natural antibodies can be determined by size exclusion chromatography (e.g., size exclusion high performance liquid chromatography [SE-HPLC]). As used herein, the phrase "an acceptable degree of stability" means that at least 90% of the natural antibodies can be detected in the formulation after storage at a given temperature for a period of time. In certain embodiments, at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% can be detected in the formulation after storage at a defined temperature for a period of time. %, 99%, or 100% natural antibodies. The defined period of time measured as stable is at least 2 weeks, at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 12 months, at least 18 months, at least 24 months, or longer. The defined temperature for a storable pharmaceutical formulation that is determined to be stable can be any temperature from about -80°C to about 45°C, such as storage at about -30°C, about -20°C, about 0°C, about 4°C~8 °C, about 5°C, about 25°C, or about 45°C. For example, if a pharmaceutical formulation is stored at 5° C. for 3 months and can be determined by SE-HPLC to be greater than about 90%, 95%, 96%, 97%, or 98% of natural antibodies, it can be considered stable. If a pharmaceutical formulation has been stored at 5°C for 6 months and can detect greater than about 90%, 95%, 96%, 97%, or 98% of natural antibodies by SE-HPLC, it can also be considered stable. If a pharmaceutical formulation has been stored at 5°C for 9 months and can detect greater than about 90%, 95%, 96%, 97%, or 98% of natural antibodies by SE-HPLC, it can also be considered stable. If a pharmaceutical formulation is stored at 25°C for 3 months and can be determined by SE-HPLC to be greater than about 90%, 95%, 96%, or 97% of natural antibodies, it can also be considered stable. If a pharmaceutical formulation has been stored at 25°C for 6 months and can detect greater than about 90%, 95%, 96%, or 97% of natural antibodies by SE-HPLC, it can also be considered stable. If a pharmaceutical formulation has been stored at 25°C for 9 months and can detect greater than about 90%, 95%, 96%, or 97% of natural antibodies by SE-HPLC, it can also be considered stable.

The stability can be determined by measuring the percentage of aggregated antibodies in the formulation after storage at a defined temperature for a period of time, where the stability is inversely proportional to the percentage of aggregated antibodies formed. Can be especially by size exclusion chromatography (e.g. size exclusion high performance liquid chromatography [SE-HPLC]) Determine the percentage of aggregated antibodies. The phrase "an acceptable degree of stability" as described herein means that up to 5% of aggregated antibodies can be detected in the formulation after storage at a given temperature for a period of time. In some embodiments, an acceptable degree of stability means that at most about 5%, 4%, 3%, 2%, 1%, 0.5% can be detected in the formulation after storage at a given temperature for a period of time. % Or 0.1% of aggregated antibody. The defined period of time measured as stable is at least 2 weeks, at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 12 months, at least 18 months, at least 24 months, or longer. The defined temperature for a storable pharmaceutical formulation that is determined to be stable can be any temperature from about -80°C to about 45°C, such as storage at about -30°C, about -20°C, about 0°C, about 4°C~8 °C, about 5°C, about 25°C, or about 45°C. For example, if a pharmaceutical formulation is stored at 5°C for 3 months if it measures less than about 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of aggregated antibodies, it can be considered stable . After a pharmaceutical formulation is stored at 5°C for 6 months, if it is measured to be less than about 5%, 4%, 3%, 2%, 1%, 0.5% or 0.1% of the antibody in the aggregate form, it can also be considered stable. After a pharmaceutical formulation is stored at 5°C for 9 months, it can also be considered stable if the concentration of antibodies in the aggregate form is less than about 5%, 4%, 3%, 2%, 1%, 0.5% or 0.1% . After a pharmaceutical formulation is stored at 25°C for 3 months, it can also be considered stable if it measures less than about 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of the antibody in the aggregate form . After a pharmaceutical formulation is stored at 25°C for 6 months, it can also be considered stable if it measures less than about 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of the antibody in the aggregate form . After a pharmaceutical formulation is stored at 25°C for 9 months, it can also be considered stable if the concentration of antibodies in the aggregate form of less than about 5%, 4%, 3%, 2%, 1%, 0.5% or 0.1% is measured .

In particular, its stability can be determined by measuring the ratio of the antibody's migration to the more acidic part ("acidic form") and the main part of the antibody ("neutral configuration") during ion exchange, wherein the stability and the acidic form of the antibody Inversely proportional. Without being bound by theory, the deamidation of antibodies can lead to antibodies with more negative charges and therefore more acidic than non-deamidated antibodies (see, for example, Robinson, N., Deamidation of proteins, PNAS , April 16, 2002, 99(8): 5283~5288). The "acidified" or "deamidated" antibody can be determined especially by ion exchange chromatography (for example, cation exchange high performance liquid chromatography [CEX-HPLC]). The phrase "an acceptable degree of stability" as described herein means that up to 45% of the acidic form of the antibody can be detected in the formulation after storage at a defined temperature for a period of time. In certain embodiments, an acceptable degree of stability means that up to about 45%, 40%, 35%, 30%, 25%, 20% can be detected in the formulation after storage at a given temperature for a period of time. %, 15%, 10%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of the acid form antibody. The defined period of time measured as stable is at least 2 weeks, at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 12 months, at least 18 months, at least 24 months, or longer. The defined temperature for a storable pharmaceutical formulation that is determined to be stable can be any temperature from about -80°C to about 45°C, such as storage at about -30°C, about -20°C, about 0°C, about 4°C~8 °C, about 5°C, about 25°C, or about 45°C. For example, if a pharmaceutical formulation is stored at 5°C for 3 months if it is measured to be less than about 15%, 14%, 13%, 12%, 10%, 9%, 8%, 7%, 6%, 5%, An acidic form antibody of 4%, 3%, 2%, 1%, 0.5%, or 0.1% can be considered stable. If a pharmaceutical formulation is stored at 25°C for 3 months if it is measured to be less than about 18%, 17%, 16%, 15%, 14%, 13%, 12%, 10%, 9%, 8%, 7% , 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of acidic form antibodies can also be considered stable. If a pharmaceutical formulation is stored at 45°C for 8 weeks if it is measured to be less than about 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 5%, 4%, 3%, Acidic form antibodies of 2%, 1%, 0.5%, or 0.1% can also be considered diazepam. If a pharmaceutical formulation is stored at 40°C for 2 weeks, if it is measured to be less than about 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of the acidic form of antibodies can also be considered stable.

Other methods can be used to determine the stability of the formulations of the present invention, such as measuring thermal stability by differential scanning calorimetry (DSC), measuring mechanical stability by controlled agitation, and measuring the absorbance of a solution at about 350 or about 405 nm. Turbidity. For example, a pharmaceutical formulation of the present invention is stored at about 25 deg.] C to about 5 ℃ after 6 months or more if the formulation is less than about 0.05 OD ₄₀₅ (e.g., 0.04,0.03,0.02 zero time OD ₄₀₅ changes from, 0.01, or lower) is considered stable.

The stability of the antibody can also be assessed by measuring its biological activity or binding affinity to its target. For example, after a formulation of the present invention is stored at 5°C, 25°C, 45°C, etc. for a period of time (for example, 1 to 12 months), the anti-IL-4Rα antibody contained in the formulation has an antibody binding affinity of at least 90 before storage. %, 95% or higher bound to IL-4Rα, it can be considered stable. By For example, ELISA or plasma resonance method to determine binding affinity. The biological activity is measured by the IL-4Rα activity detection method, for example, contacting a cell expressing IL-4Rα with a formulation containing the anti-IL-4Rα antibody. The binding of such cells to the antibody can be directly measured, for example, by FACS analysis. Alternatively, the downstream activity of the IL-4Rα system is measured in the presence of the antibody and an IL-4Rα agonist, and then the activity of the IL-4Rα system without the antibody is compared. In some specific embodiments, the IL-4Rα may be endogenous in the cell. In other specific embodiments, the IL-4Rα can be expressed ectopicly in the cell.

Other methods for determining the stability of the antibody in the formulation are described in the examples below.

In certain embodiments, the pharmaceutical formulations of the present invention exhibit low to moderate viscosity. The "viscosity" mentioned here can be "dynamic viscosity" or "absolute viscosity". "Dynamic viscosity" is the measured flow resistance of a liquid under the influence of gravity. When two equal amounts of liquids are placed in the same capillary viscometer and flow under gravity, a viscous liquid requires a longer time for a lower viscous liquid to flow through the capillary. For example, if one liquid takes 200 seconds and another liquid takes 400 seconds to complete its flow, the second liquid has twice the viscosity of the first one on a dynamic viscosity scale. "Absolute viscosity" refers to the product of dynamic viscosity and liquid density (absolute viscosity = dynamic viscosity x density), which is sometimes referred to as dynamic or simple viscosity. The dimension of dynamic viscosity is L ² /T, the length of the L system and the time of the T system. Generally, dynamic viscosity is expressed as centistokes (cSt). The SI unit of dynamic viscosity is mm ² /s, which refers to 1 cSt. The unit of absolute viscosity is centipoise (cP). The SI unit of absolute viscosity is centipa-second (mPa-s), and its 1cP=1mPa-s.

As described herein, a low viscosity liquid formulation of the present invention will exhibit an absolute viscosity of less than about 15 centipoise (cP). For example, if a liquid formulation of the present invention is considered to have "low viscosity" after being measured by a standard viscosity measurement method, the formulation will exhibit about 15cP, about 14cP, about 13cP, about 12cP, about 11cP, about 10cP, about 9cP, about 8cP, or lower absolute viscosity. As described herein, a medium viscosity liquid formulation of the present invention will exhibit an absolute viscosity ranging from about 35 cP to about 15 cP. For example, if a liquid formulation of the present invention is considered to have "medium viscosity", the formulation will exhibit about 34cP, about 33cP, about 32cP, about 31cP, about 30cP, about 29cP, about 28cP, about 27cP, about 26cP , About 25cP, about 24cP, about 23cP, about 22cP, about 21cP, about 20cP, about 19cP, about 18cP, about 17cP, about 16cP, or about 15.1cP absolute viscosity.

As described in the examples below, the inventors unexpectedly discovered that the antibody can be formulated with arginine from about 25 mM to about 100 mM to obtain a high concentration of anti-hIL-4Rα antibody (for example, from about 100 mg/ml up to at least 200 mg/ml). ) Liquid formulations of low to medium viscosity. In addition, it has been further discovered that by adjusting the sucrose content to below about 10%, the viscosity of the formulation can be reduced to even greater extent.

Container and drug administration method for the pharmaceutical preparation

The pharmaceutical formulations of the present invention can be placed in any container suitable for storing drugs and other therapeutic compositions. For example, the pharmaceutical formulation can be placed in a sealed and sterilized plastic or glass container with a limited capacity, such as a vial, ampoule, syringe, cartridge, or vial. The formulation of the present invention can be put into different types of small glass bottles such as transparent and opaque (e.g. amber) glass or plastic bottles. Likewise, any type of syringe can be used to contain or administer the pharmaceutical formulation of the present invention.

The pharmaceutical formulation of the present invention can be placed in a "normal tungsten" syringe or a "low tungsten" syringe. As understood by those skilled in the art, the process of manufacturing a glass syringe usually involves the use of a hot tungsten rod that penetrates the glass to create small holes that can suck liquid from the syringe. This process causes a trace of tungsten to be deposited on the inner surface of the barrel. Washing and other processing steps can then be used to reduce the amount of tungsten in the syringe. The term "ordinary tungsten" as described herein means that the syringe contains more than 500 parts per billion (ppb) of tungsten. The term "low tungsten" means that the syringe contains less than 500 ppb of tungsten. For example, a low tungsten syringe according to the present invention may contain less than about 490, 480, 470, 460, 450, 440, 430, 420, 410, 390, 350, 300, 250, 200, 150, 100, 90, 80 , 70, 60, 50, 40, 30, 20, 10 or less ppb tungsten.

The rubber plunger used for the syringe and the rubber stopper used to close the opening of the glass bottle can be coated to avoid contamination of the syringe or the drug in the bottle, or to preserve its stability. Therefore, the pharmaceutical formulations of the present invention according to certain specific embodiments can be placed in a syringe containing a coated plunger, or in a glass bottle sealed with a coated rubber stopper. For example, the plunger or rubber stopper may be coated with a fluorocarbon film. Examples of coating plungers or rubber stoppers suitable for glass vials and syringes containing the pharmaceutical formulations of the present invention have been described in U.S. Patent Nos. 4,997,423, 5,908,686, 6,286,699, 6,645,635 and 7,226,554, the contents of which are fully incorporated by reference this. The special coated rubber plunger and rubber stopper used in the present invention can be purchased from the commercial product "FluroTec®" of West Pharmaceutical Services (Lionville, Pennsylvania).

According to certain embodiments of the present invention, the pharmaceutical formulation can be placed in a low-tungsten syringe containing a fluorocarbon-coated piston.

The pharmaceutical formulation can be administered to the patient via parenteral routes such as injection (such as subcutaneous, intravenous, intramuscular, intraperitoneal, etc.) or transdermal, mucosal, nasal, pulmonary or oral administration. Many reusable injection pens or automatic injection delivery devices can be used for subcutaneous infusion of the pharmaceutical formulations of the present invention. Examples include, but are not limited to, AUTOPEN ^TM (Owen Mumford, Woodstock, UK), DISETRONIC ^TM pen (Disetronic Medical, Bergdorf, Switzerland), HUMALOG MIX 75/25 ^TM pen, HUMALOG ^TM pen, HUMALIN 70/30 ^TM pen (Indian Eli Lilly, Indianapolis, N.A.), NOVOPEN ^TM I, II, and III (Novo Nordisk, ^{Copenhagen, Denmark), NOVOPEN JUNIOR TM} (Novo Nordisk, Copenhagen, Denmark), BD ^TM pen (Becton Dickinson, Franklin, New Jersey) , OPTIPEN ^TM , OPTIPEN PRO ^TM , OPTIPEN STARLET ^TM , and OPTICLIK ^TM (Sanofi-Aventis, Frankfurt, Germany). Examples of disposable injection pens or automatic injection delivery devices that have been used for subcutaneous infusion of the pharmaceutical composition of the present invention include, but are not limited to, SOLOSTAR ^TM pen (Sanofi-Aventis), FLEXPEN ^TM (Novo Nordisk), and KWIKPEN ^TM (Eli Lilly ); SURECLICK ^TM auto-injector (Amgen, Thousand Oaks, California), PENLET ^TM (Haselmeier, Stuttgart, Germany), EPIPEN ^TM (Dey, LP), and HUMIRA ^TM pen (Abbott Laboratory, Abbott Park, Illinois).

A microsyringe is also used here for infusion of the pharmaceutical formulation of the present invention. As described herein, "microsyringe" means to slowly administer a large amount (e.g., up to about 2.5 ml or more) of a therapeutic formulation over a long period of time (e.g., about 10, 15, 20, 25, 30 minutes or longer) Hypodermic infusion set. Please see, for example, US 6,629,949, US 6,659,982; and Meehan et al., J. Controlled Release 46: 107~116 (1996). The micro infusion device is most suitable for infusion of high-dose therapeutic proteins or viscous solutions containing high concentrations (such as about 100, 125, 150, 175, 200 mg/ml, or higher).

In a specific embodiment, the liquid pharmaceutical formulation containing about 150±15 mg/ml anti-IL-4Rα antibody is subcutaneously administered from a pre-filled syringe in an auto-injector with a volume of about 1±0.15 ml. In another specific embodiment, the formulation is administered from a microsyringe in a volume between about 1 ml and 2.5 ml. The formulation can be pre-filled into a pocket or cartridge of the microsyringe.

Therapeutic use of pharmaceutical formulations

The pharmaceutical formulations of the present invention are particularly useful for treating, preventing or alleviating any diseases or disorders related to IL-4 activity, including diseases or disorders mediated by IL-4Rα activation. Non-limiting example diseases and disorders that can be treated or prevented by administering the pharmaceutical formulations of the present invention include various allergic diseases such as atopic dermatitis, allergic conjunctivitis, allergic rhinitis, asthma, and other IgE/Th2 mediators. Leading disease.

Therefore, the present invention includes methods of treating, preventing or alleviating any disease or disorder related to IL-4 activity or IL-4Rα activation (including any of the above-mentioned exemplary diseases, disorders, and disorders). The treatment method of the present invention comprises administering the above-mentioned anti-hIL-4Rα antibody-containing formulation to a living body. The organism to which the pharmaceutical formulation is administered can be, for example, any human or non-human animal in need of such treatment, prevention or alleviation, or that can benefit from inhibiting or alleviating IL-4 or IL-4Rα-mediated activity. For example, the organism may be an individual who has been diagnosed or thought to be at risk of suffering from the aforementioned diseases or disorders. The present invention further includes any of the pharmaceutical formulations disclosed herein in the manufacture for the treatment, prevention or alleviation of any disease or disorder related to IL-4 activity or IL-4Rα activation (including any of the above-mentioned exemplary diseases, disorders, and disorders) the use of.

The following examples provide a complete disclosure and explanation of how to make and use the method and composition of the present invention by those skilled in the art, and are not intended to limit the scope of the invention regarded by the inventors. Although efforts have been made to ensure the accuracy of the numbers used (for example, quantity, temperature, etc.), some errors and deviations may still occur in some experiments. Unless otherwise specified, the parts are mole parts, the molecular weight is the average molecular weight, the temperature is in degrees Celsius, and the pressure is at or near atmospheric pressure.

The activity of forming the preliminary formulation involves screening the organic cosolvent, thermal stabilizer and buffer in the liquid formulation of mAb1 (anti-IL-4Rα antibody of the present invention) to confirm that the excipient is compatible with the protein and helps it. Stability, while maintaining the molar osmolality and viscosity for subcutaneous injection. The buffer conditions were also checked to determine the optimum pH for the highest protein stability.

Example 1. Organic cosolvent

It has been found that mAb1 is unstable under oscillating stress. Reversed-phase high performance liquid layer Analysis method (RP-HPLC) and size exclusion high performance liquid chromatography (SE-HPLC) analysis proved that when mAb1 was shaken at room temperature, protein loss and agglutinating protein increased (Table 1, please see "No Cosolvent "data). The addition of organic co-solvents to the mAb1 solution can prevent protein degradation as measured by SE-HPLC and RP-HPLC (Table 1). However, it has been found that the addition of some organic co-solvents will reduce the thermal stability of mAbl (Table 2). It has been found that formulations containing PEG 3350 (3%) and PEG 300 (10% and 20%) will lose the recovered protein measured by RP-HPLC after thermal stress (Table 2). In addition, formulations containing PLURONIC F68 (poloxamer 181) (0.2%), PEG 300 (10% and 20%), and propylene glycol (20%) formed more aggregation measured by SE-HPLC than formulations without cosolvent. Things. Polysorbate 20 (0.2%) and polysorbate 80 (0.2%) have considerable stability to vibration and thermal stress.

According to Table 1, in a 2 ml glass bottle, 0.3 ml of mAb1 at 15 mg/ml in 10 mM phosphate pH 6.0 and various organic co-solvents were shaken for about 120 minutes. The turbidity was measured at the optical density (OD) at 405 nm and the relative change in the OD of the starting material at 405 nm was compared. The percentage of natural and aggregated mAbl was determined by size exclusion HPLC (SE-HPLC) method. The SE-HPLC results shown in the "Starting Materials" table are the average values of each formulation without shaking.

According to Table 2, 0.3 ml of mAb1 at 15 mg/ml in 10 mM phosphate pH 6.0 and various organic co-solvents in a 2 ml glass bottle were stored at about 45° C. for about 28 days. The turbidity was measured at the optical density (OD) at 405 nm and the relative change in the OD of the starting material at 405 nm was compared. The percentage of total recovered mAb1 was determined by reverse phase HPLC (RP-HPLC) method. The percentage of natural and aggregated mAbl was determined by size exclusion HPLC (SE-HPLC) method. The SE-HPLC results shown in the "Starting Materials" table are the average values of the formulations that are not stressed by heat.

Example 2. Heat stabilizer

Check the ability of various heat stabilizers, such as sugars, amino acids and inorganic salts to inhibit mAb1 degradation when stored at about 45°C. A summary of a study of heat stabilizers is shown in Table 3. The formulations containing sucrose or trehalose have a higher mAb1 stabilizing effect (determined by SE-HPLC) when cultured in a high-temperature solution. Because sucrose has a long history of safety in monoclonal antibody formulations, it was chosen as a stabilizer.

According to Table 3, 0.3 ml of mAb1 at 25 mg/ml in 10 mM acetate pH 5.3 and various heat stabilizers in a 2 ml glass bottle were stored at about 45° C. for about 28 days. The turbidity was measured at the optical density (OD) at 405 nm and the relative change in the OD of the starting material at 405 nm was compared. The turbidity difference of all samples can be ignored. The percentage of total recovered mAb1 was determined by reverse phase HPLC (RP-HPLC) method. The percentage of natural and aggregated mAbl was determined by size exclusion HPLC (SE-HPLC) method. Acidic or basic substances are defined as the total amount of mAb1 peaks dialyzed from the cation exchange (CEX-HPLC) column earlier or later than the main peak respectively. Shown in the "Starting Materials" table The SE-HPLC result is the average value of each formulation that is not stressed by heat.

Example 3. Buffer and pH

The effects of pH and buffer type on the stability of mAb1 were also examined. 15 mg/ml mAb1 was cultured with different buffers at different pH values ranging from pH 4.5 to 7.0. The protein stability was monitored by SE-HPLC and cation exchange HPLC (CEX-HPLC). When mAb1 was formulated in pH 6.0 histidine buffer or pH 5.3 acetate buffer, it had the greatest protein stability determined by SE-HPLC and CEX-HPLC (Table 4 and Table 5). The acetate buffer system has a wider pH stability range and a lower rate of variable charge formation relative to the formulation containing histidine buffer (Table 5). Therefore, an acetate buffer with pH 5.3 was selected for the formulation of mAb1 drugs.

According to Table 4, a 2ml glass bottle containing 10mM of various buffers mixed with 0.3ml of 15mg/ml mAb1 and 0.2% of polysorbate 20 was stored at about 45°C for about 14 days. The turbidity was measured at the optical density (OD) at 405 nm and the relative change in the OD of the starting material at 405 nm was compared. The turbidity difference of all samples can be ignored. The percentage of total recovered mAb1 was determined by reverse phase HPLC (RP-HPLC) method. The percentage of natural and aggregated mAbl was determined by size exclusion HPLC (SE-HPLC) method. Acidic or basic substances are defined as the total amount of mAb1 peaks dialyzed from the cation exchange (CEX-HPLC) column earlier or later than the main peak respectively. The SE-HPLC results shown in the "Starting Materials" table are the average values of the formulations that are not stressed by heat.

According to Table 5, a 2ml glass bottle containing 10mM of various buffers mixed with 0.3ml of 15mg/ml mAb1 and 0.2% of polysorbate 20 was stored at about 45°C for about 14 days. The turbidity was measured at the optical density (OD) at 405 nm and the relative change in the OD of the starting material at 405 nm was compared. The turbidity difference of all samples can be ignored. The percentage of total recovered mAb1 was determined by reverse phase HPLC (RP-HPLC) method. The percentage of natural and aggregated mAbl was determined by size exclusion HPLC (SE-HPLC) method. Acidic or basic substances are defined as the total amount of mAb1 peaks dialyzed from the cation exchange (CEX-HPLC) column earlier or later than the main peak respectively. The SE-HPLC results shown in the "Starting Materials" table are the average values of the formulations that are not stressed by heat.

6.5)下溶液內mAb1可被去醯胺。反之，低於pH 5.0時已發現可增加形成變異分子量之mAb1的速率。根據這些數據，該mAb1配製物的pH被維持在約pH 5.6和pH 6.2之間。已發現mAb1可穩定存在於此pH範圍。 The formulation formation test was shown in alkaline conditions (pH

6.5) mAb1 in the next solution can be desamidated. Conversely, it has been found to increase the rate of forming mAb1 with variable molecular weight below pH 5.0. According to these data, the pH of the mAbl formulation was maintained between about pH 5.6 and pH 6.2. It has been found that mAb1 can stably exist in this pH range.

The effects of pH and buffer types containing 20 mM histidine pH 6, 12.5 mM acetate pH 5.3 or a mixture of 20 mM histidine acid and 12.5 acetate pH 5.9 in the formulation on the stability of mAb1 were further evaluated (Table 6). Compared with individual buffer systems, mAb1 containing histidine and acetate with a pH of about 5.9 in the formulation is the most stable. When mAbl was formulated in this mixed buffer system (SE-HPLC), it had the slowest aggregation rate (Table 6).

According to Table 6, 0.4ml of 150mg/ml mAb1, 10% sucrose, 0.2% polysorbate 20 mixed with various buffers in a 2ml glass bottle was stored at about 45°C for about 14 days. The turbidity was measured at the optical density (OD) at 405 nm and the relative change in the OD of the starting material at 405 nm was compared. The turbidity difference of all samples can be ignored. The percentage of total recovered mAb1 was determined by reverse phase HPLC (RP-HPLC) method. The percentage of natural and aggregated mAbl was determined by size exclusion HPLC (SE-HPLC) method. Acidic or basic substances are defined as the total amount of mAb1 peaks dialyzed from the cation exchange (CEX-HPLC) column earlier or later than the main peak respectively. The SE-HPLC results shown in the "Starting Materials" table are the average values of the formulations that are not stressed by heat.

Example 4. Viscosity and tension control

Evaluate mixing various excipients and high-concentration mAb1 (ie 150mg/ml, 175 mg/ml and 200mg/ml) viscosity and tension (expressed as molar osmolality). The contents of sucrose, sodium chloride and L-arginine hydrochloride were adjusted to form a low viscosity and physiological isotonic formulation containing high concentration of mAb1 for easy, comfortable and rapid subcutaneous infusion of high amounts of mAb1 (Table 7). Containing 25mM arginine, 20mM histidine, 12.5mM acetate, 5% (w/v) sucrose, 0.2% (w/v) polysorbate 20 and 150mg/ml mAb1 pH 5.9 liquid formulation (formulation A ) Is the best formulation with low viscosity (about 8.5cPoise) and physiological isotonicity (about 293mOsm/kg) while still maintaining the stability of mAb1.

Example 5. Characterization of Formulation A

The main degradation pathways in the formation of mab1 liquid formulations are aggregates, fragmentation products and charge variants. The formation of these degradation products can be reduced by formulating mAb1 in a pH 5.9 formulation containing 20 mM histidine, 12.5 mM acetate, 0.2% polysorbate 20, 5% sucrose, and 25 mM L-arginine hydrochloride. It has been found that these formulated 150 mg/ml mAb1 are transparent to slightly opalescent liquid solutions with substantially no particles visible to the naked eye.

The formulated mAb1 has physical and chemical stability under various stress (cultivation at 25°C and 45°C) and immediate storage conditions (5°C) (Table 8). The mAb1 does not affect its appearance when stored at 25°C (3 months) or 5°C for 6 months. In addition, it has been found that it does not affect the solution pH, turbidity, or the amount of mAb1 recovered. After the formulated mAb1 was stored at 25°C for 3 months, the antibody determined by SE-HPLC was not significantly degraded and the degradation determined by CEX-HPLC was only 3.3% higher. It has been found that after 8 weeks of storage at 45°C, the amount of degradation determined by SE-HPLC and CEX-HPLC increases and shows signs of appearance. Aggregation and charge variation are the main degradation pathways of the mAb1 antibody molecule. The formulated mAb1 antibody was not found to be degraded when stored at 5°C for 6 months.

According to Table 8, OD=optical density; RP-HPLC=reverse phase high performance liquid chromatography; SE-HPLC=size exclusion high performance liquid chromatography; and CEX-HPLC=cation exchange high performance liquid chromatography. Acidic or basic substances are defined as the total amount of mAb1 peaks dialyzed from the cation exchange (CEX-HPLC) column earlier or later than the main peak respectively.

Example 6. Container

Filter-sterilized formulations containing mAb1 have been tested and proved to be stable. The manufacturing system for clinical supply uses the microporous MILLIPAK filter device, and the same composition filter (microporous Millex DURAPORE) is used in the research room.

Take at least 2.5ml 150mg/ml mAb1 at pH 5.9, 5%(w/v) sucrose, 25mM L-arginine hydrochloride, 0.2%(w/v) polysorbate 20, 12.5mM acetate, 20 mM histidine is filled into a 5ml glass bottle. Use 0.5ml of over-formulation in a 5ml bottle to ensure that 2.0ml of formulation can be withdrawn. This excess is not intended to compensate for losses during manufacture of mAbl or mAbl-containing formulations, degradation during manufacture, degradation during storage (shelf life), or to extend the shelf life.

Compared with storage in glass bottles, the stability of the formulated mAb1 (formulation A) is not affected when stored in polypropylene glycol tubes, polystyrene tubes, polycarbonate tubes, or glass bottles containing stainless steel blocks (Table 9).

According to Table 9, store 150mg/ml mAb1 with pH 5.9, 5% sucrose, 25mM arginine hydrochloride, 0.2% PS-20, 20mM histidine, 12.5mM acetate in various materials at 40℃ for 14 days . OD = optical density; RP-HPLC = reversed-phase high performance liquid chromatography; SE-HPLC = size exclusion high performance liquid chromatography; and CEX-HPLC = cation exchange high performance liquid chromatography. Turbidity is recorded as the difference in turbidity at 405 nm compared to the starting material. Acidic or alkaline substances are defined as the total amount of mAb1 peaks dialyzed from the CEX-HPLC column earlier or later than the main peak retention time respectively.

<110> Regeneron Pharmaceuticals, Inc.

<120> Stabilization formulation containing anti-interleukin-4 receptor (IL-4R) antibody

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Claims (39)

A liquid pharmaceutical formulation comprising: (i) a human antibody that specifically binds to human interleukin-4 receptor α (hIL-4Rα); (ii) a buffer solution at pH 5.9 ± 0.5 pH unit; (iii) ) Organic cosolvent; (iv) heat stabilizer; and (v) viscosity reducer. For example, the liquid pharmaceutical formulation of item 1 in the scope of patent application, wherein the concentration of the antibody is 150±50 mg/ml. For example, the liquid pharmaceutical formulation of item 1 or 2 of the scope of patent application, wherein the concentration of the antibody is 150±15 mg/ml. For example, the liquid pharmaceutical formulation of any one of items 1 to 3 in the scope of the patent application, wherein the buffer comprises a first buffer with an effective pH range of 3.6 to 5.6 and a second buffer with an effective pH range of 5.5 to 7.4 liquid. Such as the liquid pharmaceutical formulation of claim 4, wherein the first buffer solution has a pKa of 4.8±0.3 and the second buffer solution has a pKa of 6.0±0.3. Such as the liquid pharmaceutical formulation of item 5 in the scope of patent application, wherein the first buffer is acetate and the second buffer is histidine. For example, the liquid pharmaceutical formulation of any one of items 1 to 6 in the scope of patent application, wherein the concentration of the acetate is 12.5mM±1.85mM and the concentration of histidine is 20mM±0.3mM. Such as the liquid pharmaceutical formulation of any one of items 1 to 7 in the scope of patent application, wherein the organic cosolvent is selected from the group consisting of polysorbate 20, poloxamer 181 and polyethylene glycol 3350 . For example, the liquid pharmaceutical formulation of any one of items 1 to 8 in the scope of patent application, wherein the concentration of the organic co-solvent is about 0.085% to about 1.15% w/v. For example, the liquid pharmaceutical formulation of any one of items 1 to 9 in the scope of patent application, wherein the organic aid The solvent is polysorbate 20 with a concentration of 0.2%±0.03% w/v. Such as the liquid pharmaceutical formulation of any one of items 1 to 10 in the scope of patent application, wherein the heat stabilizer is selected from the group consisting of sucrose, mannitol and trehalose. Such as the liquid pharmaceutical formulation of any one of items 1 to 11 in the scope of patent application, wherein the concentration of the heat stabilizer is about 3.5% to about 11% w/v. For example, the liquid pharmaceutical formulation of any one of items 1-12 in the scope of patent application, wherein the heat stabilizer is sucrose with a concentration of 5%±0.75% w/v. For example, the liquid pharmaceutical formulation of any one of items 1 to 13 in the scope of the patent application, wherein the concentration of the viscosity reducing agent is less than about 100 mM. For example, the liquid pharmaceutical formulation of any one of items 1 to 14 in the scope of the patent application, wherein the viscosity reducing agent is arginine at a concentration of 25 mM ± 3.75 mM or 50 mM ± 7.5 mM. For example, the liquid pharmaceutical formulation of any one of items 1 to 15 in the scope of patent application, wherein the viscosity of the liquid is lower than or equal to 35 ± 3.5 centipoise. For example, the liquid pharmaceutical formulation of any one of items 1 to 16 in the scope of patent application, wherein the viscosity of the liquid is 21.5±13.5 centipoise. For example, the liquid pharmaceutical formulation of any one of items 1 to 16 in the scope of patent application, wherein the viscosity of the liquid is 11±1.1 centipoise or 8.5±0.85 centipoise. For example, the liquid pharmaceutical formulation of any one of items 1 to 18 in the scope of patent application, wherein the molar osmolality of the liquid is 290±20 mOsm/kg. Such as the liquid pharmaceutical formulation of any one of items 1 to 19 in the scope of the patent application, wherein when measured by size exclusion chromatography, at least about 90% of the antibody in its natural form is stored at about 5°C for about six months. Recycling. Such as the pharmaceutical formulation of any one of items 1 to 20 in the scope of the patent application, wherein at least about 95% of the antibody in its natural form is recovered after being stored at about 5°C for about six months when measured by size exclusion chromatography . Such as the pharmaceutical formulation of any one of items 1 to 21 in the scope of the patent application, wherein at least about 98% of the antibody in its natural form is recovered after being stored at about 5°C for about six months when measured by size exclusion chromatography . Such as the liquid pharmaceutical formulation of any one of items 1 to 22 in the scope of the patent application, wherein at least about 90% of the antibody in its natural form is recovered after being stored at about 45°C for about eight weeks when measured by size exclusion chromatography . Such as the liquid pharmaceutical formulation of any one of items 1 to 23 in the scope of the patent application, wherein less than about 45% of the recovered antibody is acidic after being stored at about 45°C for about eight weeks when measured by cation exchange chromatography form. Such as the liquid pharmaceutical formulation of any one of items 1 to 24 in the scope of the patent application, wherein less than about 4% of the recovered anti-system aggregates after being stored at 25°C for six months when measured by size exclusion exchange chromatography of. For example, the liquid pharmaceutical formulation of the first item in the scope of patent application, wherein the human antibody that specifically binds hIL-4R comprises a heavy chain variable region (HCVR) and a light chain variable region (LCVR), wherein (a) the HCVR contains (i) SEQ ID NO: 2, SEQ ID NO: 3, and SEQ ID NO: 4, respectively; (ii) SEQ ID NO: 10, SEQ ID NO: 11, and SEQ ID NO: 12, respectively; or (iii) The heavy chain complementarity determining regions HCDR1, HCDR2 and HCDR3 of the amino acid sequences of SEQ ID NO: 18, SEQ ID NO: 19 and SEQ ID NO: 20, respectively; and (b) the LCVR contains (i) Respectively SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8; (ii) Respectively SEQ ID NO: 14, SEQ ID NO: 15 and SEQ ID NO: 16; or (iii) Respectively SEQ ID NO: The light chain complementarity determining regions LCDR1, LCDR2 and LCDR3 of the amino acid sequence of ID NO: 22, SEQ ID NO: 23 and SEQ ID NO: 24. Such as the liquid pharmaceutical formulation of item 26 in the scope of patent application, wherein the HCVR and LCVR respectively comprise the amino acid sequences of SEQ ID NO:1 and SEQ ID NO:5. For example, the liquid pharmaceutical formulation of any one of items 1 to 27 in the scope of patent application is put into a glass bottle. For example, the liquid pharmaceutical formulation of any one of items 1 to 27 in the scope of patent application is put into a syringe. For example, the liquid pharmaceutical formulation of any one of items 1 to 27 in the scope of patent application is put into a micro syringe. Such as the liquid pharmaceutical formulation of item 29 in the scope of patent application, wherein the syringe contains a fluorocarbon coated piston. For example, the liquid pharmaceutical formulation of item 29 or 31 in the scope of patent application, wherein the syringe is a low-tungsten syringe. A liquid pharmaceutical formulation comprising: (i) a 150 mg/ml ± 50 mg/ml human antibody that specifically binds hIL-4Rα, wherein the antibody comprises the heavy chain of the amino acid sequence of SEQ ID NO: 1/5 and Light chain variable region (HCVR/LCVR); (ii) 12.5mM±2mM acetate; (iii) 20mM±3mM histidine; (iv) 5%±0.75% sucrose; (v) 0.2%±0.03% poly Sorbate 20; and (vi) 25mM±3.75mM arginine at pH 5.9±0.5. For example, the liquid pharmaceutical formulation of item 33 in the scope of patent application, wherein the viscosity of the liquid is 11±1.1 centipoise or 8.5±0.85 centipoise. For example, the liquid pharmaceutical formulation of item 33 or 34 in the scope of patent application, wherein the molar osmolality of the liquid is 290±20 mOsm/kg. Such as the liquid pharmaceutical formulation of any one of 33 to 35 of the scope of patent application, wherein at least 98% of the antibody in its natural form is recovered after storage at 5°C for six months when measured by size exclusion chromatography. Such as the liquid pharmaceutical formulation of any one of 33 to 36 in the scope of patent application, wherein at least 90% of the antibody in its natural form is recovered after being stored at 45°C for eight weeks when measured by size exclusion chromatography. Such as the liquid pharmaceutical formulation of any one of the 33 to 37 patents, wherein less than 45% of the recovered antibody is in acid form after being stored at 45°C for eight weeks when measured by cation exchange chromatography. Such as the liquid pharmaceutical formulation of any one of the 33 to 38 patents, in which less than about 4% of the recovered anti-system aggregates after being stored at 25°C for six months when measured by size exclusion exchange chromatography of.