TWI856388B - 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

Stabilized formulation containing anti-interleukin-4 receptor (IL-4R) antibodies

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 can specifically bind to human interleukin-4 receptor.

Sequence Listing

This patent specification is also accompanied by a ST.25 compliant text file with a sequence list. The contents of the text file are incorporated herein by reference. A paper copy of the sequence list with the same contents as the ST.25 compliant text file is incorporated as part of this patent specification.

Therapeutic macromolecules such as antibodies must be formulated in a manner that not only renders the molecule suitable for administration to a patient, but also maintains stability during storage and subsequent use. For example, therapeutic antibodies in liquid solutions are susceptible to degradation, aggregation, or undesirable chemical modifications unless the solution is properly formulated. The stability of antibodies in liquid formulations depends not only on the type of excipients used in the formulation, but must also depend on the relative amounts and ratios of the excipients. Furthermore, other factors besides stability must be considered when preparing a liquid antibody formulation. Examples of such additional considerations include the viscosity of the solution and the antibody concentration that can be accommodated in a given formulation, as well as the visual qualities or appeal of the formulation. Therefore, when formulating a therapeutic antibody, care must be taken to ensure that the formulation remains stable, contains the appropriate antibody, and has the appropriate viscosity and other properties to facilitate administration of the formulation to a patient.

Antibodies to human interleukin-4 receptor alpha (hIL-4Rα) are an example of therapeutically relevant macromolecules that require appropriate formulation. Anti-hIL-4Rα antibodies are used clinically to treat or prevent diseases such as atopic dermatitis and allergic asthma, among other conditions. Examples of anti-hIL-4Rα antibodies are described in particular in U.S. Patent Nos. 7,605,237, 7,608,693, 7,465,450, and 7,186,809; and U.S. Patent Application Nos. 2010-0047254 and 2010-0021476.

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

Abstract of the invention

The present invention satisfies the above needs by providing a pharmaceutical formulation containing a human antibody that can specifically bind to human interleukin-4 receptor α (hIL-4Rα).

In one embodiment, a liquid pharmaceutical formulation is provided, comprising: (i) a human antibody that can specifically bind to human interleukin-4 receptor α (hIL-4Rα); (ii) a buffer solution; (iii) an organic cosolvent; (iv) a thermal stabilizer; and (v) a viscosity reducer.

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

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

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

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

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

In one embodiment, the liquid pharmaceutical formulation comprises a buffer system comprising at least two buffers. In one embodiment, the buffer system comprises a first buffer having an effective pH range of 3.6 to 5.6 and a second buffer having an effective pH range of 5.5 to 7.4. In one 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 particular embodiment, the first buffer is an acetate buffer and the second buffer is a histidine buffer. In a specific embodiment, the concentration of acetate is 12.5±1.9 mM and the concentration of histidine is 20±3 mM.

In one embodiment, the organic cosolvent is a nonionic polymer containing polyoxyethylene groups. In some embodiments, the organic cosolvent is any one or more of polysorbate 20, poloxamer 181 and polyethylene glycol 3350. In a particular embodiment, the organic cosolvent is polysorbate 20.

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

In one embodiment, the thermal stabilizer is a sugar. In one embodiment, the sugar is selected from the group consisting of sucrose, mannose and trehalose. In a particular embodiment, the thermal stabilizer is sucrose.

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

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

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

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

In one embodiment, the molar osmotic pressure concentration of the liquid pharmaceutical formulation is less than about 450 mOsm/kg. In one embodiment, the molar osmotic pressure concentration of the liquid pharmaceutical formulation is about 290±20 mOsm/kg.

In one embodiment, at least 90% or at least 95% of the native form of the anti-hIL-4Rα antibody is recovered from the liquid pharmaceutical formulation after storage at 5°C for six months as determined by size exclusion chromatography. In a particular embodiment, at least 98% of the native form of the antibody can be recovered from the liquid pharmaceutical formulation after storage at 5°C for six months as determined by size exclusion chromatography.

In one embodiment, at least 90% of the native form of the antibody can be recovered from the liquid pharmaceutical formulation after storage at 45°C for eight weeks as determined by size exclusion chromatography.

In one embodiment, less than 45% of the antibody in the acidic form can be recovered from the liquid pharmaceutical formulation after storage at 45°C for eight weeks as determined by cation exchange chromatography.

In one embodiment, less than about 4% of the antibodies recovered from the liquid pharmaceutical formulation after storage at 25°C for six months are aggregated as determined by size exclusion exchange chromatography.

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

In one embodiment, the liquid pharmaceutical formulation has a viscosity of about 8.5±0.85 cmP to about 11±1.1 cmP. In a particular embodiment, the viscosity of the liquid pharmaceutical formulation is about 8.5±0.85 cmP.

In one embodiment, the liquid pharmaceutical formulation is physiologically isotonic. In one embodiment, the molar osmotic pressure concentration of the liquid pharmaceutical formulation is about 290±20 mOsm/kg.

In one embodiment, at least 98% of the native form of the anti-hIL-4Rα antibody is recovered from the liquid pharmaceutical formulation after storage at 5°C for six months as determined by size exclusion chromatography.

In one embodiment, at least 90% of the native form of the anti-hIL-4Rα antibody is recovered from the liquid pharmaceutical formulation after storage at 45°C for eight weeks as determined by size exclusion chromatography.

In one embodiment, less than 45% of the antibody in the acidic form can be recovered from the liquid pharmaceutical formulation after storage at 45°C for eight weeks as determined by cation exchange chromatography.

In one embodiment, the antibody recoverable from the liquid pharmaceutical formulation after storage at 25°C for six months is less than about 4% aggregation as determined by size exclusion exchange chromatography.

In one embodiment, a stable low-viscosity isotonic liquid pharmaceutical formulation containing at least 100 mg/ml of a stable anti-hIL-4Rα antibody is provided. In one 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 one embodiment, the antibody comprises any one or more amino acid sequences of SEQ ID NO: 1-8. In one embodiment, the antibody comprises a heavy chain variable region (HCVR) and a light chain variable region (LCVR), wherein the HCVR/LCVR combination comprises heavy chain and light chain complementary determining regions (HCDR1-HCDR2-HCDR3/LCDR1-LCDR2-LCDR3) containing amino acid sequences of SEQ ID NO: 2-3-4 and SEQ ID NO: 6-7-8, respectively. In a specific embodiment, an anti-hIL-4Rα antibody comprises a HCVR and a LCVR.

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

In one embodiment, the formulation has a physiologically compatible molar osmotic pressure concentration. In a particular embodiment, the formulation comprises a molar osmotic pressure concentration of 290±20 mOsm/kg.

In one embodiment, the antibody has a stability of at least about six months at about 5°C. In a particular embodiment, at least about 98% of the antibody retains its native conformation when stored at 5°C for about six months as determined by size exclusion chromatography.

In one embodiment, the antibody has a storage stability of at least about eight weeks at about 45°C. In a particular embodiment, at least about 90% of the antibody retains its native conformation when stored at 45°C for about eight weeks as measured by size exclusion chromatography. In a particular embodiment, less than about 45% of the antibody is in the acidic form when stored at 45°C for about eight weeks as measured by cation exchange chromatography.

In one embodiment, the antibody has a storage stability of at least about six months at about 25°C. In a particular embodiment, less than about 4% of the antibody comprises aggregated forms when stored at 25°C for about six months as determined by size exclusion analysis.

In one embodiment, the formulation comprises a buffer having a pH of about 5.9±0.5. In one embodiment, the buffer comprises an acetate buffer and a histidine buffer. In a particular embodiment, the concentration of the acetate is 12.5mM±1.9mM and the concentration of the histidine is 20mM±3mM.

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

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

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

In a specific embodiment, the stable low-viscosity isotonic liquid pharmaceutical formulation has a viscosity of about 8.5±2.5 mmHg and a molar osmotic pressure concentration of about 290±20 mOsm/kg, and comprises: (i) 150 mg/ml±15 mg/ml of an anti-hIL-4Rα antibody, wherein the antibody comprises a HCVR and a LCVR comprising the amino acid sequences of SEQ ID NO: 1 and SEQ ID NO: 5, respectively; (ii) 12.5 mM±1.9 mM acetate; (iii) 20 mg/ml±3 mM histidine; (iv) 0.2%±0.03% w/v polysorbate 20; (v) 5%±0.75% w/v sucrose; and (vi) 25 mM±3.75 mM L-arginine hydrochloride. According to this embodiment, (i) at least about 98% of the antibody retains its native conformation when stored at 5°C for at least about six months as determined by size exclusion chromatography; (ii) at least about 90% of the antibody retains its native conformation when stored at 45°C for at least about eight weeks as determined by size exclusion chromatography; (iii) less than about 45% of the antibody is in the acidic form when stored at 45°C for eight weeks as determined by cation exchange chromatography; and (iv) less than about 4% of the antibody comprises aggregated form when stored at 25°C for about six months as determined by size exclusion chromatography.

In one embodiment, a liquid pharmaceutical formulation of any of the above embodiments is provided in a container. In one 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 particular embodiment, the syringe includes a fluorocarbon-coated piston. In a particular embodiment, the syringe is a low-tungsten syringe.

Other specific embodiments of the present invention can be better understood from the detailed description that follows.

Detailed description of invention

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

Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art. The term "approximately" when used herein in relation to a particular stated value or range of values means that the value is not more than 1% of the stated value. For example, the expression "about 100" herein includes 99 and 101 and all values therebetween (e.g., 99.1, 99.2, 99.3, 99.4, etc.).

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

Pharmaceutical preparations

The statement "pharmaceutical formulation" herein means a combination of at least one active ingredient (e.g., a small molecule, a macromolecule, a compound, etc. that can exhibit a biological effect in a human or non-human animal) and at least one inactive ingredient, which can be therapeutically administered to a human or non-human animal when the active ingredient is mixed with one or more additional inactive ingredients. Unless otherwise expressly stated, the term "formulation" herein means a "pharmaceutical formulation." The present invention provides a pharmaceutical formulation comprising at least one therapeutic polypeptide. According to certain specific embodiments of the present invention, the therapeutic polypeptide is an antibody or an antigen-binding fragment thereof that can specifically bind to human interleukin-4 antibody α (hIL-4Rα). More specifically, the present invention includes a pharmaceutical formulation containing (i) a human antibody that can specifically bind to hIL-4Rα; (ii) an acetate/histidine buffer system; (iii) an organic cosolvent that is a nonionic surfactant; (iv) a carbohydrate thermal stabilizer; and (v) a viscosity reducer. Specific exemplary ingredients and formulations included 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 an antigen-binding fragment thereof that specifically binds to 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 antibody contained in the pharmaceutical formulation of the present invention specifically binds to the extracellular region of hIL-4Rα. An exemplary amino acid sequence of human IL-4 receptor α (hIL-4Rα) is described in SEQ ID NO: 25. Antibodies to hIL-4Rα are described in U.S. Patent Nos. 7,605,237 and 7,608,693. The extracellular region of hIL-4Rα is represented by the amino acid sequence of SEQ ID NO: 26.

The term "antibody" as used herein generally refers to immunoglobulin molecules comprising four polypeptide chains, two heavy chains (H) and two light chains (L) interconnected by disulfide chains, and multimers thereof (such as IgM); however, immunoglobulin molecules consisting only of heavy chains (i.e., without light chains) also fall within the definition of "antibody". Each heavy chain comprises a heavy chain variable region (abbreviated herein as HCVR or _VH ) and a heavy chain constant region. The heavy chain constant region comprises three domains, CH1, CH2, and CH3. Each light chain comprises a light chain variable region (abbreviated herein as LCVR or _VL ) and a light chain constant region. The light chain constant region comprises one domain (CL1). The _VH and _VL can be further divided into interspersed more conserved regions called framework regions (FRs) and hypervariable regions called complementarity determining regions (CDRs). Each _VH and _VL is composed of three CDRs and four FRs arranged from amino-terminus to carboxyl-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.

Unless otherwise specified, the term "antibody" as used herein shall be deemed to include complete antibody molecules and antigen-binding fragments thereof. The "antigen-binding region" or "antigen-binding fragment" (or simply "antibody region" or "antibody fragment") of an antibody as used herein refers to one or more fragments of an antibody that retain the ability to specifically bind to hIL-4Rα or its antigenic epitope.

The "isolated antibody" mentioned here means an antibody that is substantially free of other antibodies with different antigenic specificities (e.g., an isolated antibody that can specifically bind to hIL-4Rα is substantially free of antibodies that specifically bind to antigens other than hIL-4Rα).

The term "specifically binds" or the like means that an antibody or an antigen-binding fragment thereof forms a complex with an antigen that is relatively stable under physiological conditions. Specific binding is characterized by having a dissociation constant of at least about ^1x10-6 M or higher. Methods for determining whether two molecules specifically bind are known in the art and include, for example, dialysis equilibrium, surface plasmon resonance, etc. However, an isolated antibody that specifically binds hIL-4Rα has cross-reactivity with other antigens, such as IL-4R (homologous genes) 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. Furthermore, an isolated antibody may be substantially free of other cellular material or chemicals.

Exemplary anti-hIL-4Rα antibodies for incorporation into 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 certain specific embodiments of the present invention, the anti-hIL-4Rα antibody is a human IgG1 comprising a heavy chain variable region of the IGHV 3-9 subtype and a light chain variable region of the IGKV 2-28 subtype (see Barbie and Lefranc, Human immunoglobulin kappa variable region (IGKV) gene and joining (IGKJ) fragment, Exp. Clin. Immunogenet. 1998, 15: 171-183; and Scaviner, D. et al., Protein display of human immunoglobulin heavy chain, kappa and lambda variable regions and joining regions, Exp. Clin. Immunogenet. 1999, 16: 234-240).

In some embodiments, the anti-hIL-4Rα comprises at least one amino acid substitution that results in a charge change on the exposed surface of the antibody relative to a germline IGHV 3-9 sequence or a germline IGKV 2-28 sequence. The germline IGHV 3-9 and germline IGKV 2-28 sequences shown herein, as well as amino acid position designation numbers and the International Immunogenetics (IMGT) Information System are 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 comprises a complementary determining region (CDR). In some embodiments, the one or more substitutions of the amino acid are selected from the group consisting of (a) a basic amino acid replacing a natural amino acid in CDR2 (e.g., at position 58) of IGHV 3-9; (b) a natural amino acid replacing an amino acid in CDR3 (e.g., at position 107) of IGHV 3-9; and (c) a natural amino acid replacing a basic amino acid in CDR1 (e.g., at position 33) of IGKV 2-28. The special arrangement of the charge distribution of an antibody, especially at the environmental interface (e.g., in CDR), will cause unexpected conditions in the stability of the antibody in solution.

In some embodiments, the anti-hIL-4Rα antibody comprises at least one amino acid substitution that produces a torsional strain change in the framework region of the antibody variable region relative to the germline IGHV 3-9 sequence or the germline IGKV 2-28 sequence. In some embodiments, the one or more amino acid substitutions are selected from the combination of (a) a proline substitution in the framework region 3 (FR3) of IGHV 3-9 (such as at position 96) for a non-proline; and (b) a non-proline amino acid substitution in the framework region 2 (FR2) of IGKV 2-28 (such as at position 46) for a proline. The alteration of the peptide chain, particularly a framework region, that affects the torsional ability of the CDR and the solvent interface will cause an unexpected condition in the stability of the antibody in solution.

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

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

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

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

The non-limiting example antibody in the examples herein is referred to as "mAb1". This antibody is also referred to as H4H098P in US 7,608,693. mAb1 (H4H098P) comprises a HCVR/LCVR amino acid sequence pair having SEQ ID NO: 1/5, and HCDR1-HCDR2-HCDR3/LCDR1-LCDR2-LCDR3 domains representing SEQ ID NO: 2-3-4/SEQ ID NO: 6-7-8.

Another non-limiting example antibody used in the practice of the present invention is referred to as "mAb2". This antibody is also referred to as H4H083P in US 7,608,693. mAb2 (H4H083P) comprises a HCVR/LCVR amino acid sequence pair having SEQ ID NO: 9/13, and HCDR1-HCDR2-HCDR3/LCDR1-LCDR2-LCDR3 domains representing SEQ ID NO: 10-11-12/SEQ ID NO: 14-15-16.

Yet another non-limiting example antibody used in the practice of the present invention is referred to as "mAb3". This antibody is also referred to as H4H095P in US 7,608,693. mAb3 (H4H095P) comprises a HCVR/LCVR amino acid sequence pair having SEQ ID NO: 17/21, and HCDR1-HCDR2-HCDR3/LCDR1-LCDR2-LCDR3 domains representing SEQ ID NO: 18-19-20/SEQ ID NO: 22-23-24.

The amount of antibody or antigen-binding fragment thereof in the pharmaceutical formulation of the present invention depends on the desired specific properties of the formulation and the specific situation and purpose for which the formulation is intended to be used. In certain embodiments, the pharmaceutical formulation is a liquid formulation containing 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, about 130±13 to about 170±17 mg/ml, about 140±14 to about 160±16 mg/ml, or about 150±15 mg/ml of the antibody. For example, the formulations of the present invention contain about 90 mg/ml, about 95 mg/ml, about 100 mg/ml, about 105 mg/ml, about 110 mg/ml, about 115 mg/ml, about 120 mg/ml, about 125 mg/ml, about 130 mg/ml, about 131 mg/ml, about 132 mg/ml, about 133 mg/ml, about 134 mg/ml, about 135 mg/ml, about 140 mg/ml, about 145 mg/ml, about 150 mg/ml, about 155 mg/ml, about 160 mg/ml, about 165 mg/ml, about 170 mg/ml, about 175 mg/ml, about 180 mg/ml, about 185 mg/ml, about 190 mg/ml, about 195 mg/ml, or about 200 mg/ml of an antibody or an antigen-binding fragment thereof that specifically binds to hIL-4Rα.

Excipients and pH

The pharmaceutical formulation of the present invention comprises 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 some specific embodiments, the pharmaceutical formulation of the present invention contains at least one organic cosolvent that can stabilize the type and amount of the hIL-4Rα antibody under vigorous operation, such as oscillation. In some specific embodiments, the term "stable" means that during the vigorous operation, the total amount of antibodies (on a molar basis) can be prevented from forming more than 2% of aggregated antibodies. In some specific embodiments, the vigorous operation is to vibrate the solution containing the antibody and the organic cosolvent for about 120 minutes.

In certain embodiments, the organic cosolvent is a nonionic surfactant, such as an alkyl poly(ethylene oxide). Specific nonionic surfactants that can be incorporated into the formulations of the present invention include, for example, polysorbates 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, poloxamer 407; or polyethylene glycol (PEG). Polysorbate 20 is also known as Tween 20, sorbitan monolaurate, and polyoxyethylene sorbitan monolaurate. Poloxamer 181 is also known as PLURONIC F68.

The content of the organic cosolvent in the pharmaceutical formulation of the present invention depends on the desired specific properties of the formulation and the specific circumstances and uses in which the formulation is intended to be used. In certain specific embodiments, the surfactant content in the formulation is about 0.1±0.01% to about 2±0.2%. For example, the formulations of the 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% polysorbate 20 or poloxamer 181. For example, the formulations 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 include 0.2%±0.02% polysorbate 20, 0.2%±0.02% polyxamer 181, or 1%±0.1% PEG 3350.

The pharmaceutical formulation of the present invention also contains one or more thermal stabilizers of type and amount capable of stabilizing the hIL-4Rα antibody under heat stress. In some embodiments, the term "stable" means that when the solution containing the antibody and the thermal stabilizer is stored at about 45°C for up to about 28 days, the native conformation of the antibody can be maintained at greater than about 92%. In some embodiments, the term "stable" means that when the solution containing the antibody and the thermal stabilizer is stored at about 45°C for up to about 28 days, less than about 5% of the aggregated antibody can be maintained.

In some embodiments, the thermal stabilizer is a sugar or sugar alcohol selected from sucrose, trehalose and mannose or any combination thereof, and the amount in the formulation depends on the specific situation and the purpose for which the formulation is intended to be used. In some 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% of 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% of sugar or sugar alcohol (such as sucrose, trehalose or mannose).

The pharmaceutical formulation 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 embodiments, the term "stable" means that when the solution containing the antibody and the buffer is stored at about 45° C. for up to about 14 days, less than 3.0% ± 0.5% of aggregated antibody can be maintained. In some embodiments, the term "stable" means that when the solution containing the antibody and the buffer is stored at about 25° C. for up to about 6 months, less than 3.7% ± 0.5% of aggregated antibody can be maintained. In some embodiments, the term "stable" means that at least 95% ± 0.5% of the native conformation of the antibody can be maintained as determined by size exclusion analysis when the solution containing the antibody and the buffer is stored at about 45°C for up to about 14 days. In some embodiments, the term "stable" means that at least 96% ± 0.5% of the native conformation of the antibody can be maintained as determined by size exclusion analysis when the solution containing the antibody and the buffer is stored at about 25°C for up to about 6 months. In some embodiments, the term "stable" means that when the solution containing the antibody and the buffer is stored at about 45° C. for up to about 14 days, at least 62%±0.5% of the neutral conformation of the antibody can be maintained as determined by cation exchange chromatography. In some embodiments, the term "stable" means that when the solution containing the antibody and the buffer is stored at about 25° C. for up to about 6 months, at least 54%±0.5% of the neutral conformation of the antibody can be maintained as determined by cation exchange chromatography. "Neutral conformation" refers to the portion of the antibody that is dialyzed from the main peak in the ion exchange resin, which usually has a more "basic" peak on one side and a more "acidic" peak on the other side.

The pharmaceutical formulation of the present invention has a pH from about 5.2 to about 6.4. For example, the pharmaceutical formulation of the present invention may have a pH 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. 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 comprises at least one buffer that is completely overlapping or in a partial range of pH 5.2-6.4. In one embodiment, the buffer or buffer system comprises two buffers, the first of which is in an effective pH range of 3.6-5.6 and the second of which is in an effective pH range of 5.5-7.4. In one 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 certain embodiments, the buffer system comprises an acetate buffer and a histidine buffer. In some embodiments, each molar ratio of acetate contains about 1.3 to 1.9 parts of histidine. In some embodiments, each molar ratio of acetate contains about 1.6 ± 0.25 parts of histidine. In some 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 20.5 mM, about 5.0 mM to about 20 mM, about 5.5 mM to about 19.5 mM, about 6.0 mM to about 19.0 mM, about 6.5 mM to about 18.5 mM, about 7.0 mM to about 18.0 mM, about 7.5 mM to about 17.5 mM, 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 16 mM to about 24 mM, about 17 mM to about 23 mM, about 18 mM to about 22 mM, or about 19 mM to about 21 mM. In certain embodiments, the buffer system comprises about 12.5 mM acetate at about pH 5.9 and about 20 mM histidine.

The pharmaceutical formulations of the present invention also include one or more excipients for maintaining low viscosity or reducing the viscosity of formulations containing high concentrations of protein (e.g., typically >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 35 cPoise, less than about 30 cPoise, less than about 25 cPoise, less than about 20 cPoise, less than about 15 cPoise, less than about 14 cPoise, less than about 13 cPoise, less than about 12 cPoise, less than about 10 cPoise, or less than about 9 cPoise.

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

Representative formulations

According to one aspect of the present invention, the pharmaceutical formulation is a low-viscosity, usually physiologically isotonic liquid formulation comprising: (i) a human antibody that specifically binds to hIL-4Rα (e.g., mAb1, mAb2, or mAb3 [as described above]) at a concentration of about 100 mg/ml or more; (ii) a buffer system sufficient to buffer at about pH 5.9±0.6; (iii) a sugar specifically used as a thermal stabilizer; (iv) an organic cosolvent that protects the structural integrity of the antibody in vivo; and (v) an amino acid that maintains a viscosity that is easy to use for subcutaneous injection.

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

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

Other non-limiting examples of pharmaceutical formulations of the present invention are described elsewhere herein, including the embodiments 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" herein with respect to the pharmaceutical formulation means that the antibody in the pharmaceutical formulation maintains an acceptable degree of chemical structure or biological function under defined storage conditions. An antibody contained in a formulation is stable even if its chemical structure or biological function cannot be maintained 100% after storage for a period of time. In some cases, the antibody structure or function can be considered "stable" when it is maintained at about 90%, about 95%, about 96%, about 97%, about 98% or about 99% after storage for a period of time.

The stability can be determined by a method that specifically measures the percentage of natural antibodies retained in the formulation after storage at a defined temperature for a period of time. The percentage of natural antibodies can be determined in particular by size exclusion chromatography (e.g., size exclusion high performance liquid chromatography [SE-HPLC]). The phrase "an acceptable degree of stability" as described herein means that at least 90% of natural antibodies can be detected in the formulation after storage at a given temperature for a period of time. In certain specific embodiments, at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% of natural antibodies can be detected in the formulation after storage at a defined temperature for a period of time. The defined period of time for determining stability 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 determining stability of the storable pharmaceutical formulation 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 to 8°C, about 5°C, about 25°C, or about 45°C. For example, a pharmaceutical formulation can be considered stable if it can detect greater than about 90%, 95%, 96%, 97% or 98% of natural antibodies by SE-HPLC after storage at 5°C for 3 months. A pharmaceutical formulation may also be considered stable if it can detect greater than about 90%, 95%, 96%, 97% or 98% of natural antibodies by SE-HPLC after 6 months of storage at 5°C. A pharmaceutical formulation may also be considered stable if it can detect greater than about 90%, 95%, 96%, 97% or 98% of natural antibodies by SE-HPLC after 9 months of storage at 5°C. A pharmaceutical formulation may also be considered stable if it can detect greater than about 90%, 95%, 96% or 97% of natural antibodies by SE-HPLC after 3 months of storage at 25°C. A pharmaceutical formulation may also be considered stable if it can detect greater than about 90%, 95%, 96% or 97% of natural antibodies by SE-HPLC after 6 months of storage at 25°C. If a pharmaceutical formulation can detect greater than about 90%, 95%, 96% or 97% of natural antibodies by SE-HPLC after storage at 25°C for 9 months, it can also be considered stable.

The stability can be determined by, in particular, measuring the percentage of aggregated antibodies in the formulation after storage at a defined temperature for a period of time, wherein the stability is inversely proportional to the percentage of aggregated antibodies formed. The percentage of aggregated antibodies can be determined in particular by size exclusion chromatography (e.g., size exclusion high performance liquid chromatography [SE-HPLC]). As used herein, the phrase "an acceptable level of stability" means that up to 5% aggregated antibodies can be detected in the formulation after storage at a given temperature for a period of time. In certain embodiments, an acceptable level of stability means that up to about 5%, 4%, 3%, 2%, 1%, 0.5% or 0.1% aggregated antibodies can be detected in the formulation after storage at a given temperature for a period of time. The defined period of time for determining stability 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 at which the storable pharmaceutical formulation 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 to 8°C, about 5°C, about 25°C, or about 45°C. For example, a pharmaceutical formulation can be considered stable if less than about 5%, 4%, 3%, 2%, 1%, 0.5% or 0.1% of aggregated antibodies are determined after storage at 5°C for 3 months. A pharmaceutical formulation can also be considered stable if less than about 5%, 4%, 3%, 2%, 1%, 0.5% or 0.1% of the aggregated form of the antibody is detected after 6 months of storage at 5°C. A pharmaceutical formulation can also be considered stable if less than about 5%, 4%, 3%, 2%, 1%, 0.5% or 0.1% of the aggregated form of the antibody is detected after 9 months of storage at 5°C. A pharmaceutical formulation can also be considered stable if less than about 5%, 4%, 3%, 2%, 1%, 0.5% or 0.1% of the aggregated form of the antibody is detected after 3 months of storage at 25°C. A pharmaceutical formulation can also be considered stable if less than about 5%, 4%, 3%, 2%, 1%, 0.5% or 0.1% of the aggregated form of the antibody is detected after 6 months of storage at 25°C. A pharmaceutical formulation can also be considered stable if less than about 5%, 4%, 3%, 2%, 1%, 0.5% or 0.1% of aggregated antibodies are detected after storage at 25°C for 9 months.

The stability can be determined in particular by measuring the ratio of the antibody's migration to the more acidic part ("acidic form") to the main part of the antibody ("neutral conformation") during ion exchange, wherein the stability is inversely proportional to the acidic form of the antibody. Without being bound by theory, deamidation of an antibody can result in an antibody that has a more negative charge and is therefore more acidic than a non-deamidated antibody (see, e.g., Robinson, N., Protein deamidation, PNAS , April 16, 2002, 99(8): 5283-5288). "Acidified" or "deamidated" antibodies can be determined in particular by ion exchange chromatography (e.g., cation exchange high performance liquid chromatography [CEX-HPLC]). As used herein, the phrase "an acceptable level of stability" 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 level of stability means that up to about 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 5%, 4%, 3%, 2%, 1%, 0.5% or 0.1% of the acidic form of the antibody can be detected in the formulation after storage at a given temperature for a period of time. The defined period of time for which the test is 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 which the storable pharmaceutical formulation 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 to 8°C, about 5°C, about 25°C, or about 45°C. For example, a pharmaceutical formulation can be considered stable if less than about 15%, 14%, 13%, 12%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of the acidic form of the antibody is detected after 3 months of storage at 5°C. A pharmaceutical formulation can also be considered stable if 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 the acidic form of the antibody is detected after 3 months of storage at 25°C. A pharmaceutical formulation can also be considered stable if less than about 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of the acidic form of the antibody is detected after storage at 45°C for 8 weeks. A pharmaceutical formulation can also be considered stable if 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 the antibody is detected after storage at 40°C for 2 weeks.

Other methods can be used to determine the stability of the formulations of the present invention, such as differential scanning calorimetry (DSC) for thermal stability, controlled agitation for mechanical stability, and absorbance at about 350 or about 405 nm for solution turbidity. For example, a pharmaceutical formulation of the present invention is considered stable if the change in OD ₄₀₅ from the zero time point is less than about _0.05 (e.g., 0.04, 0.03, 0.02, 0.01, or lower) after storage at about 5° C. to about 25° C. for 6 or more months.

The stability of the antibody can also be assessed by measuring its biological activity or affinity for binding to its target. For example, after a formulation of the present invention is stored at, for example, 5°C, 25°C, 45°C, etc. for a period of time (e.g., 1 to 12 months), the anti-IL-4Rα antibody contained in the formulation can be considered stable when the anti-IL-4Rα antibody bound to IL-4Rα with at least 90%, 95% or more of the antibody binding affinity before storage. The binding affinity is measured by, for example, ELISA or plasma resonance. The biological activity is measured by an IL-4Rα activity assay, for example, by 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 compared to the activity of the IL-4Rα system without the antibody. In some embodiments, the IL-4Rα may be endogenous in the cell. In other embodiments, the IL-4Rα may be expressed heterotopically in the cell.

Other methods for determining the stability of antibodies in formulations are described in the examples below.

In certain specific embodiments, the pharmaceutical formulations of the present invention exhibit low to moderate viscosity. The "viscosity" described herein may be "dynamic viscosity" or "absolute viscosity.""Dynamicviscosity" 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 takes longer to flow through the capillary than a less viscous liquid. For example, if one liquid takes 200 seconds and another liquid takes 400 seconds to complete its flow, the second liquid is twice as viscous as the first 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 dimensions of dynamic viscosity are L ² /T, where L is length and T is time. Usually, 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 milliPascal-second (mPa-s), where 1 cP = 1 mPa-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, a liquid formulation of the present invention will exhibit an absolute viscosity of about 15 cP, about 14 cP, about 13 cP, about 12 cP, about 11 cP, about 10 cP, about 9 cP, about 8 cP, or less when measured by standard viscometry. As described herein, a medium viscosity liquid formulation of the present invention will exhibit an absolute viscosity between about 35 cP and about 15 cP. For example, if a liquid formulation of the present invention is considered to have a "medium viscosity", the formulation will exhibit an absolute viscosity of 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.

As described in the examples below, the inventors unexpectedly discovered that a low to moderate viscosity liquid formulation containing a high concentration of anti-hIL-4Rα antibody (e.g., from about 100 mg/ml up to at least 200 mg/ml) can be obtained by formulating the antibody with arginine from about 25 mM to about 100 mM. In addition, it has been further discovered that the viscosity of the formulation can be reduced to an even greater extent by adjusting the sucrose content to less than about 10%.

Container for the pharmaceutical preparation and method of administration

The pharmaceutical formulation of the present invention may be placed in any container suitable for storing drugs and other therapeutic compositions. For example, the pharmaceutical formulation may be placed in a sealed and sterilized plastic or glass container of limited capacity such as a vial, ampoule, syringe, cassette or vial. The formulation of the present invention may be placed in various types of vials such as transparent and opaque (such as amber) glass or plastic bottles. Similarly, any type of syringe may be used to contain or administer the pharmaceutical formulation of the present invention.

The pharmaceutical formulations of the present invention may be placed into a "normal tungsten" syringe or a "low tungsten" syringe. As is understood by those of ordinary skill in the art, the process of making glass syringes typically involves the use of a hot tungsten rod that penetrates the glass to create a small hole through which liquid can be drawn from the syringe. This process causes trace amounts of tungsten to be deposited on the inner surface of the syringe. Washing and other process steps may then be used to reduce the amount of tungsten in the syringe. As used herein, the term "normal tungsten" means that the syringe contains greater 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 of tungsten.

Rubber plungers for syringes and rubber stoppers for closing the openings of glass bottles can be coated to avoid contamination of the drug in the syringe or bottle, or to preserve its stability. Therefore, the pharmaceutical preparation 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 can be coated with a fluorocarbon film. Examples of coated plungers or rubber stoppers suitable for glass bottles and syringes containing the pharmaceutical preparation 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 herein by reference. Special coated rubber plungers and stoppers for use in the present invention are available from West Pharmaceutical Services, Inc. (Lionville, Pennsylvania) under the commercial trademark "FluroTec ^® ".

According to certain specific 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 a patient by, for example, injection (e.g., subcutaneous, intravenous, intramuscular, intraperitoneal, etc.) or by parenteral routes of 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 formulation of the present invention. Examples include, but are not limited to, AUTOPEN ^™ (Owen Mumford, Woodstock, England), DISETRONIC ^™ pen (Disetronic Medical, Bergdorf, Switzerland), HUMALOG MIX 75/25 ^™ pen, HUMALOG ^™ pen, HUMALIN 70/30 ^™ pen (Eli Lilly, Indianapolis, Indiana), NOVOPEN ^™ I, II and III (Novo Nordisk, Copenhagen, Denmark), NOVOPEN JUNIOR ^™ (Novo Nordisk, Copenhagen, Denmark), BD ^™ pen (Becton Dickinson, Franklin, New Jersey), OPTIPEN ^™ , OPTIPEN PRO ^™ , OPTIPEN STARLET ^™ , and OPTICLIK ^™ (Sanofi-Aventis, Frankfurt, Germany). Examples of disposable injection pens or autoinjection delivery devices that have been used for subcutaneous infusion of the pharmaceutical compositions of the present invention include, but are not limited to, the SOLOSTAR ^™ pen (Sanofi-Aventis), FLEXPEN ^™ (Novo Nordisk), and KWIKPEN ^™ (Eli Lilly); the SURECLICK ^™ autoinjector (Amgen, Thousand Oaks, CA), PENLET ^™ (Haselmeier, Stuttgart, Germany), EPIPEN ^™ (Dey, LP), and the HUMIRA ^™ pen (Abbott Laboratories, Abbott Park, IL).

Microsyringes are also used herein to infuse the pharmaceutical formulations of the present invention. As used herein, "microsyringe" means a subcutaneous infusion device that slowly administers large amounts (e.g., up to about 2.5 ml or more) of therapeutic formulations over a long period of time (e.g., about 10, 15, 20, 25, 30 minutes or more). See, e.g., US 6,629,949, US 6,659,982; and Meehan et al., J. Controlled Release 46: 107-116 (1996). Microsyringes are best suited for infusing large doses of therapeutic proteins containing high concentrations (e.g., about 100, 125, 150, 175, 200 mg/ml, or more), or viscous solutions.

In one embodiment, the liquid pharmaceutical formulation containing about 150±15 mg/ml anti-IL-4Rα antibody is administered subcutaneously from a prefilled syringe in an autoinjector in a volume of about 1±0.15 ml. In another embodiment, the formulation is administered from a microsyringe in a volume between about 1 ml and 2.5 ml. The formulation may be prefilled into a storage bag or cartridge of the microsyringe.

Therapeutic uses of pharmaceutical preparations

The pharmaceutical formulations of the present invention are particularly useful for treating, preventing or alleviating any disease or disorder associated with IL-4 activity, including diseases or disorders mediated by IL-4Rα activation. Non-limiting examples of 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-mediated diseases.

Therefore, the present invention includes methods for treating, preventing or alleviating any disease or disorder associated with IL-4 activity or IL-4Rα activation (including any of the above-mentioned exemplary diseases, disorders and conditions). The treatment method of the present invention comprises administering a formulation containing an anti-hIL-4Rα antibody as described above to an organism. The organism to which the pharmaceutical formulation is administered may be, for example, any human or non-human animal that requires such treatment, prevention or alleviation, or that may benefit from inhibiting or alleviating IL-4 or IL-4Rα-mediated activity. For example, the organism may be an individual diagnosed with or believed to be at risk of developing the above-mentioned disease or disorder. The present invention further includes the use of any of the pharmaceutical formulations disclosed herein for the manufacture of a drug for the treatment, prevention or alleviation of any disease or disorder associated with IL-4 activity or IL-4Rα activation (including any of the exemplary diseases, disorders and conditions described above).

The following examples are intended to provide a complete disclosure and description of how one skilled in the art can make and use the methods and compositions of the present invention and are not intended to limit the scope of what the inventors regard as their invention. Although efforts have been made to ensure accuracy with respect to the numbers used (e.g., amounts, temperatures, etc.), certain errors and deviations may occur in some experiments. Unless otherwise indicated, parts are molar parts, molecular weights are average molecular weights, temperatures are degrees Celsius, and pressures are at or near atmospheric pressure.

The activities to form the preliminary formulation involved screening organic cosolvents, thermal stabilizers and buffers in the liquid formulation of mAb1 (anti-IL-4Rα antibody of the present invention) to confirm that the formulation is compatible with the protein and contributes to its stability while maintaining molar osmotic pressure concentration and viscosity for subcutaneous injection. Buffer conditions were also examined to determine the optimal pH for maximum protein stability.

Example 1. Organic co-solvent

mAb1 was found to be unstable under oscillatory stress. Analysis by reverse phase high performance liquid chromatography (RP-HPLC) and size exclusion high performance liquid chromatography (SE-HPLC) demonstrated protein loss and increase in aggregated protein when mAb1 was shaken at room temperature (Table 1, see "No cosolvent" data). Addition of organic cosolvents to mAb1 solutions prevented protein degradation as measured by SE-HPLC and RP-HPLC (Table 1). However, addition of some organic cosolvents was found to reduce the thermal stability of mAb1 (Table 2). Formulations containing PEG 3350 (3%) and PEG 300 (10% and 20%) were found to lose recovered protein as measured by RP-HPLC after heat stress (Table 2). In addition, formulations containing PLURONIC F68 (poloxamer 181) (0.2%), PEG 300 (10% and 20%), and propylene glycol (20%) formed more aggregates as measured by SE-HPLC than the formulation without solvent. Polysorbate 20 (0.2%) and polysorbate 80 (0.2%) were fairly stable to shaking and heat stress.

According to Table 1, 0.3 ml of mAb1 at 15 mg/ml in 10 mM phosphate pH 6.0 and various organic cosolvents were shaken in 2 ml glass vials for about 120 minutes. The turbidity was measured by optical density (OD) at 405 nm and the relative change of OD at 405 nm was compared with the starting material. The percentage of native and aggregated mAb1 was determined by size exclusion HPLC (SE-HPLC). The SE-HPLC results shown in the "Starting Material" table are the average 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 cosolvents in 2 ml glass vials were stored at about 45°C for about 28 days. The turbidity was measured by optical density (OD) at 405 nm and the relative change of OD at 405 nm was compared with the starting material. The percentage of total recovered mAb1 was determined by reverse phase HPLC (RP-HPLC). The percentage of native and aggregated mAb1 was determined by size exclusion HPLC (SE-HPLC). The SE-HPLC results shown in the "Starting Material" table are the average of each formulation that was not heat-stressed.

Example 2. Thermal Stabilizer

Various thermal stabilizers, such as sugars, amino acids, and inorganic salts were examined for their ability to inhibit mAb1 degradation when stored at approximately 45°C. A summary of the thermal stabilizers studied is shown in Table 3. Formulations containing sucrose or trehalose had a greater mAb1 stabilizing effect when incubated in high temperature solutions (as measured by SE-HPLC). Sucrose was chosen as a stabilizer because of its long history of safety in monoclonal antibody formulations.

According to Table 3, 0.3 ml of 25 mg/ml mAb1 in 10 mM acetate pH 5.3 and various thermal stabilizers in 2 ml glass bottles were stored at about 45°C for about 28 days. The turbidity was measured by optical density (OD) at 405 nm and the relative change of OD at 405 nm of the starting material was compared. The turbidity difference was negligible for all samples. The percentage of total recovered mAb1 was determined by reverse phase HPLC (RP-HPLC). The percentage of native and aggregated mAb1 was determined by size exclusion HPLC (SE-HPLC). Acidic or basic substances were defined as the total amount of mAb1 peak dialyzed from the cation exchange (CEX-HPLC) column at an earlier or later retention time than the main peak, respectively. The SE-HPLC results shown in the "Starting Materials" table are the average values of the preparations that were not subjected to heat stress.

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. Protein stability was monitored by SE-HPLC and cation exchange HPLC (CEX-HPLC). The maximum protein stability measured by SE-HPLC and CEX-HPLC was observed when mAb1 was formulated in pH 6.0 histidine buffer or pH 5.3 acetate buffer (Tables 4 and 5). The acetate buffer system had a wider pH stability range and a lower rate of variable charge formation than the formulation containing histidine buffer (Table 5). Therefore, acetate buffer at pH 5.3 was selected for the formulation of mAb1 drug.

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

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

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。 Formulation tests showed that under alkaline conditions (pH

6.5) in solution. Conversely, below pH 5.0 it has been found that the rate of formation of mAb1 with variant molecular weights is increased. Based on these data, the pH of the mAb1 formulation is maintained between about pH 5.6 and pH 6.2. mAb1 has been found to be stable in this pH range.

The effects of pH and buffer type on the stability of mAb1 were further evaluated in formulations containing 20 mM histidine pH 6, 12.5 mM acetate pH 5.3, or a mixture of 20 mM histidine and 12.5 acetate pH 5.9 (Table 6). When comparing the individual buffer systems, mAb1 formulated with histidine and acetate at approximately pH 5.9 was the most stable. mAb1 had the slowest aggregation rate when formulated in this mixed buffer system (SE-HPLC) (Table 6).

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

Example 4. Control of viscosity and tension

The viscosity and tonicity (expressed as molar osmotic pressure concentration) of various formulations mixed with high concentrations of mAb1 (i.e., 150 mg/ml, 175 mg/ml, and 200 mg/ml) were evaluated. The levels of sucrose, sodium chloride, and L-arginine hydrochloride were adjusted to produce low viscosity and physiologically isotonic formulations containing high concentrations of mAb1 that facilitate comfortable and rapid subcutaneous infusion of high volumes of mAb1 (Table 7). The liquid formulation (Formulation A) containing 25mM arginine, 20mM histidine, 12.5mM acetate, 5% (w/v) sucrose, 0.2% (w/v) polysorbate 20 and 150mg/ml mAb1 at pH 5.9 was 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 major degradation pathways during the formation of mAb1 liquid formulations are the production of aggregates, fragmentation products, and charge variants. The formation of these degradation products can be reduced by formulating mAb1 in a formulation containing 20 mM histidine, 12.5 mM acetate, 0.2% polysorbate 20, 5% sucrose, and 25 mM L-arginine hydrochloride at pH 5.9. These 150 mg/ml mAb1 formulations were found to be clear to slightly opalescent liquid solutions with essentially no visible particles.

The formulated mAb1 was physically and chemically stable under various stress (25°C and 45°C incubation) and immediate storage conditions (5°C) (Table 8). The appearance of the mAb1 was not affected when it was stored at 25°C (3 months) or 5°C for 6 months. In addition, no effect was found on the solution pH, turbidity or the recovery of mAb1. After storage of the formulated mAb1 at 25°C for 3 months, there was no significant degradation of the antibody as determined by SE-HPLC and only 3.3% higher degradation as determined by CEX-HPLC. Storage at 45°C for 8 weeks was found to increase the amount of degradation as determined by SE-HPLC and CEX-HPLC indicating that 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 species were defined as the total amount of mAb1 peak dialyzed from the cation exchange (CEX-HPLC) column at an earlier or later retention time than the main peak, respectively.

Example 6. Container

The mAb1 formulation has been tested and confirmed to be stable after sterilization. The clinical supply is manufactured using a Millipak filter device, while the laboratory uses a filter of the same composition (Millex DURAPORE).

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

The stability of the formulated mAb1 (Formulation A) was not affected when stored in polypropylene glycol tubes, polystyrene tubes, polycarbonate tubes, or in glass vials containing stainless steel blocks compared to storage in glass vials (Table 9).

According to Table 9, 150 mg/ml mAb1 at pH 5.9, 5% sucrose, 25 mM arginine hydrochloride, 0.2% PS-20, 20 mM histidine, 12.5 mM acetate were stored in various materials at 40°C for 14 days. OD = optical density; RP-HPLC = reverse phase HPLC; SE-HPLC = size exclusion HPLC; and CEX-HPLC = cation exchange HPLC. Turbidity was recorded as the difference in OD at 405 nm compared to the starting material. Acidic or basic material was defined as the total amount of mAb1 peak dialyzed from the CEX-HPLC column at an earlier or later retention time than the main peak, respectively.

Claims (25)

A stable liquid pharmaceutical formulation for use in the preparation of a medicament for treating a disease or disorder associated with IL-4 activity or mediated by IL-4Rα activation, wherein the formulation comprises: (i) a human antibody at a concentration of 15 mg/ml to 200 mg/ml, wherein the antibody specifically binds to human interleukin-4 receptor α (hIL-4Rα) and comprises (a) a heavy chain variable region (HCVR) comprising heavy chain complementation determining regions 1, 2 and 3 (HCDR1-HCDR2-HCDR3) comprising the amino acid sequences of SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, respectively; and (b) a light chain variable region (LCVR) comprising the amino acid sequences of SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, respectively. NO: 8 amino acid sequence light chain complement determining regions 1, 2 and 3 (LCDR1-LCDR2-LCDR3); (ii) a buffer solution containing acetate at a concentration of 5mM to 20mM; (iii) a thermal stabilizer containing sucrose, trehalose or mannose at a concentration of 2.5% w/v to 10% w/v; (iv) an organic cosolvent containing polysorbate at a concentration of 0.1% w/v to 2% w/v; and (v) a viscosity reducer containing arginine at a concentration of 25mM to 100mM, wherein the formulation has a pH of 5.6 to 6.2. For use in claim 1, wherein: (a) the concentration of the antibody is 15 mg/ml to 100 mg/ml; or (b) the concentration of the antibody is 100 mg/ml; or (c) the concentration of the antibody is 150 mg/ml±15 mg/ml; or (d) the concentration of the antibody is 150 mg/ml; or (e) the concentration of the antibody is 175 mg/ml; or (f) the concentration of the antibody is 200 mg/ml. For use as in claim 1, the buffer solution contains acetate at a concentration of 10mM to 15mM. For use as in item 3 of the patent application, the buffer further comprises histidine at a concentration of 10mM to 30mM. For use as in claim 4, wherein the buffer solution contains histidine at a concentration of 15mM to 25mM. For use as in item 1 of the patent application, the buffer solution contains acetate at a concentration of 12.5mM±1.85mM and histidine at a concentration of 20mM±0.3mM. For the use in item 1 of the patent application, the polysorbate is polysorbate 20 or polysorbate 80. For use in item 7 of the patent application, the organic cosolvent is polysorbate 20 with a concentration of 0.2%±0.03% w/v. For use in item 7 of the patent application, the polysorbate is polysorbate 80 with a concentration of 0.2%±0.03% w/v. For the use in item 1 of the patent application, the thermal stabilizer is sucrose with a concentration of 5%±0.75% w/v. For use in item 1 of the patent application, the viscosity reducer is arginine at a concentration of 25mM±3.75mM. For use in item 1 of the patent application, the viscosity reducer is arginine at a concentration of 50mM±7.5mM. A stable liquid pharmaceutical formulation for use in preparing a drug for treating a disease or disorder associated with IL-4 activity or mediated by IL-4Rα activation, wherein the formulation comprises: (i) 150 mg/ml±50 mg/ml of a human antibody that specifically binds to hIL-4Rα, wherein the antibody comprises a heavy chain variable region (HCVR) amino acid sequence comprising the amino acid sequence of SEQ ID NO: 1 and a 5-mercaptoethanol residue comprising the amino acid sequence of SEQ ID NO: 2. NO:5 amino acid sequence of light chain variable region (LCVR); (ii) 12.5mM±2mM acetate; (iii) 20mM±3mM histidine; (iv) 5%±0.75% sucrose; (v) 0.2%±0.03% polysorbate 20; and (vi) 25mM±3.75mM arginine, at pH 5.9±0.5. A stable liquid pharmaceutical formulation for use in preparing a drug for treating a disease or disorder associated with IL-4 activity or mediated by IL-4Rα activation, wherein the formulation comprises: (i) 150 mg/ml±50 mg/ml of a human antibody that specifically binds to hIL-4Rα, wherein the antibody comprises a heavy chain variable region (HCVR) amino acid sequence comprising SEQ ID NO: 1 and a heavy chain variable region (HCVR) amino acid sequence comprising SEQ ID NO: NO: 5 light chain variable region (LCVR) amino acid sequence; (ii) 12.5mM±2mM acetate; (iii) 20mM±3mM histidine; (iv) 5%±0.75% sucrose; (v) 0.2%±0.03% polysorbate 80; and (vi) 25mM±3.75mM arginine, at pH 5.9±0.5. A stable liquid pharmaceutical formulation for use in preparing a drug for treating a disease or disorder associated with IL-4 activity or mediated by IL-4Rα activation, wherein the formulation comprises: (i) 175 mg/ml of a human antibody that specifically binds to hIL-4Rα, wherein the antibody comprises a heavy chain variable region (HCVR) amino acid sequence containing SEQ ID NO: 1 and a light chain variable region (LCVR) amino acid sequence containing SEQ ID NO: 5; (ii) 12.5 mM ± 2 mM acetate; (iii) 20 mM ± 3 mM histidine; (iv) 5% ± 0.75% sucrose; (v) 0.2% ± 0.03% polysorbate 20; and (vi) 50 mM ± 3.75 mM arginine, at a pH of 5.9 ± 0.5. A stable liquid pharmaceutical formulation for use in preparing a drug for treating a disease or disorder associated with IL-4 activity or mediated by IL-4Rα activation, wherein the formulation comprises: (i) 175 mg/ml of a human antibody that specifically binds to hIL-4Rα, wherein the antibody comprises a heavy chain variable region (HCVR) amino acid sequence containing SEQ ID NO: 1 and a light chain variable region (LCVR) amino acid sequence containing SEQ ID NO: 5; (ii) 12.5 mM ± 2 mM acetate; (iii) 20 mM ± 3 mM histidine; (iv) 5% ± 0.75% sucrose; (v) 0.2% ± 0.03% polysorbate 80; and (vi) 50 mM ± 3.75 mM arginine, at a pH of 5.9 ± 0.5. For use as in item 1 of the patent application, the antibody comprises a HCVR containing the amino acid sequence of SEQ ID NO: 1 and a LCVR containing the amino acid sequence of SEQ ID NO: 5. For use in any of items 1 to 17 of the patent application, the disease or disorder associated with IL-4 activity or mediated by IL-4Rα activation is an IgE/Th2-mediated disease. For use in any of items 1 to 17 of the patent application, the disease or disorder associated with IL-4 activity or mediated by IL-4Rα activation is atopic dermatitis. For use in any one of items 1 to 17 of the patent application, the disease or disorder associated with IL-4 activity or mediated by IL-4Rα activation is asthma. For the use of any of items 1 to 17 in the patent application scope, the drug is contained in a glass bottle. For use in any of items 1 to 17 of the patent application, the drug is contained in a syringe. For use in any of items 1 to 17 of the patent application, the drug is contained in a micro-syringe. For use in any of items 1 to 17 of the patent application, the drug is contained in an automatic injection delivery device. For use in any of items 1 to 17 of the patent application, the drug is contained in a pen-type delivery device.