EA042167B1 - STABILIZED COMPOSITIONS CONTAINING ANTIBODIES TO INTERLEUKIN-4 RECEPTOR (IL-4R) - Google Patents

Description

Technical field

The invention relates to the field of compositions of therapeutic antibodies. In particular, the invention relates to the field of pharmaceutical compositions containing human antibodies that specifically bind to the human interleukin-4 receptor.

Sequence list

The corresponding ST.25 sequence listing text file is filed concurrently with the present application. The contents of the text file are incorporated into the present description by reference. A paper copy of the sequence listing, which is identical in content to the corresponding ST.25 text file, is included as part of this application.

Prior Art

Therapeutic macromolecules (eg, antibodies) must be formulated in such a way as to not only make the molecules suitable for administration to a patient, but also maintain their stability during storage and subsequent use. For example, therapeutic antibodies in aqueous solution are susceptible to degradation, aggregation, or unwanted chemical modifications if the solution is not formulated correctly. The stability of antibodies in a liquid composition depends not only on the type of excipients used in the composition, but also on the amount and proportions of excipients relative to each other. Moreover, when preparing a liquid composition, it is necessary to take into account other characteristics of antibodies, in addition to stability. Examples of such additional characteristics include the viscosity of the solution and the concentration of antibodies that a given composition may contain, and the visual quality or appearance of the composition. Therefore, when formulating therapeutic antibodies, great care must be taken to obtain a composition that remains stable, contains an adequate concentration of antibodies, and has a suitable viscosity, as well as other characteristics that allow convenient administration of the composition to patients.

Antibodies to the human interleukin-4 receptor alpha (hIL-4Ra) are one example of a therapeutically important macromolecule that requires appropriate formulation. Anti-hIL-4Rα antibodies are clinically useful in the treatment or prevention of diseases such as atopic dermatitis and allergic asthma and other conditions. Examples of anti-hIL-4Rα antibodies are described in particular in US Pat. Nos. 7,605,237; 7608693; 7465450 and 7186809; and US Patent Applications No. 2010-0047254 and 2010-002147 6.

Although anti-hIL-4Rα antibodies are known, there remains a need in the art for new pharmaceutical compositions containing anti-hIL-4Rα antibodies that are sufficiently stable and suitable for administration to patients.

The essence of the invention

The invention satisfies the above need by providing pharmaceutical compositions containing human antibodies that specifically bind to the human interleukin-4 alpha receptor (hIL-4Ra).

In one aspect, a liquid pharmaceutical composition is provided comprising: (i) human antibodies that specifically bind to the human interleukin-4 receptor alpha (hIL-4Ra); (ii) buffer; (iii) an organic co-solvent; (iv) a thermal stabilizer; and (v) a viscosity reducer.

In one embodiment, the antibodies are provided at a concentration of about 150 mg/mL±50 mg/mL. In another embodiment, the antibodies are provided at a concentration of about 150 mg/ml±15 mg/ml. In a specific embodiment, the antibodies are provided at a concentration of about 150 mg/ml.

In one embodiment, the antibodies comprise any one or more amino acid sequences of SEQ ID NOs: 1-8. In one embodiment, the antibodies comprise (a) a heavy chain variable region (HCVR) comprising heavy chain complementarity determining regions 1, 2, and 3 (HCDR1-HCDR2-HCDR3), each comprising the sequence 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 complementarity determining regions 1, 2 and 3 (LCDR1LCDR2-LCDR3), each comprising the sequence of SEQ ID NO: 6, SEQ ID NO: 7, and SEQ ID NO: 8 respectively. In a specific embodiment, the antibodies include HCVR and LCVR, each of which includes the amino acid sequence of SEQ ID NO: 1 and SEQ ID NO: 5, respectively.

In one embodiment, the antibodies comprise any one or more of the amino acid sequences of SEQ ID NO: 9-16. In one embodiment, the antibodies comprise (a) a heavy chain variable region (HCVR) comprising heavy chain complementarity determining regions 1, 2, and 3 (HCDR1-HCDR2-HCDR3), each comprising the sequence 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 complementarity determining regions 1, 2, and 3 (LCDR1-LCDR2-LCDR3), each comprising the sequence of SEQ ID NO: 14, SEQ ID NO: 15, and

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SEQ ID NO: 16 respectively. In a specific embodiment, the antibodies include HCVR and LCVR, each of which includes the amino acid sequence of SEQ ID NO: 9 and SEQ ID NO: 13, respectively.

In one embodiment, the antibodies comprise any one or more of the amino acid sequences of SEQ ID NO: 17-24. In one embodiment, the antibodies comprise (a) a heavy chain variable region (HCVR) comprising heavy chain complementarity determining regions 1, 2, and 3 (HCDR1-HCDR2-HCDR3), each comprising the sequence 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 complementarity determining regions 1, 2 and 3 (LCDR1-LCDR2-LCDR3), each comprising the sequence of SEQ ID NO: 22, SEQ ID NO: 23, and SEQ ID NO : 24 respectively. In a specific embodiment, the antibodies include HCVR and LCVR, each of which includes the amino acid sequence of SEQ ID NO: 17 and SEQ ID NO: 21, respectively.

In one embodiment of the invention, the pH of the liquid composition is about pH 5.9±0.5. In a specific embodiment of the invention, the pH of the liquid composition is about pH 5.9±0.1. In one embodiment of the invention, the liquid pharmaceutical buffer includes one or more buffers that can maintain a pH of about 5.6 to about 6.2.

In one embodiment of the invention, the liquid pharmaceutical composition contains a buffer system that includes at least two buffers. In one embodiment of the invention, the buffer system includes a first buffer having an effective pH range in the range of 3.6-5.6 and a second buffer having an effective pH range in the 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 a specific embodiment of the invention, the first buffer is an acetate buffer and the second buffer is a histidine buffer. In one specific embodiment of the invention, the acetate is at a concentration of 12.5 mm ± 1.9 mm and histidine at a concentration of 20 mm ± 3 mm.

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

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

In one embodiment of the invention, the thermal stabilizer is sugar. In one embodiment of the invention, the sugar is selected from the group consisting of sucrose, mannitol and trehalose. In a specific embodiment of the invention, the thermal stabilizer is sucrose.

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

In one embodiment, the viscosity reducer 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 of the invention, the viscosity reducer is L-arginine hydrochloride.

In one embodiment of the invention, the viscosity reducer is at a concentration that is not more than 100 mM. In one embodiment of the invention, the viscosity reducer is at a concentration of 50 mM ± 7.5 mM. In another embodiment of the invention, the viscosity reducer is at a concentration of about 25 mM±3.75 mM. In a specific embodiment of the invention, the viscosity reducer is 25 mm ± 3.75 mm L-arginine hydrochloride.

In one embodiment of the invention, the viscosity of the liquid pharmaceutical composition is less than or equal to about 35±3.5 centipoise. In one 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 of the invention, the viscosity of the liquid pharmaceutical composition is about 8.5±0.85 centipoise.

In one embodiment of the invention, the osmolality of the liquid pharmaceutical composition is less than about 450 mOsm/kg. In one embodiment of the invention, the osmolality of the liquid pharmaceutical composition is about 290±20 mOsm/kg.

In one embodiment of the invention, at least 90% or at least 95% of

- 2 042167 active form of anti-hIL-4Rα antibodies is recovered from the liquid pharmaceutical composition after six months of storage of the liquid pharmaceutical composition at 5°C, as determined by size exclusion chromatography. In a specific embodiment of the invention, at least 98% of the native form of the antibody is recovered after six months of storage at 5° C., as determined by size exclusion chromatography.

In one embodiment of the invention, at least 90% of the native form of the antibodies is recovered from the liquid pharmaceutical composition after eight weeks of storage at 45° C., as determined by size exclusion chromatography.

In one embodiment of the invention, less than 45% of the antibodies that are recovered from the liquid pharmaceutical composition after eight weeks of storage at 45° C. are in the acid form, as determined by cation exchange chromatography.

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

In one aspect, a liquid pharmaceutical composition is provided containing: (i) about 150 mg/mL±50 mg/mL of human antibodies that specifically bind to hIL-4Ra, wherein the antibody comprises a heavy chain variable region (HCVR) and a light chain variable region ( LCVR), comprising the amino acid sequence of SEQ ID NO: 1 and SEQ ID NO: 5, respectively; (ii) about 12.5 mM±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 a pH of about 5.9 ± 0.5.

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

In one embodiment of the invention, the liquid pharmaceutical composition is physiologically isotonic. In one embodiment of the invention, the osmolality of the liquid pharmaceutical composition is about 290±20 mOsm/kg.

In one embodiment of the invention, at least about 98% of the native form of the anti-hIL4-Ra antibody is recovered from the liquid composition after six months of storage at 5° C., as determined by size exclusion chromatography.

In one embodiment of the invention, at least about 90% of the native form of the anti-hIL4-Ra antibody is recovered from the liquid pharmaceutical composition after eight weeks of storage at 45° C. as determined by size exclusion chromatography.

In one embodiment, less than about 45% of the antibodies that are recovered from the liquid pharmaceutical composition after eight weeks of storage at 45° C. are in the acid form as determined by cation exchange chromatography.

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

In one aspect, a stable low viscosity isotonic liquid pharmaceutical composition is provided that contains at least 100 mg/mL of a stable anti-hIL4-Ra antibody. In one embodiment, the antibodies are at a concentration of about 150 mg/ml±50 mg/ml. In a specific embodiment of the invention, the antibody concentration is about 150 mg/ml±15 mg/ml.

In one embodiment, the antibodies comprise any one or more amino acid sequences of SEQ ID NOs: 1-8. In one embodiment, the antibodies comprise a heavy chain variable region (HCVR) and a light chain variable region (LCVR), wherein the HCVR/LCVR combination comprises heavy and light chain complementarity determining regions (HCDR1-HCDR2-HCDR3/LCDR1-LCDR2-LCDR3) , which include the amino acid sequences of SEQ ID NO: 2-3-4/SEQ ID NO: 6-7-8, respectively. In a specific embodiment, the antibodies include HCVR and LCVR, each of which includes the amino acid sequence of SEQ ID NO: 1 and SEQ ID NO: 5, respectively.

In some embodiments, the composition has a viscosity of 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 of the invention, the liquid composition has a viscosity of about 8.5±2.5 centipoise.

In one embodiment of the invention, the composition has an osmolality that is physiologically compatible. In a specific embodiment of the invention, the composition has an osmolality of 290±20 mOsm/kg.

In one embodiment, the antibodies are stable for at least about six months at about 5°C. In a specific embodiment of the invention, at least about 98% of the antibodies retain their natural conformation within about six months of storage at 5°C, as determined by size exclusion chromatography.

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In one embodiment, the antibodies are stable for at least about eight weeks of storage at about 45°C. In a specific embodiment of the invention, at least about 90% of the antibodies retain their native conformation after about eight weeks of storage at 45°C, as determined by size exclusion chromatography. In a specific embodiment of the invention, less than about 45% of the antibodies are acidic after about eight weeks of storage at 45° C., as determined by cation exchange chromatography.

In one embodiment, the antibodies remain stable for at least about six months of storage at about 25°C. In a specific embodiment, less than about 4% of the antibodies are in an aggregated form after about six months of storage at 25° C., as determined by size exclusion chromatography.

In one embodiment of the invention, the composition includes a buffer and has a pH of about pH 5.9±0.5. In one embodiment of the invention, the buffer includes an acetate buffer and a histidine buffer. In a specific embodiment, the acetate is at a concentration of 12.5 mM±1.9 mM and the histidine is at a concentration of 20 mM±3 mM.

In one embodiment of the invention, the composition includes an organic co-solvent at a concentration of from about 0.2%±0.03% to about 1%±0.15% w/v. In one embodiment of the invention, the organic co-solvent is a non-ionic polymer containing a polyoxyethylene component. In some embodiments, the organic co-solvent 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 at a concentration of about 0.2%±0.03% w/v.

In one embodiment of the invention, the composition contains a thermal stabilizer at a concentration of from about 0.9%±0.135% w/v. up to about 10%±1.5% w/v. In one embodiment of the invention, the thermal stabilizer is sugar. In one embodiment of the invention, the sugar is selected from the group consisting of sucrose, mannitol and trehalose. In a specific embodiment of the invention, the thermal stabilizer is sucrose at a concentration of about 5%±0.75% w/v.

In one embodiment of the invention, the composition contains a viscosity reducer at a concentration that is not more than about 100 mm. In one embodiment of the invention, the viscosity reducer is arginine. In a specific embodiment of the invention, the viscosity reducer is L-arginine hydrochloride at a concentration of 25 mM±3.75 mM.

In a specific embodiment of the invention, the low viscosity stable isotonic liquid pharmaceutical composition has a viscosity of about 8.5±2.5 cP and an osmolality of about 290±20 mOsm/kg and comprises: (i) 150 mg/ml±15 mg/ml anti- hIL4-Rα antibodies, where the antibodies include HCVR and LCVR, each of which includes the amino acid sequence of SEQ ID NO: 1 and SEQ ID NO: 5, respectively; (ii) 12.5 mM±1.9 mM acetate; (iii) 20 mM±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. In accordance with this embodiment of the invention, (i) at least about 98% of the antibodies retain their native conformation, as determined by size exclusion chromatography when stored at 5°C for at least about six months, (ii) at least about 90% of the antibodies retain their native conformation as determined by size exclusion chromatography when stored at 45°C for at least about eight weeks, (iii) less than about 45% of the antibodies are acidic as determined by cation exchange chromatography when stored at 45°C for about eight weeks, and (iv) less than about 4% of the antibodies are aggregated as determined by size exclusion chromatography when stored at 25°C for about six months.

In one aspect, the liquid pharmaceutical composition of any of the preceding aspects is provided in a container. In one embodiment of the invention, the container is a glass vial. In another embodiment of the invention, the container is a microinfusor. In another embodiment, the container is a syringe. In one specific embodiment of the invention, the syringe includes a plunger coated with fluorocarbon. In one specific embodiment of the invention, the syringe is a low tungsten syringe.

Other embodiments of the present invention will be apparent from a review of the following detailed description.

Detailed description of the invention

Before the present invention will be described, it should be understood that the present invention is not limited to certain described methods and experimental conditions, such methods and conditions may vary. It should also be understood that the terminology used herein is for the purpose of describing certain embodiments of the invention only and is not intended to be limiting, as the scope of the present invention is only limited by the appended claims.

Unless otherwise indicated, all technical and scientific terms used in this specification are

- 4 042167 have the meanings generally understood by one of ordinary skill in the art to which the present invention pertains. As used herein, the term about, when used with reference to a specific specified numerical value or range of values, means that the value may vary from the specified value by no more than 1%. For example, as used herein, an expression near 100 includes 99 and 101 and all values in between (eg, 99.1, 99.2, 99.3, 99.4, etc.).

While all methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, the following describes the preferred methods and materials. All publications cited in the present description are incorporated into the present description by reference for the description in their entirety.

Pharmaceutical compositions

As used herein, the expression pharmaceutical composition means a combination of at least one active ingredient (e.g., a small molecule, a macromolecule, a compound, etc. that is capable of producing a biological effect in a human or non-human animal) and at least one an inactive ingredient which, when combined with the active ingredient or one or more additional inactive ingredients, is suitable for therapeutic administration to a human or non-human animal. The term composition, as used in the present description, means a pharmaceutical composition, unless otherwise specifically indicated. The present invention provides pharmaceutical compositions containing at least one therapeutic polypeptide. In accordance with certain embodiments of the present invention, the therapeutic polypeptide is an antibody, or an antigen-binding fragment thereof, that binds specifically to the human interleukin-4 receptor alpha (hIL-4Ra). More specifically, the present invention includes pharmaceutical compositions that contain: (i) human antibodies that specifically bind to hIL-4Ra; (ii) an acetate/histidine buffer system; (iii) an organic co-solvent, which is a non-ionic surfactant; (iv) a thermal stabilizer, which is a hydrocarbon; and (v) a viscosity reducer. Specific examples of the components and compositions included in the present description are described in detail below.

Antibodies that specifically bind to hIL-4R.

Pharmaceutical compositions of the present invention may include a human antibody, or antigen-binding fragment thereof, that specifically binds to hIL-4Ra. As used herein, the term hIL-4Ra means a human cytokine receptor that specifically binds to interleukin-4 (IL-4). In certain embodiments, the antibodies contained in the pharmaceutical compositions of the present invention specifically bind to the extracellular domain of hIL-4Ra. An exemplary amino acid sequence of human IL-4 receptor alpha (hIL-4Ra) is described in SEQ ID NO: 25. Antibodies to hIL-4Ra are described in US Pat. Nos. 7,605,237 and 7,608,693.

The term antibody, as used herein, generally refers to immunoglobulin molecules comprising four polypeptide chains, two heavy (H) chains, and two light (L) chains linked by disulfide bonds, as well as their multimers (eg, IgM); however, immunoglobulin molecules consisting only of heavy chains (ie, no light chains) are also covered by the definition of the term antibody. Each heavy chain includes a heavy chain variable region (abbreviated herein as HCVR or VH) and a heavy chain constant region. The heavy chain constant region includes three domains, CH1, CH2 and CH3. Each light chain includes a light chain variable region (abbreviated herein as LCVR or VL) and a light chain constant region. The light chain constant region includes one domain (CL1). The V _H and V _L regions can be further subdivided into regions of hypervariability, called complementarity determining regions (CDRs), interspersed with regions that are more conserved, called framework regions (FRs). Each V _H and VL consists of three CDRs and four FRs, arranged from amino to carboxy in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.

Unless specifically stated otherwise, the term antibody, as used herein, is to be understood as encompassing entire antibody molecules as well as antigen-binding fragments thereof. The term antigen-binding portion or antigen-binding fragment of an antibody (or simply antibody portion or antibody fragment) as used herein refers to one or more antibody fragments that retain the ability to specifically bind to hIL-4Ra or an epitope thereof.

An isolated antibody, as used herein, is intended to mean an antibody that is substantially free of other antibodies having different antigenic specificities (e.g., an isolated antibody that specifically binds hIL-4Ra is essentially free of antibodies that specifically bind with antigens other than hIL-4Ra).

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The term specifically binds or 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 can be characterized by a dissociation constant of at least about 1x10 ^-6 M or more. Methods for determining whether two molecules can specifically bind are well known in the art and include, for example, equilibrium dialysis, surface plasmon resonance, and the like. An isolated antibody that specifically binds to hIL-4Ra may, however, cross-react with other antigens such as other species of IL-4R molecules (orthologs). In the context of the present invention, multispecific (eg, bispecific) antibodies that bind to h!L-4Ra as well as one or more additional antigens are considered to specifically bind hIL-4Ra. Moreover, the isolated antibody may be substantially free of other cellular materials or chemicals.

Examples of anti-hIL-4Rα antibodies that can be included in the pharmaceutical compositions of the present invention are presented in US 7605237 and US 7608693, the description of which is incorporated by reference in their entirety.

In accordance with certain embodiments of the present invention, the anti-hIL4Ra antibody is a human IgG1 comprising a heavy chain variable region, which is the IGHV3-9 subtype, and a light chain variable region, which is the IGKV2-28 subtype (see Barbie and Lefranc, The Human Immunoglobulin Kappa Variable (IGKV) Genes and Joining (IGKJ) Segments, Exp Clin Immunogenet 1998;15:171-183 and Scaviner, D. et al Protein Displays of the Human Immunoglobulin Heavy, Kappa and Lambda Variable 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 change in charge on the exposed surface of the antibody relative to the original IGHV3-9 sequence or the original IGKV2-28 sequence. The original sequences of IGHV3-9 and IGKV2-28 and the position designations of the amino acid numbers presented herein correspond to the International Immunogenetics Information System (IMGT) as described in Lefranc, M.-P., et al. IMGT®, the 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 amino acid substitution or substitutions are selected from the group consisting of (a) a basic amino acid substituted with a neutral amino acid within CDR2 (e.g. at position 58) of IGHV3-9, (b) a neutral amino acid substituted with an acidic amino acid within CDR3 (eg at position 107) of IGHV3-9, (c) a neutral amino acid substituted with a basic amino acid within CDR1 (eg at position 33) of IGKV2-28. Unique permutations in the charge distribution of the antibody, especially at the interface with the environment (such as, for example, in the CDR) are expected to create unpredictable conditions for the stability of the antibody in solution.

In some embodiments, the anti-hIL-4Rα antibody comprises at least one amino acid substitution that creates a twist-strain change in the antibody framework relative to the original IGHV3-9 sequence or the original IGKV2-28 sequence. In some embodiments, the amino acid substitution or substitutions are selected from the group consisting of (a) a proline substituted with a non-proline amino acid in framework region 3 (FR3) (e.g., position 96) of IGHV3-9, and (b) a non-proline amino acid substituted with a proline in frame region 2 (FR2) (eg at position 46) of IGKV2-28. Changes in the ability of the peptide chain to rotate, especially in the framework region, which affects the interaction of the CDR with the solvent, can probably create unpredictable conditions for the stability of the antibody in solution.

In accordance with certain embodiments of the present invention, the anti-hIL4Ra antibodies or antigen-binding fragment thereof comprise a heavy chain complementarity determining region (HCDR) 1 SEQ ID NO: 2, HCDR2 SEQ ID NO: 3, and HCDR3 SEQ ID NO: 4. In certain embodiments, the anti-hIL-4Rα antibodies or antigen-binding fragment thereof comprise HCVD SEQ ID NO: 1.

In accordance with certain embodiments of the present invention, the anti-anti-hIL-4Ra, or antigen-binding fragment thereof, comprises a light (kappa) chain complementarity determining region (LCDR) 1 of SEQ ID NO: 6, LCDR2 of SEQ ID NO: 7, and LCDR3 of SEQ ID NO: 8 In certain embodiments, the anti-anti-hIL-4Rα antibodies, or antigen-binding fragments thereof, comprise the LCVD of SEQ ID NO: 5.

In accordance with certain other embodiments of the present invention, anti-anti-hIL-4Rα antibodies or antigen-binding fragments thereof include HCDR1 SEQ ID NO: 10, HCDR2 SEQ ID NO: 11, HCDR3 SEQ ID NO: 12, LCDR1 SEQ ID NO: 14, LCDR2 SEQ ID NO: 15 and LCDR3 SEQ ID NO: 16. In certain embodiments, anti-hIL-4Rα antibodies or antigen-binding fragments thereof include HCVD SEQ ID NO: 9 and LCVD SEQ ID NO: 13.

In accordance with certain other embodiments of the present invention, an

- 6 042167 ti-hIL-4Rα antibodies or antigen-binding fragments thereof include HCDR1 SEQ ID NO: 18, HCDR2 SEQ ID NO: 19, HCDR3 SEQ ID NO: 20, LCDR1 SEQ ID NO: 22, LCDR2 SEQ ID NO: 23 and LCDR3 SEQ ID NO: 24. In certain embodiments, anti-hIL-4Rα antibodies or antigen-binding fragments thereof include HCVD SEQ ID NO: 17 and LCVD SEQ ID NO: 21.

Non-limiting examples of antibodies used in the examples in the present description, referred to as mAb1. These antibodies are also presented in US 7608693 as H4H098P. mAb1 (H4H098P) includes a pair of amino acid sequences HCVR/LCVR, SEQ ID NO: 1/5, and HCDR1-HCDR2HCDR3/LCDR1-LCDR2-LCDR3 domains represented by SEQ ID NO:2-3-4/SEQ ID NO: 6-7 -8.

Other non-limiting examples of antibodies that can be used in the implementation of the present invention, referred to as mAb2. These antibodies are also represented in US 7608693 as H4H083P. mAb2 (H4H083P) comprises the HCVR/LCVR amino acid sequence pair having SEQ ID NO: 9/13 and HCDR1-HCDR2-HCDR3/LCDR1-LCDR2-LCDR3 domains represented by SEQ ID NO: 10-11-12/SEQ ID NO: 14-15-16.

Another non-limiting example of an antibody that can be used in the implementation of the present invention is mAb3. The specified antibody is also presented in US 7608693 as H4H095P. mAb3 (H4H095P) includes a pair of HCVR/LCVR amino acid sequences having SEQ ID NO: 17/21 and HCDR1-HCDR2-HCDR3/LCDR1-LCDR2-LCDR3 domains represented by SEQ ID NO: 18-19-20/SEQ ID NO: 22-23-24.

The amount of antibodies or antigen-binding fragments thereof contained in the pharmaceutical compositions of the present invention may vary depending on the specific desired properties of the compositions, as well as the specific circumstances and purposes for which the compositions are intended to be used. In certain embodiments of the invention, the pharmaceutical compositions are liquid compositions, which may contain from about 100±10 mg/ml to about 200±20 mg/ml antibodies; from about 110±11 mg/ml to about 190±19 mg/ml antibody; about 120±12 mg/ml to about 180±18 mg/ml antibody; from about 130±13 mg/ml to about 170±17 mg/ml antibody; from about 140±14 mg/ml to about 160±16 mg/ml antibody; about 150±15 mg/ml antibody. For example, the compositions of the present invention may include 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 antigen-binding antibody thereof that specifically binds to hIL-4Ra.

Excipients and pH.

Pharmaceutical compositions of the present invention include one or more excipients. The term excipient, as used herein, means any non-therapeutic agent added to a composition to provide the desired consistency, viscosity, and stabilizing effect.

In certain embodiments, the pharmaceutical composition of the invention contains at least one organic co-solvent of a type and amount that stabilizes hIL-4Ra antibodies under rough handling conditions, such as, for example, shaking. In some embodiments of the invention, stabilizes is understood to mean preventing the formation of more than 2% of aggregated antibodies of the total number of antibodies (on a molar basis) with careless handling. In some embodiments, the rough handling is shaking the solution containing the antibodies and the organic co-solvent for about 120 minutes.

In certain embodiments, the organic co-solvent is a non-ionic surfactant such as an alkyl poly(ethylene oxide). Specific non-ionic surfactants that may be included in the compositions 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 polysorbate 85; poloxamers such as poloxamer 181, poloxamer 188, poloxamer 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 amount of organic co-solvent contained in the pharmaceutical compositions of the present invention may vary depending on the specific properties desired for the compositions, as well as on the specific circumstances and purposes for which the compositions are intended to be used. In certain embodiments, the compositions may contain from about 0.1%±0.01% to about 2%±0.2% surfactant. For example, the compositions of the present invention may include 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%; near

- 7 042167

0.28%; about 0.29% or about 0.30% polysorbate 20 or poloxamer 181. For example, compositions of the present invention may include 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.

Examples of organic co-solvents that stabilize hIL-4Ra antibodies include 0.2%±0.02% polysorbate 20, 0.2%±0.02% poloxamer 181, or 1%±0.1% PEG 3350.

The pharmaceutical compositions of the present invention may also contain one or more type and amount of thermal stabilizers that stabilize hIL-4Ra antibodies under thermal stress conditions. In some embodiments, the term stabilizes means maintaining more than about 92% of the antibodies in native conformation when the solution containing the antibodies and thermal stabilizer is stored at about 45° C. for up to about 28 days. In some embodiments, stabilized means that less than about 5% of the antibodies are aggregated when the solution containing the antibodies and thermal stabilizer is stored at about 45° C. for up to about 28 days.

In certain embodiments of the invention, the thermal stabilizer is a sugar or sugar alcohol selected from sucrose, trehalose and mannitol, or any combination thereof, the amount of which contained in the composition may vary depending on the specific circumstances and intended use for which the composition is used. In certain embodiments, the compositions may contain from about 2.5% to about 10% sugar or sugar alcohol; about 3% to about 9.5% sugar or sugar alcohol; about 3.5% to about 9% sugar or sugar alcohol; about 4% to about 8.5% sugar or sugar alcohol; about 4.5% to about 8% sugar or sugar alcohol; about 5% to about 7.5% sugar or sugar alcohol; about 5.5% to about 7% sugar or sugar alcohol; or about 6.0% to about 6.5% sugar or sugar alcohol. For example, pharmaceutical compositions of the present invention may include about 2.5%±0.375%; about 3%±0.45%; about 3.5%±0.525%; 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%; 8.5%±1.275%; about 9.0% ± 1.35% or about 10.0% ± 1.5% sugar or sugar alcohol (eg sucrose, trehalose or mannitol).

The pharmaceutical compositions of the present invention may also include a buffer or buffer system that is designed to maintain a stable pH and to improve the stability of hIL-4Ra antibodies. In some embodiments, stabilizes is understood to mean that less than 3.0%±0.5% of the antibodies aggregate when the solution containing the antibodies and buffer is stored at about 45° C. for up to about 14 days. In some embodiments, stabilizes means that less than 3.7% ± 0.5% of antibodies are aggregated when the solution containing the antibodies and buffer is stored at about 25° C. for about 6 months. In some embodiments, stabilizes is understood to mean that at least 95% ± 0.5% of the antibodies are in their native conformation, as determined by size exclusion chromatography when the solution containing the antibodies and buffer is stored at about 45° C. for up to about 14 days. In some embodiments, stabilizes is understood to mean that at least 96% ± 0.5% of the antibodies are in their native state, as determined by size exclusion chromatography when the solution containing the antibodies and buffer is stored at about 25° C. for up to about 6 months. In some embodiments, stabilizes is understood to mean that at least 62%±0.5% of the antibodies are in their neutral conformation as determined by cation exchange chromatography when the solution containing the antibodies and buffer is stored at about 45°C for up to about 14 days. In some embodiments, stabilizes is understood to mean that at least 54%±0.5% of the antibodies are in their neutral conformation as determined by cation exchange chromatography when the solution containing the antibodies and buffer is stored at about 25°C for up to about 6 months. By neutral conformation is meant the fraction of antibodies that elutes from the ion exchange resin in the main peak, which is usually limited to more alkaline peaks on one side and more acidic peaks on the other side.

Pharmaceutical compositions of the present invention may have a pH of about 5.2 to about 6.4. For example, the compositions of the present invention may have a pH of 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 of the invention, the buffer or buffer system includes at least one buffer that has a buffer range that fully or partially covers the pH range of 5.2-6.4. In one embodiment of the invention, the buffer or buffer system comprises two buffers, the first having an effective pH range of 3.6-5.6 and the second having 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

- 8 042167 embodiments of the invention, the buffer system includes an acetate buffer and a histidine buffer. In certain embodiments, the histidine is present in an amount of 1.3-1.9 parts per 1 part mole of acetate buffer. In certain embodiments, the histidine is present in an amount of about 1.6 ± 0.25 parts per 1 mole of acetate. In certain embodiments, the acetate is present at a concentration of 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 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; from about 12.5 mM±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 histidine is present at a concentration of 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 and about 20 mM histidine, at a pH of about 5.9.

Pharmaceutical compositions of the present invention may also include one or more excipients that are designed to maintain a reduced viscosity or to reduce the viscosity of compositions containing a high concentration of protein (eg, typically >100 mg/ml of protein). In some embodiments, the composition includes an amount of arginine sufficient to maintain a viscosity of the liquid composition of less than about 35 centipoise, less than about 30 centipoise, less than about 25 centipoise, less than about 20 centipoise, less than about 15 centipoise, less than about 14 cps, less than about 13 cps, less than about 12 cps, less than about 10 cps, or less than about 9 cps.

In certain embodiments, the pharmaceutical composition of the present invention contains arginine, preferably L-arginine hydrochloride, at a concentration of about 25 mM±3.75 mM, about 50 mM±7.5 mM, or about 100 mM±15 mM. In certain embodiments, the arginine is in an amount of 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 from about 24 mM to about 26 mM.

Composition examples.

In accordance with one aspect of the present invention, the pharmaceutical composition is a generally physiologically isotonic low viscosity liquid composition that contains: (i) human antibodies that specifically bind to hIL-4Ra (e.g. mAb1, mAb2 or mAb3 [above] ), at a concentration of about 100 mg/ml or more; (ii) a buffer system that provides sufficient buffering at a level of about 5.9±0.6; (iii) sugar, which is intended, inter alia, as a thermal stabilizer; (iv) an organic co-solvent that protects the structural integrity of the antibodies; and (v) an amino acid that is designed to maintain a viscosity acceptable for subcutaneous injection.

In accordance with one embodiment of the invention, the pharmaceutical composition comprises: (i) human IgG1 antibodies that specifically bind to hIL-4Ra, and which comprise a substituted heavy chain variable region of the IGHV3-9 type and a substituted light chain variable region of the IGLV2-28 type (e.g. , mAb1) at a concentration of about 100 mg/ml to about 200 mg/ml; (ii) a buffer system comprising acetate and histidine that effectively maintains a pH of about 5.9±0.6; (iii) sucrose as a thermal stabilizer; (iv) polysorbate as an organic co-solvent; and (v) arginine as a viscosity reducer.

In accordance with one embodiment of the invention, the pharmaceutical composition contains: (i) human IgG1 antibodies that specifically bind to hIL-4Ra, and which include HCDR1 SEQ ID NO: 2, HCDR2 SEQ ID NO: 3, HCDR3 SEQ ID NO: 4, LCDR1 SEQ ID NO: 6, LCDR2 SEQ ID NO: 7, and LCDR3 SEQ ID NO: 8, at a concentration of about 150 mg/ml±25 mg/ml; (ii) acetate at a concentration of about 12.5 mM±1.9 mM and histidine at a concentration of about 20 mM±3 mM, which effectively buffer at a pH of about 5.9±0.3; (iii) sucrose at a concentration of about 5% w/v±0.75% w/v; (iv) polysorbate 20 at a concentration of about 0.2% w/v±0.03% w/v; and (v) arginine as L-arginine hydrochloride at a concentration of about 25 mM±3.75 mM.

Additional non-limiting examples of pharmaceutical compositions covered by the present invention are presented later in the present description, including working examples below.

Stability and viscosity of pharmaceutical compositions.

Pharmaceutical compositions of the present invention generally exhibit high stability. The term "stable" as used herein in relation to pharmaceutical

- 9 042167 compositions, means that the antibodies in pharmaceutical compositions retain an acceptable degree of chemical structure or biological function after storage under certain conditions. The composition may be stable even if the antibodies contained therein do not retain 100% of their chemical structure or biological function after storage for a certain amount of time. Under certain circumstances, retention of about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% of the structure or function of antibodies after storage for a certain amount of time can be regarded as stable.

Stability can be measured, among other things, by determining the percentage of native antibodies that remain in the composition after storage for a certain amount of time at a certain temperature. The percentage of native antibodies can be determined inter alia by size exclusion chromatography (eg size exclusion high performance liquid chromatography [E-HPLC]). An acceptable degree of stability, as used herein, means that at least 90% of the native form of the antibody can be detected in the composition after storage for a certain amount of time at a given temperature. In certain embodiments of the invention, at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% of native form antibodies can be detected in the composition after storage for a certain period of time at a certain temperature. The specified amount of time after which stability is measured can be 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 more. The specific temperature at which the pharmaceutical composition can be stored in stability evaluation can be any temperature from about -80°C to about 45°C, for example, storage at about -30°C, about -20°C, about 0°C, about 4°-8°C, about 5°C, about 25°C or about 45°C. For example, a pharmaceutical composition may be considered stable if, after 3 months of storage at 5° C., more than about 90%, 95%, 96%, 97%, or 98% of native antibodies are detected by E-HPLC. A pharmaceutical composition can also be considered stable if, after 6 months of storage at 5°C, more than about 90%, 95%, 96%, 97%, or 98% of native antibodies are detected by E-HPLC. A pharmaceutical composition can also be considered stable if, after 9 months of storage at 5° C., more than about 90%, 95%, 96%, 97%, or 98% of native antibodies are detected by E-HPLC. A pharmaceutical composition can also be considered stable if, after 3 months of storage at 25° C., more than about 90%, 95%, 96%, or 97% of native antibodies are detected by E-HPLC. A pharmaceutical composition can also be considered stable if, after 6 months of storage at 25° C., more than about 90%, 95%, 96%, or 97% of native antibodies are detected by E-HPLC. A pharmaceutical composition can also be considered stable if, after 9 months of storage at 25° C., more than about 90%, 95%, 96%, or 97% of native antibodies are detected by E-HPLC.

Stability can be measured, inter alia, by determining the percentage of antibodies that form aggregates in the composition after storage for a certain period of time at a certain temperature, where the stability is inversely proportional to the percentage of the formed aggregate. The percentage of aggregated antibodies can be determined inter alia by size exclusion chromatography (eg size exclusion high performance liquid chromatography [E-HPLC]). An acceptable degree of stability, as this phrase is used herein, means that no more than 5% of the antibodies are in the aggregated form defined in the composition after storage for a certain period of time at a given temperature. In certain embodiments of the invention, an acceptable degree of stability means that no more than about 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of antibodies can be detected in the aggregate in the composition after storage for a certain period of time at a given temperature. The specified amount of time after which stability is measured can be 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 more. The temperature at which the pharmaceutical composition can be stored in stability evaluation can be any temperature from about -80°C to about 45°C, for example, storage at about -30°C, about -20°C, about 0°C, about 4°-8°C, about 5°C, about 25°C or about 45°C. For example, a pharmaceutical composition may be considered stable if, after 3 months of storage at 5°C, less than about 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of antibodies are detected in aggregated form. . A pharmaceutical composition can also be considered stable if, after 6 months of storage at 5° C., less than about 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of antibodies are detected in aggregated form. A pharmaceutical composition can also be considered stable if, after 9 months of storage at 5° C., less than about 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of antibodies are detected in aggregated form. The pharmaceutical composition is also

- 10 042167 can be considered stable if, after 3 months of storage at 25°C, less than about 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of antibodies are detected in aggregated form. A pharmaceutical composition can also be considered stable if, after 6 months of storage at 25° C., less than about 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of antibodies are detected in aggregated form. A pharmaceutical composition can also be considered stable if, after 9 months of storage at 25° C., less than about 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of antibodies are detected in aggregated form.

Stability can be measured, inter alia, by determining the percentage of antibodies that migrate to a more acidic fraction during ion exchange (acidic form) than to the main antibody fraction (neutral conformation), where stability is inversely proportional to the acidic antibody fraction. Without wishing to be bound by theory, deamidation of antibodies can cause the formation of a more negatively charged antibody and therefore more acidic relative to the non-deamidated antibody (see, e.g., Robinson, N., Protein Deamidation, PNAS, April 16, 2002, 99(8) :52835288). The percentage of acidified or deamidated antibody can be determined, inter alia, by ion exchange chromatography (eg, cation exchange high performance liquid chromatography [KO-HPLC]). An acceptable degree of stability, as this phrase is used herein, means that no more than 45% of the antibodies are in the composition in a more acidic form after storage for a certain period of time at a certain temperature. In certain embodiments of the invention, an acceptable degree of stability means that no more than about 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 5%, 4%, 3%, 2%, 1%, 0.5% or 0.1% antibodies can be detected in acidic form in the composition after storage for a certain period of time at a given temperature. The specified amount of time after which stability is measured can be 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 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 2 4 months or more. The temperature at which the pharmaceutical composition can be stored for stability evaluation can be any temperature from about -80°C to about 45°C, for example, storage at about -30°C, about -20°C, about 0°C, about 4°-8°C, about 5°C, about 25°C or about 45°C. For example, a pharmaceutical composition may be considered stable if, after 3 months of storage at 5°C, there are less than about 15%, 14%, 13%, 12%, 10%, 9%, 8%, 7%, 6%, 5 %, 4%, 3%, 2%, 1%, 0.5% or 0.1% of antibodies is in a more acidic form. A pharmaceutical composition can also be considered stable if, after 3 months of storage at 25°C, there are 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 antibodies are in a more acidic form. A pharmaceutical composition can also be considered stable if, after 8 weeks of storage at 45°C, there are less than about 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 5%, 4%, 3%, 2%, 1%, 0.5% or 0.1% of the antibodies are in a more acidic form. A pharmaceutical composition can also be considered stable if, after 2 weeks of storage at 40°C, there are 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% antibodies can be detected in the more acidic form.

Other methods can be used to evaluate the stability of the compositions of the present invention, such as, for example, differential scanning calorimetry (DSC) to determine thermal stability, controlled stirring to determine mechanical stability, and absorbance at about 350 nm or about 405 nm to determine the turbidity of the solution. For example, a composition of the present invention may be considered stable if, after 6 months or more of storage at 5°C to about 25°C, the change in OD ₄₀₅ of the composition is less than about 0.05 (e.g., 0.04, 0.03, 0.02, 0.01 or less) from the composition OD ₄₀₅ at time zero.

Stability can also be assessed by measuring the biological activity or binding affinity of an antibody to its target. For example, the composition of the present invention can be regarded as stable if after storage, for example, at 5°C, 25°C, 45°C, etc., for a certain amount of time (for example, from 1 to 12 months), anti -IL-4Rα antibodies contained in the composition bind to IL-4Ra with an affinity that is at least 90%, 95% or more of the antibody binding affinity prior to said storage. Binding affinity can be determined, for example, by ELISA or plasmon resonance. Biological activity can be determined by assaying IL-4Ra activity, such as, for example, contacting a cell that expresses IL-4Ra with a composition containing anti-IL-4Rα antibodies. The binding of antibodies to such a cell can be measured directly, for example, using FACS analysis. Alternatively, the downstream activity of the IL-4Ra system can be measured in the presence of an antibody and an IL-4Ra agonist and compared to the activity of the IL-4Ra system in the absence of antibodies. In some embodiments, the IL-4Ra may be endogenous to the cell. In other embodiments, IL-4Ra may be ectopically expressed in a cell.

Additional methods for assessing the stability of antibodies in a composition are demonstrated in

- 11 042167 measures below.

Liquid pharmaceutical compositions of the present invention may, in certain embodiments, exhibit a low to moderate viscosity. Viscosity, as used herein, may be kinematic viscosity or absolute viscosity. Kinematic viscosity is a measure of the resistive flow of a fluid under the influence of gravity. When two liquids of equal volume are placed in identical capillary viscometers and allowed to flow under the influence of gravity, the viscous liquid takes longer to flow through the capillary than the less viscous one. For example, if it takes 200 seconds for one fluid to complete the current and 400 seconds for another fluid, the second fluid is twice as viscous as the first fluid on the kinematic viscosity scale. Absolute viscosity, sometimes referred to as dynamic or simple viscosity, is the product of kinematic viscosity and fluid density (Absolute Viscosity=Kinematic ViscosityxDensity). The measure of kinematic viscosity is L2/T, where L is length and T is time. Typically, kinematic viscosity is expressed in centistokes (cSt). The SI unit of kinematic viscosity is mm ² /s, which is 1 cSt. Absolute viscosity is expressed in units of centipoise (cP). The SI unit of absolute viscosity is the milliPascal second (mPa-s), where 1 cP=1 mPa-s.

As used herein, a low level of viscosity, in relation to a liquid composition of the present invention, means an absolute viscosity of less than about 15 centipoise (cps). For example, a liquid composition of the invention is considered to have a low viscosity if, when measured using standard viscosity measurement techniques, the composition exhibits 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. As used herein, a moderate level of viscosity, in relation to the liquid composition of the present invention, means an absolute viscosity of from about 35 centipoise to about 15 centipoise. For example, a liquid composition of the invention is considered to have a moderate viscosity if, when measured using standard viscosity measurement techniques, the composition exhibits an absolute viscosity of about 34 centipoise, about 33 centipoise, about 32 centipoise, about 31 centipoise, about 30 centipoise, about 29 centipoise, about 28 cP, about 27 cP, about 26 cP, about 25 cP, about 24 cP, about 23 cP, about 22 cP, about 21 cP, about 20 cP, about 19 cP, 18 cP, about 17 cP, about 16 cP or about 15.1 cP.

As illustrated in the examples below, the present inventors unexpectedly found that low to moderate viscosity liquid compositions comprising high concentrations of anti-hIL-4Rα antibodies (e.g., about 100 mg/mL to at least 200 mg/mL ) can be obtained by formulating antibodies with arginine from about 25 mM to about 100 mM. In addition, it has additionally been found that the viscosity of the composition can be reduced even to a greater extent by adjusting the sucrose content to less than about 10%.

Containers for pharmaceutical compositions and methods of administration.

The pharmaceutical compositions of the present invention may be contained in any container suitable for storing drugs and other therapeutic compositions. For example, pharmaceutical compositions may be contained in a sealed and sterilized plastic or glass container having a defined volume, such as a vial, ampoule, syringe, cartridge or bottle. Various types of vials may be used to contain the compositions of the present invention, including, for example, clear and opaque (eg, amber) glass or plastic vials. Also, any type of syringe can be used to contain or administer the pharmaceutical compositions of the present invention.

The pharmaceutical compositions of the present invention may be contained in normal tungsten syringes or low tungsten syringes. As will be appreciated by one of skill in the art, the method of making glass syringes typically involves the use of a hot tungsten rod that acts to pierce the vial, thereby creating an opening from which liquid can be drawn and expelled from the syringe. This method leads to the deposition of trace amounts of tungsten on the inner surface of the syringe. Subsequent rinsing and other processing steps can be used to reduce the amount of tungsten in the syringe. As used herein, the term normal tungsten 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 ppm. 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 ppm tungsten.

Rubber plungers used in syringes and rubber stoppers used to close vial openings may be sheathed to prevent contamination of the medical contents of the syringe or vial, or to maintain their stability. Therefore, the pharmaceutical compositions of the present invention, in accordance with certain embodiments of the invention, may be contained in a syringe that includes a coated plunger or in a vial that is closed with a coated rubber stopper. For example, piston or plug

- 12 042167 can be coated with fluorocarbon film. Examples of coated stoppers or plungers suitable for use with vials and syringes containing the pharmaceutical compositions of the present invention are given, for example, in US Pat. Nos. 4,997,423; 5908686; 6286699; 6645635 and 7226554, the contents of which are incorporated by reference into the present description in their entirety. Certain exemplary coated rubber plugs and plungers that may be used in the context of the present invention are commercially available under the tradename FluroTec® available from West Pharmaceutical Services, Inc. (Lionville, PA).

In accordance with certain embodiments of the present invention, the pharmaceutical compositions may be contained in a low tungsten syringe that includes a fluorocarbon coated plunger.

Pharmaceutical compositions can be administered to a patient by parenteral routes such as injection (eg, subcutaneous, intravenous, intramuscular, intraperitoneal, etc.) or transdermal, transmucosal, nasal, pulmonary or oral. A variety of disposable pens or auto-injection devices may be used for subcutaneous delivery of the pharmaceutical compositions of the present invention. Examples include, but are not limited to, AUTOPEN™ (Owen Mumford, Inc., Woodstock, UK), DISETRONIC™ handle (Disetronic Medical Systems, Bergdorf, Switzerland), HUMALOG MIX 75/25™ handle, HUMALOG™ handle, HUMALI N handle 70/30™ (Eli Lilly and Co., Indianapolis, IN), NOVOPEN™ I, II and III (Novo Nordisk, Copenhagen, Denmark), NOVOPEN JUNIOR™ (Novo Nordisk, Copenhagen, Denmark), BD™ pen (Becton Dickinson , Franklin Lakes, NJ), OPTI PEN™, OPTIPEN PRO™, OPTI PEN STARLET™ and OPTICLI K™ (sanofi-aventis, Frankfurt, Germany). Examples of disposable pens or auto-injection inserters having use in subcutaneous delivery of the pharmaceutical composition of the present invention include, but are not limited to, SOLOSTAR™ pen (sanofi-aventis), FLEXPEN™ (Novo Nordisk) and KWIKPEN™ (Eli Lilly) , SURECLICK™ Autoinjector (Amgen, Thousand Oaks, CA), PENLET™ (Haselmeier, Stuttgart, Germany), EPIPEN (Dey, L.P.), and HUMIRA™ Pen (Abbott Labs, Abbott Park, IL).

The use of a microinfusor to administer the pharmaceutical compositions of the present invention is also contemplated herein. As used herein, the term microinfusor means a subcutaneous device designed to slowly deliver large volumes (eg, up to about 2.5 ml or more) of a therapeutic composition over an extended period of time (eg, about 10, 15, 20, 25 , 30 minutes or more). See, for example, U.S. 6629949; US 6659982; and Meehan et al, J. Controlled Release 46:107-1 16 (1996). Microinfusors are particularly suitable for administering large doses of high concentration therapeutic proteins (eg, about 100, 125, 150, 175, 200 or more mg/mL) or viscous solutions.

In one embodiment of the invention, a liquid pharmaceutical composition containing about 150 mg/ml ± 15 mg/ml of anti-IL-4Rα antibody is administered subcutaneously in a volume of approximately 1 ml ± 0.15 ml from a pre-filled syringe in an auto-injector. In another embodiment, the composition is administered in a volume of about 1 ml to 2.5 ml from a microinfusor device. The composition may be prefilled into a bag or cartridge for use in a microinfusor. Therapeutic Use of Pharmaceutical Compositions The pharmaceutical compositions of the present invention are useful, inter alia, for the treatment, prevention or amelioration of any disease or disorder associated with IL-4 activity, including diseases or disorders mediated by IL-4Ra activation. Examples of non-limiting diseases and disorders that can be treated or prevented by administering the pharmaceutical compositions of the present invention include various atopic diseases such as, for example, atopic dermatitis, allergic conjunctivitis, allergic rhinitis, asthma, and other IgE/Th2 mediated diseases.

Therefore, the present invention includes methods of treating, preventing or alleviating any disease or disorder associated with IL-4 activity or IL-4Ra activation (including any of the above examples of diseases, disorders and conditions). Therapeutic methods of the present invention include the introduction to the patient of any composition containing anti-hIL-4Rα antibodies, as described in the present description. The patient to whom the pharmaceutical composition is administered can be, for example, any human or non-human animal that is in need of such treatment, prevention, or alleviation, or that may otherwise benefit from inhibition or suppression of IL-4 or IL-mediated activity. -4Ra. For example, a patient may be an individual who has been diagnosed with or is likely to be at risk of developing any of the above diseases or disorders. The present invention further includes the use of any of the pharmaceutical compositions described herein in the manufacture of a medicament for the treatment, prevention or amelioration of any disease or disorder associated with IL-4 activity or IL-4Ra activation (including any of the above examples of diseases, disorders and conditions).

- 13 042167

Examples

The following examples are presented to provide the ordinary person of ordinary skill in the art with complete information and a description of how to make and use the methods and compositions of the invention, and are not intended to limit the scope by which the authors evaluate their invention. Attempts have been made to ensure accuracy with respect to the numbers used (eg quantities, temperature, etc.), but some experimental errors and deviations must be taken into account. Unless otherwise indicated, parts are molar parts, molecular weight is average molecular weight, temperature is in degrees Celsius, and pressure is at or near atmospheric pressure.

Development work on the parent formulation included screening for organic co-solvents, thermal stabilizers, and buffers in mAb 1 (anti-IL-4Rα antibodies of the invention) liquid formulations to determine excipients that are compatible with the protein and improve protein stability while maintaining osmolality and subcutaneous viscosity. injections. Buffer conditions were also investigated to determine the optimum pH for maximum protein stability.

Example 1 Organic co-solvents.

mAb1s were observed to be unstable when subjected to agitation stress. Reverse phase high performance liquid chromatography (RP-HPLC) and size exclusion high performance liquid chromatography (E-HPLC) analyzes demonstrated protein loss and an increase in protein aggregates when mAb1 was shaken at room temperature (Table 1, see no cosolvent data). The addition of organic co-solvents to the mAb1 solution prevented protein degradation as measured by E-HPLC and RP-HPLC (Table 1).

However, the addition of some organic co-solvents was observed to decrease the thermal stability of mAb1 (Table 2). Loss of protein recovery was observed in compositions containing PEG 3350 (3%) and PEG 300 (10% and 20%) as determined by RP-HPLC after thermal stress (Table 2). In addition, there was more aggregate formation in formulations containing PLURONIC F68 (poloxamer 181) (0.2%), PEG 300 (10% and 20%) and propylene glycol (20%) than in formulations without co-solvent, as determined by E -HPLC. Polysorbate 20 (0.2%) and Polysorbate 80 (0.2%) provided comparable stability to shaking and thermal stress.

According to Table. 1, 0.3 ml of 15 mg/ml mAb1 in 10 mM phosphate, pH 6.0 and various organic co-solvents in a 2 ml glass vial were shaken for about 120 minutes. Haze was evaluated by optical density (OD) at 405 nm and presented as a relative change in OD at 405 nm compared to the original substance. The percentage of total mAb1 recovered was determined by reverse phase HPLC (RP-HPLC). The percentage of native and aggregated mAb1 was determined by size exclusion HPLC (E-HPLC). The E-HPLC results reported in starting material results are the average of each composition in the absence of shaking.

Table 1

Organic co-solvent Appearance Turbidity to a Total % THA (RP-HPLC) % native tay (HP-HPLC) % of aggregate tay (HR-HPLC) Starting material ² (no shaking) Satisfactory 0.00 6.0 100 96.8 1.8 No co-solvent Failed 0.87 6.0 86 95.8 3.5 0.2% polysorbate 20 Satisfactory 0.01 5, 9 98 97.0 1.7 0.2% polysorbate 80 Satisfactory 0.00 5, 9 100 96.6 2.0 0.2% Pluronic F68 Satisfactory 0.00 5, 9 99 96.9 1.7 3% PEG 3350 Satisfactory 0.00 6.0 102 96.7 2.0 1% PEG 3350 Satisfactory 0.01 6.0 99 96.8 1.8 20% PEG 300 Satisfactory 0.01 5, 9 101 96.1 2, 6 10% PEG 300 Satisfactory 0.01 6.0 100 96.7 2.0 20% propylene glycol Satisfactory 0.00 6.0 101 96.7 2.0

According to Table. 2, 0.3 ml of 15 mg/ml mAb1 in 10 mM phosphate, pH 6.0 and various organic co-solvents in a 2 ml glass vial was stored at about 45° C. for about 28 days. Haze was assessed by optical density (OD) at 405 nm and reported as relative change in OD at 405

- 14 042167 nm compared to the starting material. The percentage of all recovered mAbl was determined by reverse phase HPLC (RP-HPLC). The percentage of native and aggregated mAb1 was determined by size exclusion HPLC (E-HPLC). The E-HPLC results reported in starting material results are the average of the values of each composition in the absence of thermal stress.

table 2

Organic co-solvent Appearance Turbidity pH Total % Shay (RP-HPLC) % Native Shay (HP-HPLC) % Aggregate (HR-HPLC) Starting material (no acceleration) Satisfactory 0.00 6.0 100 96.8 18 No co-solvent Satisfactory 0.00 6, 2 98 94.9 3.5 0.2% polysorbate 20 Satisfactory 0.00 6, 3 98 94.6 3, 6 0.2% polysorbate 80 Satisfactory 0.00 6, 2 97 94.3 3, 8 0.2% Pluronic F68 Satisfactory 0.00 6, 2 96 93.0 5, 1 3% PEG 3350 Satisfactory 0.00 6, 2 73 96.5 1.4 1% PEG 3350 Satisfactory 0.01 6.0 97 94.6 3, 8 20% PEG 300 Satisfactory 0.04 4.5 74 8.5 87.5 10% PEG 300 Satisfactory 0.02 4.8 93 57.7 38.1 20% propylene glycol Satisfactory 0.00 6, 3 97 93.6 4.7

Example 2. Thermal stabilizers.

Various thermal stabilizers such as sugars, amino acids and inorganic salts have been investigated for their ability to inhibit degradation of mAb1 when stored at about 45°C. A summary of the studied thermal stabilizers is presented in Table. 3. Compositions containing either sucrose or trehalose had the greatest stabilizing effect on mAb1 in solution when incubated at elevated temperature (determined by E-HPLC). Sucrose was chosen as the stabilizer because it has a history of safe use 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 a 2 ml glass vial was stored at about 45° C. for about 28 days. Haze was assessed by optical density (OD) at 405 nm and reported as a relative change in OD at 405 nm compared to the original substance. Haze was negligible for all samples. The percentage of all recovered mAb1 was determined by reverse phase HPLC (RP-HPLC). The percentage of native and aggregated mAb1 was determined by size exclusion HPLC (E-HPLC). Acid and basic species were defined as the sum of mAb1 peaks that elute from a cation exchange column (RT-HPLC) with an earlier or later retention time than the main peak, respectively. The E-HPLC results reported in starting material results are the average of the values of each composition in the absence of thermal stress.

- 15 042167

Table 3

Buffer and pH pH Total % THA (RP-HPLC) % native tay (E-HPLC) % of aggregate tay (E-HPLC) % Tab2 (KO-HPLC) sour peak Main Peak alkali peak Starting material (no incubation at 4-5°C) 5, 3 100 97.8 1.2 17, 6 68.2 13.2 No thermal stabilizer 5, 4 106 91.9 5, 8 28.1 56.5 15, 4 8.5% sucrose 5, 4 105 93, 3 4, 6 29.5 54.7 15, 8 4.5% sorbitol 5, 3 105 91.2 6, 6 34.4 51.5 14.1 4.5% mannitol 5, 3 104 92.6 5, 2 28.4 56.0 15, 6 9.4% trehalose dihydrate 5, 4 103 93.4 4.5 29, 1 55.6 15, 3 2.2% glycine 5, 4 104 86.6 10.6 33.5 50.7 15, 8 0.9% NaCl 5, 4 98 85.0 8.7 25.2 56.0 18.7 2.5% glycerin 5, 4 104 91.9 6.0 29, 7 56.1 14.3 5% arginine 5, 4 97 83, 2 11.4 25, 3 57, 1 17, 6

Example 3 Buffers and pH.

The effect of pH and buffer variants on mAbl stability was also investigated. 15 mg/ml mAbl was incubated in various buffers at various pH values ranging from pH 4.5 to 7.0. Protein stability was monitored by E-HPLC and cation exchange HPLC (C-HPLC). Maximum protein stability was observed as determined by both E-HPLC and RT-HPLC when mAb1 was formulated at pH 6.0 in histidine buffer or at pH 5.3 in acetate buffer (Table 4 and Table 5). The acetate buffer system provided a wider range of pH stability and a slower rate of change of formulation variants relative to a formulation containing a histidine buffer (Table 5). Therefore, an acetate buffer at pH 5.3 was chosen in part to formulate the mAb1 drug substance.

According to Table. 4, 0.3 ml 15 mg/ml mAb1, 0.2% polysorbate 20, mixed with 10 mM different buffers in a 2 ml glass vial, stored at about 45° C. for about 14 days. Haze was assessed by optical density (OD) at 405 nm and reported as a relative change in OD at 405 nm compared to the original substance. Haze was negligible for all samples. The percentage of all recovered mAb1 was determined by reverse phase HPLC (RP-HPLC). The percentage of native and aggregated mAb1 was determined by size exclusion HPLC (E-HPLC). Acid and basic species were defined as the sum of mAb1 peaks that elute from a cation exchange column (RT-HPLC) with an earlier or later retention time than the main peak, respectively. The E-HPLC results reported in starting material results are the average of the values of each composition in the absence of thermal stress.

Table 4

incubation at 45°C) pH 7.0, Phosphate____ pH 6.5, Phosphate____ pH 6.0, Phosphate____ pH 6.0, Histidine pH 6.0, Succinate pH 6.0, Citrate pH 5.5, Citrate pH 5, 0, citrate____ pH 5.0, acetate____ pH 4.5, acetate

According to Table. 5, 0.3 ml 15 mg/ml mAb1, 0.2% polysorbate 20 combined with 10 mM diff.

- 16 042167 buffers in a 2 ml glass vial were stored at about 45°C for about 14 days. Haze was assessed by optical density (OD) at 405 nm and reported as a relative change in OD at 405 nm compared to the original substance. Haze was negligible for all samples. The percentage of all recovered mAb1 was determined by reverse phase HPLC (RP-HPLC). The percentage of native and aggregated mAb1 was determined by size exclusion HPLC (E-HPLC). Acid and basic species were defined as the sum of the mAb1 peaks that elute from the cation exchange column (RT-HPLC) with an earlier or later retention time than the main peak, respectively. The EHPLC results reported in starting material results are the average of the values of each composition in the absence of thermal stress.

Composition development studies have shown that under alkaline conditions (pH>6.5), mAb1 in solution can be deamidated. Conversely, below pH 5.0, an increase in the rate of formation of mAb1 molecular weight variants was observed. Based on the data obtained, the pH of the mAb1 composition was maintained from pH 5.6 to pH 6.2. mAb1 were stable in the specified pH range.

Table 5

Buffer and pH Total % TAA Recovered (RP-HPLC) % native reduced tAA (E-HPLC) % Aggregate Recovered TAN (E-HPLC) % native recovered tAA (KO-HPLC) sour peak Main Peak alkali peak Starting material ³ (no incubation at 4-5°C) 100 96.5 2, 1 18.7 66, 7 14.6 pH 5.5, histidine 94 87.5 9, 1 22.7 58.7 18.6 pH 6.0, histidine 100 96.6 2.4 22.7 63.0 14.2 pH 6.5, histidine 97 89, 8 7.7 32.1 43, 8 24.0 pH 4.7, acetate 90 90.1 6, 4 18.4 66.1 15.5 pH 5.0, acetate 100 93.7 4.3 18.0 67.0 15.0 pH 5.3, acetate 99 95.2 thirty 18.1 67.5 14.5 pH 5.6, acetate 100 93.6 5, 3 22.1 61.7 14.3

The effect of pH and buffer variants on mAb1 stability was further evaluated in formulations containing either 20 mM histidine pH 6, 12.5 mM acetate pH 5.3, or a combination of 20 mM histidine and 12.5 acetate pH 5.9 (Table 6) . Compared to a single buffer system, mAb1 were most stable in a formulation containing both histidine and acetate at approximately pH 5.9. The slowest aggregation rate was determined when mAb1 was formulated in the indicated combined buffer system (E-HPLC) (Table 6).

Table 6

Buffer and pH Total % TAA Recovered (RP-HPLC) % native reduced tAA (E-HPLC) % Aggregate Recovered TAN (E-HPLC) % recovered ² tay (KO-HPLC) sour peak Main Peak alkali peak Starting material (no incubation at 45°C) 100 97 0 2, 6 27.4 62.1 10.5 20 mM histidine, pH 5.9 100 95 , 2 4.3 34.8 53, 9 11.4 12.5 mM acetate, pH 5, 3 103 94 , θ 4.8 30, 9 56.0 13, 1 Combination of 20 mM histidine and 12.5 mM acetate, pH 5.9 104 95 , 9 3.7 33.7 54.1 12.1

According to Table. 6, 0.4 ml 150 mg/ml mAb1, 10% sucrose, 0.2% polysorbate 20 combined with various buffers in a 2 ml glass vial was stored at about 45°C for about 14 days. Haze was measured by optical density (OD) at 405 nm and reported as a relative change in OD at 405 nm compared to the original substance. Haze was negligible for all samples. The percentage of all recovered mAb1 was determined by reverse phase HPLC (RP-HPLC). The percentage of native and aggregated mAb1 was determined by size exclusion HPLC (E-HPLC). Acidic and basic species were defined as the sum of the mAb1 peaks that elute from the cation exchange column.

- 17 042167 (RT-HPLC) with an earlier or later retention time than the main peak, respectively. The E-HPLC results reported in starting material results are the average of the values of each composition in the absence of thermal stress.

Example 4 Viscosity and Tonicity Control

Combinations of different excipients with high concentrations of mAb1 (ie 150 mg/ml, 175 mg/ml and 200 mg/ml) were evaluated for viscosity and tonicity (expressed in osmolality). The levels of sucrose, sodium chloride and L-arginine hydrochloride were adjusted to provide a composition containing a high concentration of mAb1 with low viscosity and physiological tonicity to allow easy, convenient and rapid subcutaneous administration of large amounts of mAb1 (Table 7). Liquid composition containing 25 mm arginine, 20 mm histidine, 12.5 mm acetate, 5% wt./about. sucrose, 0.2% wt./about. polysorbate 20, and 150 mg/ml mAb1, at pH 5.9 (composition A) is an optimized formulation having a low viscosity (about 8.5 cP) and being physiologically isotonic (about 293 mOsm/kg) while maintaining mAb1 stability .

Table 7

SHAI (mg/ml) Histidine (mM) Acetate (mM) Arginine (mM) NaCl (mM) Sucrose (% w/v) K ίλ Viscosity (cPoise) Osmolarity (mOsm/kg) A 150 20 12.5 25 0 5 5, 9 8.5 293 IN 150 20 12.5 0 0 10 5, 9 eleven 448 WITH 175 20 12.5 100 0 1 5, 9 -8.0 -290 D 175 20 12.5 50 0 5 5, 9 -9.5 -370 E 175 20 12.5 0 0 10 5, 9 -20 -440 F 200 20 12.5 100 0 1 5, 9 -15 -290 G 200 20 12.5 0 100 5 5, 9 -19, 2 -430 H 200 20 12.5 100 0 5 5, 9 -17 -430 I 200 20 12.5 50 0 5 5, 9 -18 -330 J 200 20 12.5 25 0 5 5, 9 -23 -290 TO 200 20 12.5 0 0 10 5, 9 -35 -440

Example 5 Characterization of composition A.

The main degradation pathways identified during the development of the mAb1 liquid formulation were the formation of aggregates, cleavage products, and charge variants. The formation of such degradation products was minimized 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. It was observed that the formulated 150 mg/ml mAb1 remained a clear to slightly opalescent liquid solution substantially free of visible particles.

The formulated mAb1s were physically and chemically stable under various stress (25°C and 45°C incubation) and real-time storage conditions (5°C) (Table 8). Appearance did not change when mAb1 was incubated at 25°C (3 months) or stored at 5°C for 6 months. In addition, no effect was observed on solution pH, turbidity or on the amount of mAb1 recovered. After incubation of the formulated mAb1 for 3 months at 25°C, the antibodies were not significantly destroyed, as determined by E-HPLC, and there was no more than 3.3% destroyed, as determined by RO-HPLC. Observed increased destruction after incubation at 45°C for 8 weeks, which was determined by E-HPLC and RO-HPLC, showing that the formation of aggregates and charge variants are the main ways of destruction of the mAb1 antibody molecule. No degradation was observed when the formulated mAb1 was stored for 6 months at 5°C.

- 18 042167

Table 8

stress test Lack of storage 5°С 25°С Storage duration - 2 months 3 months 6 months 1 month 3 months Appearance Satisfactory Satisfactory Satisfactory Satisfactory Satisfactory Satisfactory Haze (OD 405 nm) 0.00 0.00 0.00 0.00 0.01 0.01 pH 6.0 6.0 5, 9 5, 9 6.0 6.0 % mAbl (RPHPLC) 100 97 104 97 102 % native mAbl (HP-HPLC) 98.1 98.2 98.2 98.1 97.8 % mAbl (peaks from Q-HPLC) Sour 14.6 14.7 14.7 16.0 17, 6 Main Peak 70.7 70.5 70.4 69.8 67.4 Alkaline 14.7 14.8 14.9 14.3 15.0 stress test Lack of storage 45°С Storage duration - 2 weeks 3 weeks 6 weeks Appearance Satisfactory Satisfactory Satisfactory Satisfactory Haze (OD 405 nm) 0.00 0.02 0.03 0.05 pH 6.0 6.0 6.0 6.0 % mAbl (RP-HPLC) 100 102 98 100 % native mAbl (HP-HPLC) 98.1 95.9 94.2 90.5 % mAbl (peaks from Q-HPLC) Sour 14.6 20, 8 29, 9 44.0 Main Peak 70.7 64.5 56, 7 45.1 Alkaline 14.7 14.7 13.4 10, 9

According to Table. 8, OD=optical density; RP-HPLC = reverse phase high performance liquid chromatography; E-HPLC = Size Exclusion High Performance Liquid Chromatography; and KO-HPLC=cation exchange high performance liquid chromatography. Acid and alkaline species were defined as the sum of the mAbl peaks that elute from the KO-HPLC column with an earlier or later retention time than the main peak, respectively.

Example 6. Containers.

Compositions containing sNAI were determined to be stable upon filtration sterilization. The Millipore MILLIPAK Filtration Kit was used in the preparation of clinical batches while filtration of identical compositions was used in scientific studies (Millipore MILLEX DURAPORE).

A 5 ml glass vial was filled with a minimum of 2.5 ml of 150 mg/ml mAbl, 5% w/v. sucrose, 25 mm L-arginine hydrochloride, 0.2% w/v. polysorbate 20, 12.5 mM acetate, 20 mM histidine, pH 5.9. An excess of 0.5 ml of the composition was placed in a 5 ml vial to ensure that 2.0 ml of the composition could be withdrawn. Such an excess was not created to compensate for losses during the production of mAbl or composition containing mAbl, degradation during manufacture, degradation during storage (shelf life), or to extend the shelf life.

Compared to storage in glass vials, the stability of formulated mAbl (composition A) was not affected by storage in either a polypropylene tube, or a polystyrene tube, or a polycarbonate tube, or in a glass vial containing pieces of stainless steel (Table 9).

- 19042167

Table 9

Storage temperature Lack of storage 40°C for 14 days storage container Glass Glass Polypropylene Polystyrene Polycarbonate Stainless steel Appearance Satisfactory Satisfactory Satisfactory Satisfactory Satisfactory Satisfactory Haze (OD at 405 nm) 0.00 0.01 0.01 0.02 0.02 0.01 pH 5, 9 5, 9 5, 7 5, 8 5, 8 5, 9 % mAbl (RPHPLC) 100 102 103 107 106 102 % native mAbl (HP-HPLC) 98.4 97.6 97.4 97.5 97.5 96.1 % peak mAbl (KOHPLC) Sour 14.8 18.4 19, 1 18.4 18.4 20.3 Main 70.7 65, 8 65.5 66.0 66.5 65.0 Alkaline 14.5 15, 8 15, 3 15, 6 15, 1 14.7

According to Table. 9, 150 mg/ml mAbl, 5% sucrose, 25 mM arginine hydrochloride, 0.2% PS-20, 20 mM histidine, 12.5 mM acetate, pH 5.9 incubated with/in various substances at 40°C in within 14 days. OD=optical density; RP-HPLC = reverse phase high performance liquid chromatography; E-HPLC = Size Exclusion High Performance Liquid Chromatography; and KOHPLC=cation exchange high performance liquid chromatography. Haze is presented as the relative change in OD at 405 nm compared to the starting material. Acid or basic species are defined as the sum of the mAb1 peaks that elute from the KO-HPLC column with an earlier or later retention time than the main peak, respectively.

Claims (51)

CLAIM 1. The use of a stable liquid pharmaceutical composition, including: (i) a human antibody that specifically binds to the human interleukin-4 receptor alpha (hIL-4Ra) and contains a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 1 and a light chain variable region containing the amino acid sequence of SEQ ID NO : 5; (ii) acetate at a concentration of 10 mM to 15 mM; (iii) histidine at a concentration of 15 mM to 25 mM; (iv) sucrose at a concentration of 2.5% wt./about. up to 10% w/v; (v) polysorbate at a concentration of 0.1% wt./about. up to 0.3% w/v; and (vi) arginine at a concentration of 20 mM to 100 mM; wherein the composition has a pH of 5.6 to 6.2, in a pen or auto-injection device for administration to treat, prevent or alleviate a disease or disorder associated with IL-4 activity or IL-4Ra activation. 2. Use according to claim 1, wherein the human antibody is present at a concentration of 15 mg/ml to 200 mg/ml. 3. Use according to claim 2, wherein the human antibody is present at a concentration of 150 mg/ml±25 mg/ml. 4. Use according to claim 2, wherein the human antibody is present at a concentration of about 150 mg/ml. 5. Use according to claim 2, wherein the human antibody is present at a concentration of about 175 mg/ml. 6. Use according to any one of claims 1 to 5, wherein the acetate is present at a concentration of 12.5 mM±1.85 mM and the histidine is present at a concentration of 20 mM±0.3 mM. 7. Use according to any one of claims 1 to 6, wherein the polysorbate is polysorbate 20 or poly - 20 042167 sorbate 80. 8. Use according to any one of claims 1 to 6, where the polysorbate is polysorbate 20 at a concentration of 0.2%±0.03% w/v. 9. Use according to any one of claims 1 to 6, where the polysorbate is polysorbate 80 at a concentration of 0.2%±0.03% w/v. 10. Use according to any one of claims 1 to 9, where sucrose is present at a concentration of 5%±0.75% w/v. 11. Use according to any one of claims 1 to 10, wherein the arginine is present at a concentration of 25 mM±3.75 mM. 12. Use according to any one of claims 1 to 10, wherein the arginine is present at a concentration of 50 mM ± 75 mM. 13. Use according to any one of claims 1 to 12, wherein the viscosity of the fluid is less than 15 centipoise. 14. Use according to claim 13, where the viscosity of the liquid is 11±1.1 cps or 8.5±0.85 cps. 15. Use according to any one of claims 1 to 14, wherein the osmolality of the liquid is 290±20 mOsm/kg. 16. Use according to any one of claims 1-15, containing from about 1 ml to about 2.5 ml of a stable liquid pharmaceutical composition. 17. Use according to any one of claims 1 to 15, containing 1 ml ± 0.15 ml of a stable liquid pharmaceutical composition. 18. The use of a stable liquid pharmaceutical composition, including: (i) a human antibody at a concentration of 15 mg/mL to 200 mg/mL, wherein the antibody specifically binds to hIL-4Ra and contains a pair of heavy chain variable region and light chain variable region (HCVR/LCVR) amino acid sequences SEQ ID NO: 1 /5; (ii) 12.5 mM±2 mM acetate; (iii) 20 mM±3 mM histidine; (iv) 5%±0.75% w/v sucrose; (v) 0.2%±0.03% polysorbate 80; and (vi) 25 mM±3.75 mM arginine; at pH 5.9±0.5, in a pen or auto-injection device for administration to treat, prevent, or alleviate a disease or disorder associated with IL-4 activity or IL-4Ra activation. 19. Use according to claim 18, wherein the human antibody is present at a concentration of about 150 mg/ml. 20. The use of a stable liquid pharmaceutical composition, including: (i) a human antibody at a concentration of 15 mg/mL to 200 mg/mL, wherein the antibody specifically binds to hIL-4Ra and contains a pair of heavy chain variable region and light chain variable region (HCVR/LCVR) amino acid sequences SEQ ID NO: 1 /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 pH 5.9±0.5, in a pen or auto-injection device for administration to treat, prevent, or alleviate a disease or disorder associated with IL-4 activity or IL-4Ra activation. 21. Use according to claim 20, wherein the human antibody is present at a concentration of about 175 mg/ml. 22. Use according to any one of claims 1 to 21, wherein the stable liquid pharmaceutical composition is contained in an administration pen. 23. Use according to claim 22, wherein the injection pen is a disposable injection pen that is pre-filled with a stable liquid pharmaceutical composition. 24. Use according to any one of claims 1 to 21, wherein the stable liquid pharmaceutical composition is contained in an auto-injection device for administration. 25. Use according to any one of claims 1 to 24, wherein the disease or disorder is a Th2 mediated disease. 26. Use according to any one of claims 1 to 25, wherein the disease or disorder is atopic dermatitis. 27. Use according to any one of claims 1 to 25, wherein the disease or disorder is asthma. 28. Dosage form of a stable liquid pharmaceutical composition, which is a pen or auto-injection device for administration, where the pharmaceutical composition contains: (i) a human antibody that specifically binds to the human interleukin-4 receptor alpha (hIL-4Ra) and contains a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 1 and a light chain variable region containing the amino acid sequence of SEQ ID NO : 5; (ii) acetate at a concentration of 10 mM to 15 mM; (iii) histidine at a concentration of 15 mM to 25 mM; - 21 042167 (iv) sucrose in a concentration of 2.5% wt./about. up to 10% w/v; (v) polysorbate at a concentration of 0.1% wt./about. up to 0.3% w/v; and (vi) arginine at a concentration of 20 mM to 100 mM; where the composition has a pH of 5.6 to 6.2. 29. The dosage form of claim 28, wherein the human antibody is present at a concentration of 15 mg/mL to 200 mg/mL. 30. The dosage form of claim 29, wherein the human antibody is present at a concentration of 150 mg/ml ± 25 mg/ml. 31. The dosage form of claim 29, wherein the human antibody is present at a concentration of about 150 mg/mL. 32. The dosage form of claim 29, wherein the human antibody is present at a concentration of about 175 mg/mL. 33. A dosage form according to any one of claims 29-32, wherein the acetate is present at a concentration of 12.5 mM±1.85 mM and the histidine is present at a concentration of 20 mM±0.3 mM. 34. A dosage form according to any one of claims 29-33, wherein the polysorbate is polysorbate 20 or polysorbate 80. 35. The dosage form according to any one of claims 29-34, where the polysorbate is polysorbate 20 at a concentration of 0.2%±0.03% w/v. 36. Dosage form according to any one of claims 29-34, where the polysorbate is polysorbate 80 at a concentration of 0.2%±0.03% w/v. 37. Dosage form according to any one of paragraphs.29-36, where sucrose is present at a concentration of 5%±0.75% wt./about. 38. A dosage form according to any one of claims 29-37, wherein the arginine is present at a concentration of 25 mM ± 3.75 mM. 39. A dosage form according to any one of claims 29-37, wherein the arginine is present at a concentration of 50 mM ± 7.5 mM. 40. A dosage form according to any one of claims 29-39, wherein the viscosity of the liquid is less than 15 centipoise. 41. The dosage form of claim 40, wherein the viscosity of the liquid is 11 ± 1.1 centipoise or 8.5 ± 0.85 centipoise. 42. A dosage form according to any one of claims 29-41, wherein the osmolality of the liquid is 290±20 mOsm/kg. 43. A dosage form according to any one of claims 29-42, containing from about 1 ml to about 2.5 ml of a stable liquid pharmaceutical composition. 44. A dosage form according to any one of claims 29-42, containing 1 ml ± 0.15 ml of a stable liquid pharmaceutical composition. 45. Dosage form of a stable liquid pharmaceutical composition, which is a pen or auto-injection device for administration, where the pharmaceutical composition contains: (i) a human antibody at a concentration of 15 mg/mL to 200 mg/mL, wherein the antibody specifically binds to hIL-4Ra and contains a pair of heavy chain variable region and light chain variable region (HCVR/LCVR) amino acid sequences SEQ ID NO: 1 /5; (ii) 12.5 mM±2 mM acetate; (iii) 20 mM±3 mM histidine; (iv) 5%±0.75% w/v sucrose; (v) 0.2%±0.03% polysorbate 80; and (vi) 25 mM±3.75 mM arginine; at pH 5.9±0.5, in a pen or auto-injection device for administration to treat, prevent, or alleviate a disease or disorder associated with IL-4 activity or IL-4Ra activation. 46. The dosage form of claim 45, wherein the human antibody is present at a concentration of about 150 mg/mL. 47. Dosage form of a stable liquid pharmaceutical composition, which is a pen or auto-injection device for administration, where the pharmaceutical composition contains: (i) a human antibody at a concentration of 15 mg/mL to 200 mg/mL, wherein the antibody specifically binds to hIL-4Ra and contains a pair of heavy chain variable region and light chain variable region (HCVR/LCVR) amino acid sequences SEQ ID NO: 1 /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 pH 5.9±0.5. - 22 042167 48. The dosage form of claim 47, wherein the human antibody is present at a concentration of about 175 mg/ml. 49. A dosage form according to any one of claims 29-4 8, wherein the stable liquid pharmaceutical composition is contained in an administration pen. 50. The dosage form of claim 49, wherein the injection pen is a disposable injection pen that is pre-filled with a stable liquid pharmaceutical composition. 51. A dosage form according to any one of claims 29-48, wherein the stable liquid pharmaceutical composition is contained in an auto-injection device for administration.