A stabilized preparation containing an anti-IL-33 antibody

Abstract

The present invention provides pharmaceutical formulations containing an antibody that specifically binds to human interleukin-33 (hIL-33). In addition to the anti-IL-33 antibody, the formulation may contain a buffer, at least one amino acid, at least one sugar, or at least one non-ionic surfactant. The pharmaceutical formulations of the present invention exhibit a significant degree of antibody stability after storage for several months and after exposure to thermal and other physical stresses.

Description

Technical Field

[0001] Reference to Sequence Listing This application incorporates by reference a Sequence Listing submitted in computer-readable form as file 10516WO01-Sequence.txt, created on Feb. 20, 2020, and containing 13,659 bytes.

[0002] The present invention relates to the field of therapeutic antibody formulations. More specifically, the present invention relates to the field of pharmaceutical formulations comprising human antibodies that specifically bind to human interleukin-33.

Background Art

[0003] Interleukin-33 (IL-33) is a ligand for ST2, a toll-like / interleukin-1 receptor superfamily member that associates with the accessory protein IL-1RAcP (see, for example, Non-Patent Document 1, Non-Patent Document 2, Non-Patent Document 3, Patent Document 1, Patent Document 2 for reviews). Upon activation of ST2 / IL-1RAcP by IL-33, a signaling cascade is triggered through downstream molecules such as MyD88 (myeloid differentiation factor 88) and TRAF6 (TNF receptor-associated factor 6), resulting in, in particular, activation of NFκB (nuclear factor-κB). IL-33 signaling is involved as a factor in a variety of diseases and disorders. (Non-Patent Document 3).

[0004] Therapeutic polymers (e.g., antibodies) must be formulated in a manner that not only makes the molecules suitable for administration to a patient but also maintains their stability during storage. For example, therapeutic antibodies in solution are prone to degradation, aggregation, and / or unwanted chemical modifications unless the solution is properly formulated. The stability of an antibody in a liquid formulation depends not only on the type of excipients used in the formulation but also on the amounts and ratios of the excipients relative to each other. Furthermore, when preparing a liquid antibody formulation, other considerations besides stability must be taken into account. Examples of such additional considerations include the viscosity of the solution and the concentration of antibody that can be accommodated by a given formulation. Therefore, when formulating a therapeutic antibody, great care must be taken to arrive at a formulation that maintains stability, contains an appropriate concentration of antibody, and has a suitable viscosity as well as other properties that enable the formulation to be conveniently administered to a patient.

[0005] Antibodies against human interleukin-33 (hIL-33) are an example of a therapeutic-related polymer that requires an appropriate formulation.

[0006] Anti-hIL-33 antibodies are known in the art (see, e.g., Patent Document 3), but there remains a need for pharmaceutical formulations containing anti-hIL-33 antibodies that are sufficiently stable and suitable for administration to patients.

Prior Art Documents

Patent Documents

[0007]

Patent Document 1

Patent Document 2

Patent Document 3

Non-Patent Documents

[0008]

Non-Patent Document 1

[0009] Provided are stable liquid pharmaceutical formulations comprising an anti-IL-33 antibody and one or more excipients, as well as kits comprising such formulations and their uses.

[0010] In one aspect, provided is a stable liquid pharmaceutical formulation comprising (i) a human antibody that specifically binds to human interleukin-33 (hIL-33), (ii) a buffer, (iii) an amino acid, (iv) a thermal stabilizer, and (v) an organic co-solvent. In some embodiments, the buffer is acetate or histidine at a concentration of 1 mM to 40 mM. In some embodiments, the buffer is acetate or histidine at a concentration of 1 mM to 20 mM. In some embodiments, the amino acid is arginine or glutamic acid at a concentration of 30 mM to 110 mM. In some embodiments, the thermal stabilizer is sucrose at a concentration of 1% w / v to 20% w / v. Optionally, the thermal stabilizer is sucrose at a concentration of 1% w / v to 10% w / v. Optionally, the organic co-solvent is a surfactant at a concentration of 0.01% w / v to 0.15% w / v. In some embodiments, the surfactant is polysorbate 80. In some embodiments, the antibody is present at a concentration of 1 mg / ml to 200 mg / ml. Optionally, the antibody is present at a concentration of 15 mg / ml to 150 mg / ml.

[0011] In various embodiments of the formulation, the antibody comprises the complementarity determining regions (HCDR1-HCDR2-HCDR3) of the heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 2, and the complementarity determining regions (LCDR1-LCDR2-LCDR3) of the light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 10. Optionally, the antibody comprises the HCDR1-HCDR2-HCDR3 regions comprising the amino acid sequences of SEQ ID NOs: 4-6-8, respectively, and the LCDR1-LCDR2-LCDR3 regions comprising the amino acid sequences of SEQ ID NOs: 12-14-16, respectively. In some embodiments, the antibody comprises a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 2, and a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 10. In some embodiments, the antibody has a human IgG heavy chain constant region. In some embodiments, the heavy chain constant region is of isotype IgG1. In some embodiments, the heavy chain constant region is of isotype IgG4. In some embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 18, and a light chain comprising the amino acid sequence of SEQ ID NO: 20.

[0012] In some embodiments, the stable liquid pharmaceutical formulation comprises: (i) a human antibody that specifically binds to hIL-33 at about 15 mg / ml to about 150 mg / mL; (ii) about 5 mM to about 15 mM acetate; (iii) about 60 mM to about 80 mM arginine hydrochloride; (iv) about 3% w / v to about 7% w / v sucrose; and (v) about 0.06% w / v to about 0.1% w / v polysorbate 80. Optionally, the formulation has a pH of about 5 to about 5.6.

[0013] In some embodiments, the stable liquid pharmaceutical formulation comprises (i) an antibody at about 15 mg / ml ± 1.5 mg / ml, (ii) acetate at about 10 mM ± 2 mM, (iii) arginine hydrochloride at about 70 mM ± 14 mM, (iv) sucrose at about 5% w / v ± 1% w / v, and (iv) polysorbate 80 at about 0.08% ± 0.016% w / v. In some embodiments, the stable liquid pharmaceutical formulation comprises (i) an antibody at about 75 mg / ml ± 5 mg / ml, (ii) acetate at about 10 mM ± 2 mM, (iii) arginine hydrochloride at about 70 mM ± 14 mM, (iv) sucrose at about 5% w / v ± 1% w / v, and (iv) polysorbate 80 at about 0.08% ± 0.016% w / v. In some embodiments, the stable liquid pharmaceutical formulation comprises (i) an antibody at about 150 mg / ml ± 15 mg / ml, (ii) acetate at about 10 mM ± 2 mM, (iii) arginine hydrochloride at about 70 mM ± 14 mM, (iv) sucrose at about 5% w / v ± 1% w / v, and (iv) polysorbate 80 at about 0.08% ± 0.016% w / v. In some embodiments, the stable liquid pharmaceutical formulation comprises (i) an antibody at about 15 mg / ml ± 1.5 mg / ml, (ii) acetate at about 10 mM ± 1 mM, (iii) arginine hydrochloride at about 70 mM ± 7 mM, (iv) sucrose at about 5% w / v ± 0.5% w / v, and (iv) polysorbate 80 at about 0.08% ± 0.008% w / v. In some embodiments, the stable liquid pharmaceutical formulation comprises (i) an antibody at about 75 mg / ml ± 5 mg / ml, (ii) acetate at about 10 mM ± 1 mM, (iii) arginine hydrochloride at about 70 mM ± 7 mM, (iv) sucrose at about 5% w / v ± 0.5% w / v, and (iv) polysorbate 80 at about 0.08% ± 0.008% w / v. In some embodiments, the stable liquid pharmaceutical formulation comprises (i) an antibody at about 150 mg / ml ± 15 mg / ml, (ii) acetate at about 10 mM ± 1 mM, (iii) arginine hydrochloride at about 70 mM ± 7 mM, (iv) sucrose at about 5% w / v ± 0.5% w / v, and (iv) polysorbate 80 at about 0.08% ± 0.008% w / v. Optionally, the pH of the formulation is between 5.2 and 5.4. In some embodiments, the pH of the formulation is about 5.3.In some embodiments, the stable liquid pharmaceutical formulation comprises (i) an antibody at about 15 mg / ml ± 1.5 mg / ml, (ii) acetate at about 10 mM ± 2 mM, (iii) arginine hydrochloride at about 70 mM ± 14 mM, (iv) sucrose at about 5% w / v ± 1% w / v, and (iv) polysorbate 80 at about 0.08% ± 0.04% w / v. In some embodiments, the stable liquid pharmaceutical formulation comprises (i) an antibody at about 75 mg / ml ± 8 mg / ml, (ii) acetate at about 10 mM ± 2 mM, (iii) arginine hydrochloride at about 70 mM ± 14 mM, (iv) sucrose at about 5% w / v ± 1% w / v, and (iv) polysorbate 80 at about 0.08% ± 0.04% w / v. In some embodiments, the stable liquid pharmaceutical formulation comprises (i) an antibody at about 150 mg / ml ± 15 mg / ml, (ii) acetate at about 10 mM ± 2 mM, (iii) arginine hydrochloride at about 70 mM ± 14 mM, (iv) sucrose at about 5% w / v ± 1% w / v, and (iv) polysorbate 80 at about 0.08% ± 0.04% w / v. Optionally, the pH of the formulation is from 5.2 to 5.4. In some embodiments, the pH of the formulation is about 5.3.

[0014] In some embodiments, a stable liquid pharmaceutical formulation comprises (i) a human antibody that specifically binds to human interleukin-33 (hIL-33) at a concentration of 15 ± 1.5 mg / ml to 150 ± 15 mg / ml, wherein the antibody comprises an HCVR comprising HCDR1, HCDR2, and HCDR3 regions comprising the amino acid sequences of SEQ ID NOs: 4, 6, and 8, respectively, and an LCDR1, LCDR2, and LCDR3 regions comprising the amino acid sequences of SEQ ID NOs: 12, 14, and 16, respectively, a human antibody, (ii) 10 mM ± 2 mM acetate, (iii) 70 mM ± 14 mM arginine hydrochloride, (iv) 5% w / v ± 1% w / v sucrose, (iv) about 0.08% ± 0.016% w / v polysorbate 80, and the formulation has a pH of 5.1 to 5.5. In some embodiments, a stable liquid pharmaceutical formulation comprises (i) a human antibody that specifically binds to human interleukin-33 (hIL-33) at a concentration of 15 ± 1.5 mg / ml to 150 ± 15 mg / ml, wherein the antibody comprises an HCVR comprising HCDR1, HCDR2, and HCDR3 regions comprising the amino acid sequences of SEQ ID NOs: 4, 6, and 8, respectively, and an LCDR1, LCDR2, and LCDR3 regions comprising the amino acid sequences of SEQ ID NOs: 12, 14, and 16, respectively, a human antibody, (ii) 10 mM ± 1 mM acetate, (iii) 70 mM ± 7 mM arginine hydrochloride, (iv) 5% w / v ± 0.5% w / v sucrose, (iv) about 0.08% ± 0.008% w / v polysorbate 80, and the formulation has a pH of 5.1 to 5.5. Optionally, the antibody comprises a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 2 and a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 10. In some embodiments, the antibody has a human IgG heavy chain constant region. In some embodiments, the heavy chain constant region is of isotype IgG1. In some embodiments, the heavy chain constant region is of isotype IgG4.

[0015] In some embodiments, a stable liquid pharmaceutical formulation comprises: (i) a human antibody that specifically binds to human interleukin-33 (hIL-33) at a concentration of 15 ± 1.5 mg / ml to 150 ± 15 mg / ml, wherein the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 18 and a light chain comprising the amino acid sequence of SEQ ID NO: 20; (ii) 10 mM ± 2 mM acetate; (iii) 70 mM ± 14 mM arginine hydrochloride; (iv) 5% w / v ± 1% w / v sucrose; and (iv) about 0.08% ± 0.016% w / v polysorbate 80, and the formulation has a pH of 5.1 to 5.5. In some embodiments, a stable liquid pharmaceutical formulation comprises: (i) a human antibody that specifically binds to human interleukin-33 (hIL-33) at a concentration of 15 ± 1.5 mg / ml to 150 ± 15 mg / ml, wherein the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 18 and a light chain comprising the amino acid sequence of SEQ ID NO: 20; (ii) 10 mM ± 1 mM acetate; (iii) 70 mM ± 7 mM arginine hydrochloride; (iv) 5% w / v ± 0.5% w / v sucrose; and (iv) about 0.08% ± 0.008% w / v polysorbate 80, and the formulation has a pH of 5.1 to 5.5.

[0016] In some embodiments, a stable liquid pharmaceutical formulation contains at least 90% of the native form of the antibody after storage at 5°C for 2 months as determined by size exclusion - ultra performance liquid chromatography (SE-UPLC). Optionally, the formulation contains at least 95% of the native form of the antibody after storage at 5°C for 2 months as determined by SE-UPLC. Optionally, the formulation contains at least 99% of the native form of the antibody after storage at 5°C for 2 months as determined by SE-UPLC.

[0017] In some cases, the formulation contains at least 95% of the native form of the antibody after 9 months of storage at -20°C as determined by SE-UPLC. In some embodiments, the stable liquid pharmaceutical formulation contains at least 97.5% of the native form of the antibody after 9 months of storage at -20°C as determined by SE-UPLC. In some cases, the formulation contains at least 99% of the native form of the antibody after 9 months of storage at -20°C as determined by SE-UPLC. In some cases, the formulation contains at least 95% of the native form of the antibody after 12 months of storage at -20°C as determined by SE-UPLC. In some embodiments, the stable liquid pharmaceutical formulation contains at least 97.5% of the native form of the antibody after 12 months of storage at -20°C as determined by SE-UPLC. In some cases, the formulation contains at least 99% of the native form of the antibody after 12 months of storage at -20°C as determined by SE-UPLC. In some cases, the formulation contains at least 95% of the native form of the antibody after 18 months of storage at -20°C as determined by SE-UPLC. In some embodiments, the stable liquid pharmaceutical formulation contains at least 97.5% of the native form of the antibody after 18 months of storage at -20°C as determined by SE-UPLC. In some cases, the formulation contains at least 99% of the native form of the antibody after 18 months of storage at -20°C as determined by SE-UPLC.

[0018] In some cases, the formulation contains at least 95% of the native form of the antibody after storage at 2 - 8°C for 9 months as determined by SE-UPLC. In some embodiments, the stable liquid pharmaceutical formulation contains at least 97.5% of the native form of the antibody after storage at 2 - 8°C for 9 months as determined by SE-UPLC. In some cases, the formulation contains at least 99% of the native form of the antibody after storage at 2 - 8°C for 9 months as determined by SE-UPLC. In some cases, the formulation contains at least 95% of the native form of the antibody after storage at 2 - 8°C for 12 months as determined by SE-UPLC. In some embodiments, the stable liquid pharmaceutical formulation contains at least 97.5% of the native form of the antibody after storage at 2 - 8°C for 12 months as determined by SE-UPLC. In some cases, the formulation contains at least 99% of the native form of the antibody after storage at 2 - 8°C for 12 months as determined by SE-UPLC. In some cases, the formulation contains at least 95% of the native form of the antibody after storage at 2 - 8°C for 18 months as determined by SE-UPLC. In some embodiments, the stable liquid pharmaceutical formulation contains at least 97.5% of the native form of the antibody after storage at 2 - 8°C for 18 months as determined by SE-UPLC. In some cases, the formulation contains at least 99% of the native form of the antibody after storage at 2 - 8°C for 18 months as determined by SE-UPLC.

[0019] In some embodiments, the stable liquid pharmaceutical formulation contains 2% or less of high molecular weight (HMW) species after storage at 5°C for 2 months as determined by SE-UPLC. In some cases, the formulation contains 1% or less of HMW species after storage at 5°C for 2 months as determined by SE-UPLC. In some cases, the formulation contains 0.6% or less of HMW species after storage at 5°C for 2 months as determined by SE-UPLC.

[0020] In some embodiments, the stable liquid pharmaceutical formulation contains 2% or less of HMW species after storage at -20°C for 9 months as determined by SE-UPLC. Optionally, the formulation contains 1% or less of HMW species after storage at -20°C for 9 months as determined by SE-UPLC. Optionally, the formulation contains 0.5% or less of HMW species after storage at -20°C for 9 months as determined by SE-UPLC. In some embodiments, the stable liquid pharmaceutical formulation contains 2% or less of HMW species after storage at -20°C for 12 months as determined by SE-UPLC. Optionally, the formulation contains 1% or less of HMW species after storage at -20°C for 12 months as determined by SE-UPLC. Optionally, the formulation contains 0.5% or less of HMW species after storage at -20°C for 12 months as determined by SE-UPLC. In some embodiments, the stable liquid pharmaceutical formulation contains 2% or less of HMW species after storage at -20°C for 18 months as determined by SE-UPLC. Optionally, the formulation contains 1% or less of HMW species after storage at -20°C for 18 months as determined by SE-UPLC. Optionally, the formulation contains 0.5% or less of HMW species after storage at -20°C for 18 months as determined by SE-UPLC.

[0021] In some embodiments, a stable liquid pharmaceutical formulation contains 2% or less HMW species after storage at 2-8°C for 9 months as determined by SE-UPLC. Optionally, the formulation contains 1% or less HMW species after storage at 2-8°C for 9 months as determined by SE-UPLC. Optionally, the formulation contains 0.7% or less HMW species after storage at 2-8°C for 9 months as determined by SE-UPLC. In some embodiments, a stable liquid pharmaceutical formulation contains 2% or less HMW species after storage at 2-8°C for 12 months as determined by SE-UPLC. Optionally, the formulation contains 1% or less HMW species after storage at 2-8°C for 12 months as determined by SE-UPLC. Optionally, the formulation contains 0.7% or less HMW species after storage at 2-8°C for 12 months as determined by SE-UPLC. In some embodiments, a stable liquid pharmaceutical formulation contains 2% or less HMW species after storage at 2-8°C for 18 months as determined by SE-UPLC. Optionally, the formulation contains 1% or less HMW species after storage at 2-8°C for 18 months as determined by SE-UPLC. Optionally, the formulation contains 0.7% or less HMW species after storage at 2-8°C for 18 months as determined by SE-UPLC.

[0022] In some embodiments, the pharmaceutical formulation exhibits a viscosity of less than about 15 cP, less than about 12 cP, or less than about 10 cP when measured at 20°C.

[0023] In some embodiments, the stable liquid pharmaceutical formulation is contained in a glass vial, syringe, or large volume device or bolus syringe. In some embodiments, the syringe includes a fluorocarbon-coated plunger. In some embodiments, the syringe is a low tungsten syringe. In some embodiments, the syringe contains up to 2500 ppb of tungsten. In some embodiments, the syringe contains about 250 - 750 ppb of tungsten. In some embodiments, the syringe is a pre-filled syringe. In some embodiments, the syringe is a pre-filled staked needle syringe.

[0024] In another aspect, in any of the embodiments described above or herein, a pen or auto-injector delivery device containing a stable liquid pharmaceutical formulation is provided. Optionally, the delivery device is a disposable pen delivery device. Optionally, the delivery device is a reusable pen delivery device.

[0025] In another aspect, in any of the embodiments described above or herein, a container containing a stable liquid pharmaceutical formulation is provided.

[0026] In another aspect, in any of the embodiments described above or herein, a safety system delivery device containing a stable liquid pharmaceutical formulation is provided. In some embodiments, the safety system delivery device includes a safety sleeve configured to extend by manual operation. In some embodiments, the safety system delivery device includes a safety sleeve configured to automatically extend after injection of the stable liquid pharmaceutical formulation.

[0027] In another aspect, a kit is provided that includes (i) a container containing a stable liquid pharmaceutical formulation as described above or herein, and (ii) labeling for use of the pharmaceutical formulation. In some embodiments, the labeling lists subcutaneous administration of the pharmaceutical formulation. In some embodiments, the labeling lists intravenous administration of the pharmaceutical formulation.

[0028] In another aspect, the present invention provides a unit dosage form comprising the stable liquid pharmaceutical formulation described above or herein, wherein the anti-IL-33 antibody is present in an amount of 1 mg to 500 mg. In some cases, the anti-IL-33 antibody is present in an amount of about 150 mg. In some cases, the anti-IL-33 antibody is present in an amount of about 300 mg. In some embodiments of the unit dosage form, the formulation is contained in a syringe. In some cases, the syringe is a pre-filled syringe.

[0029] In various embodiments, any of the features or components of the embodiments described above or herein may be combined, and such combinations are encompassed within the scope of the present disclosure. Any specific value described above or herein may be combined with another related value described above or herein to enumerate a range with values representing the upper and lower limits of the range, and such ranges are encompassed within the scope of the present disclosure. Each of the values described above or herein may be represented with a variation of 1%, 5%, 10%, or 20%. For example, a concentration of 10 mM may be represented as 10 mM ± 0.1 mM (1% variation), 10 mM ± 0.5 mM (5% variation), 10 mM ± 1 mM (10% variation), or 10 mM ± 2 mM (20% variation).

[0030] Other embodiments will be apparent from a review of the manner of making and using the invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0031]

Figure 1

Figure 2

Figure 3

Figure 4A

Figure 4B

Mode for Carrying Out the Invention

[0032] Before the present invention is described, it should be understood that, since the specific methods and experimental conditions described may vary, the present invention is not limited to such methods and conditions. It should also be understood that the terms used herein are for the purpose of describing only specific embodiments and are not intended to be limiting, as the scope of the present invention is limited only by the appended claims.

[0033] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. As used herein, the term "about" when used in relation to a particular recited numerical value means that the value can vary by up to 1% from the recited value. For example, as used herein, the expression "about 100" includes 99 and 101 and all values in between (e.g., 99.1, 99.2, 99.3, 99.4, etc.).

[0034] Any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, but next, exemplary methods and materials will be described. All patents, applications, and non-patent publications mentioned herein are incorporated herein by reference in their entirety.

[0035] Pharmaceutical formulation As used herein, the expression "pharmaceutical formulation" means a combination of at least one active ingredient (e.g., an anti-IL-33 antibody capable of exerting a biological effect in a human or non-human animal) and at least one inactive ingredient suitable for therapeutic administration to a human or non-human animal when combined with the active ingredient or one or more additional inactive ingredients. As used herein, the term "formulation" means "pharmaceutical formulation" unless specifically indicated otherwise. The present invention provides a pharmaceutical formulation comprising at least one therapeutic polypeptide. According to certain embodiments of the present invention, the therapeutic polypeptide is an antibody that specifically binds to human interleukin-33 (hIL-33) or an antigen-binding fragment thereof. More specifically, the present invention includes a formulation comprising (i) a human antibody that specifically binds to hIL-33, (ii) a buffer, (iii) a heat stabilizer, (iv) a surfactant (which is also an organic co-solvent or an interfacial stabilizer), and (v) a viscosity modifier. Additional components may be included in the formulations of the present invention if such components do not significantly interfere with the viscosity and stability of the formulation. Specific exemplary components and formulations included in the present invention are described in detail below.

[0036] In certain embodiments, the pharmaceutical formulation of the present invention can be a fluid formulation. As used herein, the expression "fluid formulation" means a mixture of at least two components that are predominantly in a fluid state at about 2°C to about 45°C. Fluid formulations include, among other things, liquid formulations. Fluid formulations can be of low, medium, or high viscosity depending on their specific components.

[0037] An antibody that specifically binds to human IL-33 The pharmaceutical formulation of the present invention may comprise a human antibody that specifically binds to hIL-33 or an antigen-binding fragment thereof. As used herein, the term "hIL-33" refers to the human IL-33 protein.

[0038] As used herein, the term "antibody" is generally intended to refer to an immunoglobulin molecule comprising four polypeptide chains, two heavy (H) chains and two light (L) chains interconnected by disulfide bonds, as well as multimers thereof (e.g., IgM), but immunoglobulin molecules consisting of only heavy chains (i.e., lacking light chains) are also included within the definition of the term "antibody". Each heavy chain comprises a heavy chain variable region (abbreviated herein as HCVR or VH) and a heavy chain constant region. The heavy chain constant region comprises three domains, CH1, CH2 and CH3. Each light chain comprises a light chain variable region (abbreviated herein as LCVR or VL) and a light chain constant region. The light chain constant region comprises one domain (CL1). The VH and VL regions can be further subdivided into hypervariable regions called complementarity determining regions (CDRs) interspersed with more conserved regions called framework regions (FRs). Each VH and VL consists, in order from the amino terminus to the carboxy terminus, of three CDRs and four FRs arranged as follows: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.

[0039] In certain embodiments of the present invention, the anti-IL-33 antibody of the present invention is a human antibody. As used in the present invention, the term "human antibody" is intended to include antibodies having variable and constant regions derived from human germline immunoglobulin sequences. The human antibodies of the present invention may include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by in vitro random or site-directed mutagenesis or by somatic mutations in vivo), for example, in the CDRs, particularly CDR3. However, as used herein, the term "human antibody" is not intended to include antibodies in which CDR sequences derived from the germline of another mammalian species, such as a mouse, have been transplanted into a human framework sequence. In various embodiments, the anti-IL-33 antibody is a human IgG antibody. In various embodiments, the anti-IL-33 antibody is a human antibody of isotype IgG1, IgG2, IgG3 or IgG4, or a mixed isotype. In some embodiments, the anti-IL-33 antibody is a human IgG1 antibody. In some embodiments, the anti-IL-33 antibody is a human IgG4 antibody. In any of the embodiments described above or herein, the anti-IL-33 antibody may include a human kappa light chain. In any of the embodiments described above or herein, the anti-IL-33 antibody may include a human lambda light chain.

[0040] In some embodiments, the antibodies of the invention can be recombinant human antibodies. As used herein, the term "recombinant human antibody" refers to all human antibodies prepared, expressed, made, or isolated by recombinant means, such as antibodies expressed using a recombinant expression vector transfected into a host cell, antibodies isolated from a recombinant combinatorial human antibody library, antibodies isolated from an animal that is transgenic for human immunoglobulin genes (e.g., a mouse; see, e.g., Taylor et al. (1992) Nucl. Acids Res. 20:6287-6295), or antibodies prepared, expressed, made, or isolated by any other means that involves splicing of human immunoglobulin gene sequences to other DNA sequences. Such recombinant human antibodies have variable and constant regions derived from human germline immunoglobulin sequences. However, in certain embodiments, such recombinant human antibodies are subjected to in vitro mutagenesis (or in vivo somatic mutagenesis when transgenic animals for human Ig sequences are used), and thus the amino acid sequences of the V H and V L regions of the recombinant antibody are related to and derived from human germline V H and V L sequences but are sequences that may not naturally occur within the human antibody germline repertoire in vivo.

[0041] As used herein, the term "antigen-binding portion" or "antibody fragment" of an antibody refers to one or more fragments of the antibody that retain the ability to specifically bind to hIL-33.

[0042] As used herein, an "isolated antibody" is intended to refer to an antibody that is substantially free of other antibodies having different antigen specificities (e.g., an isolated antibody that specifically binds to hIL-33 is substantially free of antibodies that specifically bind to antigens other than hIL-33).

[0043] "Specifically binds" or similar terms mean that an antibody or an 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 1×10 -6 M or greater. Methods for determining whether two molecules specifically bind are well known in the art and include, for example, equilibrium dialysis, surface plasmon resonance, and the like. However, an isolated antibody that specifically binds to hIL-33 may have cross-reactivity to other antigens, such as IL-33 molecules from other species (orthologs). In the context of the present invention, a multispecific (e.g., bispecific) antibody that binds to hIL-33, as well as one or more additional antigens, is considered to "specifically bind" to hIL-33. Furthermore, an isolated antibody may be substantially free of other cellular materials and / or chemicals.

[0044] Exemplary anti-hIL-33 antibodies that may be included in the pharmaceutical formulations of the present invention are defined in WO2014 / 164959, the disclosure of which is incorporated herein by reference in its entirety.

[0045] According to certain embodiments of the present invention, an anti-hIL-33 antibody or an antigen-binding fragment thereof comprises heavy chain complementarity determining regions HCDR1-HCDR2-HCDR3 comprising the amino acid sequences of SEQ ID NOs: 4-6-8, respectively. According to certain embodiments of the present invention, an anti-hIL-33 antibody or an antigen-binding fragment thereof comprises light chain complementarity determining regions LCDR1-LCDR2-LCDR3 comprising the amino acid sequences of SEQ ID NOs: 12-14-16, respectively.

[0046] In certain embodiments, the anti-hIL-33 antibody or antigen-binding fragment thereof comprises a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 2. In certain embodiments, the anti-hIL-33 antibody or antigen-binding fragment thereof comprises a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 10. In certain embodiments, the anti-hIL-33 antibody or antigen-binding fragment thereof comprises a HCVR / LCVR amino acid sequence pair comprising the amino acid sequence of SEQ ID NO: 2 / 10. In some embodiments, the anti-IL-33 antibody comprises a HCVR / LCVR comprising the amino acid sequence of SEQ ID NO: 2 / 10, respectively, and a human IgG1 heavy chain constant region. In some embodiments, the anti-IL-33 antibody comprises a HCVR / LCVR comprising the amino acid sequence of SEQ ID NO: 2 / 10, respectively, and a human IgG4 heavy chain constant region. In some embodiments, the anti-IL-33 antibody comprises a HCVR / LCVR comprising the amino acid sequence of SEQ ID NO: 2 / 10, respectively, and a human IgG heavy chain constant region. In some embodiments, the anti-IL-33 antibody comprises a HCVR / LCVR comprising the amino acid sequence of SEQ ID NO: 2 / 10, respectively, and a human IgG1 or IgG4 heavy chain constant region. In some embodiments, the anti-IL-33 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 18 and a light chain comprising the amino acid sequence of SEQ ID NO: 20. An anti-IL-33 antibody having a HCVR comprising the amino acid sequence of SEQ ID NO: 2 and a LCVR comprising the amino acid sequence of SEQ ID NO: 10 is referred to herein as mAb1. This antibody has a heavy chain comprising the amino acid sequence of SEQ ID NO: 18 and a light chain comprising the amino acid sequence of SEQ ID NO: 20.

[0047] The amount of the antibody or its antigen-binding fragment contained within the pharmaceutical formulation of the present invention can vary depending on the desired specific characteristics of the formulation, as well as the specific circumstances and purposes for which the formulation is intended to be used. In certain embodiments, the pharmaceutical formulation can contain from about 1 mg / mL to about 500 mg / mL of the antibody, from about 5 mg / mL to about 400 mg / mL of the antibody, from about 5 mg / mL to about 200 mg / mL of the antibody, from about 15 mg / mL to about 150 mg / mL, from about 25 mg / mL to about 180 mg / mL of the antibody, from about 25 mg / mL to about 150 mg / mL of the antibody, from about 50 mg / mL to about 100 mg / mL, from about 50 mg / mL to about 150 mg / mL, or from about 140 mg / mL to about 160 mg / mL of the antibody. For example, the formulation of the present invention can be a liquid formulation containing an antibody or its antigen-binding fragment that specifically binds to hIL-33 at about 1 mg / mL, about 2 mg / mL, about 5 mg / mL, about 10 mg / mL, about 15 mg / mL, about 20 mg / mL, about 25 mg / mL, about 30 mg / mL, about 35 mg / mL, about 40 mg / mL, about 45 mg / mL, about 50 mg / mL, about 55 mg / mL, about 60 mg / mL, about 65 mg / mL, about 70 mg / mL, about 75 mg / mL, about 80 mg / mL, about 85 mg / mL, 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.In certain embodiments, the pharmaceutical formulation may be a liquid formulation containing an antibody at 5 ± 0.75 mg / mL to 150 ± 22.5 mg / mL, 7.5 ± 1.125 mg / mL to 140 ± 21 mg / mL, 10 ± 1.5 mg / mL to 130 ± 19.5 mg / mL, 12.5 ± 1.875 mg / mL to 120 ± 18 mg / mL, 15 ± 2.25 mg / mL to 110 ± 16.5 mg / mL, 17.5 ± 2.625 mg / mL to 100 ± 15 mg / mL, 20 ± 3 mg / mL to 90 ± 13.5 mg / mL, 22.5 ± 3.375 mg / mL to 80 ± 12 mg / mL, 25 ± 3.75 mg / mL to 70 ± 10.5 mg / mL, 27.5 ± 4.125 mg / mL to 60 ± 9 mg / mL, 30 ± 4.5 mg / mL to 50 ± 7.5 mg / mL, 25 ± 3.75 mg / mL, or 50 ± 7.5 mg / ml. In some embodiments, the pharmaceutical formulation contains an anti-IL-33 antibody at 15 ± 0.15 mg / ml to 150 ± 1.5 mg / ml. Optionally, the pharmaceutical formulation contains an anti-IL-33 antibody at 75 mg / mL ± 3.75 mg / mL. Optionally, the pharmaceutical formulation contains an anti-IL-33 antibody at 150 mg / mL ± 7.5 mg / mL.

[0048] Biological equivalents The present invention includes antibodies having an amino acid sequence that differs from those of the exemplary molecules disclosed herein but retains the ability to bind to hIL-33. Such variant molecules may contain one or more additions, deletions, or substitutions of amino acids when compared to the parent sequence, but may exhibit biological activity that is essentially equivalent to that of the antibodies discussed herein.

[0049] The present invention includes antigen-binding molecules that are biologically equivalent to any of the exemplary antibodies defined herein. For example, two antibodies are considered biologically equivalent if, when administered at the same molar dose under similar experimental conditions, whether as a single dose or multiple doses, they are pharmaceutical equivalents or alternatives such that they do not show significant differences in the rate and extent of absorption. Some antibodies will be considered equivalents or alternatives if they are equivalent in the extent of their absorption but not in their rate of absorption, provided that such differences in the rate of absorption are intentional and reflected in the labeling and are considered not essential for achieving an effective body drug concentration upon chronic use and not medically significant for the particular drug being studied, and thus can be considered biologically equivalent.

[0050] In one embodiment, two antibodies are biologically equivalent if there are no clinically meaningful differences in their safety, purity, and potency.

[0051] In one embodiment, two antibodies are biologically equivalent if a patient can be switched one or more times between a reference product and a biological product without an expected increase in the risk of adverse effects, including a clinically significant change in immunogenicity or attenuation of efficacy, compared to continuous therapy without such switching.

[0052] Biological equivalence can be demonstrated by in vivo and in vitro methods. Biological equivalence measurements include, for example, (a) in vivo tests in humans or other mammals in which the concentration of the antibody or its metabolites is measured as a function of time in blood, plasma, serum, or other biological fluids, (b) in vitro tests that correlate with and reasonably predict human in vivo bioavailability data, (c) in vivo tests in humans or other mammals in which an appropriate acute pharmacological effect of the antibody (or its target) is measured as a function of time, and (d) well-controlled clinical trials that establish the safety, efficacy, or bioavailability or biological equivalence of the antigen-binding protein.

[0053] Formulation Excipients and pH The pharmaceutical formulations of the present invention include one or more excipients. As used herein, the term "excipient" means any non-therapeutic agent added to a formulation to provide a desired consistency, viscosity or stabilizing effect.

[0054] In certain embodiments, the pharmaceutical formulations of the present invention include at least one amino acid (e.g., arginine, histidine or glutamic acid). In some embodiments, the amino acid is arginine. In some embodiments, arginine is provided in the form of arginine hydrochloride. In some embodiments, the amino acid is a combination of arginine and glutamic acid. Optionally, the amino acid (e.g., arginine) functions as a viscosity modifier for the anti-IL-33 antibody formulation.

[0055] The amount of amino acid contained within the pharmaceutical formulations of the present invention can vary depending on the desired specific characteristics of the formulation, as well as the particular circumstances and purposes for which the formulation is intended to be used. In certain embodiments, the formulation can include from 1 mM to about 200 mM amino acid, from about 5 mM to about 150 mM, from about 25 mM to about 125 mM amino acid, from about 50 mM to about 100 mM amino acid, from about 50 mM to about 90 mM amino acid, from about 60 mM to about 80 mM amino acid, or from about 65 mM to about 75 mM amino acid. For example, the pharmaceutical formulations of the present invention can include about 1 mM, about 5 mM, about 10 mM, about 15 mM, about 20 mM, about 25 mM, about 30 mM, about 35 mM, about 40 mM, about 45 mM, about 50 mM, about 55 mM, about 60 mM, about 65 mM, about 70 mM, about 75 mM, about 80 mM, about 85 mM, about 90 mM, about 95 mM, about 100 mM, about 105 mM, about 110 mM, about 115 mM, about 120 mM, or about 125 mM amino acid (e.g., arginine). In some embodiments, the formulation contains about 70 mM amino acid (e.g., arginine).

[0056] The pharmaceutical formulation of the present invention may also contain one or more carbohydrates, for example, one or more sugars. The sugar can be a reducing sugar or a non-reducing sugar. A "reducing sugar" includes, for example, sugars having a ketone or aldehyde group and includes a reactive hemiacetal group that enables the sugar to function as a reducing agent. Specific examples of reducing sugars include fructose, glucose, glyceraldehyde, lactose, arabinose, mannose, xylose, ribose, rhamnose, galactose, and maltose. Non-reducing sugars are acetals and can contain anomeric carbons that do not substantially react with amino acids or polypeptides to initiate the Maillard reaction. Specific examples of non-reducing sugars include sucrose, trehalose, sorbose, sucralose, melezitose, and raffinose. Sugar acids include, for example, saccharic acid, gluconates, and other polyhydroxy sugars and their salts. In some embodiments, the sugar is sucrose. In some cases, the sugar (e.g., sucrose) functions as a heat stabilizer for the anti-IL-33 antibody.

[0057] The amount of sugar (e.g., sucrose) contained within the pharmaceutical formulation of the present invention will vary depending on the specific circumstances in which the formulation is used and the intended purpose. In certain embodiments, the formulation can contain from 0.1% to about 20% sugar, from about 0.5% to about 20% sugar, from about 1% to about 20% sugar, from about 2% to about 15% sugar, from about 3% to about 10% sugar, from about 3% to about 7% sugar, or from about 4% to about 6% sugar. For example, the pharmaceutical formulation of the present invention can contain about 0.5%, about 1.0%, about 1.5%, about 2.0%, about 2.5%, about 3.0%, about 3.5%, about 4.0%, about 4.5%, about 5.0%, about 5.5%, about 6.0%, about 6.5%, about 7.0%, about 7.5%, about 8.0%, about 8.5%, about 9.0%, about 9.5%, about 10.0%, about 15%, or about 20% sugar (e.g., sucrose). In some embodiments, the formulation contains about 5% sugar (e.g., sucrose).

[0058] The pharmaceutical formulation of the present invention may also contain one or more organic co-solvents (or interfacial stabilizers) in types and amounts that stabilize the anti-IL-33 antibody under conditions of rough handling or agitation such as orbital shaking. In some embodiments, the organic co-solvent is a surfactant. As used herein, the term "surfactant" means a substance that reduces the surface tension of the fluid to be dissolved and / or reduces the interfacial tension between oil and water. The surfactant can be ionic or non-ionic. Exemplary non-ionic surfactants that can be included in the formulations of the present invention include, for example, alkyl poly(ethylene oxide), alkyl polyglucosides (such as octyl glucoside and decyl maltoside), fatty alcohols such as cetyl alcohol and oleyl alcohol, cocamide MEA, cocamide DEA, and cocamide TEA. Specific non-ionic surfactants that can be included in the formulations of the present invention include, for example, polysorbates such as polysorbate 20, polysorbate 28, polysorbate 40, polysorbate 60, polysorbate 65, polysorbate 80, polysorbate 81, and polysorbate 85, poloxamers such as poloxamer 188 (also known as pluronic F68), poloxamer 407, polyethylene-polypropylene glycol, or polyethylene glycol (PEG). Polysorbate 20 is also known as TWEEN 20, sorbitan monolaurate, and polyoxyethylene sorbitan monolaurate. In some embodiments, the surfactant is polysorbate 80.

[0059] The amount of surfactant contained within the pharmaceutical formulation of the present invention can vary depending on the desired specific characteristics of the formulation, as well as the particular circumstances and purposes for which the formulation is intended to be used. In certain embodiments, the formulation can contain from 0.05% to about 5% surfactant, from about 0.05% to about 0.15% surfactant, from about 0.04% to about 0.12%, from about 0.05% to about 0.11% surfactant, from about 0.06% to about 0.1% surfactant, or from about 0.07% to about 0.09% surfactant. For example, the formulation of the present invention can include about 0.05%, about 0.06%, about 0.07%, about 0.08%, about 0.09%, about 0.10%, about 0.11%, about 0.12%, about 0.13%, about 0.14%, about 0.15%, about 0.16%, about 0.17%, about 0.18%, about 0.19%, about 0.20%, about 0.21%, about 0.22%, about 0.23%, about 0.24%, about 0.25%, about 0.26%, about 0.27%, about 0.28%, about 0.29%, or about 0.30% surfactant (e.g., polysorbate 80). In some embodiments, the formulation contains about 0.08% surfactant (e.g., polysorbate 80). Each of the above percentages corresponds to weight / volume (w / v) percent.

[0060] The pharmaceutical formulation of the present invention may also include a buffer or buffer system that helps maintain a stable pH and stabilize the anti-IL-33 antibody. In some embodiments, the buffer or buffer system includes at least one buffer having a buffering range that completely or partially overlaps in the range of pH 4.9 to 5.7. In certain embodiments, the buffer includes a histidine buffer. In certain embodiments, the buffer is an acetate buffer. In certain embodiments, the buffer (e.g., acetate) is present at a concentration of about 1 mM to about 40 mM, about 1 mM to about 30 mM, about 1 mM to about 20 mM, about 3 mM to about 18 mM, about 5 mM to about 15 mM, or about 8 mM to about 12 mM. In some embodiments, the buffer (e.g., acetate) is present at a concentration of 5.3 mM ± 0.3 mM, 5.3 mM ± 0.2 mM, or 5.3 mM ± 0.1 mM. In some embodiments, the buffer is present at a concentration of about 4.6 mM, about 4.7 mM, about 4.8 mM, about 4.9 mM, about 5.0 mM, about 5.1 mM, about 5.2 mM, about 5.3 mM, about 5.4 mM, about 5.5 mM, about 5.6 mM, about 5.7 mM, about 5.8 mM, about 5.9 mM, or about 6.0 mM.

[0061] Exemplary formulations According to one aspect of the present invention, a pharmaceutical formulation includes (i) a human antibody (e.g., mAb1) that specifically binds to hIL-33, (ii) a buffer (e.g., acetate), (iii) an amino acid (e.g., arginine), (iv) a heat stabilizer (e.g., sucrose), and (v) an organic co-solvent (e.g., polysorbate 80).

[0062] Optionally, a stable liquid pharmaceutical formulation includes (i) a human antibody that specifically binds to hIL-33 (e.g., mAb1) at a concentration of about 1 mg / ml to about 200 mg / ml, (ii) a buffer (e.g., acetate) at a concentration of about 1 mM to about 20 mM, (iii) an amino acid (e.g., arginine) at a concentration of about 30 mM to about 110 mM, (iv) a heat stabilizer (e.g., sucrose) at a concentration of about 1% w / v to about 10% w / v, and (v) an organic co-solvent (e.g., polysorbate 80) at a concentration of about 0.01% w / v to about 0.15% w / v.

[0063] In some cases, a stable liquid pharmaceutical formulation comprises: (i) a human antibody that specifically binds to hIL-33 (e.g., mAb1) at a concentration of about 15 mg / ml to about 150 mg / ml; (ii) a buffer (e.g., acetate) at a concentration of about 5 mM to about 15 mM; (iii) an amino acid (e.g., arginine) at a concentration of about 50 mM to about 90 mM; (iv) a thermal stabilizer (e.g., sucrose) at a concentration of about 3% w / v to about 7% w / v; and (v) an organic co-solvent (e.g., polysorbate 80) at a concentration of about 0.05% w / v to about 0.11% w / v.

[0064] In some cases, a stable liquid pharmaceutical formulation comprises: (i) a human antibody that specifically binds to hIL-33 at a concentration of about 15 mg / ml to about 150 mg / ml and comprises a HCVR containing the amino acid sequence of SEQ ID NO: 2 and a LCVR containing the amino acid sequence of SEQ ID NO: 10; (ii) a buffer (e.g., acetate) at a concentration of about 5 mM to about 15 mM; (iii) an amino acid (e.g., arginine) at a concentration of about 50 mM to about 90 mM; (iv) a thermal stabilizer (e.g., sucrose) at a concentration of about 3% w / v to about 7% w / v; and (v) an organic co-solvent (e.g., polysorbate 80) at a concentration of about 0.05% w / v to about 0.11% w / v.

[0065] In some cases, a stable liquid pharmaceutical formulation comprises: (i) a human antibody that specifically binds to hIL-33 at a concentration of about 15 mg / ml to about 150 mg / ml and comprises a HCVR containing the amino acid sequence of SEQ ID NO: 2 and a LCVR containing the amino acid sequence of SEQ ID NO: 10, wherein the antibody has a heavy chain constant region of isotype IgG1; (ii) a buffer (e.g., acetate) at a concentration of about 5 mM to about 15 mM; (iii) an amino acid (e.g., arginine) at a concentration of about 50 mM to about 90 mM; (iv) a thermal stabilizer (e.g., sucrose) at a concentration of about 3% w / v to about 7% w / v; and (v) an organic co-solvent (e.g., polysorbate 80) at a concentration of about 0.05% w / v to about 0.11% w / v.

[0066] In some cases, a stable liquid pharmaceutical formulation comprises: (i) a human antibody that specifically binds to hIL-33 at a concentration of about 15 mg / ml to about 150 mg / ml and comprises a HCVR comprising the amino acid sequence of SEQ ID NO: 2 and a LCVR comprising the amino acid sequence of SEQ ID NO: 10, wherein the antibody has a heavy chain constant region of isotype IgG4; (ii) a buffer (e.g., acetate) at a concentration of about 5 mM to about 15 mM; (iii) an amino acid (e.g., arginine) at a concentration of about 50 mM to about 90 mM; (iv) a heat stabilizer (e.g., sucrose) at a concentration of about 3% w / v to about 7% w / v; and (v) an organic co-solvent (e.g., polysorbate 80) at a concentration of about 0.05% w / v to about 0.11% w / v.

[0067] In some cases, a stable liquid pharmaceutical formulation comprises: (i) a human antibody that specifically binds to hIL-33 at a concentration of about 15 mg / ml to about 150 mg / ml and comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 18 and a light chain comprising the amino acid sequence of SEQ ID NO: 20; (ii) a buffer (e.g., acetate) at a concentration of about 5 mM to about 15 mM; (iii) an amino acid (e.g., arginine) at a concentration of about 50 mM to about 90 mM; (iv) a heat stabilizer (e.g., sucrose) at a concentration of about 3% w / v to about 7% w / v; and (v) an organic co-solvent (e.g., polysorbate 80) at a concentration of about 0.05% w / v to about 0.11% w / v.

[0068] In some cases, a stable liquid pharmaceutical formulation comprises: (i) a human antibody that specifically binds to hIL-33 at a concentration of about 15 mg / ml to about 150 mg / ml and comprises a HCVR comprising the amino acid sequence of SEQ ID NO: 2 and a LCVR comprising the amino acid sequence of SEQ ID NO: 10; (ii) acetate at a concentration of about 5 mM to about 15 mM; (iii) arginine (e.g., arginine hydrochloride) at a concentration of about 50 mM to about 90 mM; (iv) sucrose at a concentration of about 3% w / v to about 7% w / v; and (v) polysorbate 80 at a concentration of about 0.05% w / v to about 0.11% w / v.

[0069] In some cases, a stable liquid pharmaceutical formulation comprises: (i) a human antibody that specifically binds to hIL-33 at a concentration of about 15 mg / ml to about 150 mg / ml and comprises a heavy chain variable region (HCVR) containing the amino acid sequence of SEQ ID NO: 2 and a light chain variable region (LCVR) containing the amino acid sequence of SEQ ID NO: 10, wherein the antibody has a heavy chain constant region of isotype IgG1; (ii) acetate at a concentration of about 5 mM to about 15 mM; (iii) arginine (e.g., arginine hydrochloride) at a concentration of about 50 mM to about 90 mM; (iv) sucrose at a concentration of about 3% w / v to about 7% w / v; and (v) polysorbate 80 at a concentration of about 0.05% w / v to about 0.11% w / v.

[0070] In some cases, a stable liquid pharmaceutical formulation comprises: (i) a human antibody that specifically binds to hIL-33 at a concentration of about 15 mg / ml to about 150 mg / ml and comprises a heavy chain variable region (HCVR) containing the amino acid sequence of SEQ ID NO: 2 and a light chain variable region (LCVR) containing the amino acid sequence of SEQ ID NO: 10, wherein the antibody has a heavy chain constant region of isotype IgG4; (ii) acetate at a concentration of about 5 mM to about 15 mM; (iii) arginine (e.g., arginine hydrochloride) at a concentration of about 50 mM to about 90 mM; (iv) sucrose at a concentration of about 3% w / v to about 7% w / v; and (v) polysorbate 80 at a concentration of about 0.05% w / v to about 0.11% w / v.

[0071] In some cases, a stable liquid pharmaceutical formulation comprises: (i) a human antibody that specifically binds to hIL-33 at a concentration of about 15 mg / ml to about 150 mg / ml and comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 18 and a light chain containing the amino acid sequence of SEQ ID NO: 20; (ii) acetate at a concentration of about 5 mM to about 15 mM; (iii) arginine (e.g., arginine hydrochloride) at a concentration of about 50 mM to about 90 mM; (iv) sucrose at a concentration of about 3% w / v to about 7% w / v; and (v) polysorbate 80 at a concentration of about 0.05% w / v to about 0.11% w / v.

[0072] In some cases, a stable liquid pharmaceutical formulation comprises: (i) a human antibody that specifically binds to hIL-33 and comprises a HCVR comprising the amino acid sequence of SEQ ID NO: 2 and a LCVR comprising the amino acid sequence of SEQ ID NO: 10, at a concentration of about 15 mg / ml to about 150 mg / ml; (ii) acetate at a concentration of about 8 mM to about 12 mM; (iii) arginine (e.g., arginine hydrochloride) at a concentration of about 65 mM to about 75 mM; (iv) sucrose at a concentration of about 4% w / v to about 6% w / v; and (v) polysorbate 80 at a concentration of about 0.07% w / v to about 0.09% w / v.

[0073] In some cases, a stable liquid pharmaceutical formulation comprises: (i) a human antibody that specifically binds to hIL-33 and comprises a HCVR comprising the amino acid sequence of SEQ ID NO: 2 and a LCVR comprising the amino acid sequence of SEQ ID NO: 10, at a concentration of about 15 mg / ml to about 150 mg / ml, wherein the antibody has a heavy chain constant region of isotype IgG1; (ii) acetate at a concentration of about 8 mM to about 12 mM; (iii) arginine (e.g., arginine hydrochloride) at a concentration of about 65 mM to about 75 mM; (iv) sucrose at a concentration of about 4% w / v to about 6% w / v; and (v) polysorbate 80 at a concentration of about 0.07% w / v to about 0.09% w / v.

[0074] In some cases, a stable liquid pharmaceutical formulation comprises: (i) a human antibody that specifically binds to hIL-33 and comprises a HCVR comprising the amino acid sequence of SEQ ID NO: 2 and a LCVR comprising the amino acid sequence of SEQ ID NO: 10, at a concentration of about 15 mg / ml to about 150 mg / ml, wherein the antibody has a heavy chain constant region of isotype IgG4; (ii) acetate at a concentration of about 8 mM to about 12 mM; (iii) arginine (e.g., arginine hydrochloride) at a concentration of about 65 mM to about 75 mM; (iv) sucrose at a concentration of about 4% w / v to about 6% w / v; and (v) polysorbate 80 at a concentration of about 0.07% w / v to about 0.09% w / v.

[0075] In some cases, a stable liquid pharmaceutical formulation comprises: (i) a human antibody that specifically binds to hIL-33 and comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 18 and a light chain comprising the amino acid sequence of SEQ ID NO: 20, at a concentration of about 15 mg / ml to about 150 mg / ml; (ii) acetate at a concentration of about 8 mM to about 12 mM; (iii) arginine (e.g., arginine hydrochloride) at a concentration of about 65 mM to about 75 mM; (iv) sucrose at a concentration of about 4% w / v to about 6% w / v; and (v) polysorbate 80 at a concentration of about 0.07% w / v to about 0.09% w / v.

[0076] In some cases, a stable liquid pharmaceutical formulation comprises: (i) a human antibody that specifically binds to hIL-33 and comprises a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 2 and a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 10, at a concentration of about 15 mg / ml to about 150 mg / ml; (ii) acetate at a concentration of about 10 mM; (iii) arginine (e.g., arginine hydrochloride) at a concentration of about 70 mM; (iv) sucrose at a concentration of about 5% w / v; and (v) polysorbate 80 at a concentration of about 0.08% w / v.

[0077] In some cases, a stable liquid pharmaceutical formulation comprises: (i) a human antibody that specifically binds to hIL-33 and comprises a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 2 and a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 10, at a concentration of about 15 mg / ml to about 150 mg / ml, wherein the antibody has a heavy chain constant region of isotype IgG1; (ii) acetate at a concentration of about 10 mM; (iii) arginine (e.g., arginine hydrochloride) at a concentration of about 70 mM; (iv) sucrose at a concentration of about 5% w / v; and (v) polysorbate 80 at a concentration of about 0.08% w / v.

[0078] In some cases, a stable liquid pharmaceutical formulation comprises: (i) a human antibody that specifically binds to hIL-33 at a concentration of about 15 mg / ml to about 150 mg / ml and comprises a HCVR comprising the amino acid sequence of SEQ ID NO: 2 and a LCVR comprising the amino acid sequence of SEQ ID NO: 10, wherein the antibody has a heavy chain constant region of isotype IgG4; (ii) acetate at a concentration of about 10 mM; (iii) arginine (e.g., arginine hydrochloride) at a concentration of about 70 mM; (iv) sucrose at a concentration of about 5% w / v; and (v) polysorbate 80 at a concentration of about 0.08% w / v.

[0079] In some cases, a stable liquid pharmaceutical formulation comprises: (i) a human antibody that specifically binds to hIL-33 at a concentration of about 15 mg / ml to about 150 mg / ml and comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 18 and a light chain comprising the amino acid sequence of SEQ ID NO: 20; (ii) acetate at a concentration of about 10 mM; (iii) arginine (e.g., arginine hydrochloride) at a concentration of about 70 mM; (iv) sucrose at a concentration of about 5% w / v; and (v) polysorbate 80 at a concentration of about 0.08% w / v.

[0080] In some cases, a stable liquid pharmaceutical formulation comprises: (i) a human antibody that specifically binds to hIL-33 at a concentration of about 15 mg / ml to about 150 mg / ml and comprises a HCVR comprising the amino acid sequence of SEQ ID NO: 2 and a LCVR comprising the amino acid sequence of SEQ ID NO: 10; (ii) acetate at a concentration of about 10 mM ± 1 mM; (iii) arginine hydrochloride at a concentration of about 70 mM ± 7 mM; (iv) sucrose at a concentration of 5% w / v ± 0.5% w / v; and (iv) polysorbate 80 at a concentration of about 0.08% ± 0.008% w / v, and the formulation has a pH of 5.3 ± 0.1.

[0081] In some cases, a stable liquid pharmaceutical formulation comprises: (i) a human antibody that specifically binds to hIL-33 and comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 2 and an LCVR comprising the amino acid sequence of SEQ ID NO: 10, at a concentration of about 15 mg / ml ± 1.5 mg / ml; (ii) about 10 mM ± 1 mM acetate; (iii) about 70 mM ± 7 mM arginine hydrochloride; (iv) 5% w / v ± 0.5% w / v sucrose; and (iv) about 0.08% ± 0.008% w / v polysorbate 80, and the formulation has a pH of 5.3 ± 0.1.

[0082] In some cases, a stable liquid pharmaceutical formulation comprises: (i) a human antibody that specifically binds to hIL-33 and comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 2 and an LCVR comprising the amino acid sequence of SEQ ID NO: 10, at a concentration of about 75 mg / ml ± 5 mg / ml; (ii) about 10 mM ± 1 mM acetate; (iii) about 70 mM ± 7 mM arginine hydrochloride; (iv) 5% w / v ± 0.5% w / v sucrose; and (iv) about 0.08% ± 0.008% w / v polysorbate 80, and the formulation has a pH of 5.3 ± 0.1.

[0083] In some cases, a stable liquid pharmaceutical formulation comprises: (i) a human antibody that specifically binds to hIL-33 and comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 2 and an LCVR comprising the amino acid sequence of SEQ ID NO: 10, at a concentration of about 150 mg / ml ± 15 mg / ml; (ii) about 10 mM ± 1 mM acetate; (iii) about 70 mM ± 7 mM arginine hydrochloride; (iv) 5% w / v ± 0.5% w / v sucrose; and (iv) about 0.08% ± 0.008% w / v polysorbate 80, and the formulation has a pH of 5.3 ± 0.1.

[0084] In some cases, a stable liquid pharmaceutical formulation comprises: (i) a human antibody that specifically binds to hIL-33, has an HCVR comprising the amino acid sequence of SEQ ID NO: 2, an LCVR comprising the amino acid sequence of SEQ ID NO: 10, and a human IgG4 heavy chain constant region, at a concentration of about 15 mg / ml to about 150 mg / ml; (ii) about 10 mM ± 1 mM acetate; (iii) about 70 mM ± 7 mM arginine hydrochloride; (iv) 5% w / v ± 0.5% w / v sucrose; and (iv) about 0.08% ± 0.008% w / v polysorbate 80, and the formulation has a pH of 5.3 ± 0.1.

[0085] In some cases, a stable liquid pharmaceutical formulation comprises: (i) a human antibody that specifically binds to hIL-33, has an HCVR comprising the amino acid sequence of SEQ ID NO: 2, an LCVR comprising the amino acid sequence of SEQ ID NO: 10, and a human IgG4 heavy chain constant region, at a concentration of about 15 mg / ml ± 1.5 mg / ml; (ii) about 10 mM ± 1 mM acetate; (iii) about 70 mM ± 7 mM arginine hydrochloride; (iv) 5% w / v ± 0.5% w / v sucrose; and (iv) about 0.08% ± 0.008% w / v polysorbate 80, and the formulation has a pH of 5.3 ± 0.1.

[0086] In some cases, a stable liquid pharmaceutical formulation comprises: (i) a human antibody that specifically binds to hIL-33, has an HCVR comprising the amino acid sequence of SEQ ID NO: 2, an LCVR comprising the amino acid sequence of SEQ ID NO: 10, and a human IgG4 heavy chain constant region, at a concentration of about 75 mg / ml ± 5 mg / ml; (ii) about 10 mM ± 1 mM acetate; (iii) about 70 mM ± 7 mM arginine hydrochloride; (iv) 5% w / v ± 0.5% w / v sucrose; and (iv) about 0.08% ± 0.008% w / v polysorbate 80, and the formulation has a pH of 5.3 ± 0.1.

[0087] In some cases, a stable liquid pharmaceutical formulation comprises: (i) a human antibody that specifically binds to hIL-33, contains an HCVR comprising the amino acid sequence of SEQ ID NO: 2, an LCVR comprising the amino acid sequence of SEQ ID NO: 10, and a human IgG4 heavy chain constant region, at a concentration of about 150 mg / ml ± 15 mg / ml; (ii) about 10 mM ± 1 mM acetate; (iii) about 70 mM ± 7 mM arginine hydrochloride; (iv) 5% w / v ± 0.5% w / v sucrose; and (iv) about 0.08% ± 0.008% w / v polysorbate 80, and the formulation has a pH of 5.3 ± 0.1.

[0088] In some cases, a stable liquid pharmaceutical formulation comprises: (i) a human antibody that specifically binds to hIL-33, contains a heavy chain comprising the amino acid sequence of SEQ ID NO: 18 and a light chain comprising the amino acid sequence of SEQ ID NO: 20, at a concentration of about 15 mg / ml to about 150 mg / ml; (ii) about 10 mM ± 1 mM acetate; (iii) about 70 mM ± 7 mM arginine hydrochloride; (iv) 5% w / v ± 0.5% w / v sucrose; and (iv) about 0.08% ± 0.008% w / v polysorbate 80, and the formulation has a pH of 5.3 ± 0.1.

[0089] In some cases, a stable liquid pharmaceutical formulation comprises: (i) a human antibody that specifically binds to hIL-33, contains a heavy chain comprising the amino acid sequence of SEQ ID NO: 18 and a light chain comprising the amino acid sequence of SEQ ID NO: 20, at a concentration of about 15 mg / ml ± 1.5 mg / ml; (ii) about 10 mM ± 1 mM acetate; (iii) about 70 mM ± 7 mM arginine hydrochloride; (iv) 5% w / v ± 0.5% w / v sucrose; and (iv) about 0.08% ± 0.008% w / v polysorbate 80, and the formulation has a pH of 5.3 ± 0.1.

[0090] In some cases, a stable liquid pharmaceutical formulation comprises (i) a human antibody that specifically binds to hIL-33 and comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 18 and a light chain comprising the amino acid sequence of SEQ ID NO: 20 at a concentration of about 75 mg / ml ± 5 mg / ml, (ii) about 10 mM ± 1 mM acetate, (iii) about 70 mM ± 7 mM arginine hydrochloride, (iv) 5% w / v ± 0.5% w / v sucrose, and (iv) about 0.08% ± 0.008% w / v polysorbate 80, and the formulation has a pH of 5.3 ± 0.1.

[0091] In some cases, a stable liquid pharmaceutical formulation comprises (i) a human antibody that specifically binds to hIL-33 and comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 18 and a light chain comprising the amino acid sequence of SEQ ID NO: 20 at a concentration of about 150 mg / ml ± 15 mg / ml, (ii) about 10 mM ± 1 mM acetate, (iii) about 70 mM ± 7 mM arginine hydrochloride, (iv) 5% w / v ± 0.5% w / v sucrose, and (iv) about 0.08% ± 0.008% w / v polysorbate 80, and the formulation has a pH of 5.3 ± 0.1.

[0092] Additional non-limiting examples of pharmaceutical formulations encompassed by the present invention are defined elsewhere in this specification, including the examples presented below.

[0093] Stability and Viscosity of Pharmaceutical Formulations The pharmaceutical formulations of the present invention exhibit a high level of stability. As used herein with respect to pharmaceutical formulations, the term "stable" means that the antibody within the pharmaceutical formulation retains an acceptable degree of structure and / or function and / or biological activity after storage for a defined amount of time. The formulation can be stable even if the antibody contained therein does not maintain 100% of its structure and / or function and / or biological activity after storage for a defined amount of time. Under certain circumstances, maintenance of about 90%, about 95%, about 96%, about 97%, about 98% or about 99% of the structure and / or function and / or biological activity of the antibody after storage for a defined amount of time can be considered "stable".

[0094] Stability can be measured, inter alia, by determining the percentage of native antibody remaining in the formulation after storage for a defined amount of time, defined at a given temperature. The percentage of native antibody can be determined, inter alia, by size exclusion chromatography (e.g., size exclusion high performance liquid chromatography [SE-HPLC]). "Acceptable level of stability", when the phrase is used herein, means that at least 90% of the native form of the antibody can be detected in the formulation after storage for a defined amount of time, defined at a given temperature. In certain embodiments, at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% of the native form of the antibody can be detected in the formulation after storage for a defined amount of time, defined at a given temperature. The defined amount of time after which stability is measured can be 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, at least 30 months, at least 36 months, or longer. The temperature at which the pharmaceutical formulation can be stored when evaluating stability can be any temperature from about -80°C to about 45°C, e.g., storage at about -80°C, about -30°C, about -20°C, about 0°C, about 4°C to 8°C, about 5°C, about 25°C, about 35°C, about 37°C, or about 45°C. For example, if more than about 90%, 95%, 96%, or 97% of the native antibody is detected by SE-HPLC after storage at 5°C for 3 months, the pharmaceutical formulation can be considered stable. If more than about 90%, 95%, 96%, or 97% of the native antibody is detected by SE-HPLC after storage at 5°C for 6 months, the pharmaceutical formulation can also be considered stable. If more than about 90%, 95%, 96%, 96.5%, 97%, 97.5%, 98%, 98.5%, 99% or 99.5% of the native antibody is detected by SE-HPLC after storage at 5°C for 9 months, the pharmaceutical formulation can also be considered stable.If more than about 90%, 95%, 96%, 96.5%, 97%, 97.5%, 98%, 98.5%, 99% or 99.5% of the native antibody is detected by SE-HPLC after storage at 25°C for 3 months, the pharmaceutical formulation may also be considered stable. If more than about 90%, 95%, 96%, 96.5%, 97%, 97.5%, 98%, 98.5%, 99% or 99.5% of the native antibody is detected by SE-HPLC after storage at 25°C for 6 months, the pharmaceutical formulation may also be considered stable. If more than about 90%, 95%, 96%, 96.5%, 97%, 97.5%, 98%, 98.5%, 99% or 99.5% of the native antibody is detected by SE-HPLC after storage at 25°C for 9 months, the pharmaceutical formulation may also be considered stable.

[0095] For example, other methods such as differential scanning calorimetry (DSC) to determine thermal stability, controlled agitation to determine mechanical stability, and absorbance at about 350 nm or about 405 nm to determine solution turbidity can be used to evaluate the stability of the formulations of the present invention. For example, after storage at about 5°C to about 25°C for 6 months or more, the change in OD 405 of the formulation is less than about 0.05 (e.g., 0.04, 0.03, 0.02, 0.01, or less) from the OD 405 of the formulation at t = 0, the formulation of the present invention may be considered stable.

[0096] Measuring the binding affinity of an antibody to a target can also be used to evaluate stability. For example, the formulation of the present invention, after storage for a defined amount of time (e.g., 14 days to 9 months) at, for example, -80°C, -30°C, -20°C, 5°C, 25°C, 37°C, 45°C, etc., the anti-IL-33 antibody contained in the formulation binds to hIL-33 with an affinity that is at least 80%, 85%, 90%, 95%, or more of the binding affinity of the antibody before such storage. The binding affinity can be determined by any method, such as ELISA or surface plasmon resonance. Biological activity can be determined by an IL-33 activity assay, such as by contacting cells expressing IL-33 with a formulation containing the anti-IL-33 antibody. Binding of the antibody to such cells can be measured directly, such as via FACS analysis. Alternatively, the activity downstream of the IL-33 system can be measured in the presence of the antibody and compared to the activity of the IL-33 system in the absence of the antibody.

[0097] Stability can be measured, inter alia, by determining the percentage of antibody that forms aggregates within the formulation after storage for a defined amount of time at a defined temperature, and stability is inversely proportional to the percentage of aggregates formed. The percentage of aggregated antibody can be determined, inter alia, by size exclusion chromatography (e.g., size exclusion high performance liquid chromatography [SE-HPLC] or size exclusion ultra-high performance liquid chromatography [SE-UPLC]). "Acceptable level of stability", when the phrase is used herein, means that a maximum of 6% of the antibody is in an aggregated form detected in the formulation after storage for a defined amount of time at a given temperature. In certain embodiments, an acceptable level of stability means that a maximum of about 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of the antibody can be detected in aggregates in the formulation after storage for a defined amount of time at a given temperature. The defined amount of time after which stability is measured can be at least 2 weeks, at least 28 days, 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, at least 30 months, at least 36 months, or more. The temperature at which the pharmaceutical formulation can be stored when evaluating stability can be any temperature from about -80°C to about 45°C, e.g., storage at about -80°C, about -30°C, about -20°C, about 0°C, about 4°C to 8°C, about 5°C, about 25°C, about 35°C, about 37°C, or about 45°C. For example, if less than about 2%, 1.75%, 1.5%, 1.25%, 1%, 0.75%, 0.5%, 0.25%, or 0.1% of the antibody is detected in an aggregated form after storage at 5°C for 9 months, the pharmaceutical formulation can be considered stable. If less than about 2%, 1.75%, 1.5%, 1.25%, 1%, 0.75%, 0.5%, 0.25%, or 0.1% of the antibody is detected in an aggregated form after storage at 25°C for 6 months, the pharmaceutical formulation can also be considered stable.If less than about 4%, 3.5%, 3%, 2.5%, 2%, 1.5%, 1%, 0.5%, or 0.1% of the antibody is detected in an aggregated form after storage at 45°C for 28 days, the pharmaceutical formulation may also be considered stable. If less than about 2%, 1.9%, 1.8%, 1.7%, 1.6%, 1.5%, 1%, 0.5%, or 0.1% of the antibody is detected in an aggregated form after storage at -20°C, -30°C, or -80°C for 3 months, the pharmaceutical formulation may also be considered stable.

[0098] Stability can be measured, inter alia, by determining the percentage of the antibody that migrates in the acidic fraction (the "acidic form") during ion exchange rather than in the major fraction of the antibody (the "main charge form"), and stability is inversely proportional to the fraction of the antibody in the acidic form. Without wishing to be bound by theory, deamidation of the antibody can make the antibody more negatively charged and thus more acidic relative to the non-deamidated antibody (e.g., Robinson, N., Protein Deamidation, PNAS, April 16, 2002, 99(8):5283-5288). The percentage of "acidified" antibody can be determined by ion exchange chromatography (e.g., cation exchange high performance liquid chromatography [CEX-HPLC] or cation exchange ultra-high performance liquid chromatography [CEX-UPLC]). "Acceptable level of stability", when the phrase is used herein, means that up to 52% of the antibody is in a more acidic form detected in the formulation after storage for a defined amount of time at a defined temperature. In certain embodiments, an acceptable level of stability means that up to about 52%, 50%, 45%, 40%, 35%, 30%, 29%, 28%, 27%, 26%, 25%, 20%, 15%, 10%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of the antibody can be detected in the acidic form in the formulation after storage for a defined amount of time at a given temperature. The defined amount of time after stability is measured can be at least 2 weeks, at least 28 days, 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, at least 30 months, at least 36 months, or more. The temperature at which the pharmaceutical formulation can be stored when evaluating stability can be any temperature from about -80°C to about 45°C, e.g., storage at about -80°C, about -30°C, about -20°C, about 0°C, about 4°C to 8°C, about 5°C, about 25°C, or about 45°C.For example, if less than about 29%, 28%, 27%, 26%, 25%, 24%, 23%, 22%, 21%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of the antibody is in a more acidic form after storage at -80°C, -30°C, or -20°C for 3 months, the pharmaceutical formulation is considered stable. If less than about 28%, 27%, 26%, 25%, 24%, 23%, 22%, 21%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of the antibody is in a more acidic form after storage at 5°C for 9 months, the pharmaceutical formulation is also considered stable. If less than about 30%, 29%, 28%, 27%, 26%, 25%, 24%, 23%, 22%, 21%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of the antibody is in a more acidic form after storage at 25°C for 28 days, the pharmaceutical formulation is also considered stable. If less than about 37%, 36%, 35%, 34%, 33%, 32%, 31%, 30%, 29%, 28%, 27%, 26%, 25%, 24%, 23%, 22%, 21%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of the antibody is in a more acidic form after storage at 37°C for 28 days, the pharmaceutical formulation is also considered stable. If less than about 52%, 51%, 50%, 49%, 48%, 47%, 46%, 45%, 44%, 43%, 42%, 41%, 40%, 39%, 38%, 37%, 36%, 35%, 34%, 33%, 32%, 31%, 30%, 29%, 28%, 27%, 26%, 25%, 24%, 23%, 22%, 21%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of the antibody can be detected in a more acidic form after storage at 45°C for 28 days, the pharmaceutical formulation is also considered stable.

[0099] In certain embodiments, the "stable" pharmaceutical composition or formulation of the present invention contains HMW species of 2% or less, 1.9% or less, 1.8% or less, 1.7% or less, 1.6% or less, or 1.5% or less as measured by size exclusion ultra-high performance liquid chromatography (SE-UPLC) after storage at -80°C, -30°C, or -20°C for 24 months. In certain embodiments, the "stable" pharmaceutical composition or formulation of the present invention contains HMW species of 1% or less, 0.9% or less, 0.8% or less, 0.7% or less, 0.6% or less, or 0.5% or less as measured by SE-UPLC after storage at -80°C, -30°C, or -20°C for 9 months. In certain embodiments, the "stable" pharmaceutical composition or formulation of the present invention contains HMW species of 1% or less, 0.9% or less, 0.8% or less, 0.7% or less, or 0.6% or less as measured by SE-UPLC after storage at 5°C for 2 months. In certain embodiments, the "stable" pharmaceutical composition or formulation of the present invention contains HMW species of 2% or less, 1.8% or less, 1.6% or less, 1.4% or less, 1.2% or less, or 1.0% or less as measured by SE-UPLC after storage at 25°C and 60% relative humidity for 2 months. In certain embodiments, the "stable" pharmaceutical composition or formulation of the present invention contains HMW species of 6% or less, 5.5% or less, 5.4% or less, 5.3% or less, 5.2% or less, or 5.1% or less as measured by SE-UPLC after storage at 40°C and 75% relative humidity for 2 months. In certain embodiments, the "stable" pharmaceutical composition or formulation of the present invention contains HMW species of 1% or less, 0.9% or less, 0.8% or less, or 0.7% or less as measured by SE-UPLC after storage at 2-8°C for 9 months. In certain embodiments, the "stable" pharmaceutical composition or formulation of the present invention contains HMW species of 3% or less, 2.8% or less, 2.6% or less, 2.4% or less, 2.2% or less, or 2.0% or less as measured by SE-UPLC after storage at 2-8°C for 24 months. In certain embodiments, the "stable" pharmaceutical composition or formulation of the present invention contains HMW species of 2% or less, 1.8% or less, 1.6% or less, 1.5% or less, 1.4% or less, or 1.3% or less as measured by SE-UPLC after storage at 25°C and 60% relative humidity for 6 months.In certain embodiments, the “stable” pharmaceutical composition or pharmaceutical formulation of the present invention comprises HMW species of 1% or less, 0.9% or less, 0.8% or less, or 0.7% or less as measured by SE-UPLC after storage at 2-8°C for 6 months. In certain embodiments, the “stable” pharmaceutical composition or pharmaceutical formulation of the present invention comprises HMW species of 1% or less, 0.9% or less, 0.8% or less, 0.7% or less, 0.6% or less, or 0.5% or less as measured by SE-UPLC after storage at 2-8°C for 3 months.

[0100] In certain embodiments, a “stable” pharmaceutical composition or formulation of the present invention comprises at least 95%, at least 96%, at least 97%, at least 97.5%, or at least 97.9% of the native form of the antibody as measured by size exclusion ultra-high performance liquid chromatography (SE-UPLC) after storage at -80°C, -30°C, or -20°C for 24 months. In certain embodiments, a “stable” pharmaceutical composition or formulation of the present invention comprises at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of the native form of the antibody as measured by size exclusion ultra-high performance liquid chromatography (SE-UPLC) after storage at -80°C, -30°C, or -20°C for 9 months. In certain embodiments, a “stable” pharmaceutical composition or formulation of the present invention comprises at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of the native form of the antibody as measured by size exclusion ultra-high performance liquid chromatography (SE-UPLC) after storage at 5°C for 2 months. In certain embodiments, a “stable” pharmaceutical composition or formulation of the present invention comprises at least 95%, at least 96%, at least 97%, at least 98%, at least 98.5%, or at least 98.8% of the native form of the antibody as measured by size exclusion ultra-high performance liquid chromatography (SE-UPLC) after storage at 25°C and 60% relative humidity for 2 months. In certain embodiments, a “stable” pharmaceutical composition or formulation of the present invention comprises at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, or at least 94.1% of the native form of the antibody as measured by size exclusion ultra-high performance liquid chromatography (SE-UPLC) after storage at 40°C and 75% relative humidity for 2 months. In certain embodiments, a “stable” pharmaceutical composition or formulation of the present invention comprises at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of the native form of the antibody as measured by size exclusion ultra-high performance liquid chromatography (SE-UPLC) after storage at 2-8°C for 9 months.In certain embodiments, a “stable” pharmaceutical composition or formulation of the invention comprises at least 95%, at least 96%, at least 97%, at least 97.5%, or at least 97.6% of the native form of the antibody as measured by size exclusion ultra performance liquid chromatography (SE-UPLC) after storage at 2-8° C. for 24 months. In certain embodiments, a “stable” pharmaceutical composition or formulation of the invention comprises at least 95%, at least 96%, at least 97%, at least 98%, or at least 98.4% of the native form of the antibody as measured by size exclusion ultra performance liquid chromatography (SE-UPLC) after storage at 25° C. and 60% relative humidity for 6 months. In certain embodiments, a “stable” pharmaceutical composition or formulation of the invention comprises at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of the native form of the antibody as measured by size exclusion ultra performance liquid chromatography (SE-UPLC) after storage at 2-8° C. for 3 months or 6 months.

[0101] References to the stability of a pharmaceutical formulation “after” a specific period are intended to mean that the measurement of the stability parameter (e.g., % native form, % HMW species, or % acidic form) is made at or around the end of the specific period and that the pharmaceutical formulation is not necessarily intended to maintain the same degree of stability with respect to the parameter measured thereafter. For example, a reference to a specific stability after 9 months means that the measurement of stability was made at the start of the study or around 9 months later. Additional methods for assessing the stability of the antibody in the formulation are demonstrated in the examples presented below.

[0102] In the liquid form, the pharmaceutical formulation of the present invention may exhibit a low to medium level of viscosity in certain embodiments. As used herein, "viscosity" 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 fluids of equal volume are placed in the same capillary viscometer and can flow by gravity, the more viscous fluid takes longer to flow through the capillary than the less viscous fluid. For example, if one fluid takes 200 seconds to complete its flow and another fluid takes 400 seconds, the second fluid has twice the viscosity of the first on the kinematic viscosity scale. "Absolute viscosity", also sometimes called dynamic or simple viscosity, is the product of kinematic viscosity and fluid density (absolute viscosity = kinematic viscosity × density). The dimension of kinematic viscosity is L 2 / T, where L is length and T is time. Generally, 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 millipascal second (mPa-s), and 1 cP = 1 mPa-s.

[0103] As used herein, with respect to the fluid formulations of the present invention, a low level of viscosity will exhibit an absolute viscosity of less than about 20 centipoise (cP) at 20°C. For example, when measured using standard viscosity measurement techniques, if the formulation exhibits an absolute viscosity of about 19 cP, about 18 cP, about 17 cP, about 16 cP, about 15 cP, about 14 cP, about 13 cP, about 12 cP, about 11 cP, about 10 cP, about 9 cP, about 8 cP, about 7 cP, about 6 cP, about 5 cP, about 4 cP, or less, the fluid formulation of the present invention will be considered to have a "low viscosity". As used herein, with respect to the fluid formulations of the present invention, a medium level of viscosity will exhibit an absolute viscosity of from about 30 cP to about 20 cP. For example, when measured using standard viscosity measurement techniques, if the formulation exhibits an absolute viscosity of about 30 cP, about 29 cP, about 28 cP, about 27 cP, about 26 cP, about 25 cP, about 24 cP, about 23 cP, about 22 cP, about 21 cP, or about 20 cP, the fluid formulation of the present invention will be considered to have a "medium viscosity". Each of these values refers to measurements taken at 20°C.

[0104] As shown in the following examples, the present invention is based in part on the discovery that the combination of the claimed excipients with the anti-IL-33 antibody produces a formulation that is stable and has a desirable viscosity.

[0105] Containers and Methods of Administration The pharmaceutical formulations of the present invention can be contained within any container suitable for the storage of pharmaceuticals and other therapeutic compositions. For example, the pharmaceutical formulation can be contained within a sealed and sterilized plastic or glass container having a defined volume, such as a vial, ampoule, syringe, cartridge, bottle or IV bag. For example, different types of vials, including transparent and opaque (e.g., amber-colored) glass or plastic vials, can be used to contain the formulations of the present invention. Similarly, any type of syringe can contain and / or be used to administer the pharmaceutical formulations of the present invention. In some embodiments, the pharmaceutical formulation is contained within a pre-filled syringe. In some embodiments, the pharmaceutical formulation is contained within a pre-filled fixed-needle syringe.

[0106] The pharmaceutical formulation of the present invention can be contained within a "normal tungsten" syringe or a "low tungsten" syringe. As will be understood by those skilled in the art, the process of making a glass syringe generally involves the use of a high-temperature tungsten rod that functions to create a hole in the glass, thereby creating a hole through which liquid can be drawn out and discharged from the syringe. This process results in the deposition of trace amounts of tungsten on the inner surface of the syringe. Subsequent washing 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 ppb of tungsten. For example, a low tungsten syringe according to the present invention can contain about 490, 480, 470, 460, 450, 440, 430, 420, 410, 390, 350, 300, 250, 200, 150, 100, 90, 80, 70, 60, 50, 40, 30, 20, 10 or less ppb of tungsten, or even less.

[0107] The rubber plunger used in a syringe and the rubber stopper used to close the opening of a vial can be coated to prevent contamination of the pharmaceutical contents of the syringe or vial and / or to maintain their stability. Thus, according to certain embodiments of the present invention, the pharmaceutical formulation can be contained within a syringe containing a coated plunger or within a vial sealed with a coated rubber stopper. For example, the plunger or stopper can be coated with a fluorocarbon film. Examples of coated stoppers and / or plungers suitable for use with vials and syringes containing the pharmaceutical formulations of the present disclosure are mentioned, for example, in U.S. Patent Nos. 4,997,423, 5,908,686, 6,286,699, 6,645,635, and 7,226,554, the contents of which are incorporated herein by reference in their entirety. Specific exemplary coated rubber stoppers and plungers that can be used in the practice of the present invention are commercially available under the trade name "FluroTec®" from West Pharmaceutical Services, Inc. (Lionville, PA). According to certain embodiments of the present invention, the pharmaceutical formulation can be contained within a low tungsten syringe containing a fluorocarbon-coated plunger. In some embodiments, the container is a syringe such as an Ompi EZ-Fill™ syringe or a BD Neopak™ syringe. In some cases, the syringe is a 1 mL long glass syringe having a 1 mL iWest piston, a 27G thin wall needle, and an FM30 needle shield or a BD260 needle shield. In some cases, the syringe is a 2.25 mL glass syringe having a West NovaPure™ 1-3 mL piston, a 27G thin wall needle, and an FM30 needle shield or a BD260 needle shield.In various embodiments, the syringe is a 0.5 mL, 0.6 mL, 0.7 mL, 0.8 mL, 0.9 mL, 1.0 mL, 1.1 mL, 1.2 mL, 1.3 mL, 1.4 mL, 1.5 mL, 1.6 mL, 1.7 mL, 1.8 mL, 1.9 mL, 2.0 mL, 2.1 mL, 2.2 mL, 2.3 mL, 2.4 mL, 2.5 mL, 2.6 mL, 2.7 mL, 2.8 mL, 2.9 mL, 3.0 mL, 3.5 mL, 4.0 mL, 4.5 mL, 5.0 mL, 5.5 mL, 6.0 mL, 6.5 mL, 7.0 mL, 7.5 mL, 8.0 mL, 8.5 mL, 9.0 mL, 9.5 mL, or 10 mL syringe (e.g., a glass syringe).

[0108] The pharmaceutical formulation can be administered to a subject by injection (e.g., subcutaneous, intravenous, intramuscular, intraperitoneal, etc.) or via an oral route such as transdermal, mucosal, nasal, pulmonary, and / or oral administration. A number of reusable pens and / or auto-injector delivery devices can be used to deliver the pharmaceutical formulation of the present invention subcutaneously. Examples include, but are not limited to, AUTOPEN™ (Owen Mumford, Inc., Woodstock, UK), DISETRONIC™ pen (Disetronic Medical Systems, Bergdorf, Switzerland), HUMALOG MIX 75 / 25™ pen, HUMALOG™ pen, HUMALIN 70 / 30™ pen (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), OPTIPEN™, OPTIPEN PRO™, OPTIPEN STARLET™, and OPTICLIK™ (sanofi-aventis, Frankfurt, Germany), and only a few of these are listed. Examples of disposable pens and / or auto-injector delivery devices 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™ auto-injector (Amgen, Thousand Oaks, CA), PENLET™ (Haselmeier, Stuttgart, Germany), EPIPEN (Dey, L.P.), and HUMIRA™ pen (Abbott Labs, Abbott Park, IL), and only a few of these are listed. In some cases, the pharmaceutical formulation is contained in a syringe specifically adapted for use with an auto-injector.Subcutaneous injections can be administered using a 20 - 30 gauge needle or a 25 - 30 gauge needle. In some cases, subcutaneous injections can be administered using a 25 gauge needle. In some cases, subcutaneous injections can be administered using a 27 gauge needle. In some cases, subcutaneous injections can be administered using a 29 gauge needle.

[0109] Another type of delivery device can include a safety system. Such devices are relatively inexpensive and, upon completion of the injection, operate to extend a safety sleeve manually or automatically over the needle. Examples of safety systems can include the ERIS device by West Pharmaceutical, or the UltraSafe device by Becton Dickinson. Additionally, the use of a large volume device ("LVD") or a bolus syringe for delivering the pharmaceutical formulations of the present invention is also contemplated herein. In some cases, the LVD or bolus syringe can be configured to inject the drug into a patient. For example, the LVD or bolus syringe can be configured to deliver a "large volume" drug (typically, about 2 ml to about 10 ml).

[0110] The pharmaceutical formulations of the present invention can also be included in unit dosage forms. As used herein, the term "unit dosage form" refers to a physically discrete unit suitable as a unit dosage for the patient to be treated, each unit being calculated to contain a predetermined quantity of the active compound so as to produce the desired therapeutic effect in relation to the required pharmaceutical carrier, diluent, or excipient. In various embodiments, the unit dosage form is contained within a container, as discussed herein. The actual dosage level of the active ingredient (e.g., anti-IL-33 antibody) in the formulations of the present invention can vary so as to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration without attendant adverse effects on the patient. The dosage level selected will depend on a variety of pharmacokinetic factors including the activity of the particular composition of the invention employed, the route of administration, the time of administration, the rate of excretion of the particular compound being used, the duration of the treatment employed in combination with the particular composition being used, other drugs, compounds and / or materials, the age, sex, weight, condition, general health, and prior medical history of the patient being treated, as well as similar factors well known in the medical arts. As used herein, the term "diluent" refers to a solution suitable for modifying or achieving the exemplified or appropriate concentrations described herein.

[0111] In various embodiments, the unit dosage form contains an amount of active ingredient (e.g., anti-IL-33 antibody) intended for single use. In various embodiments, the amount of active ingredient in the unit dosage form is from about 0.1 mg to about 5000 mg, from about 100 mg to about 1000 mg, and from about 100 mg to about 500 mg, from about 100 mg to about 400 mg, from about 100 mg to about 200 mg, from about 250 mg to about 350 mg, from about 125 mg to about 175 mg, from about 275 mg to about 325 mg, or ranges or intervals thereof. Intermediate ranges of the amounts listed above, such as from about 135 mg to about 165 mg or 285 mg to 315 mg, are also intended to be part of the present invention. For example, ranges of values using any combination of the values listed above (or values included within the ranges listed above) as upper and / or lower limits are intended to be included. In certain embodiments, the formulation is often supplied as a liquid in unit dosage form. In some embodiments, the unit dosage form contains about 150 mg. In some embodiments, the unit dosage form contains about 300 mg. In some embodiments, the unit dosage form according to the present invention is suitable for subcutaneous administration to a patient.

[0112] The present invention also includes a method of preparing a unit dosage form. In an exemplary embodiment, a method for preparing a pharmaceutical unit dosage form includes combining a formulation of any of the foregoing embodiments within a suitable container (e.g., those containers discussed herein).

[0113] Therapeutic Use of Pharmaceutical Formulations The pharmaceutical formulations of the present invention are useful, inter alia, for the treatment, prevention, and / or amelioration of any disease or disorder associated with IL-33 activity. In particular, the pharmaceutical formulations of the present invention are useful for the treatment, prevention, and / or amelioration of any disease or disorder that is treatable by modulating IL-33 expression, signaling, or activity, or by blocking the interaction between IL-33 and an IL-33 ligand (e.g., ST2), or otherwise inhibiting IL-33 activity and / or signaling.

[0114] The treatment method of the present invention includes administering to a subject any formulation comprising an anti-hIL-33 antibody as disclosed herein. The subject to whom the pharmaceutical formulation is administered can be any human or non-human animal that, for example, requires such treatment, prevention, and / or amelioration, or would otherwise benefit from the inhibition or attenuation of IL-33 and / or IL-33-mediated activity. For example, the subject can be an individual diagnosed with or at risk of developing any of the aforementioned diseases or disorders. The present invention further includes the use of any of the pharmaceutical formulations disclosed herein in the manufacture of a medicament for the treatment, prevention, and / or amelioration of any disease or disorder associated with IL-33 activity, including any of the exemplary diseases, disorders, and conditions described above.

[0115] In some embodiments, the present invention provides a kit comprising a pharmaceutical formulation (e.g., a container having a formulation or unit dosage form) as discussed herein, and a package or labeling (e.g., a package insert) having instructions for using the pharmaceutical formulation for the treatment of a disease or disorder as discussed above. Optionally, the instructions provide for the use of the unit dosage form for the treatment of a disease or disorder as discussed herein.

[0116] The sequences discussed herein and shown in the accompanying Sequence Listing correspond to mAb1, a fully human antibody having an IgG4 heavy chain constant region, which is used through the following examples. The sequence numbers are shown below. [Table 1]

[0117] The following examples are presented to provide those skilled in the art with a complete disclosure and description of how to make and use the methods and compositions of the present invention and are not intended to limit the scope which the inventors regard as the present invention. Efforts have been made to ensure accuracy with respect to the numerical values used (e.g., amounts, temperatures, etc.), but some experimental errors and deviations should be accounted for. Unless otherwise indicated, parts are parts by weight, molecular weights are average molecular weights, temperatures are in degrees Celsius, and pressures are at or near atmospheric pressure.

[0118] Example 1: Effects of Buffer and pH on the Stability of Anti-IL-33 Antibody The effects of buffer and pH on the thermal stability of mAb1 were examined in a liquid formulation by incubating 5 mg / mL of mAb1 at 45 °C for 28 days in a series of buffer systems over a varying pH range. The pH and buffer systems of acetate (pH 4.5 - 5.5), L-histidine (pH 5.5 - 6.5), and phosphate (pH 6.0 - 7.0) were studied. Based on the results from SE-UPLC analysis, the maximum protein stability was observed when mAb1 was formulated at pH 6.0 - pH 6.5 in L-histidine buffer. Based on the results from CEX-UPLC analysis, the maximum protein stability was observed when mAb1 was formulated at pH 5.0 - pH 6.0 in L-histidine or acetate buffer. These analyses also revealed that the formation of HMW species and charge variants are the major degradation pathways. The results are shown in Table 1.

[0119] [Table 2]

[0120] The effects of pH and buffer on the thermal stability of mAb1 were examined in liquid formulations by incubating 150 mg / mL of mAb1 at 37 °C for 28 days in a series of L-histidine and acetate buffers in the range of pH 4.5, 5.0, and 6.0 in the presence of 5% sucrose as a stabilizer. Based on the results of SE-UPLC analysis for molecular weight species and CEX-UPLC analysis for charge variants, aggregation (i.e., formation of HMW species) and formation of charge variants were the major degradation pathways. The mAb1 stability at pH 5.0 and 6.0 in L-histidine buffer and at pH 5.0 in acetate buffer was comparable as shown in Table 2.

[0121]

Table 3

[0122] Example 2: Effects of Surfactants and Thermal Stabilizers on the Stability of Anti-IL-33 Antibody The effects of two surfactants, 0.1% polysorbate 20 and 0.1% polysorbate 80, on the thermal stability of 5 mg / mL of mAb1 were first examined in liquid formulations using a research lot of the material. The results of the thermal stability study are summarized in Table 3. When incubated at 45 °C, the addition of either polysorbate 20 or polysorbate 80 had a detrimental effect on the thermal stability of mAb1 compared to the control formulation lacking any surfactant. An increase in high molecular weight species and charge variants was observed. The addition of 0.1% polysorbate 80 caused less relative increase in HMW species and formation of charge variant forms compared to the addition of 0.1% polysorbate 20.

[0123] Additional studies were conducted to investigate the polysorbate 80 concentration using a representative research lot of the material at a mAb1 concentration of 50 mg / mL. The polysorbate 80 concentrations included in the study were 0%, 0.01%, 0.02%, 0.04%, 0.06%, 0.08%, and 0.1%. The results are summarized in Tables 4 and 5. When stirred for 60 minutes, an increase in the relative percentage of HMW of 0.8% was observed for the sample without polysorbate 80 (Table 4). Examination of the SEC-UPLC chromatogram showed that this increase was due to the formation of higher-order aggregation peaks that were not present in the starting material. The addition of 0.02% or more polysorbate 80 prevented the formation of this aggregated species after 60 minutes of stirring. When incubated at 45 °C for 28 days, the thermal stability of mAb1 was not affected by the polysorbate concentrations studied (Table 5).

[0124] 5 mg / mL of mAb1 in the liquid formulation was formulated with 5% sucrose and showed improved stability when incubated under accelerated conditions, as shown in Table 3. After incubation at 45 °C for 28 days, the relative amount of HMW species increased by 0.7% in the formulation containing 5% sucrose compared to a 1.2% increase in the sucrose-free control formulation.

[0125] [Table 4]

[0126] [Table 5]

[0127] [Table 6]

[0128] Example 3: Effect of Viscosity Modifiers on the Stability of Anti-IL-33 Antibody mAb1 is contained in a pre-filled syringe (PFS) and is delivered through a delivery device such as an auto-injector. Viscosity correlates with the ease of injection through a pre-filled syringe (PFS). Maintaining a moderately low viscosity is advantageous for the development of delivery devices such as auto-injectors.

[0129] The effect of pH on the viscosity of 150 mg / mL mAb1 in 10 mM L-histidine buffer was investigated. As shown in Figure 1, mAb1 viscosity strongly depends on pH in the pH range of 4.8 - 6.7. The lower the pH, the lower the mAb1 viscosity. The effect of excipients on formulation viscosity was also investigated in mAb1 liquid formulations with the following potential viscosity modifiers: sodium acetate, L-arginine hydrochloride, sodium L-glutamate, and magnesium chloride up to 200 mM. The base formulation contained 150 mg / mL mAb1, 27 mM acetate, and 5% sucrose at pH 5.3. The results are shown in Figure 2. All of these additives at 25 - 150 mM decreased the viscosity of 150 mg / mL mAb1 at pH 5.3 by 2 - 5 cP.

[0130] The effect of viscosity modifiers on the stability of 150 mg / mL mAb1 was also investigated by incubating the formulation at 37 °C for 34 days. HMW species formation is the main degradation pathway, and the results are shown in Figure 3. The addition of magnesium chloride or sodium acetate promoted HMW formation after incubation at 37 °C compared to a control formulation containing no viscosity modifier. On the other hand, L-arginine hydrochloride and sodium L-glutamate did not have a detrimental effect on mAb1 stability. Instead, 75 mM L-arginine hydrochloride improved mAb1 stability against HMW formation.

[0131] Based on the viscosity-lowering effect and the impact on thermal stability, L-arginine hydrochloride was selected as a viscosity modifier for the mAb1 formulation.

[0132] Example 4: Influence of formulation parameters on viscosity and stability A custom experimental design was applied to understand the effects of each formulation component, as well as the interactions of parameters on the stability and viscosity of mAb1, similar to pH. The formulation parameters in this study included protein concentration (135 - 165 mg / mL), sucrose concentration (5 - 9%), L-arginine hydrochloride concentration (0 - 75 mM), and pH. For all formulations in this study, the surfactant remained constant at 0.1%.

[0133] The viscosity of the formulation remained unchanged before and after incubation at 40°C / 75% RH for 28 days.

[0134] The viscosity against the main formulation parameters was analyzed by a fitted model using JMP12 with standard least squares personality and effect leverage emphasis. The main factors affecting the formulation viscosity were mAb1 concentration, pH, and L-arginine hydrochloride concentration, as shown in Figure 4A.

[0135] The stability of the formulation after 28 days of culture at 40°C / 75% RH was also investigated. No significant differences in color and appearance, turbidity, pH change, protein recovery, or charge variant change were observed for the 14 formulations after 28 days of incubation at 40°C / 75% RH. The main stability indicating the attribute was the formation of HMW species.

[0136] The rate of HMW formation against the main formulation parameters was analyzed by a fitted model using JMP12 with standard least squares personality and effect leverage emphasis. The main factors affecting HMW formation were pH, L-arginine hydrochloride concentration, and sucrose concentration, as shown in Figure 4B.

[0137] The selection of the formulation composition was based on minimizing the formulation viscosity and the HMW formation rate using the JMP12 desirability function. Multi-parametric analysis was used to generate a composition containing 5% sucrose, and 150 mg / ml of antibody (mAb1) concentration with 70 mM of L-arginine hydrochloride at pH 5.3. This formulation minimized the formulation viscosity as well as HMW formation. On the other hand, it reduced the sensitivity of the viscosity and stability variations to the variations in the excipient composition.

[0138] Example 5: Effect of Surfactant Concentration on the Stability of Anti-IL-33 Antibody Polysorbate 80 was identified as the stabilizing surfactant in the study considered in Example 2 above. The basic formulation for this study was 150 mg / mL of mAb1, 10 mM acetate buffer, 5% (w / v) sucrose, and 70 mM L-arginine hydrochloride. The polysorbate 80 concentrations in the initial evaluation were 0.01%, 0.02%, 0.04%, 0.05%, 0.06%, 0.08%, and 0.1%. The results are summarized in Table 6. After orbital shaking at 250 rpm for 48 hours, an increase in the relative percentage of HMW was observed for the samples containing low levels of polysorbate 80. Polysorbate 80 at 0.055% or more prevented the formation of this aggregated species after 48 hours of orbital shaking.

[0139] [Table 7]

[0140] The acceptable range of polysorbate 80 in the 150 mg / mL mAb1 formulation was further investigated by studying the agitation stability as well as the thermal stability. Using the same basic formulation, the polysorbate 80 concentrations in the studies covering this range were 0.02%, 0.05%, 0.08%, and 0.12%. The formulations were subjected to orbital shaking at 250 rpm for 48 hours and thermal stress at 40 °C / 75% RH for 1 month. The results of orbital shaking confirmed that polysorbate 80 at 0.05% and above prevented the formation of these aggregated species after 48 hours of orbital shaking (see Table 7). The thermal stability of mAb1 was not affected by the polysorbate concentrations studied (see Table 7). These results indicated that 0.05 - 0.12% of polysorbate 80 provided sufficient stabilization to prevent the formation of aggregates under agitation stress without having a detrimental effect on the stability of the formulation. Based on these results, a target concentration of 0.08% of polysorbate 80 was selected for the 150 mg / mL mAb1 formulation.

[0141]

Table 8

[0142] Example 6: Stability of Liquid-Formulated Anti-IL-33 Antibody Prodrug For the formulation of mAb1, a continuous long-term storage stability study has been conducted to evaluate its stability through storage at -80°C, -30°C, and -20°C for 36 months. As shown in Tables 9 - 10, the composition containing 150 mg / ml of mAb1 was physically and chemically stable when stored at -80°C, -30°C, and -20°C for 24 months. No recognizable changes in physical or chemical stability were detected in any of the monitored attributes. Furthermore, the composition containing 150 mg / ml of mAb1 has been shown to be stable when stored at -80°C, -30°C, and -20°C for at least 9 months (see Tables 11 - 13). Finally, the composition containing 15 mg / ml of mAb1 has been shown to be stable when stored at -80°C, -30°C, and -20°C for at least 9 months (see Tables 16 - 18). All the results collected so far are shown in Tables 8 - 20 below.

[0143]

Table 9

[0144]

Table 10

[0145]

Table 11

[0146]

Table 12

[0147]

Table 13

[0148]

Table 14

[0149]

Table 15

[0150]

Table 16

[0151]

Table 17

[0152]

Table 18

[0153]

Table 19

[0154]

Table 20

[0155]

Table 21

[0156] Example 7: Stability of Liquid-Formulated Anti-IL-33 Antibody Drug For the 15 mg / mL and 150 mg / mL drug products of mAb1 in glass vials, 9-month study stability data have been available to date. Both antibody concentrations were physically and chemically stable when stored at 2 - 8 °C for 9 months (see Tables 21 and 22). Additional stability studies of the 150 mg / ml mAb1 drug product were observed to be physically and chemically stable when stored at 2 - 8 °C for 24 months (see Table 23). No recognizable changes in physical or chemical stability were detected in any of the monitored attributes.

[0157] Results from the analysis of the mAb1 drug product formulations at 15 mg / mL and 150 mg / mL after incubation under accelerated and stress conditions are provided in Tables 24 and 25, respectively. The mAb1 drug product was physically and chemically stable when agitated for 48 hours (orbital shaking at 250 rpm at ambient temperature). No recognizable changes in physical or chemical stability were detected in any of the monitored attributes. For both the 15 mg / mL and 150 mg / mL drug product formulations, no recognizable changes in HMW and LMW species were observed when incubated at 25 °C for 1 month, indicating that the mAb1 drug product can be exposed to room temperature for short periods. After incubation at 40 °C / 75% RH for 2 months, recognizable formation of HMW species and charge variants (increase in the relative proportion of acidic species) was detected. The results from this accelerated condition demonstrated that the increase in HMW species and the formation of charge variants are the major degradation pathways for the drug product.

[0158] Additionally, 6-month study stability data for the 150 mg / mL drug product formulation in pre-filled syringes (PFS) have been available to date. Stability has been tested in five PFS. The stability data are provided in Tables 26 - 34. 3-month study stability data for the 75 mg / mL drug product formulation in glass vials are available (see Table 36).

[0159]

Table 22

[0160]

Table 23

[0161]

Table 24

[0162]

Table 25

[0163]

Table 26

[0164]

Table 27

[0165]

Table 28

[0166]

Table 29

[0167]

Table 30

[0168]

Table 31

[0169]

Table 32

[0170]

Table 33

[0171]

Table 34

[0172]

Table 35

[0173]

Table 36

[0174]

Table 37

[0175] Results from studies on long-term storage, acceleration, and stress stability indicate that the mAb1 formulation is stable during manufacture (formulation, filling / finishing, and labeling operations) and can withstand short-term exposure to room temperature without compromising physical or chemical stability.

[0176] The present invention should not be limited in scope by the specific embodiments described herein. Indeed, various modifications of the invention in addition to those described herein will be apparent to those skilled in the art from the foregoing description. Such modifications are intended to be included within the scope of the appended claims.

Claims

1. A stable liquid pharmaceutical formulation comprising: (i) a human antibody that specifically binds to human interleukin-33 (hIL-33), said antibody comprising a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 2 and a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 10, said antibody comprising a heavy chain constant region of isotype human IgG4, wherein the antibody is at a concentration of 1 mg / ml to 200 mg / ml; (ii) 5 mM to 15 mM acetate; (iii) 60 mM to 80 mM arginine hydrochloride; (iv) sucrose at a concentration of 3% w / v to 7% w / v; and (v) polysorbate 20 or polysorbate 80 at a concentration of 0.06% w / v to 0.1% w / v, comprising wherein the pH of said formulation is from about 5 to about 5.6, said stable liquid pharmaceutical formulation.

2. The stable liquid pharmaceutical formulation according to claim 1, wherein the antibody is present at a concentration of 15 mg / ml to 150 mg / ml.

3. The stable liquid pharmaceutical formulation according to claim 1, wherein the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 18 and a light chain comprising the amino acid sequence of SEQ ID NO:

20.

4. (i) about 15 mg / ml to about 150 mg / mL of said antibody, (ii) about 5 mM to about 15 mM acetate, (iii) about 60 mM to about 80 mM arginine hydrochloride, (iv) about 3% w / v to about 7% w / v sucrose, and (v) about 0.06% w / v to about 0.1% w / v polysorbate 80, the stable liquid pharmaceutical formulation according to claim 1.

5. (i) about 15 mg / ml ± 1.5 mg / ml of said antibody, (ii) about 10 mM ± 2 mM acetate, (iii) about 70 mM ± 14 mM arginine hydrochloride, (iv) about 5% w / v ± 1% w / v sucrose, and (iv) about 0.08% ± 0.016% w / v polysorbate The stable liquid pharmaceutical preparation according to claim 1, comprising lubelate 80.

6. (i) about 75 mg / ml ± 5 mg / ml of said antibody, (ii) about 10 mM ± 2 mM of acetate, (iii) about 70 mM ± 14 mM of arginine hydrochloride (iv) about 5% w / v ± 1% w / v of sucrose, and (iv) about 0.08% ± 0.016% w / v of polysorbate 80, the stable liquid pharmaceutical preparation according to claim 1.

7. (i) about 150 mg / ml ± 15 mg / ml of said antibody, (ii) about 10 mM ± 2 mM of acetate, (iii) about 70 mM ± 14 mM of arginine hydrochloride (iv) about 5% w / v ± 1% w / v of sucrose, and (v) about 0.08% ± 0.016% w / v of polysorbate 80, the stable liquid pharmaceutical preparation according to claim 1.

8. The stable liquid pharmaceutical preparation according to claim 1, wherein the pH of said preparation is 5.2 to 5.

4.

9. The stable liquid pharmaceutical preparation according to claim 8, wherein the pH of said preparation is about 5.

3.

10. (i) A human antibody that specifically binds to human interleukin-33 (hIL-33) at a concentration of 15 ± 1.5 mg / ml to 150 ± 15 mg / ml, said antibody comprising a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 2 and a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 10, wherein said antibody comprises a heavy chain constant region of isotype human IgG4, (ii) 10 mM ± 2 mM of acetate, and (iii) 70 mM ± 14 mM of arginine hydrochloride, and (iv) 5% w / v ± 1% w / v of sucrose, and (iv) about 0.08% ± 0.016% w / v of polysorbate 80, a stable liquid pharmaceutical preparation having a pH of 5.1 to 5.

5. The stable liquid pharmaceutical preparation.

11. The stable liquid pharmaceutical preparation according to claim 10, wherein the antibody is present at a concentration of 75 mg / ml ± 5 mg / ml.

12. The stable liquid pharmaceutical preparation according to claim 10, wherein the antibody is present at a concentration of 150 mg / ml ± 15 mg / ml.

13. The stable liquid pharmaceutical preparation according to claim 10, wherein the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 18 and a light chain comprising the amino acid sequence of SEQ ID NO:

20.

14. (i) A human antibody that specifically binds to human interleukin-33 (hIL-33) at a concentration of 15 ± 1.5 mg / ml to 150 ± 15 mg / ml, the human antibody comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 18 and a light chain comprising the amino acid sequence of SEQ ID NO: 20, and (ii) 10 mM ± 2 mM acetate, and (iii) 70 mM ± 14 mM arginine hydrochloride, and (iv) 5% w / v ± 1% w / v sucrose, and (iv) about 0.08% ± 0.016% w / v polysorbate 80, a stable liquid pharmaceutical preparation having a pH of 5.1 to 5.

5. The stable liquid pharmaceutical preparation according to claim 1, wherein the antibody has a pH of 5.1 to 5.

5.

15. At least 90% of the native form of the antibody is recovered after storage at 5°C for 2 months as determined by size exclusion - ultra performance liquid chromatography (SE-UPLC), the stable liquid pharmaceutical preparation according to claim 1.

16. At least 95% of the native form of the antibody is recovered after storage at 5°C for 2 months as determined by SE-UPLC, the stable liquid pharmaceutical preparation according to claim 14.

17. At least 99% of the native form of the antibody is determined by SE-UPLC If so, it is recovered after storage at 5°C for 2 months, the stable liquid pharmaceutical preparation according to claim 16.

18. The stable liquid pharmaceutical preparation according to claim 1, wherein at least 95% of the native form of the antibody is recovered after storage at -20°C for 9 months as determined by SE-UPLC.

19. The stable liquid pharmaceutical preparation according to claim 18, wherein at least 97.5% of the native form of the antibody is recovered after storage at -20°C for 9 months as determined by SE-UPLC.

20. The stable liquid pharmaceutical preparation according to claim 19, wherein at least 99% of the native form of the antibody is recovered after storage at -20°C for 9 months as determined by SE-UPLC.

21. The stable liquid pharmaceutical preparation according to claim 1, wherein at least 95% of the native form of the antibody is recovered after storage at 2-8°C for 9 months as determined by SE-UPLC.

22. The stable liquid pharmaceutical preparation according to claim 21, wherein at least 97.5% of the native form of the antibody is recovered after storage at 2-8°C for 9 months as determined by SE-UPLC.

23. The stable liquid pharmaceutical preparation according to claim 22, wherein at least 99% of the native form of the antibody is recovered after storage at 2-8°C for 9 months as determined by SE-UPLC.

24. The stable liquid pharmaceutical preparation according to claim 1, wherein the preparation contains 2% or less of high molecular weight (HMW) species after storage at 5°C for 2 months as determined by SE-UPLC.

25. The stable liquid pharmaceutical preparation according to claim 24, wherein the preparation contains 1% or less of HMW species after storage at 5°C for 2 months as determined by SE-UPLC.

26. The stable liquid pharmaceutical preparation according to claim 25, wherein the preparation contains 0.6% or less of HMW species after storage at 5°C for 2 months as determined by SE-UPLC. Claim 27 The stable liquid pharmaceutical preparation according to claim 1, wherein when the preparation is determined by SE - UPLC, it contains HMW species of 2% or less after storage at - 20°C for 9 months. Claim 28 The stable liquid pharmaceutical preparation according to claim 27, wherein when the preparation is determined by SE - UPLC, it contains HMW species of 1% or less after storage at - 20°C for 9 months. Claim 29 The stable liquid pharmaceutical preparation according to claim 28, wherein when the preparation is determined by SE - UPLC, it contains HMW species of 0.5% or less after storage at - 20°C for 9 months. Claim 30 The stable liquid pharmaceutical preparation according to claim 1, wherein when the preparation is determined by SE - UPLC, it contains HMW species of 2% or less after storage at 2 - 8°C for 9 months. Claim 31 The stable liquid pharmaceutical preparation according to claim 30, wherein when the preparation is determined by SE - UPLC, it contains HMW species of 1% or less after storage at 2 - 8°C for 9 months. Claim 32 The stable liquid pharmaceutical preparation according to claim 31, wherein when the preparation is determined by SE - UPLC, it contains HMW species of 0.7% or less after storage at 2 - 8°C for 9 months. Claim 33 The stable liquid pharmaceutical preparation according to claim 1, wherein the preparation exhibits a viscosity of less than about 15 cP. Claim 34 The stable liquid pharmaceutical preparation according to any one of claims 1 - 33, contained in a glass vial. Claim 35 The stable liquid pharmaceutical preparation according to any one of claims 1 - 33, contained in a syringe. Claim 36 The stable liquid pharmaceutical preparation according to claim 35, wherein the syringe contains a fluorocarbon - coated plunger. Claim 37 The stable liquid pharmaceutical preparation according to claim 35, wherein the syringe is a low-tungsten syringe.

38. The stable liquid pharmaceutical preparation according to claim 35, which is a pre-filled syringe.

39. The stable liquid pharmaceutical preparation according to claim 35, which is a pre-filled stacked needle syringe.

40. The stable liquid pharmaceutical preparation according to any one of claims 1 to 33, which is included in a large-volume device or a bolus syringe.

41. The stable liquid pharmaceutical preparation according to any one of claims 1 to 33, which is included in a pen delivery device.

42. The stable liquid pharmaceutical preparation according to any one of claims 1 to 33, which is included in an auto-injector delivery device.

43. A pen or auto-injector delivery device comprising the stable liquid pharmaceutical preparation according to any one of claims 1 to 33.

44. The delivery device according to claim 43, which is a disposable pen delivery device.

45. The delivery device according to claim 43, which is a reusable pen delivery device.

46. A container comprising the stable liquid pharmaceutical preparation according to any one of claims 1 to 33.

47. A kit comprising (i) a container comprising the stable liquid pharmaceutical preparation according to any one of claims 1 to 33, and (ii) labeling for use of the stable liquid pharmaceutical preparation.

48. The kit according to claim 47, wherein the labeling lists subcutaneous administration of the stable liquid pharmaceutical preparation. of the kit.

49. The kit according to claim 47, wherein the labeling lists intravenous administration of the stable liquid pharmaceutical preparation. Claim 50 A unit dosage form comprising the stable liquid pharmaceutical formulation according to claim 6, wherein the anti-IL-33 antibody is present in an amount of about 150 mg. Claim 51 A unit dosage form comprising the stable liquid pharmaceutical formulation according to claim 7, wherein the anti-IL-33 antibody is present in an amount of about 300 mg. Claim 52 The unit dosage form according to claim 50 or 51, wherein the stable liquid pharmaceutical formulation is contained in a syringe. Claim 53 The unit dosage form according to claim 52, wherein the syringe is a pre-filled syringe. Claim 54 A safety system delivery device comprising the stable liquid pharmaceutical formulation according to any one of claims 1 to 33. Claim 55 The safety system delivery device according to claim 54, comprising a safety sleeve configured to extend by manual operation. Claim 56 The safety system delivery device according to claim 54, comprising a safety sleeve configured to automatically extend after injection of the stable liquid pharmaceutical formulation.

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