Anti-IL-33 antibody formulations

Abstract

The present disclosure is directed to a composition comprising greater than about 100 mg / ml of an anti-IL-33 antibody, a surfactant, arginine, and a buffer. Methods for making the composition and methods for treating a disease in a subject are also disclosed.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to Greek Patent Application No. 20200100239, filed May 11, 2020, the contents of which are incorporated herein by reference in their entirety.

[0002] The present disclosure relates to anti-IL-33 antibodies, including highly concentrated aqueous formulations of 33_640087-7B and biosimilars thereof. [Background technology]

[0003] 33_640087-7B is a human immunoglobulin (Ig) G1 monoclonal antibody (mAb) that binds to human interleukin (IL)-33, prevents IL-33 from binding to its receptor ST2, and inhibits the conversion of disulfide-bonded (DSB) IL-33. 33_640087-7B has therapeutic potential in many diseases and is currently being developed for the treatment of moderate to severe chronic obstructive pulmonary disease (COPD), asthma, and atopic dermatitis (AD); and diabetic nephropathy (DKD).

[0004] 33_640087-7B is intended to be administered via subcutaneous injection in the future. The present disclosure addresses this need. Summary of the Invention [Means for solving the problem]

[0005] A Phase 1 clinical trial (Study D9180C00001) of 33_640087-7B has been completed. Study D9180C00001 was a first-in-human, randomized, placebo-controlled, blinded (blinded to investigators and participants; open to sponsor) clinical trial in 88 participants (Part I: single ascending dose (SAD) study in 56 healthy volunteers with a history of mild atopy; Part II: multiple ascending dose (MAD) study in 24 participants with mild chronic obstructive pulmonary disease; Part III: single dose in 8 healthy Japanese volunteers) to evaluate the safety, tolerability, PK, and immunogenicity of 33_640087-7B. 33_640087-7B was found to be generally safe and well-tolerated, with no safety concerns following intravenous (IV) administration of up to 300 mg of 33_640087-7B in Parts I and III of this study. In Part 2 of this study, patients with COPD received three subcutaneous doses of 33_640087-7B, 14 days apart.

[0006] For Phase 1 clinical trials, 33_640087-7B was supplied in vials as a sterile, white to off-white, lyophilized powder. Each vial contained a nominal 50 mg of active 33_640087-7B intended for IV or SC administration. Upon reconstitution with 1.2 mL of sterile water for injection, the solution contained 50 mg / mL 33_640087-7B in 20 mM L-histidine / L-histidine hydrochloride, 80 mM L-arginine hydrochloride, 120 mM sucrose, 0.02% (weight / volume [w / v]) polysorbate 80, pH 6.0.

[0007] The expected effective dose of 33_640087-7B may be as much as (or greater than) 300 mg for certain conditions. Therefore, the low-concentration formulation may pose a barrier to subcutaneous administration, especially for use in chronic conditions. It is unrealistic to expect patients suffering from chronic disorders to routinely receive subcutaneous administration of 6 ml or more of drug product to receive a therapeutic dose.

[0008] As such, there is a need to increase the concentration of 33_640087-7B in drug formulations, especially for subcutaneous administration.

[0009] However, increasing the protein concentration in a drug formulation can cause stability problems, such as the formation of high molecular weight species (HMWS) as a result of protein aggregation. HMWS, especially those that retain most of the native conformation of antibodies, can be of particular concern in some protein formulations. High protein concentrations can induce reversible self-association (RSA), which can lead to the formation of soluble higher-order species. Aggregation can also affect the subcutaneous bioavailability and pharmacokinetics of therapeutic proteins. Furthermore, the formation of soluble higher-order species at high concentrations can increase the viscosity of the solution, making it difficult to deliver the drug product, especially from devices with high backpressure, such as pre-filled syringes.

[0010] Provided herein are improved formulations for anti-IL-33 antibodies that contain high concentrations of antibody while having low viscosity and reduced reversible self-association characteristics.

[0011] In one aspect, the disclosure provides a composition comprising greater than about 100 mg / ml of an anti-IL-33 antibody, at least about 170 mM arginine, and a buffer, wherein the anti-IL-33 antibody comprises a heavy chain variable domain comprising a VHCDR1 having the sequence of SEQ ID NO: 1, a VHCDR2 having the sequence of SEQ ID NO: 2, a VHCDR3 having the sequence of SEQ ID NO: 3; and a heavy chain variable domain comprising a VLCDR1 having the sequence of SEQ ID NO: 5, a VLCDR2 having the sequence of SEQ ID NO: 6, and a VLCDR3 having the sequence of SEQ ID NO: 7. In some examples, the composition further comprises a surfactant.

[0012] In another aspect, the disclosure provides a composition comprising greater than about 100 mg / ml of an anti-IL-33 antibody, at least about 150 mM lysine, and a buffer, wherein the anti-IL-33 antibody comprises a heavy chain variable domain comprising a VHCDR1 having the sequence of SEQ ID NO: 1, a VHCDR2 having the sequence of SEQ ID NO: 2, a VHCDR3 having the sequence of SEQ ID NO: 3; and a heavy chain variable domain comprising a VLCDR1 having the sequence of SEQ ID NO: 5, a VLCDR2 having the sequence of SEQ ID NO: 6, and a VLCDR3 having the sequence of SEQ ID NO: 7. In some examples, the composition further comprises a surfactant.

[0013] In some examples, the composition is liquid. In some examples, the composition is characterized by a reduced viscosity compared to a composition comprising a lower concentration of an anti-IL-33 antibody, 20 mM histidine, 80 mM arginine, 120 mM sucrose, and 0.02% (w / v) polysorbate 80, pH 6.0. In some examples, the composition is characterized by a reduced reversible self-association of the anti-IL-33 antibody compared to the reversible self-association of the anti-IL-33 antibody in a composition comprising 20 mM histidine, 80 mM arginine, 120 mM sucrose, and 0.02% (w / v) polysorbate 80, pH 6.0 and a lower concentration of the anti-IL-33 antibody.

[0014] In a further aspect, the disclosure provides a composition comprising about 130 mg / ml to about 170 mg / ml of 33_640087-7B, about 0.03% (w / v) ±0.015% polysorbate 80, about 220 mM arginine, and about 16 to about 24 mM histidine buffer, wherein the pH is pH 5.5 ±0.5.

[0015] In a further aspect, the present disclosure provides a composition comprising about 130 mg / ml to about 170 mg / ml of an anti-IL-33 antibody, about 0.03% (w / v) ± 0.015% polysorbate 80, about 220 mM arginine, and about 16 to about 24 mM histidine buffer, wherein the pH is 5.5 ± 0.5.

[0016] In a further aspect, the present disclosure provides an article of manufacture comprising a composition disclosed herein, for example, comprising 0.5 ml to about 5 ml (eg, 1 ml to about 3 ml) of the composition.

[0017] In a further aspect, the present disclosure provides an article of manufacture comprising a composition disclosed herein, for example, comprising about 0.5 ml to about 5 ml (eg, 1 ml to about 3 ml) of the composition.

[0018] In a further aspect, the present disclosure provides a method of treating an IL-33-mediated disorder in a subject, comprising administering to the subject a therapeutically effective amount of a composition disclosed herein.

[0019] In a further aspect, the disclosure provides a method for making a stable liquid composition having a viscosity of less than about 10 cP and comprising greater than about 100 mg / mL of an anti-IL-33 antibody, at least about 170 mM arginine, and optionally a surfactant and a buffer. The method includes the steps of (i) combining a first solution comprising a first concentration of antibody and a buffer with arginine to obtain a solution comprising about 110 mg / mL to about 200 mg / mL of an anti-IL-33 antibody, at least about 170 mM arginine, and a buffer; and optionally (ii) adding a surfactant to the solution to achieve a final concentration of surfactant of about 0.03% (w / v) ± 0.015% (w / v).

[0020] In a further aspect, the disclosure provides a method for making a stable liquid composition having a viscosity of less than about 10 cP and comprising greater than about 100 mg / mL of an anti-IL-33 antibody, at least about 150 mM lysine, and optionally a surfactant and a buffer. The method includes (i) combining a first solution comprising a first concentration of antibody and a buffer with arginine to obtain a solution comprising about 110 mg / mL to about 200 mg / mL of an anti-IL-33 antibody, at least about 150 mM lysine, and a buffer, and optionally (ii) adding a surfactant to the solution to achieve a final concentration of about 0.02% (w / v) ± 0.015% (w / v) surfactant.

[0021] Embodiments of the present invention will now be described, by way of example only, with reference to the following drawings: [Brief explanation of the drawings]

[0022] [Figure 1] FIG. 1 shows the stability of the % distribution of soluble higher order species of a composition of the present disclosure, 33_640087-7B, when stored at 2° C.-8° C. over a 6-month period. [Figure 2] Figure 2 shows the long-term stability of 33_640087-7B in compositions of the present disclosure, measured as a function of the formation of insoluble aggregates over time (months). Aggregation levels were measured during storage in glass vials and pre-filled syringes (PFS). The compositions were stored at 2-8°C. [Figure 3] Figure 3 shows the stability of 33_640087-7B in compositions of the present disclosure, measured as a function of the formation of insoluble aggregates over time (months-M). Aggregation levels were measured during storage in glass vials and pre-filled syringes (PFS). The compositions were stored at 25°C. [Figure 4] Figure 4 shows the stability of 33_640087-7B in compositions of the present disclosure, measured as a function of the formation of insoluble aggregates over time (months-M). Aggregation levels were measured during storage in glass vials and pre-filled syringes (PFS). The compositions were stored at 40°C. [Figure 5] FIG. 5 shows the effect of arginine and temperature on formulation viscosity. [Figure 6] Figure 6 shows the effect of 33-640087_7B concentration, arginine, and temperature on formulation viscosity. Viscosity (Cp) was measured at each arginine concentration and each 33-640087_7B concentration at 5°C, 18°C, 25°C, and 30°C. DETAILED DESCRIPTION OF THE INVENTION

[0023] definition It should be understood that the specific implementations shown and described herein are examples and are not intended to otherwise limit the scope of the application in any way.

[0024] Published patents, patent applications, websites, company names, and scientific literature mentioned in this specification are incorporated by reference in their entirety into this specification to the same extent as if each was specifically and individually indicated to be incorporated by reference.

[0025] As used herein, the singular forms "a," "an," and "the" specifically include plural forms of the terms they refer to, unless the content clearly dictates otherwise.

[0026] The term "about" or "approximately" refers to an acceptable error for a particular value as determined by one of ordinary skill in the art, and depends in part on how the value is measured or determined. In certain embodiments, the term "about" or "approximately" means within 1, 2, 3, or 4 standard deviations. In certain embodiments, the term "about" or "approximately" means within 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.05% of a given value or range. Whenever the term "about" or "approximately" appears before the first number in a series of two or more numbers, it should be understood that the term "about" or "approximately" applies to each and every one of the numbers in the series.

[0027] The methods disclosed herein and their individual steps can be performed manually and / or with assistance provided by electronic devices or automated. While steps have been described with reference to specific examples, those skilled in the art will readily recognize that other methods of performing the acts associated with the methods can be used. For example, unless otherwise noted, the order of various steps can be changed without departing from the scope or spirit of the methods. Furthermore, some of the individual steps can be combined, omitted, or further subdivided into additional steps.

[0028] The present compositions and methods are intended to include embodiments that include any combination of one or more of the additional optional elements, features, and steps further described below (including those shown in the drawings), unless otherwise stated.

[0029] 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 disclosure belongs. References, including but not limited to, the following, provide those skilled in the art with general definitions of many of the terms used in this disclosure: Singleton et al., DICTIONARY OF MICROBIOLOGY AND MOLECULAR BIOLOGY (2nd Ed. 1994); THE CAMBRIDGE DICTIONARY OF SCIENCE AND TECHNOLOGY (Walker Ed., 1988); THE GLOSSARY OF GENETICS, 5th Ed., R. Rieger et al. (Eds.), Springer Verlag (1991); and Hale & Marham, THE HARPER COLLINS DICTIONARY OF BIOLOGY (1991).

[0030] The term "antibody" or "immunoglobulin" refers to a tetrameric glycoprotein consisting of two heavy chains and two light chains, each containing a variable region and a constant region. Antigen-binding portions may be produced by recombinant DNA techniques or by enzymatic or chemical cleavage of intact antibodies. The term "antibody" includes monoclonal antibodies, polyclonal antibodies, chimeric antibodies, human antibodies, and humanized antibodies.

[0031] Antibody variants include antibody fragments and antibody-like proteins that have changes to the structure of a canonical tetrameric antibody. Typically, antibody variants include V regions with changes to the constant region, or alternatively, optionally non-canonically, the addition of V regions to the constant region. Examples include multispecific antibodies (e.g., bispecific antibodies with extra V regions), antibody fragments that can bind to antigen (e.g., Fab', F'(ab)2, Fv, single-chain antibodies, diabodies), biparatopic and recombinant peptides containing the foregoing, so long as they exhibit the desired biological activity.

[0032] Antibody fragments include antigen-binding portions of antibodies (i.e., antigen-binding fragments"), including Fab, Fab', F(ab'), Fv, domain antibodies (dAbs), complementarity-determining region (CDR) fragments, CDR-grafted antibodies, single-chain antibodies (scFv), single-chain antibody fragments, chimeric antibodies, diabodies, triabodies, tetrabodies, minibodies, linear antibodies; chelating recombinant antibodies, tribodies or bibodies, intrabodies, nanobodies, small molecule modular immunopharmaceuticals (SMIPs), antigen-binding domain immunoglobulin fusion proteins, single-domain antibodies (including camelized antibodies), VHH-containing antibodies or variants or derivatives thereof, and polypeptides containing at least a portion of an immunoglobulin sufficient to confer specific antigen binding to the polypeptide, such as one, two, three, four, five, or six CDR sequences, so long as the antibody retains the desired biological activity.

[0033] The terms "treat," "treating," and "treatment" refer to eliminating, alleviating, suppressing, or ameliorating, either temporarily or permanently, either partially or completely, the clinical symptoms, signs, or progression of an event, disease, or condition associated with a disorder described herein. As recognized in the relevant art, a drug used as a therapeutic agent may reduce the severity of certain disease states, but need not eliminate all signs of the disease to be considered a useful therapeutic agent. Similarly, a treatment administered prophylactically need not be completely effective in preventing the onset of the condition to constitute a viable prophylactic agent. Simple reduction in the effects of the disease (e.g., by reducing the number or severity of its symptoms, or by improving the effectiveness of another treatment, or by producing another beneficial effect), or reducing the likelihood that the disease will occur or worsen in a subject, is sufficient. One embodiment of the present disclosure is directed to a method for determining the efficacy of treatment, the method comprising administering to a patient a therapeutic agent in an amount and for a time sufficient to induce a sustained improvement over baseline in an indicator reflective of the severity of a particular disorder.

[0034] As used herein, "IL-33" protein refers to interleukin-33, specifically mammalian interleukin-33 protein, e.g., the human protein deposited under UniProt number 095760. IL-33 exists in reduced and oxidized forms. The terms "IL-33" and "IL-33 polypeptide" are used interchangeably. In certain embodiments, the IL-33 is full-length. In other embodiments, the IL-33 is mature truncated IL-33 (amino acids 112-270). Recent studies suggest that full-length IL-33 is active (Cayrol and Girard, Proc Natl Acad Sci USA 106(22):9021-6(2009); Hayakawa et al., Biochem Biophys Res Commun. 387(1):218-22(2009); Talabot-Ayer et al., J Biol Chem. 284(29):19420-6(2009)). However, N-terminally processed or truncated forms of IL-33, including but not limited to aa 72-270, 79-270, 95-270, 99-270, 107-270, 109-270, 111-270, and 112-270, may have enhanced activity (Lefrancais 2012, 2014).

[0035] "Oxidized IL-33" or "oxIL-33," as used herein, refers to a form of IL-33 that binds to RAGE and initiates RAGE-mediated signaling. Oxidized IL-33 refers to a protein that appears as a characteristic band, for example, by Western blot analysis under non-reducing conditions, and in particular has a mass 4 Da less than the corresponding reduced form. Specifically, it refers to a protein that has one or two disulfide bonds between cysteines independently selected from cysteines 208, 227, 232, and 259. In one embodiment, oxidized IL-33 does not exhibit binding to ST2.

[0036] "Reduced IL-33" or "redIL-33," as used herein, refers to a form of IL-33 that binds to ST2 and initiates ST2-mediated signaling. In particular, cysteines 208, 227, 232, and 259 in the reduced form are not disulfide bonded. In one embodiment, reduced IL-33 does not exhibit binding to RAGE.

[0037] The term "IL-33-mediated disorder," as used herein, refers to any disorder or condition mediated by or associated with the IL-33 system. In some examples, an IL-33-mediated disorder is associated with excessive IL-33 levels or activity, which may manifest atypical symptoms locally and / or systemically in the body due to IL-33 levels or activity. Exemplary IL-33-mediated disorders include inflammatory conditions, immune disorders, fibrotic disorders, eosinophilic disorders, infections, pain, central nervous system disorders, solid tumors, and ophthalmological disorders. IL-33-mediated disorders are described, for example, in Liew et al. Nature Reviews Immunology 10:103-110, 2010, which is incorporated herein by reference in its entirety. "IL-33-mediated disorder" may also refer to "IL33-driven disorders" herein.

[0038] In some examples, the IL-33 mediated inflammatory disease can be any of asthma, sepsis, septic shock, atopic dermatitis, allergic rhinitis, rheumatoid arthritis, chronic obstructive pulmonary disease (COPD), asthma, COPD overlap syndrome (ACOS), chronic bronchitis, emphysema, chronic sinusitis with or without nasal polyps, vasculitis, GvHD, uveitis, chronic spontaneous urticaria, sinusitis, or pancreatitis.

[0039] In some examples, the IL-33-mediated disorder can be diabetic nephropathy. Diabetic nephropathy, as defined herein, refers to a diagnosis of type 2 diabetes and an estimated glomerular filtration rate (eGFR) of 30-75 ml / min. Typically, DKD is further defined as a diagnosis of a UACR ratio of 100-3000 mg albumin to 1 g creatinine. The term "therapeutically effective amount" refers to an amount of a therapeutic agent that is effective to ameliorate or alleviate the symptoms or signs of a disease associated with a disease or disorder.

[0040] Compositions with low reversible self-association 33_640087-7B has been shown to be safe and generally well tolerated in humans at doses up to 300 mg administered parenterally. Depending on the disease and the local biology of interleukin-33, a therapeutically effective amount of 33_640087-7B can be equal to or exceed 300 mg.

[0041] Many conditions believed to be mediated by IL-33 are chronic and long-lasting. This means that patients may need to be administered anti-IL-33-based therapeutics over a long period of time to effectively manage the symptoms of the disease. Therefore, compositions containing 33_640087-7B that are suitable for therapeutic use should make long-term treatment as comfortable as possible for the patient.

[0042] Therefore, high-concentration formulations suitable for parenteral administration (e.g., subcutaneous or intravenous administration) are desirable. However, formulations with high protein concentrations can be challenging from a developability perspective. For example, 33_640087-7B has been shown to reversibly self-associate at moderately high concentrations (e.g., approximately 50 mg / ml). Reversible self-association (RSA) is an important developability parameter for high-concentration formulations. RSA often manifests in the form of soluble, reversible higher-order species (e.g., noncovalent dimers), which can cause problems during manufacturing and drug administration. For example, the presence of RSA can lead to increased viscosity. High viscosity can make manufacturing very difficult, for example, by clogging filters during the filtration process. This, in turn, can lead to reduced yields if significant amounts of drug substance are lost during the purification process as the higher-order species are purified from the monomer. Increased viscosity can also negatively impact drug administration by reducing the functionality of the device used to administer the antibody, especially when the drug is administered parenterally, such as by subcutaneous administration. In such situations, high viscosity can impair the ability of the end user (e.g., patient or medical professional) to manually inject.

[0043] Provided herein are stable liquid compositions (i.e., "liquid formulations") suitable for parenteral administration that can be stored for long or short periods and contain a high concentration of an anti-IL-33 antibody (meaning greater than 100 mg / ml), optionally a surfactant, a high concentration of arginine or lysine, and a buffer. As used herein, "high concentration of arginine" refers to a concentration of arginine greater than about 170 mM, such as greater than about 190 mM. "High concentration of lysine" refers to a concentration of lysine greater than about 150 mM. The compositions disclosed herein have surprisingly been shown to reduce reversible self-association (RSA) of 33_640087-7B in high concentration liquid compositions compared to RSA previously observed at lower 33_640087-7B concentrations in Phase I formulations (see Example 1).

[0044] In certain examples, the anti-IL-33 antibody is present in the composition at a concentration greater than about 100 mg / ml and optionally less than about 200 mg / ml. In some examples, the anti-IL-33 antibody is present in the composition at a concentration of 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 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 190 mg / ml, or about 195 mg / ml. In some examples, the anti-IL-33 antibody is present in the composition at a concentration of about 105 mg / ml to about 190 mg / ml, about 110 mg / ml to about 180 mg / ml, about 110 mg / ml to about 170 mg / ml, about 110 mg / ml to about 165 mg / ml, about 110 mg / ml to about 160 mg / ml, about 120 mg / ml to about 160 mg / ml, about 130 mg / ml to about 160 mg / ml, or 140 mg / ml to about 160 mg / ml. In some examples, the anti-IL-33 antibody is present in the composition at a concentration of about 110 mg / ml ± 10%, about 115 mg / ml ± 10%, about 120 mg / ml ± 10%, about 125 mg / ml ± 10%, about 130 mg / ml ± 10%, about 135 mg / ml ± 10%, about 140 mg / ml ± 10%, about 145 mg / ml ± 10%, about 150 mg / ml ± 10%, about 155 mg / ml ± 10%, about 160 mg / ml ± 10%, about 165 mg / ml ± 10%, about 170 mg / ml ± 10%, about 175 mg / ml ± 10%, about 180 mg / ml ± 10%, about 185 mg / ml ± 10%, about 190 mg / ml ± 10%, or about 195 mg / ml ± 10%. In one example, the anti-IL-33 antibody is present in the composition at a concentration of about 150 mg / ml.

[0045] In some examples, the compositions of the present disclosure include a surfactant. A surfactant is an amphiphilic, surface-active substance (the head is polar and the tail is hydrophobic). Surfactants preferentially accumulate at interfaces, resulting in a decrease in interfacial tension. The use of a surfactant can also help mitigate the formation of large proteinaceous particles. In some examples, the surfactant present in the compositions of the present disclosure is an amphiphilic and / or nonionic surfactant. Exemplary surfactants, either within or between surfactant classes, include polyoxyethylene sorbitan fatty acid esters (e.g., polysorbate 20, polysorbate 80), alkylaryl polyethers such as oxyethylated alkylphenols (e.g., Triton™ X-100), and poloxamers (e.g., Pluronics®, e.g., Pluronic® F68), and combinations of any of the foregoing. Polysorbate 20 and polysorbate 80 (and optionally mixtures thereof) are specifically contemplated. In representative examples, the surfactant is present in the composition at a concentration of about 0.050% (w / v) ± 0.015% (w / v) or less. For example, the composition may contain about 0.005% (w / v) to about 0.05% (w / v) of the surfactant, such as about 0.005% (w / v), about 0.015% (w / v), about 0.02% (w / v), about 0.025% (w / v), about 0.03% (w / v), about 0.035% (w / v), about 0.04% (w / v), about 0.045% (w / v), or about 0.05% (w / v). In some examples, the surfactant in the composition is at a concentration of 0.03% (w / v) ± 0.015% (w / v). In some examples, the surfactant in the composition is at a concentration of 0.03% (w / v) ± 0.01% (w / v). In some examples, the surfactant in the composition is at a concentration of about 0.02% (w / v) to about 0.04% (w / v). In some embodiments, the surfactant comprises polysorbate 80.

[0046] The compositions of the present disclosure also include at least about 170 mM arginine. In some examples, the compositions include at least about 190 mM arginine. In some examples, the compositions of the present disclosure include L-arginine. In some examples, the compositions of the present disclosure include L-arginine hydrochloride. In some examples, the compositions include more than 190 mM arginine. In some examples, the compositions include less than about 500 mM arginine. For example, the compositions include about 200 mM, about 210 mM, about 220 mM, about 240 mM, about 260 mM, about 280 mM, about 300 mM, about 350 mM, about 400 mM, or about 450 mM arginine. In some examples, the compositions include about 220 mM arginine. High concentrations of arginine (i.e., greater than about 190 mM) have been found to contribute to a surprising reduction in reversible self-association of 33_640087-7B in stable liquid compositions compared to the reversible self-association observed in a composition comprising 20 mM histidine, 80 mM arginine, 120 mM sucrose, 0.02% (w / v) polysorbate 80, pH 6.0, and a lower concentration of 33_640087-7B (about 50 mg / ml). In some examples, compositions of the present disclosure comprise about 220 mM arginine when the composition comprises about 150 mg / ml of an anti-IL-33 antibody.

[0047] In another embodiment, the compositions of the present disclosure alternatively comprise at least about 150 mM lysine. In some examples, the compositions of the present disclosure comprise L-lysine. In some examples, the compositions of the present disclosure comprise L-lysine hydrochloride. In some examples, the compositions comprise more than 150 mM lysine. In some examples, the compositions comprise less than about 500 mM lysine. For example, the compositions comprise about 160 mM, about 170 mM, about 180 mM, about 200 mM, about 220 mM, about 240 mM, about 260 mM, about 280 mM, about 300 mM, about 350 mM, about 400 mM, or about 450 mM lysine. In some examples, the compositions comprise about 170 mM lysine. High concentrations of lysine (i.e., concentrations greater than about 150 mM) have been found to contribute to a surprising decrease in the viscosity of 33_640087-7B in stable liquid compositions compared to the viscosity observed in a composition comprising 20 mM histidine, 80 mM arginine, 120 mM sucrose, 0.02% (w / v) polysorbate 80, pH 6.0, and a lower concentration of 33_640087-7B (about 50 mg / ml). In some examples, compositions of the present disclosure comprise about 170 mM lysine when the composition comprises about 150 mg / ml of an anti-IL-33 antibody.

[0048] In some examples, the compositions of the present disclosure are characterized by reduced reversible self-association of the anti-IL-33 antibody compared to a composition comprising 20 mM histidine, 80 mM arginine, 120 mM sucrose, 0.02% (w / v) polysorbate 80, pH 6.0, and a lower concentration of the anti-IL-33 antibody. In some examples, this RSA is reduced by two-fold compared to the RSA of the anti-IL-33 antibody in a liquid composition comprising a lower concentration of the anti-IL-33 antibody in 20 mM histidine, 80 mM arginine, 120 mM sucrose, 0.02% (w / v) polysorbate 80, pH 6.0. In some examples, the lower concentration of the anti-IL-33 antibody is 50 mg / ml. RSA is positively correlated with protein concentration. Therefore, it is expected that increasing the concentration of the therapeutic protein will lead to an increased RSA of the therapeutic protein. RSA can be expressed as a function of the % monomer of soluble species in the liquid composition. This example unexpectedly demonstrates that the novel formulations can achieve a two-fold increase in weight percent monomer when the concentration of a therapeutic protein is increased three-fold compared to the weight percent monomer observed in a composition comprising 20 mM histidine, 80 mM arginine, 120 mM sucrose, 0.02% (w / v) polysorbate 80, pH 6.0. In some examples, the compositions of the present disclosure comprise at least about 30%, e.g., about 30%, about 31%, about 32%, about 33%, about 34%, about 35%, about 36%, about 37%, about 38%, about 39%, about 40%, about 41%, about 42%, about 43%, about 44%, about 45%, about 46%, about 47%, about 48%, about 49%, or about 50% weight percent monomer. In some examples, the compositions of the present disclosure comprise between about 35% and about 50% weight percent monomer. In some examples, compositions of the present disclosure comprise about 40% to about 45% by weight of monomer. In some examples, compositions of the present disclosure comprise about 35% to about 50% by weight of monomer after about 3 months of storage at a temperature of about 2°C to about 8°C. In some examples, compositions of the present disclosure comprise about 40% by weight of monomer after about 3 months of storage at a temperature of about 2°C to about 8°C. In some examples, compositions of the present disclosure comprise about 35% to about 50% by weight of monomer after about 6 months of storage at a temperature of about 2°C to about 8°C. In some examples, compositions of the present disclosure comprise from about 40% to about 60% by weight of monomer after about 6 months of storage at a temperature of about 2°C to about 8°C.RSA and % monomer can be calculated using static light scattering intensity in volts measured as a function of concentration (mg / mL). This can then be converted to apparent molecular weight (kDa) at each concentration using the Rayleigh equation. Unless otherwise stated, this method is used to measure RSA in the stable liquid compositions disclosed herein.

[0049] The compositions of the present disclosure include a buffer. The buffer can be, for example, an organic buffer. In some examples, the buffer is centered around pH 5 to pH 6.5 or between pH 5 and pH 6 at 25°C. In some embodiments, the buffer can have a pKa within 1 pH unit of pH 5.4 to pH 5.6 at 25°C. An exemplary buffer is histidine / histidine hydrochloride, which has a pKa of approximately pH 6.09 at 25°C. Another such buffer is acetic acid / acetate, which has a pKa of approximately 4.75 at 25°C. Other contemplated alternative buffers include ion-based buffers including propionate (pKa 4.87 at 25°C), malate (pKa 5.13 at 25°C), pyridine (pKa 5.23 at 25°C), piperazine (pKa 5.33 at 25°C), and succinate (pKa 5.40 at 25°C). Histidine-based buffers are particularly contemplated. In some examples, the buffer is histidine.

[0050] In some examples, the buffer in the composition is present at a concentration of about 1 mM to about 50 mM, about 1 to 40 mM, or about 1 mM to about 30 mM. In some examples, the composition includes about 5 mM to about 50 mM, about 10 mM to about 40 mM, about 15 to about 30 mM buffer, or about 15 to about 25 mM buffer. In some examples, the buffer in the composition is present at a concentration of about 11 mM, about 12 mM, about 13 mM, about 14 mM, about 15 mM, about 16 mM, about 17 mM, about 18 mM, about 19 mM, about 20 mM, about 21 mM, about 22 mM, about 23 mM, about 24 mM, about 25 mM, about 26 mM, about 27 mM, about 28 mM, about 29 mM, or about 30 mM. In some examples, the buffer is at a concentration of about 16 mM to about 24 mM, about 17 mM to about 24 mM, about 18 mM to about 24 mM, or about 19 mM to about 21 mM. In some examples, the composition comprises about 20 mM ± 10% buffer.

[0051] In examples, the compositions of the present disclosure may include additional components, such as acidifiers, additives, adsorbents, aerosol propellants, air displacement agents, etc., in various embodiments. The pharmaceutical composition may comprise any pharmaceutically acceptable ingredient, including an aqueous solution of a pharmaceutical composition comprising an aqueous solution of a pharmaceutical agent, an alkalizing agent, an anti-caking agent, an anticoagulant, an antimicrobial agent, a preservative, an antioxidant, an antiseptic, a base, a binder, a buffering agent, a chelating agent, a coating agent, a coloring agent, a drying agent, a detergent, a diluent, a disinfectant, a disintegrating agent, a dispersing agent, a dissolution enhancer, a dye, an emollient, an emulsifier, an emulsion stabilizer, a filler, a film-forming agent, a flavor enhancer, a flavoring agent, a flow enhancer, a gelling agent, a granulating agent, a humectant, a lubricant, a mucoadhesive base, an ointment, an oily vehicle, an organic base, a lozenge base, a pigment, a plasticizer, an abrasive, a preservative, a sequestering agent, a skin penetration agent, a solubilizer, a solvent, a stabilizer, a suppository base, a surface-active agent, a surfactant, a suspending agent, a sweetener, a therapeutic agent, a thickener, an isotonic agent, a toxicant, a viscosity-enhancing agent, a water-absorbing agent, a water-miscible co-solvent, a water softener, or a humectant. See, e.g., Handbook of Pharmaceutical Excipients, Third Edition, A.H. Kibbe (Pharmaceutical Press, London, UK, 2000), which is incorporated by reference in its entirety; Remington's Pharmaceutical Sciences, Sixteenth Edition, E.W. Martin (Mack Publishing Co., Easton, Pa., 1980), which is incorporated by reference in its entirety.

[0052] In some examples, the compositions of the present disclosure are liquid. In particular examples, the pH of the liquid is less than about 6.0, optionally about 5.5. In some examples, the pH is about 5.0 to about 6.0 or about 5.3 to about 5.8, e.g., about 5.3, about 5.4, about 5.5, about 5.6, about 5.7, or about 5.8, or about 5.4. In some examples, the pH is about 5.5.

[0053] In some examples, the composition is characterized by a reduced viscosity compared to a formulation comprising a lower concentration of anti-IL-33 antibody, 20 mM histidine, 80 mM arginine, 120 mM sucrose, 0.02% (w / v) polysorbate 80, pH 6.0. In some examples, the composition is characterized by a viscosity of less than about 25 centipoise (cP) at 23° C. when the concentration of the anti-IL-33 antibody is less than 165 mg / mL, optionally a viscosity of about 9 cP when the concentration of the anti-IL-33 antibody is about 150 mg / mL, or a viscosity of about 5 cP when the concentration of the anti-IL-33 antibody is about 130 mg / mL. In certain aspects, the composition is characterized in that when the concentration of the anti-IL-33 antibody is less than about 150 mg / mL (e.g., about 130 mg / mL, about 140 mg / mL, about 150 mg / mL), the viscosity is about 5 cP to about 20 cP, e.g., about 5 cP to about 15 cP, about 5 cP to about 10 cP, about 10 cP to about 20 cP, about 15 cP to about 20 cP, or about 5 cP, about 6 cP, about 7 cP, about 8 cP, about 9 cP, about 10 cP, about 11 cP, about 12 cP, about 13 cP, about 14 cP, about 15 cP, about 16 cP, about 17 cP, about 18 cP, about 19 cP, or about 20 cP. In exemplary embodiments, the composition has a viscosity of about 10 cP±5 cP when the antibody concentration is about 130 mg / mL to about 170 mg / mL. In some examples, the composition is characterized by a viscosity of less than about 10 centipoise (cP) at 23°C when the anti-IL-33 antibody concentration is about 150 mg / mL. In some examples, the viscosity is about 5 cP to about 20 cP, optionally less than about 10 cP, such as about 9 cP. Unless otherwise stated, all viscosities disclosed herein refer to viscosities measured using a rotational viscometer at 23°C and a shear rate of about 1000 l / s.

[0054] In a typical embodiment, the composition is intended for subcutaneous administration to a subject, and therefore is isotonic with the intended administration site. For example, the osmolality of the composition ranges from about 340 to about 520 mOsm / kg, or from about 344 to about 516 mOsm / kg, or from about 400 to about 500 mOsm / kg. In a typical example, the liquid pharmaceutical composition has an osmolality ranging from about 300 mOsm / kg to about 600 mOsm / kg, or from about 340 mOsm / kg to about 520 mOsm / kg, or from about 360 mOsm / kg to about 500 mOsm / kg. In some examples, the osmolality is about 452 mOsm / kg.

[0055] Compositions of the present disclosure are advantageously suitable for long-term or short-term storage. In some examples, the compositions are suitable for long-term or short-term storage at freezing or refrigeration temperatures or higher. Thus, compositions of the present disclosure can be stored at temperatures below 0°C (e.g., about -80°C to about -10°C, about -60°C to about -20°C, or about -30°C) or at temperatures of about 1°C to about 10°C (e.g., about 2°C to about 8°C). Optionally, storage at these temperatures (below 10°C) can be for long-term storage, e.g., at least 6 months, at least 12 months, at least 18 months, at least 24 months, at least 30 months, or at least 36 months. Compositions of the present disclosure can be stored at room temperature (e.g., about 20°C to about 30°C, about 23°C to about 27°C, about 25°C, or about 30°C). In some examples, compositions of the present disclosure can be stored at temperatures above room temperature (e.g., above 30°C (e.g., about 35°C to about 45°C, about 40°C)).

[0056] In some examples, the compositions of the present disclosure are highly stable and can withstand long-term storage at refrigerated or frozen temperatures. The compositions of the present disclosure are highly stable as liquids or solids. Optionally, less than about 5% (e.g., less than about 4%, less than about 3%, less than about 2%, less than about 1%) of the anti-IL-33 antibodies aggregate after about 1 month to about 3 months of storage at about 2°C to about 8°C (e.g., about 2°C, about 4°C, about 8°C, about -20°C). As used herein, "aggregates" refers to insoluble aggregates of anti-IL-33 antibodies. In some examples, less than about 5% (e.g., less than about 4%, less than about 3%, less than about 2%, less than about 1%) of the anti-IL-33 antibodies aggregate after 6 or 12 months of storage at about 2°C to about 8°C (e.g., about 2°C, about 4°C, about 8°C, about 10°C), as measured by SEC. In some examples, less than about 5% (e.g., less than about 4%, less than about 3%, less than about 2%, less than about 1%) of the anti-IL-33 antibody degrades after 18 months of storage at about 2°C to about 8°C (e.g., about 2°C, about 4°C, about 8°C) as measured by SEC. In some examples, greater than 95% of the anti-IL-33 antibody is intact in a glass vial or syringe after 18 months of storage at about 2°C to about 8°C (e.g., about 2°C, about 4°C, about 8°C) as measured by SEC. In some examples, less than about 5% (e.g., less than about 4%, less than about 3%, less than about 2%, less than about 1%) of the antibody in a composition of the disclosure aggregates after about 18 months of storage at about 2°C to about 8°C (e.g., about 2°C, about 4°C, about 8°C) as measured by SEC.

[0057] In some examples, the compositions of the present disclosure are highly stable and can withstand long-term storage at room temperature. Optionally, after about 1 month to about 3 months of storage at about 23°C to about 27°C (e.g., about 23°C, about 24°C, about 25°C, about 26°C, about 27°C), less than about 5% (e.g., less than about 4%, less than about 3%, less than about 2%, less than about 1%) of the anti-IL-33 antibodies aggregate. In some examples, after about 6 months of storage at about 23°C to about 27°C, less than about 5% (e.g., less than about 4%, less than about 3%, less than about 2%, less than about 1%) of the anti-IL-33 antibodies aggregate as measured by SEC. In examples, after about 6 months of storage in glass vials or syringes at about 23°C to about 27°C, more than 95% of the anti-IL-33 antibodies are intact as measured by SEC. In some examples, less than about 5% (e.g., less than about 4%, less than about 3%, less than about 2%, less than about 1%) of the anti-IL-33 antibodies in the compositions of the disclosure degrade after about 6 months of storage at about 23°C to about 27°C, as measured by SEC.

[0058] In various examples, compositions of the present disclosure are highly stable and can withstand short-term storage under stressful storage conditions. In some examples, less than about 5% (e.g., less than about 4%, less than about 3%, less than about 2%, less than about 1%) of the anti-IL-33 antibodies aggregate after about 1 month to about 3 months of storage at about 38°C to about 42°C (e.g., about 38°C, about 39°C, about 40°C, about 41°C, about 42°C). In some examples, greater than 95% of the anti-IL-33 antibodies remain intact in glass vials or syringes after about 3 months of storage at about 38°C to about 42°C (e.g., about 38°C, about 39°C, about 40°C, about 41°C, about 42°C) as measured by SEC. In some examples, less than about 5% (e.g., less than about 4%, less than about 3%, less than about 2%, less than about 1%) of the anti-IL-33 antibodies in the compositions of the disclosure aggregate after about 3 months of storage at about 38°C to about 42°C (e.g., about 38°C, about 39°C, about 40°C, about 41°C, about 42°C) as measured by SEC.

[0059] In particular examples, the compositions are provided for storage or use in, for example, single-use vials, single-use syringes, or glass, glass-lined, or glass-coated primary containers. In a representative example, the compositions are provided in single-use system bags or polycarbonate carboys for frozen storage. In alternative examples, the compositions are contained in glass vials or syringes for long-term storage at about 2°C to about 8°C or for storage at higher temperatures (e.g., about 25°C, about 30°C, about 40°C).

[0060] In certain instances, the compositions are provided for use in ready-to-use and / or self-administered delivery systems. In certain instances, the compositions are provided in pre-filled syringes or autoinjectors, pen injectors, dual-chamber pens, etc. Such products are known in the art and commercially available. See, for example, Shire, Steven, "Monoclonal Antibodies: Meeting the Challenges in Manufacturing, Formulation, Delivery and Stability of Final Drug Product," Chapter 8: "Development of delivery device technology to deal with the challenges of highly viscous mAb formulations at high concentration," Woodhead Publishing, Cambridge, UK, pages 153-162 (2015).

[0061] The compositions of the present disclosure may be suitable for administration by any acceptable route, including parenteral and, in particular, subcutaneous. For example, subcutaneous administration may be into the upper arm, upper thigh, or abdomen. Other routes include, for example, intravenous, intradermal, intramuscular, intraperitoneal, intranodal, and intrasplenic. The subcutaneous route is preferred. In some instances, the intravenous route is preferred. For example, the stable liquid composition may be diluted in IV fluid before delivery via the intravenous route.

[0062] The compositions disclosed herein include anti-IL-33 antibodies. Interleukin-33 (IL-33), also known as IL-1F11, is a member of the IL-1 cytokine family that stimulates the production of cells, cytokines, and immunoglobulins characteristic of type 2 immune responses. IL-33 is a 270-amino acid protein consisting of two domains: a homeodomain and a cytokine (IL-1-like) domain. The homeodomain contains a nuclear localization signal (NLS). IL-33 mediates signaling through ST2, a receptor expressed on Th2 cells, mast cells, and a wide variety of other cell types.

[0063] Extracellular IL-33 stimulates target cells by binding to ST2, subsequently activating the NFKB and MAP kinase pathways, leading to various functional responses, including the production of cytokines and chemokines. Soluble ST2 (sST2) is thought to be a decoy receptor that blocks IL-33 signaling.

[0064] In humans, IL-33 has been found to be constitutively expressed in smooth muscle and bronchial epithelium. Its expression can be induced by IL-Iβ and TNF-α in lung and skin fibroblasts (Schmitz et al. (2005)). Levels of soluble ST2 protein and IL-33 mRNA / protein are elevated in the serum and tissues of asthmatic patients (Oboki et al., Allergology International 59:143-160 (2010)).

[0065] In vivo, IL-33 induces the expression of IL-4, IL-5, and IL-13, causing severe lesions in mucosal organs. Administration of IL-33 to mice has potent inflammatory effects, including severe blood eosinophilia, elevated serum levels of IL-5 and IgE, and goblet cell hyperplasia at mucosal surfaces (Schmitz et al. (2005)). Intraperitoneal or intranasal administration of IL-33 to mice led to the induction of eosinophilic inflammation in the lung and intestinal mucosa through IL-13- and STAT6-dependent pathways (Oboki et al. (2010)). Therefore, IL-33 may be involved in allergic diseases such as asthma and inflammatory airway diseases such as chronic obstructive pulmonary disorder (COPD).

[0066] It is therefore contemplated that compositions comprising anti-IL-33 antibodies may be useful in the treatment of IL-33-mediated diseases such as asthma or COPD.

[0067] In some examples, the antibody present in the composition of the present invention comprises a heavy chain variable domain comprising a VHCDR1 having the sequence of SEQ ID NO: 1, a VHCDR2 having the sequence of SEQ ID NO: 2, and a VHCDR3 having the sequence of SEQ ID NO: 3; and a light chain variable domain comprising a VLCDR1 having the sequence of SEQ ID NO: 5, a VLCDR2 having the sequence of SEQ ID NO: 6, and a VLCDR3 having the sequence of SEQ ID NO: 7.

[0068] In some examples, the antibody present in the composition of the invention comprises (a) a heavy chain variable domain that is at least 95%, 90%, 85%, or 80% identical to SEQ ID NO:4 or a sequence of amino acids encoded by a polynucleotide sequence that is at least 95%, 90%, 85%, or 80% identical to SEQ ID NO:11; (b) a light chain variable domain that is at least 80% identical to SEQ ID NO:8 or a sequence of amino acids encoded by a polynucleotide sequence that is at least 95%, 90%, 85%, or 80% identical to SEQ ID NO:12; or (c) the heavy chain variable domain of (a) and the light chain variable domain of (b).

[0069] In some examples, the antibody is a human antibody, a humanized antibody, a chimeric antibody, a monoclonal antibody, a recombinant antibody, an antigen-binding antibody fragment, a single-chain antibody, a monomeric antibody, a diabody, a triabody, a tetrabody, a Fab fragment, an IgG1 antibody, an IgG2 antibody, an IgG3 antibody, or an IgG4 antibody. In some embodiments, the anti-IL-33 antibody is an IgG1 antibody.

[0070] In some examples, the anti-IL-33 antibody comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 4. In some examples, the anti-IL-33 antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 8. In some examples, the anti-IL-33 antibody comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 4 and a light chain comprising the amino acid sequence of SEQ ID NO: 8.

[0071] In some examples, the anti-IL-33 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 9. In some examples, the anti-IL-33 antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 10. In some examples, the anti-IL-33 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 9 and a light chain comprising the amino acid sequence of SEQ ID NO: 10.

[0072] In some examples, the composition comprises an anti-IL-33 antibody that competes with 33_640087-7B for binding to IL-33 in an in vitro HTRF competitive binding assay. An antibody is said to competitively inhibit the binding of a reference antibody to a given epitope if it specifically binds to that epitope to the extent that it blocks, to some extent, the binding of the reference antibody to that epitope. Competitive inhibition can be determined by any method known in the art, for example, solid-phase assays such as competitive ELISA assays, dissociation-promoted lanthanide fluorescent immunoassays (DELFIA®, Perkin Elmer), and radioligand binding assays. For example, one skilled in the art can determine whether an antibody competes for binding to IL-33 by using an in vitro competitive binding assay, such as the HTRF assay described in paragraphs 881-886 of WO 2016 / 156440, which is incorporated herein by reference. For example, one skilled in the art could label a first anti-IL-33 antibody with a donor fluorophore and mix multiple concentrations with a fixed concentration sample of redIL-33 labeled with an acceptor fluorophore. Subsequently, fluorescence resonance energy transfer between the donor and acceptor fluorophores in each sample could be measured to confirm binding characteristics. To identify competitively binding anti-IL-33 antibodies, one skilled in the art could first mix various concentrations of the test antibody with a fixed concentration of labeled antibody from Table 6. A reduction in the FRET signal when the mixture is incubated with labeled IL-33 compared to a positive control of labeled antibody alone indicates competitive binding to IL-33. A binding molecule or fragment thereof can be said to competitively inhibit binding of a reference antibody to a given epitope by at least 90%, at least 80%, at least 70%, at least 60%, or at least 50%.

[0073] In some examples, the composition comprises the anti-IL-33 antibody 33_640087-7B (as described in WO 2016 / 156440, incorporated herein by reference). WO 2016 / 156440 discloses that 33_640087-7B binds to redIL-33 with particularly high affinity and attenuates both ST-2- and RAGE-dependent IL-33 signaling.

[0074] Other representative anti-IL-33 antibodies include ANB020, also known as ethoximab (as described in WO 2015 / 106080), 9675P (as described in U.S. Patent Application Publication No. 2014 / 0271658), A25-3H04 (as described in U.S. Patent Application Publication No. 2017 / 0283494), Ab43 (as described in WO 2018 / 081075), IL33-158 (as described in U.S. Patent Application Publication No. 2018 / 0037644), 10C12.38.H6.87Y.581 IgG4 (as described in WO 2016 / 077381), or binding fragments thereof. Other representative anti-IL-33 antibodies or antigen-binding fragments thereof include any of the other anti-IL-33 antibodies described in WO 2016 / 156440, WO 2015 / 106080, U.S. Patent Application Publication Nos. 2014 / 0271658, 2017 / 0283494, WO 2018 / 081075, U.S. Patent Application Publication No. 2018 / 0037644, or WO 2016 / 077381, all of which are incorporated herein by reference.

[0075] Production method Further provided herein are methods of making the compositions of the present disclosure. Accordingly, further provided are methods of making a stable liquid composition having a viscosity of less than about 10 cP and comprising greater than about 100 mg / mL of an anti-IL-33 antibody, at least about 170 mM arginine, such as at least about 190 mM arginine, optionally a surfactant and a buffer. In some examples, the method includes: (i) combining the antibody, arginine, and a buffer in a solution to obtain a solution comprising about 100 mg / mL to about 200 mg / mL of an anti-IL-33 antibody, at least about 170 mM arginine, such as at least about 170 mM arginine, such as at least about 190 mM arginine, and a buffer; and (ii) adding a surfactant to the solution to achieve a final surfactant concentration of about 0.03% (w / v) ± 0.015% (w / v).

[0076] Further provided are methods for making a stable liquid composition having a viscosity of less than about 10 cP and comprising greater than about 100 mg / mL of an anti-IL-33 antibody, at least about 150 mM lysine, optionally a surfactant and a buffer. In some examples, the method includes (i) combining the antibody, arginine, and a buffer in solution to obtain a solution comprising about 100 mg / mL to about 200 mg / mL of an anti-IL-33 antibody, at least about 170 mM arginine, such as at least about 150 mM lysine, and a buffer; and (ii) adding a surfactant to the solution to achieve a final surfactant concentration of about 0.03% (w / v) ± 0.015% (w / v).

[0077] In some examples, the stable liquid composition comprises a surfactant at a concentration of 0.03% (w / v) ± 0.01% (w / v), hi some examples, the stable liquid composition comprises a surfactant at a concentration of about 0.02% (w / v) to about 0.04% (w / v).

[0078] In some examples, the stable liquid composition comprises a surfactant at a concentration of 0.02% (w / v) ± 0.01% (w / v), hi some examples, the stable liquid composition comprises a surfactant at a concentration of about 0.01% (w / v) to about 0.02% (w / v).

[0079] In some examples, the stable liquid composition comprises about 150 mg / mL of the anti-IL-33 antibody.

[0080] In some examples, the stable liquid composition comprises more than 170 mM arginine. In some examples, the stable liquid composition comprises more than 190 mM arginine. In some examples, the composition comprises less than about 500 mM arginine. For example, the composition comprises about 200 mM, about 210 mM, about 220 mM, about 240 mM, about 260 mM, about 280 mM, about 300 mM, about 350 mM, about 400 mM, or about 450 mM arginine. In some examples, the composition comprises about 220 mM arginine.

[0081] In some examples, the stable liquid composition comprises at least about 150 mM lysine. In some examples, the composition comprises less than about 500 mM lysine. For example, the composition comprises about 160 mM, about 170 mM, about 180 mM, about 200 mM, about 220 mM, about 240 mM, about 260 mM, about 280 mM, about 300 mM, about 350 mM, about 400 mM, or about 450 mM lysine. In some examples, the composition comprises about 170 mM lysine.

[0082] In some examples, the viscosity of a stable liquid composition with a high concentration of arginine is reduced compared to a liquid composition comprising a lower concentration of anti-IL-33 antibody, 20 mM histidine, 80 mM arginine, 120 mM sucrose, 0.02% (w / v) polysorbate 80, pH 6.0, where the lower concentration is compared to the concentration of anti-IL-33 antibody in the stable liquid composition.

[0083] In some examples, the viscosity of the stable liquid composition is less than about 10 cP. In some examples, the viscosity of the stable liquid composition is about 9 cP. In some examples, the viscosity is measured at 23°C.

[0084] In some examples, the reversible self-association (RSA) of the anti-IL-33 antibody in the stable liquid composition is reduced compared to the RSA of the anti-IL-33 antibody in a liquid composition comprising a lower concentration of the anti-IL-33 antibody in 20 mM histidine, 80 mM arginine, 120 mM sucrose, 0.02% (w / v) polysorbate 80, pH 6.0, where the lower concentration is compared to the concentration of the anti-IL-33 antibody in the stable liquid composition.

[0085] In some examples, the RSA is reduced by 2-fold compared to the RSA of the anti-IL-33 antibody in a liquid composition comprising a lower concentration of the anti-IL-33 antibody in 20 mM histidine, 80 mM arginine, 120 mM sucrose, 0.02% (w / v) polysorbate 80, pH 6.0.

[0086] In some examples, the stable liquid composition comprises at least about 30% by weight of monomer, e.g., about 30%, about 31%, about 32%, about 33%, about 34%, about 35%, about 36%, about 37%, about 38%, about 39%, about 40%, about 41%, about 42%, about 43%, about 44%, about 45%, about 46%, about 47%, about 48%, about 49%, or about 50%. In some examples, the stable liquid composition comprises about 35% to about 50% by weight of monomer. In some examples, the stable liquid composition comprises about 40% to about 45% by weight of monomer. In some examples, the stable liquid composition comprises about 35% to about 50% by weight of monomer after about 3 months of storage at a temperature of about 2°C to about 8°C. In some examples, the stable liquid composition comprises about 35% to about 50% weight percent monomer after about 3 months of storage at a temperature of about 2° C. to about 8° C. In some examples, the stable liquid composition comprises about 40% weight percent monomer after about 3 months of storage at a temperature of about 2° C. to about 8° C. In some examples, the stable liquid composition comprises about 35% to about 50% weight percent monomer after about 6 months of storage at a temperature of about 2° C. to about 8° C. In some examples, the stable liquid composition comprises about 40% weight percent monomer after about 6 months of storage at a temperature of about 2° C. to about 8° C.

[0087] In some examples, the surfactant is polysorbate 80 or polysorbate 20. In some examples, the surfactant is polysorbate 80.

[0088] In some examples, the buffer is made from histidine. In some examples, the stable liquid composition has a final buffer concentration of about 16 mM to about 24 mM, optionally about 17 mM to about 24 mM, optionally about 18 mM to about 24 mM.

[0089] In some examples, the stable liquid composition has a pH of about pH 5.5.

[0090] In some examples, the anti-IL-33 antibody is any of those described herein. In some examples, the anti-IL-33 antibody is 33_640087-7B.

[0091] Product, syringe and vial The present disclosure provides a product comprising any one of the disclosed compositions, optionally about 0.5 mL to about 5 mL (e.g., about 0.5 mL to about 4.5 mL, about 0.5 mL to about 4 mL, about 0.5 mL to about 3.5 mL, about 0.5 mL to about 3 mL, about 0.5 mL to about 2.5 mL, about 0.5 mL to about 2 mL, about 0.5 mL to about 1.5 mL, about 0.5 mL to about 1 mL, about 1 mL to about 5 mL, about 1.5 mL to about 5 mL, about 2 mL to about 5 mL, about 2.5 mL to about 5 mL, about 3 mL to about 5 mL, about 3.5 mL to about 5 mL, about 4 mL to about 5 mL, about 4.5 mL to about 5 mL). In some examples, the composition comprises greater than about 100 mg / mL of an anti-IL33 antibody (e.g., 33_640087-7B). In some examples, the composition comprises about 130 mg / ml to about 170 mg / ml of an anti-IL-33 antibody (e.g., 33_640087-7B), about 0.03% (w / v) ±0.015% (w / v) polysorbate 80, about 220 mM arginine, and about 16 mM to about 24 mM histidine, wherein the pH is less than about 6, optionally about pH 5.5. Optionally, the pH is about pH 5.0 to about pH 6.0.

[0092] The present disclosure also provides a pre-filled syringe (PFS) comprising any one of the disclosed compositions, optionally containing about 0.5 mL to about 5 mL (e.g., about 0.5 mL to about 4.5 mL, about 0.5 mL to about 4 mL, about 0.5 mL to about 3.5 mL, about 0.5 mL to about 3 mL, about 0.5 mL to about 2.5 mL, about 0.5 mL to about 2 mL, about 0.5 mL to about 1.5 mL, about 0.5 mL to about 1 mL, about 1 mL to about 5 mL, about 1.5 mL to about 5 mL, about 2 mL to about 5 mL, about 2.5 mL to about 5 mL, about 3 mL to about 5 mL, about 3.5 mL to about 5 mL, about 4 mL to about 5 mL, or about 4.5 mL to about 5 mL). In some examples, the composition comprises greater than about 100 mg / mL of an anti-IL33 antibody (e.g., 33_640087-7B). In some examples, the composition comprises about 130 mg / ml to about 170 mg / ml of an anti-IL-33 antibody (e.g., 33_640087-7B), about 0.03% (w / v) ±0.015% (w / v) polysorbate 80, about 220 mM arginine, and about 16 mM to about 24 mM histidine, wherein the pH is less than about 6, optionally about pH 5.5.

[0093] Also provided are vials containing any one of the disclosed compositions, optionally containing about 0.5 mL to about 5 mL (e.g., about 0.5 mL to about 4.5 mL, about 0.5 mL to about 4 mL, about 0.5 mL to about 3.5 mL, about 0.5 mL to about 3 mL, about 0.5 mL to about 2.5 mL, about 0.5 mL to about 2 mL, about 0.5 mL to about 1.5 mL, about 0.5 mL to about 1 mL, about 1 mL to about 5 mL, about 1.5 mL to about 5 mL, about 2 mL to about 5 mL, about 2.5 mL to about 5 mL, about 3 mL to about 5 mL, about 3.5 mL to about 5 mL, about 4 mL to about 5 mL, about 4.5 mL to about 5 mL). In some examples, the composition contains greater than about 100 mg / mL of an anti-IL33 antibody (e.g., 33_640087-7B). In some examples, the composition comprises about 130 mg / ml to about 170 mg / ml of an anti-IL-33 antibody (e.g., 33_640087-7B), about 0.03% (w / v) ±0.015% (w / v) polysorbate 80, about 220 mM arginine, and about 16 mM to about 24 mM histidine, wherein the pH is less than about 6, optionally about pH 5.5.

[0094] kit The present disclosure also provides a kit comprising the composition described herein together with a package insert, package label, instruction manual or other labeling that indicates or discloses any of the methods disclosed herein.In certain examples, the present disclosure provides a kit for making a single-dose dosage unit.In certain examples of the present disclosure, the kit includes a single-chamber and multi-chamber pre-filled syringe (for example, liquid syringe).

[0095] Treatment method The disclosure also provides the use of 33_640087-7B or another anti-IL-33 antibody, or antigen-binding portion thereof, disclosed herein in the manufacture of a medicament for treating a subject with an IL-33-mediated disorder.

[0096] Provided herein are methods for treating an IL-33-mediated disease in a subject. In some examples, the present disclosure provides a composition disclosed herein for use in treating an IL-33-mediated disease in a subject. As provided herein, the present disclosure also provides the use of an anti-IL-33 antibody in the manufacture of a medicament for treating a subject with an IL-33-mediated disease, wherein the medicament comprises any of the compositions disclosed herein.

[0097] In some examples, the methods, compositions for use, or uses provided herein are for the treatment of an IL-33 mediated disorder selected from asthma, atopic dermatitis, and chronic obstructive pulmonary disorder.

[0098] In some examples, the methods, compositions for use or uses provided herein are for the treatment of diabetic nephropathy.

[0099] In some examples, the subject is a human.

[0100] Embodiment 1. A composition comprising greater than about 100 mg / ml of an anti-IL-33 antibody, at least 170 mM arginine or at least 150 mM lysine, and a buffer, wherein the anti-IL-33 antibody is i. a heavy chain variable domain comprising a VHCDR1 having the sequence of SEQ ID NO: 1, a VHCDR2 having the sequence of SEQ ID NO: 2, and a VHCDR3 having the sequence of SEQ ID NO: 3; ii. a light chain variable domain comprising a VLCDR1 having the sequence of SEQ ID NO: 5, a VLCDR2 having the sequence of SEQ ID NO: 6, and a VLCDR3 having the sequence of SEQ ID NO: 7; A composition comprising: 2. The anti-IL-33 antibody is i. i. a sequence of amino acids that is at least 95%, 90%, 85%, or 80% identical to SEQ ID NO:4; or ii. A sequence of amino acids encoded by a polynucleotide sequence that is at least 80% identical to SEQ ID NO:11 a heavy chain variable domain which is ii. i. a sequence of amino acids that is at least 95%, 90%, 85%, or 80% identical to SEQ ID NO: 8; or ii. A sequence of amino acids encoded by a polynucleotide sequence that is at least 80% identical to SEQ ID NO: 12 a light chain variable domain which is iii. (a) a heavy chain variable domain and (b) a light chain variable domain 2. The composition of embodiment 1, comprising: 3. The composition of embodiment 1 or 2, wherein said anti-IL-33 antibody is an IgG1 antibody. 4. The composition of any of embodiments 1 to 3, wherein the anti-IL-33 antibody comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 4, a light chain comprising the amino acid sequence of SEQ ID NO: 8, or a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 4 and a light chain comprising the amino acid sequence of SEQ ID NO: 8. 5. The composition of any of embodiments 1 to 4, wherein the anti-IL-33 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 9, a light chain comprising the amino acid sequence of SEQ ID NO: 10, or a heavy chain comprising the amino acid sequence of SEQ ID NO: 9 and a light chain comprising the amino acid sequence of SEQ ID NO: 10. 6. The composition of any of embodiments 1-5, wherein said anti-IL-33 antibody competes with 33_640087-7B for binding to IL-33 in an in vitro HTRF competitive binding assay. 7. The composition of any of embodiments 1-6, wherein said anti-IL-33 antibody is present at a concentration of less than about 200 mg / ml. 8. The composition of any of embodiments 1-7, wherein said anti-IL-33 antibody is present at a concentration of less than about 180 mg / ml. 9. The composition of any of embodiments 1-8, wherein said anti-IL-33 antibody is present at a concentration of less than about 160 mg / ml. 10. The composition of any of embodiments 1 to 9, wherein the anti-IL-33 antibody is present at a concentration of about 100 mg / ml to about 200 mg / ml. 11. The composition of any of embodiments 1 to 10, wherein the anti-IL-33 antibody is present at a concentration of about 130 mg / ml to about 170 mg / ml. 12. The composition of any of embodiments 1 to 11, wherein said anti-IL-33 antibody is present at a concentration of 150 mg / ml ± 10%. 13. The composition of any one of embodiments 1 to 12, further comprising a surfactant. 14. The composition of any one of embodiments 13, wherein the surfactant is amphiphilic and anionic. 15. The composition of embodiment 14, wherein the surfactant is a polysorbate. 16. The composition of embodiment 15, wherein the surfactant is polysorbate 20 or polysorbate 80 or a mixture thereof. 17. The composition of any one of embodiments 13 to 16, wherein the surfactant is at a concentration of about 0.005% (w / v) to about 0.05% (w / v). 18. The composition of embodiment 17, comprising 0.03% (w / v) ± 0.010% (w / v) of a surfactant. 19. The composition of embodiment 17, comprising about 0.015% (w / v), 0.03% (w / v), or 0.045% (w / v) of a surfactant. 20. The composition of any one of embodiments 16-19, wherein the surfactant is polysorbate 80. 21. The composition of any of embodiments 1-20, comprising at least about 190 mM arginine. 22. The composition of any of embodiments 1-21, comprising about 190 mM to about 250 mM arginine. 23. The composition of any of embodiments 1-22, comprising about 220 mM arginine. 24. The composition of any of embodiments 1-23, wherein the arginine is L-arginine hydrochloride. 25. The composition of any of embodiments 1-20, comprising about 150 mM to about 250 mM lysine. 26. The composition of embodiment 25, comprising about 170 mM lysine. 27. The composition of any of embodiments 1-20, 25, or 26, wherein the lysine is L-lysine. 28. The composition of any of embodiments 1-27, wherein the buffer is succinate, histidine, or acetate. 29. The composition of embodiment 28, wherein the buffer is histidine. 30. The composition of embodiment 29, wherein the buffer is L-histidine / L-histidine hydrochloride. 31. The composition of any of embodiments 1 to 30, wherein the buffer is at a concentration of about 10 mM to about 30 mM. 32. The composition of embodiment 31, wherein the buffer is at a concentration of about 16 mM to about 24 mM, optionally about 17 mM to about 24 mM, optionally about 18 mM to about 24 mM. 33. The composition of any of embodiments 31 or 32, wherein the buffer is at a concentration of about 19 mM to about 21 mM. 34. The composition of any of embodiments 1-33, comprising 20 mM±10% buffer. 35. The composition of any one of embodiments 1 to 34, which is a liquid. 36. The composition of any of embodiments 1-35, wherein the pH is less than about pH 6.0. 37. The composition of any one of embodiments 1 to 36, having a pH of about pH 5.0 to about pH 6.0. 38. A composition described in any of embodiments 1 to 37, wherein the pH is about pH 5.2, about pH 5.5, or about pH 5.8. 39. A composition described in any of embodiments 1 to 38, wherein the pH is about pH 5.5. 40. The composition of any one of embodiments 35 to 39, characterized in that the composition has a reduced viscosity compared to a composition comprising 40.80 mM or less of arginine. 41. Lower concentrations of anti-IL-33 antibodies, For example, 50 mg / ml of anti-IL-33 antibody, 40. The composition of any one of embodiments 35 to 39, characterized in that it has a reduced viscosity compared to a composition comprising 20 mM histidine, 80 mM arginine, 120 mM sucrose, 0.02% (w / v) polysorbate 80, pH 6.0. 42. The composition according to any one of embodiments 35 to 41, characterized in that the viscosity is less than about 10 cP at 23°C and the concentration of the anti-IL-33 antibody is about 150 mg / ml ± 10%. 43. The composition of any of embodiments 35-41, wherein the viscosity is from about 5 cP to about 20 cP, optionally less than about 10 cP, such as about 9 cP. 44. The composition of any one of embodiments 35 to 43, wherein the reversible self-association of the anti-IL-33 antibody is reduced compared to the reversible self-association of the anti-IL-33 antibody in a composition comprising 20 mM histidine, 80 mM arginine, 120 mM sucrose, 0.02% (w / v) polysorbate 80, pH 6.0 and a lower concentration of the anti-IL-33 antibody, e.g., 50 mg / ml. 45. The composition of any of embodiments 35-44, comprising about 35% to about 50% by weight of the monomer. 46. ​​The composition of any of embodiments 1 to 45, wherein less than about 5% of the antibody, optionally less than about 2%, aggregates after about 12 to about 18 months of storage at about 2°C to about 8°C, as measured by size exclusion chromatography (SEC). 47. i. The solution contains about 130 mg / ml to about 170 mg / ml of 33_640087-7B, about 0.03% (w / v) ±0.015% polysorbate 80, about 220 mM arginine, and about 16 to about 24 mM histidine buffer, and has a pH of 5.5 ±0.5. ; ii. about 130 mg / ml to about 170 mg / ml of 33_640087-7B, about 0.02% (w / v) ± 0.015% polysorbate 80, about 170 mM lysine, and about 16 to about 24 mM histidine buffer, having a pH of 5.5 ± 0.5; or iii. A solution containing about 130 mg / ml to about 170 mg / ml of 33_640087-7B, about 0.02% (w / v) ± 0.015% polysorbate 80, about 190 mM arginine, and about 16 to about 24 mM histidine buffer, having a pH of 5.5 ± 0.5; composition. 48. The composition of embodiment 47, comprising about 150 mg / ml of 33_640087-7B. 49. The composition of any of embodiments 47 or 48, comprising about 18 mM to about 22 mM histidine buffer, optionally about 20 mM histidine buffer. 50. The composition of any one of embodiments 47-49, wherein the arginine is L-arginine hydrochloride. 51. A composition comprising about 130 mg / ml to about 170 mg / ml of an anti-IL-33 antibody, about 0.03% (w / v) ± 0.015% polysorbate 80, about 220 mM arginine, and about 16 to about 24 mM histidine buffer, having a pH of 5.5 ± 0.5. 52. The composition of any one of embodiments 47 to 51, which is a liquid. 53. A product comprising the composition of any one of embodiments 1-52, optionally comprising 0.5 mL to about 5 ml (e.g., 1 mL to about 3 ml) of said composition. 54. A vial comprising the composition of any of embodiments 1-52, optionally containing about 0.5 mL to about 5 ml (e.g., 1 mL to about 3 ml) of said composition. 55. A method of treating an IL-33-mediated disorder in a subject, comprising administering to the subject a therapeutically effective amount of the composition of any one of embodiments 1-52. 56. The method of embodiment 55, wherein said IL-33 mediated disorder is asthma, atopic dermatitis, or chronic obstructive pulmonary disorder. 57. The method of embodiment 55, wherein the IL-33-mediated disorder is diabetic nephropathy. 58. A method of making a stable liquid composition having a viscosity of less than about 10 cP and comprising greater than about 100 mg / ml of an anti-IL-33 antibody, at least about 170 mM arginine or about 150 mM lysine, a surfactant, and a buffer, comprising: i. combining in solution an anti-IL-33 antibody, arginine, and buffer to obtain a solution comprising about 100 mg / ml to about 200 mg / ml of the antibody, at least about 170 mM arginine or about 150 mM lysine, and buffer; ii. adding a surfactant to the solution to achieve a final surfactant concentration of about 0.03% (w / v) ± 0.015% (w / v); A method comprising the steps of: 59. The method of embodiment 58, wherein the stable liquid composition comprises more than 190 mM arginine. 60. The method of any of embodiments 58 or 59, wherein the stable liquid composition comprises about 220 mM arginine. 61. The composition of any one of embodiments 58-60, wherein the arginine is L-arginine hydrochloride. 62. The method of any one of embodiments 58 to 61, wherein the reversible self-association (RSA) of the anti-IL-33 antibody in the stable liquid composition is reduced compared to the RSA of the anti-IL-33 antibody in a liquid composition comprising a lower concentration of the anti-IL-33 antibody in 20 mM histidine, 80 mM arginine, 120 mM sucrose, 0.02% (w / v) polysorbate 80, pH 6.0. 63. The method of any of embodiments 58-62, wherein the viscosity of the liquid formulation is reduced compared to a liquid formulation comprising a lower concentration of anti-IL-33 antibody, 20 mM histidine, 80 mM arginine, 120 mM sucrose, 0.02% (w / v) polysorbate 80, pH 6.0. 64. The method of any of embodiments 58-63, wherein the surfactant is polysorbate 80. 65. The method of any of embodiments 58-64, wherein the buffer is made from histidine. 66. The method of embodiment 65, resulting in a stable liquid formulation having a final buffer concentration of about 16 mM to about 24 mM, optionally about 17 mM to about 24 mM, optionally about 18 mM to about 24 mM. 67. The method of any of embodiments 58-66, which results in a stable liquid formulation having a pH of about pH 5 to about pH 6, optionally about pH 5.5. 68. The method according to any one of embodiments 58 to 67, wherein said anti-IL-33 antibody is as defined in any one of embodiments 1 to 5. 69. The method of any one of embodiments 58-68, wherein cP is measured at 23°C. [Example]

[0101] Creating formulations with improved characteristics 33_640087-7B is a human immunoglobulin (Ig) G1 monoclonal antibody (mAb) that binds to human interleukin (IL)-33, preventing the binding of IL-33 to its receptor ST2 and inhibiting the conversion of disulfide-bonded (DSB) IL33.

[0102] A Phase 1 clinical trial (Study D9180C00001) of 33_640087-7B has been completed. 33_640087-7B was found to be generally safe and well-tolerated, with no safety concerns, following intravenous (IV) administration of up to 300 mg of 33_640087-7B. 33_640087-7B was supplied as a lyophilized powder in vials. Each vial contained a nominal amount of 50 mg of 33_640087-7B. Upon reconstitution with 1.2 mL of sterile water for injection, the solution contained 50 mg / mL 33_640087-7B in 20 mM L-histidine / L-histidine-hydrochloride, 80 mM L-arginine-hydrochloride, 120 mM sucrose, 0.02% (weight / volume [w / v]) polysorbate 80, pH 6.0 ("Phase I formulation").

[0103] This example describes the surprising results of an attempt to reformulate 33_640087-7B to achieve a higher unit dose / ml composition for subsequent clinical trials.

[0104] The reversible self-association characteristics of 33_640087-7B in Phase I formulations were measured using static light scattering to obtain the weight percent monomer and soluble reversible higher-order species. Static light scattering intensity in volts was measured as a function of concentration (mg / mL), which was converted to apparent molecular weight (kDa) using the Rayleigh equation. The weight fractions of monomer and soluble reversible higher-order species were extracted from the measured apparent molecular weight using an effective rigid particle model (Fernandez and Minton (2009) Biophys J 96:1992-8).

[0105] Table 1 shows the reversible self-association characteristics of aqueous Phase I compositions:

[0106] [Table 1]

[0107] The concentration of 33_640087-7B was then increased three-fold to 150 mg / ml and the viscosity was measured at 23° C. The viscosity was measured to be approximately 24 centipoise.

[0108] Next, we attempted to reconstitute 33_640087-7B. Unexpectedly, we found that varying the amount of excipients in this composition led to improved RSA characteristics of the formulation compared to the Phase 1 study formulation.

[0109] Table 2 shows the reversible self-association characteristics of the aqueous first phase and the next generation compositions.

[0110] [Table 2]

[0111] The reduction in RSA led to a significant improvement in viscosity (approximately 9 cP) measured at 23° C. when compared to the viscosity of a Phase 1 test composition containing an equivalent antibody concentration (approximately 24 cP). Notably, this improvement was observed at three times the protein concentration.

[0112] The RSA profile was also found to be stable over time. The percent distribution of soluble higher order species did not change significantly over a 6-month storage period at 2-8°C. RSA stability was observed in the next-generation formulation at multiple antibody concentrations (Figure 3).

[0113] Table 3 shows the weight percent distribution of monomer and trimers and hexamers (ie, soluble higher order species) over time.

[0114] [Table 3]

[0115] The long-term stability of the next generation composition was tested at multiple time points and multiple concentrations. Stability data was generated for compositions stored in either glass vials or pre-filled syringes.

[0116] Table 4 shows the long-term stability characteristics as a function of insoluble aggregate formation expressed as a percentage of the original antibody concentration (approximately 150 mg / ml).

[0117] [Table 4]

[0118] This data is illustrated in Figures 2, 3 and 4.

[0119] Next, a univariate analysis of viscosity (Cp) was performed across multiple 33-640087_7B and arginine concentrations (holding the remaining formulation components fixed).

[0120] The results are shown in Figure 5. At several high arginine concentrations (150 mM, 190 mM, 220 mM, and 250 mM), the Cp at 25°C was significantly lower than the Cp (24 Cp) calculated for the Phase 1 study formulation at 23°C. The Cp at 5°C improved with increasing concentrations of arginine.

[0121] Cp was also tested at multiple protein concentrations (135 mg / ml, 150 mg / ml, and 165 mg / ml). The results are shown in Figure 6. Cp was measured at multiple temperatures (5°C, 18°C, 25°C, and 30°C). The analysis reveals that increasing arginine concentration decreases Cp, even at high protein concentrations, especially at temperatures above 18°C. Generally, when at least 190 mM arginine is used, the viscosity at about 25°C is below the desired Cp value of 10 when the formulation contains 165 mg / ml of 33-640087_7B.

[0122] The robustness of the 33-640087_7B formulation was also analyzed at several temperatures, pHs, arginine concentrations, and excipient concentrations. Formulations were stored at either 40°C, 25°C, or 2-8°C for 1, 6, or 11 months, respectively. Samples were taken at multiple time points under each condition and examined for aggregate formation using standard analytical techniques. The percent aggregate formation per month was calculated. The slopes from the resulting plots are shown in Table 5. The results indicate that the aggregation rate is stable within the range of pH, surfactant, or arginine concentrations under the conditions tested. A slight increase in aggregation rate was observed at pH 5.0 during the 1-month accelerated stability conditions (40°C).

[0123] [Table 5]

[0124] Several additional formulations were tested. Of particular interest, a formulation containing 20 mM histidine / histidine-HCl, 170 mM lysine-HCl, 0.02% PS80, pH 5.5, and approximately 160 mg / ml 33-640087_7B also showed a significant improvement in Cp (8.7 Cp) at 23° C. Analysis of the Cp at 23° C. of a formulation containing 150 mg / ml 33-640087_7B and either 150 mM or 190 mM lysine (and 20 mM histidine / histidine-HCl, 0.02% PS80, pH 5.5) also showed a significant improvement in viscosity, i.e., a Cp of less than 10, compared to the Cp of the Phase 1 study formulation.

[0125] conclusion A next-generation formulation has been developed that leads to a surprising reduction in RSA at high antibody concentrations. This next-generation formulation may enable the administration of a higher unit dose / volume of anti-IL-33 antibody, allowing for the administration of higher therapeutic doses. This may reduce the volume of drug product delivered to the injection site, for example for subcutaneous delivery, thereby reducing patient discomfort and improving patient compliance. This may also enable the exploration of a wider dynamic range of doses in the clinic, improving the likelihood of finding the most therapeutically effective dose.

[0126] Furthermore, the formulation has an acceptable long-term stability profile and reduced viscosity, which may have a positive impact on drug administration by improving the functionality of devices used to administer the antibody. Similarly, low viscosity may improve the ability of healthcare professionals or patients to manually inject the drug into a patient.

[0127] array SEQ ID NO: 1:33_640087-7B VH CDR1 Ser Tyr Ala Met Ser SEQ ID NO:2:33_640087-7B VH CDR2 Gly Ile Ser Ala Ile Asp Gln Ser Thr Tyr Tyr Ala Asp Ser Val Lys Gly SEQ ID NO:3:33_640087-7B VH CDR3 Gln Lys Phe Met Gln Leu Trp Gly Gly Gly Leu Arg Tyr Pro Phe Gly Tyr SEQ ID NO: 4:33_640087-7B VH [ka] SEQ ID NO: 5:33_640087-7B VL CDR1 Ser Gly Glu Gly Met Gly Asp Lys Tyr Ala Ala SEQ ID NO: 6:33_640087-7B VL CDR2 Arg Asp Thr Lys Arg Pro Ser SEQ ID NO: 7:33_640087-7B VL CDR3 Gly Val Ile Gln Asp Asn Thr Gly Val SEQ ID NO: 8:33_640087-7B VL [ka] SEQ ID NO: 9:33_640087-7B HC [ka] SEQ ID NO: 10:33_640087-7B LC [ka] SEQ ID NO: 11:33_640087-7B VH DNA [ka] SEQ ID NO: 12:33_640087-7B VL DNA [ka]

Claims

1. A composition comprising 130 mg / ml to 170 mg / ml of an anti-IL-33 human IgG1 antibody, about 190 mM to about 250 mM arginine, polysorbate 80 at a concentration of about 0.005% (w / v) to about 0.05% (w / v), and a buffer solution, wherein the buffer solution is histidine and has a pH of about pH 5.0 to about pH 6.0, and the anti-IL-33 human IgG1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:9 and a light chain comprising the amino acid sequence of SEQ ID NO:

10.

2. 2. The composition of claim 1, wherein the anti-IL-33 human IgG1 antibody is present at a concentration of 150 mg / ml ± 10%.

3. 3. The composition of claim 1, comprising 0.03% (w / v) ± 0.010% (w / v) polysorbate 80.

4. 4. The composition of claim 1, comprising about 220 mM arginine.

5. The composition according to any one of claims 1 to 4, wherein the arginine is L-arginine hydrochloride.

6. 6. The composition according to claim 1, wherein the buffer is L-histidine / L-histidine hydrochloride.

7. 7. The composition of claim 6, wherein the buffer is at a concentration of about 19 mM to about 21 mM, optionally 20 mM ± 10% buffer.

8. The composition according to any one of claims 1 to 7, which is a liquid.

9. A composition described in any of claims 1 to 8, wherein the pH is about pH 5.

5.

10. 10. The composition of claim 8 or 9, wherein the anti-IL-33 antibody has reduced reversible self-association compared to the reversible self-association of the anti-IL-33 antibody in a composition comprising 20 mM histidine, 80 mM arginine, 120 mM sucrose, 0.02% (w / v) polysorbate 80, pH 6.0 and a lower concentration of anti-IL-33 antibody, e.g., 50 mg / ml.

11. A composition comprising about 130 mg / ml to about 170 mg / ml of an anti-IL-33 human IgG1 antibody comprising a heavy chain consisting of the amino acid sequence of SEQ ID NO: 9 and a light chain consisting of the amino acid sequence of SEQ ID NO: 10, about 0.03% (w / v) ± 0.015% polysorbate 80, about 220 mM arginine, and about 16 to about 24 mM histidine buffer (pH is pH 5.5 ± 0.5).

12. A composition comprising 150 mg / ml of an anti-IL-33 human IgG1 antibody comprising a heavy chain consisting of the amino acid sequence of SEQ ID NO: 9 and a light chain consisting of the amino acid sequence of SEQ ID NO: 10, 20 mM His / His-HCl, 220 mM Arg-HCl, 0.03% (w / v) polysorbate 80, and having a pH of 5.5±0.

5.

13. A product comprising a composition according to any one of claims 1 to 12, optionally containing 1 ml to 3 ml of said composition.

14. A vial containing a composition according to any one of claims 1 to 12, optionally containing 1 ml to 3 ml of said composition.

15. The composition according to any one of claims 1 to 12, for treating an IL-33-mediated disorder, wherein the IL-33-mediated disorder is asthma, atopic dermatitis, chronic obstructive pulmonary disorder, or diabetic nephropathy.

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