Anti-TL1A antibody preparation

Abstract

Pharmaceutical formulations comprising an antibody or antigen-binding fragment thereof that specifically binds to TL1A are provided, which formulations have advantageous properties including, for example, stability.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This international application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 369,638, filed July 27, 2022, the entire contents of which are incorporated herein by reference.

[0002] Reference to an electronically submitted sequence listing The contents of the electronically submitted sequence listing (Name: 2873_369PC02_SequenceListing_St26.xml, Size: 67,054 bytes, and Creation Date: July 20, 2023) are incorporated herein by reference in their entirety. [Background technology]

[0003] TNF-like ligand 1A (TL1A, synonyms TNF superfamily member 15 (TNFSF15), TL1, and VEGI) is a member of the tumor necrosis factor superfamily expressed by antigen-presenting cells (including dendritic cells, B cells, and macrophages), CD4+ and CD8+ T cells, and endothelial cells. TL1A is expressed on the cell surface or secreted as a soluble cytokine. The receptor for TL1A, death receptor 3 (DR3), is expressed by a variety of cells, including CD4+ and CD8+ T cells, NK cells, NKT cells, FOXP3+ regulatory T (Treg) cells, and type 2 and type 3 innate lymphoid cells (ILC2s and ILC3s).

[0004] TL1A can also bind to the decoy receptor (DcR3), which competitively inhibits DR3. DcR3 also serves as a decoy receptor for Fas-ligand (Fas-L) and lymphotoxin-like-inducible protein, and competes with glycoprotein D for binding to herpesvirus entry mediator (LIGHT) on T cells. Therefore, DcR3 is a key regulator of several signaling pathways.

[0005] The TL1A / DR3 signaling pathway is involved in several biological systems related to human diseases. For example, TL1A has been shown to be involved in immunity, angiogenesis, and barrier tissue homeostasis. Blocking the interaction of TL1A with DR3 has also been shown to enhance the therapeutic effects of several immune-mediated diseases, including experimental autoimmune encephalomyelitis (EAE; a model of multiple sclerosis), colitis, ulcerative colitis, Crohn's disease, inflammatory bowel disease, skin diseases, asthma, and arthritis. Therefore, antibodies that bind to TL1A have been proposed as therapeutic agents for these diseases. Summary of the Invention [Problem to be solved by the invention]

[0006] However, the use of such antibodies to provide treatment for these diseases requires stable formulations of the antibodies. [Means for solving the problem]

[0007] The present specification provides a pharmaceutical formulation, comprising: (a) about 100 mg / mL to about 250 mg / mL of an antibody or antigen-binding fragment thereof that specifically binds to TNF-like ligand 1A (TL1A), the antibody or antigen-binding fragment thereof comprising a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 2, a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 3, a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 4, and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 5; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 6; (b) about 5 mM to about 15 mM histidine; (c) about 50 mM to about 150 mM arginine hydrochloride (Arg-HCl); (d) about 2.5% (w / v) to about 7.5% (w / v) sucrose; and (e) about 0.01% (w / v) to about 0.03% (w / v) polysorbate-80. In some embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 7 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 8. In some embodiments, the antibody or antigen-binding fragment thereof comprises an IgG1 constant region. In some embodiments, the antibody or antigen-binding fragment thereof 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.

[0008] In some embodiments, the pharmaceutical formulations disclosed herein contain about 100, about 150, about 200, about 225, or about 250 mg / mL of an antibody or antigen-binding fragment thereof. In some embodiments, the pharmaceutical formulations disclosed herein contain about 5 mM, about 10 mM, or about 15 mM histidine. In some embodiments, the pharmaceutical formulations disclosed herein contain about 50 mM, about 100 mM, or about 150 mM arginine hydrochloride (Arg-HCl). In some embodiments, the pharmaceutical formulations disclosed herein contain about 2.5%, about 5%, or about 7.5% (w / v) sucrose. In some embodiments, the pharmaceutical formulations disclosed herein contain about 0.01%, about 0.02%, or about 0.03% (w / v) polysorbate-80.

[0009] In some aspects, the pharmaceutical formulations disclosed herein comprise about 250 mg / mL of an antibody or antigen-binding fragment thereof, about 10 mM histidine, about 100 mM arginine hydrochloride (Arg-HCl), about 5% (w / v) sucrose, and about 0.02% (w / v) polysorbate-80.

[0010] In some aspects, the pharmaceutical formulations disclosed herein comprise about 200 mg / mL of an antibody or antigen-binding fragment thereof, about 10 mM histidine, about 100 mM arginine hydrochloride (Arg-HCl), about 5% (w / v) sucrose, and about 0.02% (w / v) polysorbate-80.

[0011] In some aspects, the pharmaceutical formulations disclosed herein comprise about 150 mg / mL of an antibody or antigen-binding fragment thereof, about 10 mM histidine, about 100 mM arginine hydrochloride (Arg-HCl), about 5% (w / v) sucrose, and about 0.02% (w / v) polysorbate-80.

[0012] In some embodiments, the pharmaceutical formulation is lyophilized. In some embodiments, the pharmaceutical formulation is liquid. In some embodiments, the pharmaceutical formulation has a pH of 6.0±0.5 after storage at room temperature for 24 hours or at 2-8° C. for 24 hours, 72 hours, or 10 days.

[0013] In some embodiments, the pharmaceutical formulation has an osmolality of 200 mOsm / kg to 500 mOsm / kg after storage at room temperature for 24 hours or at 2-8°C for 24 hours, 72 hours, or 10 days. In some embodiments, the pharmaceutical formulation has at least 99% antibody monomer content after storage at 2-8°C for 24 hours, 72 hours, or 10 days. In some embodiments, the pharmaceutical formulation has no significant change in charge heterogeneity profile after storage at 2-8°C for 24 hours, 72 hours, or 10 days. In some embodiments, the pharmaceutical formulation has no significant change in purity after storage at room temperature for 24 hours or at 2-8°C for 24 hours, 72 hours, or 10 days. In some embodiments, the pharmaceutical formulation has at least 90% purity after storage at room temperature for 24 hours or at 2-8°C for 24 hours, 72 hours, or 10 days. In some embodiments, the pharmaceutical formulations have no significant change in particle concentration after storage at room temperature for 24 hours or at 2-8°C for 24 hours, 72 hours, or 10 days. In some embodiments, the pharmaceutical formulations have no significant difference in visual appearance after storage at 2-8°C for up to 36 months. In some embodiments, the pharmaceutical formulations disclosed herein have no significant difference in protein concentration, osmolality, or viscosity after storage at 2-8°C, 25°C, or 40°C for up to 36 months. In some embodiments, the pharmaceutical formulations disclosed herein have a monomer content of 95% or greater, a dimer content of 5.0% or less, or no significant difference in low molecular weight species content after storage at 2-8°C for up to 36 months. In some embodiments, the pharmaceutical formulations disclosed herein have a purity of 90% or greater after storage at 2-8°C for up to 36 months. In some embodiments, the pharmaceutical formulations disclosed herein have a major species content of 50% to 90%, an acidic species content of 10% to 40%, and / or a basic species content of 0% to 20% after storage for up to 36 months at 2-8° C. In some embodiments, the pharmaceutical formulations disclosed herein do not have a significant difference in subvisible particle content after storage for up to 36 months at 2-8° C., 25° C., or 40° C.In some embodiments, the pharmaceutical formulations disclosed herein have no significant difference in oxidation of methionine 81 and / or methionine 254 of TEV-48574 and / or deamidation of asparagine 317 of TEV-48574 after storage for up to 36 months at 2-8°C, 25°C, or 40°C. In some embodiments, the pharmaceutical formulations disclosed herein have a relative potency of 70% to 135% as measured by enzyme-linked immunosorbent assay (ELISA) after storage for up to 36 months at 2-8°C. In some embodiments, the pharmaceutical formulations disclosed herein have no significant difference in thermal stability after storage for up to 6 months at 2-8°C, 25°C, or 40°C. In some embodiments, the pharmaceutical formulations disclosed herein have no significant difference in thermal stability after storage for up to 36 months at 2-8°C. In some embodiments, the pharmaceutical formulations disclosed herein have no significant difference in secondary and / or tertiary protein structure after storage for up to 3 months at 2-8° C., 25° C., or 40° C. In some embodiments, the pharmaceutical formulations disclosed herein have no significant difference in secondary protein structure after storage for up to 36 months at 2-8° C. In some embodiments, the pharmaceutical formulations disclosed herein have no significant difference in polysorbate-80 concentration after storage for up to 24 months at 2-8° C.

[0014] Also provided is a container that contains the pharmaceutical preparation disclosed herein.In some embodiments, the container is a glass vial.In some embodiments, the container is a glass vial with a filling volume of 3mL.In some embodiments, the container is a glass vial with a filling volume of 3mL or less.In some embodiments, the container is a syringe, for example, a pre-filled syringe.In some embodiments, the pre-filled syringe has a volume of 2mL or less.

[0015] In some embodiments, the antibody or antigen-binding fragment thereof is present in a pharmaceutical formulation or container disclosed herein. In some embodiments, the antibody or antigen-binding fragment thereof is formulated in a volume of 3 mL or less. In some embodiments, the antibody or antigen-binding fragment thereof is formulated in a volume of 2 mL or less.

[0016] In some aspects, provided herein are methods of treating a disease in a subject in need thereof, the method comprising administering to the subject any of the pharmaceutical formulations provided herein, including the pharmaceutical formulation in any of the containers provided herein. In some aspects, the disease is a respiratory disease, a gastrointestinal disease, a skin disease, or arthritis.

[0017] In some embodiments, the airway disease is asthma, chronic obstructive pulmonary disease (COPD), pulmonary fibrosis, pulmonary sarcoidosis, allergic rhinitis, or cystic fibrosis.

[0018] In some aspects, the gastrointestinal condition is inflammatory bowel disease, Crohn's disease, colitis, ulcerative colitis, eosinophilic esophagitis, or irritable bowel syndrome.

[0019] In some embodiments, the arthritis is rheumatoid arthritis.

[0020] In some embodiments, the skin disorder is atopic dermatitis, eczema, or scleroderma.

[0021] In some embodiments, the pharmaceutical formulation is administered intravenously. In some embodiments, the pharmaceutical formulation is administered subcutaneously.

[0022] Compositions for use in accordance with the methods disclosed herein are also provided.

[0023] The present specification provides a pharmaceutical formulation, comprising: (a) 150 mg / mL of an antibody or antigen-binding fragment thereof that specifically binds to TNF-like ligand 1A (TL1A), the antibody or antigen-binding fragment thereof comprising: (i) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 7 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 8; (ii) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 2, a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 3, and a light chain variable region CDR4 comprising the amino acid sequence of SEQ ID NO: 4; Provided is a pharmaceutical formulation comprising: (i) an antibody or antigen-binding fragment thereof comprising a heavy chain having a CDR1 region, a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 5, and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 6; or (ii) a heavy chain having a CDR1 region, a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 6; (iii) an antibody or antigen-binding fragment thereof comprising a heavy chain having a CDR1 region, a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 9, and a light chain having the amino acid sequence of SEQ ID NO: 10; (b) 10 mM histidine; (c) 100 mM arginine hydrochloride (Arg-HCl); (d) 5% (w / v) sucrose; and (e) 0.02% (w / v) polysorbate-80. In some embodiments, the antibody or antigen-binding fragment of (i) or (ii) comprises an IgG1 constant region.

[0024] In some embodiments, the pharmaceutical formulation is lyophilized. In some embodiments, the pharmaceutical formulation is liquid. In some embodiments, the pharmaceutical formulation has a pH of 6.0±0.5 after storage at room temperature for 24 hours or at 2-8° C. for 24 hours, 72 hours, or 10 days.

[0025] In some embodiments, the pharmaceutical formulation has an osmolality of 200 mOsm / kg to 500 mOsm / kg after storage at room temperature for 24 hours or at 2-8°C for 24 hours, 72 hours, or 10 days. In some embodiments, the pharmaceutical formulation has at least 99% antibody monomer content after storage at 2-8°C for 24 hours, 72 hours, or 10 days. In some embodiments, the pharmaceutical formulation has no significant change in charge heterogeneity profile after storage at 2-8°C for 24 hours, 72 hours, or 10 days. In some embodiments, the pharmaceutical formulation has no significant change in purity after storage at room temperature for 24 hours or at 2-8°C for 24 hours, 72 hours, or 10 days. In some embodiments, the pharmaceutical formulation has at least 90% purity after storage at room temperature for 24 hours or at 2-8°C for 24 hours, 72 hours, or 10 days. In some embodiments, the pharmaceutical formulations have no significant change in particle concentration after storage at room temperature for 24 hours or at 2-8°C for 24 hours, 72 hours, or 10 days. In some embodiments, the pharmaceutical formulations have no significant difference in visual appearance after storage at 2-8°C for up to 36 months. In some embodiments, the pharmaceutical formulations disclosed herein have no significant difference in protein concentration, osmolality, or viscosity after storage at 2-8°C, 25°C, or 40°C for up to 36 months. In some embodiments, the pharmaceutical formulations disclosed herein have a monomer content of 95% or greater, a dimer content of 5.0% or less, or no significant difference in low molecular weight species content after storage at 2-8°C for up to 36 months. In some embodiments, the pharmaceutical formulations disclosed herein have a purity of 90% or greater after storage at 2-8°C for up to 36 months. In some embodiments, the pharmaceutical formulations disclosed herein have a major species content of 50% to 90%, an acidic species content of 10% to 40%, and / or a basic species content of 0% to 20% after storage for up to 36 months at 2-8° C. In some embodiments, the pharmaceutical formulations disclosed herein do not have a significant difference in subvisible particle content after storage for up to 36 months at 2-8° C., 25° C., or 40° C.In some embodiments, the pharmaceutical formulations disclosed herein have no significant difference in oxidation of methionine 81 and / or methionine 254 of TEV-48574 and / or deamidation of asparagine 317 of TEV-48574 after storage for up to 36 months at 2-8°C, 25°C, or 40°C. In some embodiments, the pharmaceutical formulations disclosed herein have a relative potency of 70% to 135% as measured by enzyme-linked immunosorbent assay (ELISA) after storage for up to 36 months at 2-8°C. In some embodiments, the pharmaceutical formulations disclosed herein have no significant difference in thermal stability after storage for up to 6 months at 2-8°C, 25°C, or 40°C. In some embodiments, the pharmaceutical formulations disclosed herein have no significant difference in thermal stability after storage for up to 36 months at 2-8°C. In some embodiments, the pharmaceutical formulations disclosed herein have no significant difference in secondary and / or tertiary protein structure after storage for up to 3 months at 2-8° C., 25° C., or 40° C. In some embodiments, the pharmaceutical formulations disclosed herein have no significant difference in secondary protein structure after storage for up to 36 months at 2-8° C. In some embodiments, the pharmaceutical formulations disclosed herein have no significant difference in polysorbate-80 concentration after storage for up to 24 months at 2-8° C.

[0026] Also provided is a container containing the pharmaceutical formulation disclosed herein. In some embodiments, the container is a glass vial. In some embodiments, the container is a glass vial with a fill volume of 3 mL.

[0027] In some aspects, the antibody or antigen-binding fragment thereof is present in a pharmaceutical formulation or container disclosed herein.

[0028] In some aspects, provided herein are methods of treating a disease in a subject in need thereof, the method comprising administering to the subject any of the pharmaceutical formulations provided herein, including the pharmaceutical formulation in any of the containers provided herein. In some aspects, the disease is a respiratory disease, a gastrointestinal disease, a skin disease, or arthritis.

[0029] In some embodiments, the airway disease is asthma, chronic obstructive pulmonary disease (COPD), pulmonary fibrosis, pulmonary sarcoidosis, allergic rhinitis, or cystic fibrosis.

[0030] In some aspects, the gastrointestinal condition is inflammatory bowel disease, Crohn's disease, colitis, ulcerative colitis, eosinophilic esophagitis, or irritable bowel syndrome.

[0031] In some embodiments, the arthritis is rheumatoid arthritis.

[0032] In some embodiments, the skin disorder is atopic dermatitis, eczema, or scleroderma.

[0033] Compositions for use in accordance with the methods disclosed herein are also provided. [Brief explanation of the drawings]

[0034] [Figures 1A-1C] The percent (%) of monomer composition of three TEV-48574 formulations (F1-F3) after storage at 2-8°C (Figure 1A), 25°C (Figure 1B), and 40°C (Figure 1C) is shown. [Figures 2A-2C] The percent (%) dimer composition of three TEV-48574 formulations (F1-F3) after storage at 2-8°C (Figure 2A), 25°C (Figure 2B), and 40°C (Figure 2C) is shown. [Figure 3A-3C] Figure 3 shows the percent (%) low molecular weight (LWM) protein fragment composition of three TEV-48574 formulations (F1-F3) after storage at 2-8°C (Figure 3A), 25°C (Figure 3B), and 40°C (Figure 3C). [Figures 4A-4C] The purity of three TEV-48574 formulations (F1-F3) after storage at 2-8°C (Figure 4A), 25°C (Figure 4B), and 40°C (Figure 4C) as measured using CGE-non-reducing conditions (IgG+ 125 kDa peak (%)). [Figures 5A-5C]The purity of three TEV-48574 formulations (F1-F3) after storage at 2-8°C (Figure 5A), 25°C (Figure 5B), and 40°C (Figure 5C), as measured using CGE-reducing conditions (heavy chain + light chain (%)). [Figures 6A-6C] Charge heterogeneity profiles of Formulations 1-3 (F1-F3) after storage at 2-8°C (Figure 6A), 25°C (Figure 6B), and 40°C (Figure 6C) are shown as percent (%) of the major species (main peak). [Figure 7A-7C] Charge heterogeneity profiles of Formulations 1-3 (F1-F3) after storage at 2-8°C (Figure 7A), 25°C (Figure 7B), and 40°C (Figure 7C) are shown as percent (%) of acidic species (acidic peak). [Figures 8A-8C] Charge heterogeneity profiles of Formulations 1-3 (F1-F3) after storage at 2-8°C (Figure 8A), 25°C (Figure 8B), and 40°C (Figure 8C) are shown as percent (%) of basic species (basic peak). [Figure 9A-9B] (FIG. 9A) Thermal stability of Formulations 1 to 3 (F1 to F3) at time zero (T0) and after 3 months of storage at 2 to 8°C, 25°C, and 40°C. (FIG. 9B) Thermal stability of Formulations 1 to 3 (F1 to F3) at time zero (T0) and after 6 months of storage at 2 to 8°C, 25°C, and 40°C. [Figure 9C] 1 shows the thermal stability of Formulations 1-3 (F1-F3) at time zero (T0) and after 36 months of storage at 2-8°C. [Figures 10A-10B] (Figure 10A) Far-ultraviolet (UV) circular dichroism analysis (CD) is used to show the secondary structure of Formulations 1-3 at time zero (T0) and after 3 months of storage at 2-8°C, 25°C, and 40°C. (Figure 10B) Far-UV CD is used to show the secondary structure of Formulations 1-3 at time zero (T0) and after 24 months of storage at 2-8°C. [Figures 10C-10D] (Figure 10C) Far-UV CD is used to show the secondary structure of Formulations 1-3 at time zero (T0) and after 36 months of storage at 2-8°C. (Figure 10D) Far-UV CD is used to show the tertiary structure of Formulations 1-3 at time zero (T0) and after 6 months of storage at 2-8°C, 25°C, and 40°C. [Figures 10E-10F](Figure 10E) Far-UV CD is used to show the tertiary structure of Formulations 1-3 at time zero (T0) and after 24 months of storage at 2-8°C. (Figure 10F) Far-UV CD is used to show the tertiary structure of Formulations 1-3 at time zero (T0) and after 36 months of storage at 2-8°C. [Figures 11A-11B] (FIG. 11A) The secondary structure of Formulation 3 stored in Nipro prefilled syringes (PFS) is shown using far-UV CD at time zero (T0) and after 3 months (3M), 6 months (6M), and 24 months (24M) of storage at 2-8° C. and after 3 months (3M) and 6 months (6M) of storage at 25° C. and 40° C. (FIG. 11B) The tertiary structure of Formulation 3 stored in Nipro prefilled syringes (PFS) is shown using near-UV CD at time zero (T0) and after 3 months (3M), 6 months (6M), and 24 months (24M) of storage at 2-8° C. and after 3 months (3M) and 6 months (6M) of storage at 25° C. and 40° C. [Figures 12A-12C] (Figure 12A) Percent monomer composition (%) for a 100 mg / mL lyophilized TEV-48574 formulation (F4) and a 200 mg / mL liquid TEV-48574 formulation (F5) after storage for up to 24 months at 2-8°C. (Figure 12B) Percent dimer composition (%) for a 100 mg / mL lyophilized TEV-48574 formulation (F4) and a 200 mg / mL liquid TEV-48574 formulation (F5) after storage for up to 24 months at 2-8°C. (Figure 12C) Percent low molecular weight species (%) for a 100 mg / mL lyophilized TEV-48574 formulation (F4) and a 200 mg / mL liquid TEV-48574 formulation (F5) after storage for up to 24 months at 2-8°C. [Figures 13A-13B] (Figure 13A) Purity (%) measured using CGE-non-reducing conditions (IgG + 125 kDa peak (%)) for two TEV-48574 formulations (F4 and F5) after up to 24 months of storage at 2-8°C. (Figure 13B) Purity (%) measured using CGE-reducing conditions (heavy chain (HC) + light chain (LC) (%)) for two TEV-48574 formulations (F4 and F5) after up to 24 months of storage at 2-8°C. [Figures 14A-14C](Figure 14A) Charge heterogeneity profiles of Formulations 4 and 5 (F4 and F5) after storage at 2-8°C for up to 24 months are shown as percent (%) of the major species (main peak). (Figure 14B) Charge heterogeneity profiles of Formulations 4 and 5 (F4 and F5) after storage at 2-8°C for up to 24 months are shown as percent (%) of the acidic species (acidic peak). (Figure 14C) Charge heterogeneity profiles of Formulations 4 and 5 (F4 and F5) after storage at 2-8°C for up to 24 months are shown as percent (%) of the basic species (basic peak). [Figures 15A-15B] (FIG. 15A) The secondary structure of Formulation 4 at time zero (T0) and after 3 months (3M), 6 months (6M), and 12 months (12M) storage at 25° C., and Formulation 5 at T0 and after 3M and 12M storage at 25° C. are shown using far-UV CD. (FIG. 15B) The tertiary structure of Formulation 4 at time zero (T0) and after 3 months (3M) and 12 months (12M) storage at 25° C., and Formulation 5 at T0 and after 3M, 6 months (6M), and 12M storage at 25° C. are shown using near-UV CD. [Figure 16] Viscosity at 20°C is shown for liquid formulations containing increasing concentrations of TEV-48574 (0-150 mg / mL) with (1A) or without (1B) 100 mM arginine-HCl as an excipient. [Figures 17A-17C](FIG. 17A) Percent (%) monomer composition of two liquid TEV-48574 formulations, 150 mg / mL (1A) and 100 mg / mL (2A), with 100 mM arginine-HCl and 150 mg / mL (1B) and 100 mg / mL (2B), without arginine-HCl, after storage at 40° C. (FIG. 17B) Percent (%) dimer composition of two liquid TEV-48574 formulations, 150 mg / mL (1A) and 100 mg / mL (2A), with 100 mM arginine-HCl and 150 mg / mL (1B) and 100 mg / mL (2B), without arginine-HCl, after storage at 40° C. for up to 8 weeks. (FIG. 17C) Percent fragments (%) of two liquid TEV-48574 formulations at 150 mg / mL (1A) and 100 mg / mL (2A) with 100 mM arginine-HCl and at 150 mg / mL (1B) and 100 mg / mL (2B) without arginine-HCl after storage at 40° C. for up to 8 weeks. [Figures 18A-18B] (FIG. 18A) Percent purity (%) of two liquid TEV-48574 formulations at 150 mg / mL (1A) and 100 mg / mL (2A) with and without 100 mM arginine-HCl after up to 8 weeks of storage at 40° C., as measured using CGE-non-reducing conditions (IgG+125 kDa peak (%)). (FIG. 18B) Percent purity (%) of two liquid TEV-48574 formulations at 150 mg / mL (1A) and 100 mg / mL (2A) with and without 100 mM arginine-HCl after up to 8 weeks of storage at 40° C., as measured using CGE-reducing conditions (heavy chain (HC) + light chain (LC) (%)). [Figures 19A-19C](FIG. 19A) Charge heterogeneity profiles are shown as percent (%) of the major species (main peak) for two liquid TEV-48574 formulations at 150 mg / mL (1A) and 100 mg / mL (2A) with and without 100 mM arginine-HCl after storage at 40° C. for up to 8 weeks. (FIG. 19B) Charge heterogeneity profiles, expressed as percent (%) of acidic species (acidic peak), of two liquid TEV-48574 formulations at 150 mg / mL (1A) and 100 mg / mL (2A) with and without 100 mM arginine-HCl after storage at 40° C. for up to 8 weeks. (FIG. 19C) Charge heterogeneity profiles, expressed as percent (%) of basic species (basic peak), of two liquid TEV-48574 formulations at 150 mg / mL (1A) and 100 mg / mL (2A) with and without 100 mM arginine-HCl after storage at 40° C. for up to 8 weeks. [Figure 20] Figure 1 shows the thermal stability at time zero (T0) of two liquid TEV-48574 formulations at 150 mg / mL (1A) and 100 mg / mL (2A) with 100 mM arginine-HCl and at 150 mg / mL (1B) and 100 mg / mL (2B) without arginine-HCl. DETAILED DESCRIPTION OF THE INVENTION

[0035] In order that this disclosure may be more readily understood, certain terms are first defined. As used in this application, each of the following terms shall have the meaning set forth below, unless expressly defined otherwise herein. Additional definitions are set forth throughout this application.

[0036] definition Various terms relating to aspects of the present disclosure are used throughout the specification and claims. Such terms shall have their ordinary meaning in the art unless otherwise specified. Other specifically defined terms shall be interpreted consistent with the definitions provided herein.

[0037] The term "antibody" refers to an immunoglobulin molecule that recognizes and specifically binds to a target (e.g., a protein, polypeptide, peptide, carbohydrate, polynucleotide, lipid, or a combination of the above) via at least one antigen recognition site within the variable region of the immunoglobulin molecule. As used herein, the term "antibody" encompasses intact polyclonal antibodies, intact monoclonal antibodies, chimeric antibodies, humanized antibodies, human antibodies, antibody-containing fusion proteins, and any other modified immunoglobulin molecule, so long as the antibody exhibits the desired biological activity. Antibodies may belong to any of the five major classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, or their subclasses (isotypes) (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2) (based on the identity of the heavy chain constant domains, designated alpha, delta, epsilon, gamma, and mu, respectively). Different classes of immunoglobulins have different and well-known subunit structures and three-dimensional configurations. The antibody may be a naked antibody or may be conjugated to another molecule, such as a radioisotope.

[0038] Unless expressly stated or the context dictates otherwise, the term "antibody" includes monospecific antibodies, bispecific antibodies, or multispecific antibodies. In some embodiments, an antibody is a bispecific antibody. The term "bispecific antibody" refers to an antibody that binds to two different epitopes.

[0039] The term "antibody fragment" refers to a portion of an intact antibody. An "antigen-binding fragment," "antigen-binding domain," or "antigen-binding region" refers to a portion of an intact antibody that binds to an antigen. In the case of a bispecific antibody, the "antigen-binding fragment" binds to two antigens. An antigen-binding fragment may contain the antigen recognition site of the intact antibody (e.g., sufficient complementarity-determining regions (CDRs) to specifically bind to the antigen). Examples of antigen-binding fragments of antibodies include, but are not limited to, Fab, Fab', F(ab')2, and Fv fragments, linear antibodies, and single-chain antibodies. Antigen-binding fragments of antibodies may be derived from any animal species, including rodents (e.g., mice, rats, or hamsters) and humans, or may be artificially generated.

[0040] A "monoclonal" antibody or antigen-binding fragment thereof refers to a population of homogeneous antibodies or antigen-binding fragments responsible for highly specific binding to a single antigenic determinant or epitope. This is in contrast to polyclonal antibodies, which typically contain different antibodies directed against different antigenic determinants. The term "monoclonal" antibody or antigen-binding fragment thereof encompasses both intact and full-length monoclonal antibodies, as well as antibody fragments (Fab, Fab', F(ab')2, Fv, etc.), single-chain (scFv) variants, fusion proteins containing antibody portions, and any other modified immunoglobulin molecule containing an antigen-recognition site. Furthermore, a "monoclonal" antibody or antigen-binding fragment thereof refers to antibodies and antigen-binding fragments thereof produced by any number of methods, including, but not limited to, by hybridoma, phage selection, recombinant expression, and transgenic animals.

[0041] As used herein, the terms "variable region" and "variable domain" are used interchangeably and are common in the art. A variable region typically refers to a portion of an antibody, generally a light or heavy chain, typically the amino-terminal 110-120 or 110-125 amino acids in a mature heavy chain and approximately 90-115 amino acids in a mature light chain, which differ in sequence among antibodies and are responsible for the binding and specificity of a particular antibody to its particular antigen. Sequence variability is concentrated in regions called complementarity-determining regions (CDRs), while the more highly conserved regions in the variable domain are called framework regions (FRs). While not wishing to be bound by any particular mechanism or theory, it is believed that the CDRs of the light and heavy chains are primarily responsible for the interaction and specificity of the antibody with the antigen. In some embodiments of the present disclosure, the variable region is a human variable region. In some embodiments of the present disclosure, the variable region comprises rodent or murine CDRs and human framework regions (FRs). In certain aspects of the present disclosure, the variable region is a primate (e.g., non-human primate) variable region. In some aspects of the present disclosure, the variable region comprises rodent or murine CDRs and primate (e.g., non-human primate) framework regions (FRs).

[0042] The terms "VL" and "VL domain" are used interchangeably and refer to the light chain variable region of an antibody.

[0043] The terms "VH" and "VH domain" are used interchangeably and refer to the heavy chain variable region of an antibody.

[0044] The term "Kabat numbering" and similar terms are art-recognized and refer to a system for numbering amino acid residues in the heavy and light chain variable regions of an antibody or antigen-binding fragment thereof. In some embodiments, CDRs can be determined according to the Kabat numbering system (see, e.g., Kabat EA & Wu TT (1971) Ann NY Acad Sci 190:382-391 and Kabat EA et al., (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, USDapartment of Health and Human Services, NIH Publication No. 91-3242). Using the Kabat numbering system, the CDRs in an antibody heavy chain molecule are typically located at amino acid positions 31-35, and may optionally include one or two additional amino acids following position 35 (designated 35A and 35B in the Kabat numbering scheme) (CDR1), amino acid positions 50-65 (CDR2), and amino acid positions 95-102 (CDR3). Using the Kabat numbering system, the CDRs in an antibody light chain molecule are typically located at amino acid positions 24-34 (CDR1), amino acid positions 50-56 (CDR2), and amino acid positions 89-97 (CDR3).

[0045] Chothia instead refers to the location of the structural loops (Chothia and Lesk, J. Mol. Biol. 196:901-917 (1987)). The end of the Chothia CDR-H1 loop, when numbered using the Kabat numbering convention, varies from H32 to H34 depending on the length of the loop (this is because the Kabat numbering scheme places insertions at H35A and H35B; if neither 35A nor 35B are present, the loop ends at 32; if only 35A is present, the loop ends at 33; and if both 35A and 35B are present, the loop ends at 34). The AbM hypervariable regions represent a compromise between the Kabat CDRs and the Chothia structural loops and are used by Oxford Molecular's AbM antibody modeling software.

[0046] [Table 1]

[0047] As used herein, the terms "constant region" and "constant domain" are interchangeable and have their common meaning in the art. The constant region is the carboxyl-terminal portion of an antibody portion, e.g., the light chain and / or heavy chain, that is not directly involved in binding the antibody to an antigen but may exhibit various effector functions, such as interacting with Fc receptors. The constant region of an immunoglobulin molecule generally has a conserved amino acid sequence compared to the immunoglobulin variable domain. In some embodiments, the antibody or antigen-binding fragment comprises a constant region or portion thereof sufficient for antibody-dependent cell-mediated cytotoxicity (ADCC).

[0048] As used herein, the term "heavy chain," when used in reference to an antibody, can refer to any distinct type, e.g., alpha (α), delta (δ), epsilon (ε), gamma (γ), and mu (μ), based on the amino acid sequence of the constant domain, which give rise to the IgA, IgD, IgE, IgG, and IgM classes of antibodies, respectively, including subclasses of IgG, e.g., IgG1, IgG2, IgG3, and IgG4. Heavy chain amino acid sequences are well known in the art. In some aspects of the present disclosure, the heavy chain is a human heavy chain.

[0049] As used herein, the term "light chain," when used in reference to an antibody, can refer to any distinct type, e.g., κ (kappa) or λ (lambda), based on the amino acid sequence of the constant domain. Light chain amino acid sequences are well known in the art. In some aspects of the present disclosure, the light chain is a human light chain.

[0050] The term "chimeric" antibody or antigen-binding fragment thereof refers to an antibody or antigen-binding fragment thereof whose amino acid sequence is derived from two or more species. Typically, the variable regions of both the light and heavy chains correspond to the variable regions of an antibody or antigen-binding fragment thereof from one species of mammal (e.g., mouse, rat, rabbit, etc.) having the desired specificity, affinity, and capacity, while the constant regions are homologous to the sequences of an antibody or antigen-binding fragment thereof from another (usually human) so as not to elicit an immune response in that species.

[0051] The term "humanized" antibody or antigen-binding fragment thereof refers to forms of non-human (e.g., murine) antibodies or antigen-binding fragments that are specific immunoglobulin chains, chimeric immunoglobulins, or fragments thereof that contain minimal non-human (e.g., murine) sequence. Typically, humanized antibodies or antigen-binding fragments thereof are human immunoglobulins in which residues of the complementarity-determining regions (CDRs) are replaced with residues from CDRs of a non-human species (e.g., mouse, rat, rabbit, hamster) having the desired specificity, affinity, and capacity ("CDR-grafted") (Jones et al., Nature, 321:522-525 (1986); Riechmann et al., Nature, 332:323-327 (1988); Verhoeyen et al., Science, 239:1534-1536 (1988)). In some instances, particular Fv framework region (FR) residues of a human immunoglobulin are substituted with corresponding residues in an antibody or fragment from a non-human species having the desired specificity, affinity, and capacity. Humanized antibodies or antigen-binding fragments thereof can be further modified by substitution of additional residues in the Fv framework regions and / or within non-human CDR residues to refine and optimize the specificity, affinity, and / or capacity of the antibody or antigen-binding fragment. Generally, a humanized antibody or antigen-binding fragment thereof will comprise variable domains that include all or substantially all of the CDR regions corresponding to those of a non-human immunoglobulin, while all or substantially all of the FR regions are of human immunoglobulin consensus sequences. Humanized antibodies or antigen-binding fragments thereof may also comprise at least a portion of an immunoglobulin constant region or domain (Fc), typically that of a human immunoglobulin. Examples of methods used to generate humanized antibodies are described in U.S. Patent No. 5,225,539, Roguska et al., Proc. Natl. Acad. Sci., USA, 91(3):969-73 (1994), and Roguska et al., Protein Eng. 9(10):895-904 (1996). In some aspects of the present disclosure, the "humanized antibody" is a resurfaced antibody.

[0052] The term "human" antibody or antigen-binding fragment thereof means an antibody or antigen-binding fragment thereof having an amino acid sequence derived from the human immunoglobulin locus, and such antibody or antigen-binding fragment is produced using any technique known in the art. This definition of human antibody or antigen-binding fragment thereof includes intact or full-length antibodies and fragments thereof.

[0053] "Binding affinity" generally refers to the overall strength of non-covalent interactions between a single binding site of a molecule (e.g., an antibody or antigen-binding fragment thereof) and its binding partner (e.g., an antigen). Unless otherwise specified, "binding affinity" as used herein refers to the intrinsic binding affinity, which indicates a 1:1 interaction between members of a binding pair (e.g., an antibody or antigen-binding fragment thereof and an antigen). The affinity of a molecule X for its partner Y can generally be expressed by a dissociation constant (KD). Affinity can be measured and / or expressed in several ways known in the art, including, but not limited to, the equilibrium dissociation constant (KD) and the equilibrium association constant (KA). KD is determined by the k off / k on KA is calculated from the quotient of k off / k on It is calculated from the quotient of K on means, for example, the association rate constant of an antibody or antigen-binding fragment thereof to an antigen, and k off k means, for example, dissociation of an antibody or antigen-binding fragment thereof from an antigen. on and k off can be determined by techniques known to those skilled in the art, such as BIAcore® or KinExA.

[0054] As used herein, "epitope" is a term of the art and refers to a localized region of an antigen to which an antibody or antigen-binding fragment thereof can specifically bind. An epitope can be, for example, consecutive amino acids of a polypeptide (a linear or continuous epitope), or can be joined together, for example, by two or more non-contiguous regions of a polypeptide (a conformational, non-linear, discontinuous, or discontinuous epitope). In some aspects of the present disclosure, the epitope to which an antibody or antigen-binding fragment thereof specifically binds can be determined by, for example, NMR spectroscopy, X-ray diffraction crystallography studies, ELISA assays, hydrogen / deuterium exchange mass spectrometry (e.g., liquid chromatography electrospray mass spectrometry), array-based oligopeptide scanning assays, and / or mutagenesis mapping (e.g., site-directed mutagenesis mapping). For X-ray crystallography, crystallization can be achieved using any of the methods known in the art (e.g., Giege R et al., (1994) Acta Crystallogr D Biol Crystallogr 50(Pt 4):339-350; McPherson A (1990) Eur J Biochem 189:1-23; Chayen NE (1997) Structure 5:1269-1274; McPherson A (1976) J Biol Chem 251:6300-6303).Crystal structures containing complexes of antigen and antibody or antigen-binding fragments can be studied using well-known X-ray diffraction techniques and refined using computer software such as X-PLOR (Yale University, 1992, Molecular Simulations, Inc.; see, e.g., Meth Enzymol (1985) volumes 114&115, eds. Wyckoff HW et al.; U.S. Patent Application Publication No. 2004 / 0014194) and BUSTER (Bricogne G (1993) Acta Crystallogr D Biol Crystallogr 49(Pt 1):37-60; Bricogne G (1997) Meth Enzymol 276A:361-423, ed. Carter CW; Roversi P et al., (2000) Acta Crystallogr D Biol Crystallogr 56(Pt 10):1316-1323). Mutagenesis mapping studies can be accomplished using any method known to those of skill in the art. For example, for a description of mutagenesis techniques, including alanine scanning mutagenesis techniques, see Champe M et al., (1995) J Biol Chem 270:1388-1394 and Cunningham BC & Wells JA (1989) Science 244:1081-1085.

[0055] An "isolated" polypeptide, antibody, polynucleotide, vector, cell, or composition is a polypeptide, antibody, polynucleotide, vector, cell, or composition in a form not found in nature. Isolated polypeptides, antibodies, polynucleotides, vectors, cells, or compositions include those that have been purified to the extent that they are no longer in a form found in nature. In some aspects of the present disclosure, an antibody, polynucleotide, vector, cell, or composition that is isolated is substantially pure. As used herein, "substantially pure" refers to material that is at least 50% pure (i.e., free from contaminants), at least 90% pure, at least 95% pure, at least 98% pure, or at least 99% pure.

[0056] The terms "polypeptide," "peptide," and "protein" are used interchangeably herein to refer to a polymer of amino acids of any length. The polymer can be linear or branched, it can comprise modified amino acids, and it can be interrupted by non-amino acids. The term also includes amino acid polymers that are modified, naturally or by intervention, for example, by disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or any other manipulation or modification, such as conjugation with a labeling component. Also included within the definition are polypeptides containing one or more analogs of an amino acid (including, for example, unnatural amino acids), as well as other modifications known in the art. Because the polypeptides of this disclosure are based on antibodies, it is understood that in some aspects of this disclosure, the polypeptides can occur as single chains or associated chains.

[0057] As used herein, the term "pharmaceutical formulation" refers to a preparation that is in a form that allows the biological activity of the active ingredient to be effective and does not contain additional ingredients that are unacceptably toxic to the subject to which the formulation is administered. The formulation may be sterile. In some aspects, the formulation is suitable for therapeutic use in human subjects.

[0058] As used herein, the terms "administer," "administering," "administration," and the like refer to methods that can be used to deliver a drug, such as an anti-TL1A antibody or an antigen-binding fragment thereof, to a desired site of biological action (e.g., intravenous administration). Administration techniques that can be used with the agents and methods described herein are described, for example, in Goodman and Gilman, The Pharmacological Basis of Therapeutics, current edition, Pergamon; and Remington's, Pharmaceutical Sciences, current edition, Mack Publishing Co., Easton, Pa. and Matucci, A. et al., Respiratory Research, 19(1):154 (2018).

[0059] As used herein, the term "combination" or "administered in combination" means that an antibody or antigen-binding fragment thereof described herein can be administered with one or more additional therapeutic agents. In some aspects of the combinations provided herein, the antibody or antigen-binding fragment thereof can be administered simultaneously or sequentially with the one or more additional therapeutic agents. In some aspects, the antibody or antigen-binding fragment thereof described herein can be administered with one or more additional therapeutic agents in the same composition or in different compositions.

[0060] The terms "subject" and "patient" are used interchangeably and include any animal. In some embodiments, the subject is a mammal, including companion animals (e.g., cats, dogs) and farm mammals (e.g., pigs, horses, cows), as well as mice, rabbits, and rats, guinea pigs, and other rodents. In some embodiments, the subject is a non-human primate, such as a cynomolgus monkey. In some embodiments, the subject is a human.

[0061] A "therapeutically effective amount" refers to an amount of an agent, such as an anti-TL1A antibody (e.g., TEV-48574) or an antigen-binding fragment thereof, that is effective in treating a disease or disorder of interest. Terms such as "treating," "treatment," "treat," "alleviating," and "alleviating" refer to utilizing an approach to obtain such beneficial or desired clinical results, including, but not limited to, approaches that achieve such beneficial or desired clinical results, which may include therapeutic measures that result in improvement, cure, slowing, alleviation of symptoms, and / or halt of progression of a pathological condition or disorder. Accordingly, those in need of treatment include those already diagnosed with a disorder or those suspected of having a disorder.

[0062] In the context of antibody-antigen interactions, "specificity" is not necessarily an absolute designation, but rather a relative term that refers to the degree of selectivity of an antibody for antigen-positive cells compared to antigen-negative cells. The specificity of an antibody for antigen-positive cells is mediated by the antibody's variable region and, typically, its complementarity-determining region (CDR). Constructs may have approximately 100- to 1000-fold greater specificity for antigen-positive cells compared to antigen-negative cells.

[0063] As used herein, the term "recombinant" includes expression from genes produced by genetic engineering or otherwise by laboratory manipulation.

[0064] As used herein, the term "TEV 48574" or "TEV-48574" refers to a highly potent, fully human immunoglobulin G (IgG) subclass 1 (IgG1) (lambda) monoclonal antibody (mAb) that binds with high affinity to human, cynomolgus monkey, and rat TL1A. TEV-48574 comprises a light chain of SEQ ID NO: 9 and a heavy chain of SEQ ID NO: 10. TEV-48574 is a blocking antibody that acts by competitively inhibiting the interaction of TL1A with its cognate signaling receptor, DR3. By competitively inhibiting the binding of TL1A to DR3, this antibody prevents activation of the DR3 signaling pathway. Although TEV-48574 inhibits the interaction of TL1A with DR3 more than the interaction of TL1A with DcR3, TEV-48574 also inhibits the binding of TL1A to DcR3. TEV-48574 has demonstrated anti-inflammatory and anti-fibrotic effects in animal models of colitis. TEV-48574 was safe and well tolerated in the Phase 1 study TV48574-SAD-10126. TEV-48574 is disclosed in U.S. Patent No. 10,138,296 as "320-587," the entire contents of which are incorporated herein by reference.

[0065] As used herein, the term "TL1A" refers to tumor necrosis factor (TNF)-like ligand 1A, also known as TNF superfamily member 15 (TNFSF15) and vascular endothelial growth inhibitory factor (VEGI), a member of the TNF superfamily expressed by antigen-presenting cells (including dendritic cells, B cells, and macrophages), CD4+ and CD8+ T cells, and endothelial cells. TL1A is expressed on the cell surface or secreted as a soluble cytokine. Its cognate signaling receptor, death receptor 3 (DR3), is expressed by a variety of cells, including CD4+ and CD8+ T cells, NK cells, NKT cells, FOXP3+ regulatory T (Treg) cells, and type 2 and type 3 innate lymphoid cells (ILC2s and ILC3s). TL1A can also bind to decoy receptors (DcR3s), which competitively inhibit DR3. DcR3 also serves as a decoy receptor for Fas-ligand (Fas-L) and lymphotoxin-like inducer protein, and competes with glycoprotein D for binding to herpesvirus entry mediator (LIGHT) on T cells. As such, DcR3 is a key regulator of several signaling pathways.

[0066] As used in this disclosure and claims, the singular forms "a," "an," and "the" include the plural forms unless the context clearly dictates otherwise.

[0067] Whenever aspects of the present disclosure are described herein using the term "comprising," it is understood that other similar embodiments described using the terms "consisting of" and / or "consisting essentially of" are also provided.

[0068] Unless otherwise specified or clear from the context, as used herein, the term "or" is understood to be inclusive. When used herein in phrases such as "A and / or B," the term "and / or" is intended to include both "A and B," "A or B," "A" and "B." Similarly, when used in phrases such as "A, B and / or C," the term "and / or" is intended to include each of the following aspects: A, B and C; A, B or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone).

[0069] As used herein, the terms "about" and "approximately," when used to modify a numerical value or numerical range, indicate that a deviation of ±10% of the numerical value or numerical range remains within the intended meaning of the recited value or range. As will be understood by those skilled in the art, reference herein to a value or range as "about" includes (and describes) the case itself directed to that value or range. For example, a reference to "about X" includes the description of "X."

[0070] Any composition or method provided herein can be combined with one or more of any of the other compositions and methods provided herein.

[0071] Units, prefixes, and symbols are shown in their International System of Units (SI) recognized form. Numerical ranges are inclusive of the numbers defining the range. The headings provided herein are not intended to limit the various aspects of this disclosure, but may be had by reference to the specification as a whole. As such, the terms defined immediately below are more fully defined by reference to the specification as a whole.

[0072] Pharmaceutical Composition The present disclosure provides pharmaceutical compositions comprising an antibody or antigen-binding fragment that binds to TL1A. The pharmaceutical composition may comprise any of the antibodies or antigen-binding fragments thereof described and / or exemplified herein and an acceptable carrier, such as a pharmaceutically acceptable carrier. Suitable carriers include any medium that does not interfere with the biological activity of the antibody or antigen-binding fragment thereof and is not toxic to the host to which it is administered. The pharmaceutical composition may be formulated for administration to a subject in any suitable dosage form.

[0073] Pharmaceutical compositions suitable for administration to human patients are typically formulated, for example, with a liquid carrier for parenteral administration, or are suitable for reconstitution into a liquid solution or suspension for intravenous or subcutaneous administration.

[0074] In general, such compositions typically include a pharmaceutically acceptable carrier. As used herein, the term "pharmaceutically acceptable" means approved by a government regulatory agency for use in animals, particularly humans, or listed in the United States Pharmacopoeia or other generally recognized pharmacopeia. The term "carrier" refers to a diluent, adjuvant, excipient, or vehicle with which a compound is administered. Such pharmaceutical carriers may be sterile liquids. Water or aqueous solutions such as saline, dextrose, and glycerol solutions may be employed as carriers, particularly for injectable solutions. Liquid compositions for parenteral administration may be formulated for administration by injection or continuous infusion. Routes of administration by injection or infusion include intravenous and subcutaneous.

[0075] In some embodiments, the pharmaceutical composition comprises: (a) about 100 mg / mL of an antibody or antigen-binding fragment thereof that specifically binds to TNF-like ligand 1A (TL1A), the antibody or antigen-binding fragment thereof comprising: (i) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 7 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 8; (ii) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 1; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 2; a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 3; and a light chain variable region CDR4 comprising the amino acid sequence of SEQ ID NO: 4. (b) about 10 mM histidine; (c) about 100 mM arginine hydrochloride (Arg-HCl); (d) about 5% (w / v) sucrose; and (e) about 0.02% (w / v) polysorbate-80. In some embodiments, the pharmaceutical formulation is lyophilized. In some embodiments, the pharmaceutical formulation is liquid.

[0076] In some embodiments, the pharmaceutical formulation comprises: (a) about 150 mg / mL of an antibody or antigen-binding fragment thereof that specifically binds to TNF-like ligand 1A (TL1A), the antibody or antigen-binding fragment thereof comprising: (i) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 7 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 8; (ii) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 1; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 2; a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 3; and a light chain variable region CDR4 comprising the amino acid sequence of SEQ ID NO: 4. (b) about 10 mM histidine; (c) about 100 mM arginine hydrochloride (Arg-HCl); (d) about 5% (w / v) sucrose; and (e) about 0.02% (w / v) polysorbate-80. In some embodiments, the pharmaceutical formulation is lyophilized. In some embodiments, the pharmaceutical formulation is liquid.

[0077] In some embodiments, the pharmaceutical formulation comprises: (a) about 100 mg / mL of an antibody or antigen-binding fragment thereof that specifically binds to TNF-like ligand 1A (TL1A), the antibody or antigen-binding fragment thereof comprising: (i) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 7 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 8; (ii) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 1; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 2; a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 3; and a light chain variable region CDR4 comprising the amino acid sequence of SEQ ID NO: 4. (b) about 10 mM histidine; (c) about 100 mM arginine hydrochloride (Arg-HCl); (d) about 5% (w / v) sucrose; and (e) about 0.02% (w / v) polysorbate-80.

[0078] In some embodiments, the pharmaceutical formulation comprises: (a) about 150 mg / mL of an antibody or antigen-binding fragment thereof that specifically binds to TNF-like ligand 1A (TL1A), the antibody or antigen-binding fragment thereof comprising: (i) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 7 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 8; (ii) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 1; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 2; a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 3; and a light chain variable region CDR4 comprising the amino acid sequence of SEQ ID NO: 4. (b) about 10 mM histidine; (c) about 100 mM arginine hydrochloride (Arg-HCl); (d) about 5% (w / v) sucrose; and (e) about 0.02% (w / v) polysorbate-80.

[0079] In some embodiments, the pharmaceutical formulations provided herein have a pH of 6.0±0.5 after storage at room temperature for 24 hours or at 2-8°C for 24 hours, 72 hours, or 10 days. In some embodiments, the pharmaceutical formulations provided herein have an osmolality of 200 mOsm / kg to 500 mOsm / kg after storage at room temperature for 24 hours or at 2-8°C for 24 hours, 72 hours, or 10 days. In some embodiments, the pharmaceutical formulations provided herein have an antibody monomer content of at least 99% after storage at 2-8°C for 24 hours, 72 hours, or 10 days. In some embodiments, the pharmaceutical formulations provided herein do not have a significant change in charge heterogeneity profile after storage at 2-8°C for 24 hours, 72 hours, or 10 days. In some embodiments, the pharmaceutical formulations provided herein do not have a significant change in purity after storage at room temperature for 24 hours or at 2-8°C for 24 hours, 72 hours, or 10 days. In some embodiments, the pharmaceutical formulations provided herein have a purity of at least 90% after storage at room temperature for 24 hours or at 2-8°C for 24 hours, 72 hours, or 10 days. In some embodiments, the pharmaceutical formulations provided herein have no significant change in particle concentration after storage at room temperature for 24 hours or at 2-8°C for 24 hours, 72 hours, or 10 days. In some embodiments, the pharmaceutical formulations provided herein have no significant difference in visual appearance after storage at 2-8°C for up to 24 months. In some embodiments, the pharmaceutical formulations provided herein have no significant difference in visual appearance after storage at 2-8°C for up to 36 months. In some embodiments, the pharmaceutical formulations provided herein have no significant difference in protein concentration, osmolality, or viscosity after storage at 2-8°C, 25°C, or 40°C for up to 24 months. In some embodiments, the pharmaceutical formulations provided herein have no significant difference in protein concentration, osmolality, or viscosity after storage at 2-8°C, 25°C, or 40°C for up to 36 months. In some embodiments, the pharmaceutical formulations provided herein have a monomer content of 95% or greater, a dimer content of 5.0% or less, or no significant difference in low molecular weight species content after storage at 2-8° C. for up to 24 months.In some embodiments, the pharmaceutical formulations provided herein have a monomer content of 95% or greater, a dimer content of 5.0% or less, or no significant difference in low molecular weight species content after storage for up to 36 months at 2-8°C. In some embodiments, the pharmaceutical formulations provided herein have a purity of 90% or greater after storage for up to 24 months at 2-8°C. In some embodiments, the pharmaceutical formulations provided herein have a purity of 90% or greater after storage for up to 36 months at 2-8°C. In some embodiments, the pharmaceutical formulations provided herein have a major species content of 50%-90%, an acidic species content of 10%-40%, and / or a basic species content of 0%-20% after storage for up to 24 months at 2-8°C. In some embodiments, the pharmaceutical formulations provided herein have a major species content of 50%-90%, an acidic species content of 10%-40%, and / or a basic species content of 0%-20% after storage for up to 36 months at 2-8°C. In some embodiments, the pharmaceutical formulations provided herein have no significant difference in subvisible particle content after storage for up to 24 months at 2-8° C., 25° C., or 40° C. In some embodiments, the pharmaceutical formulations provided herein have no significant difference in subvisible particle content after storage for up to 36 months at 2-8° C., 25° C., or 40° C. In some embodiments, the pharmaceutical formulations disclosed herein have no significant difference in oxidation of methionine 81 and / or methionine 254 of TEV-48574 and / or deamidation of asparagine 317 of TEV-48574 after storage for up to 24 months at 2-8° C., 25° C., or 40° C. In some aspects, the pharmaceutical formulations disclosed herein have no significant difference in oxidation of methionine 81 and / or methionine 254 of TEV-48574 and / or deamidation of asparagine 317 of TEV-48574 after storage for up to 36 months at 2-8° C., 25° C., or 40° C. In some aspects, the pharmaceutical formulations provided herein have a relative potency of 70% to 135% as measured by enzyme-linked immunosorbent assay (ELISA) after storage for up to 24 months at 2-8° C.In some embodiments, the pharmaceutical formulations provided herein have a relative potency of 70% to 135% as measured by enzyme-linked immunosorbent assay (ELISA) after storage at 2-8°C for up to 36 months. In some embodiments, the pharmaceutical formulations provided herein have no significant difference in thermal stability after storage at 2-8°C, 25°C, or 40°C for up to 6 months. In some embodiments, the pharmaceutical formulations provided herein have no significant difference in thermal stability after storage at 2-8°C for up to 36 months. In some embodiments, the pharmaceutical formulations provided herein have no significant difference in secondary and / or tertiary protein structure after storage at 2-8°C, 25°C, or 40°C for up to 3 months. In some embodiments, the pharmaceutical formulations provided herein have no significant difference in secondary protein structure after storage at 2-8°C for up to 36 months. In some embodiments, the pharmaceutical formulations provided herein have no significant difference in polysorbate-80 concentration after storage at 2-8°C for up to 24 months.

[0080] method The present disclosure provides methods of treating a disease in a subject in need thereof, the methods comprising administering to the subject any of the pharmaceutical formulations provided herein or containers provided herein. In some aspects, the disease is a respiratory disease, a gastrointestinal disease, a skin disease, or arthritis.

[0081] In some embodiments, the airway disease is asthma, chronic obstructive pulmonary disease (COPD), pulmonary fibrosis, pulmonary sarcoidosis, allergic rhinitis, or cystic fibrosis.

[0082] In some aspects, the gastrointestinal condition is inflammatory bowel disease, Crohn's disease, colitis, ulcerative colitis, eosinophilic esophagitis, or irritable bowel syndrome.

[0083] In some embodiments, the arthritis is rheumatoid arthritis.

[0084] In some embodiments, the skin disorder is atopic dermatitis, eczema, or scleroderma.

[0085] In some embodiments, the subject is a human subject. In some embodiments, the subject is a non-human primate, such as a cynomolgus monkey. In some embodiments, the subject is a non-human mammal, such as a mouse, rat, guinea pig, cat, pig, rabbit, or dog.

[0086] Anti-TL1A antibody In some embodiments, the formulations provided herein comprise an anti-TL1A antibody or antigen-binding fragment thereof. In some embodiments, the antibody or antigen-binding fragment thereof inhibits the ability of TL1A to interact with DR3, and in some embodiments also interacts with DcR3, further inhibiting signaling induced by the interaction of TL1A with DR3. In some embodiments, the antibody or antigen-binding fragment thereof has enhanced potency relative to antibody 320-179. In some embodiments, the antibody or antigen-binding fragment thereof has enhanced affinity for TL1A compared to antibody 320-179.

[0087] In some aspects, the antibody or antigen-binding fragment thereof disclosed herein comprises a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 2, a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 3, a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 5, and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 6. In some aspects, the antibody or antigen-binding fragment thereof can inhibit the interaction of TL1A and DR3.

[0088] In some embodiments, the antibodies or antigen-binding fragments thereof disclosed herein comprise a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 7. In some embodiments, the antibodies or antigen-binding fragments thereof comprise a light chain variable region comprising the amino acid sequence of SEQ ID NO: 8. In some embodiments, the antibodies or antigen-binding fragments thereof comprise a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 7 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 8.

[0089] In some embodiments, the heavy chain variable region is appended to a heavy chain constant region. In some embodiments, the heavy chain constant region is an IgG1, IgG2, or IgG4 heavy chain constant region. In some embodiments, the heavy chain variable region of SEQ ID NO: 7 is appended to a human IgG1(ΔK) heavy chain constant region (e.g., SEQ ID NO: 14) such that the heavy chain comprises SEQ ID NO: 9. In some embodiments, the heavy chain constant regions provided herein do not include a C-terminal lysine. In some embodiments, the light chain variable region of SEQ ID NO: 8 is appended to a lambda human light chain constant region (e.g., SEQ ID NO: 29) such that the light chain comprises SEQ ID NO: 10.

[0090] In some aspects, the antibodies or antigen-binding fragments thereof disclosed herein are monoclonal antibodies or antigen-binding fragments thereof.

[0091] In some aspects, the antibodies or antigen-binding fragments thereof disclosed herein are recombinant antibodies or antigen-binding fragments thereof. In some aspects, the recombinant antibodies are full-length antibodies. In some aspects, the recombinant antibodies are monoclonal antibodies.

[0092] In some embodiments, the antibodies or antigen-binding fragments thereof disclosed herein bind to TL1A with enhanced affinity relative to a 320-179 anti-TL1A antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 11 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 12. In some embodiments, the antibodies or antigen-binding fragments thereof disclosed herein have enhanced potency relative to a 320-179 anti-TL1A antibody. The enhanced potency may be at least about 10-fold, at least about 12-fold, at least about 13-fold, at least about 15-fold, at least about 20-fold, at least about 25-fold, at least about 27-fold, at least about 40-fold, about 10-fold to about 40-fold, and may be about 12-fold to about 40-fold, about 13-fold to about 40-fold, about 15-fold to about 40-fold, about 20-fold to about 40-fold, about 25-fold to about 40-fold, or about 27-fold to about 40-fold relative to a 320-179 anti-TL1A antibody. The fold increase in potency can be measured by a TL1A-induced caspase activity assay using TF-1 cells. See, e.g., U.S. Patent No. 10,138,296. The 320-179 antibody has favorable biophysical properties, potently inhibits TL1A, and has a predicted low immunogenicity profile. U.S. Patent No. 10,138,296 and U.S. Patent Application Publication No. 2014 / 0255302 are incorporated herein in their entireties.

[0093] In some embodiments, the antibody disclosed herein is TEV-48574, which is also referred to as the 320-587 antibody in U.S. Pat. No. 10,138,296 (in which the VH is SEQ ID NO: 3 and the VL is SEQ ID NO: 4).

[0094] In some aspects, the antibodies or antigen-binding fragments thereof disclosed herein comprise a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:7 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:8, and bind to TL1A with enhanced affinity relative to the anti-TL1A 320-179 antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:11 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:12. The anti-TL1A 320-179 antibody was previously described in U.S. Patent No. 10,138,296 (in which the VH is SEQ ID NO:1 and the VL is SEQ ID NO:2) and U.S. Patent Application Publication No. 2014 / 0255302 (in which the VH is SEQ ID NO:186 and the VL is SEQ ID NO:199). The 320-179 antibody had favorable biophysical properties, potently inhibited TL1A, and had a predicted low immunogenicity profile. U.S. Patent No. 10,138,296 and U.S. Patent Application Publication No. 2014 / 0255302 are incorporated herein in their entireties.

[0095] In some embodiments, the antibodies or antigen-binding fragments thereof disclosed herein comprise a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:7 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:8, and have enhanced potency relative to the 320-179 anti-TL1A antibody. The enhanced potency can be at least about 10-fold, at least about 12-fold, at least about 13-fold, at least about 15-fold, at least about 20-fold, at least about 25-fold, at least about 27-fold, or at least about 40-fold compared to the 320-179 anti-TL1A antibody. The fold enhancement in potency can be measured according to a TL1A-induced caspase potency assay using TF-1 human erythroleukemia cells (ATCC). See, e.g., U.S. Patent No. 10,138,296.

[0096] In some aspects, the antibodies or antigen-binding fragments thereof disclosed herein are full-length antibodies. In some aspects, the antibodies disclosed herein are monoclonal antibodies.

[0097] In some embodiments, the antibodies or antigen-binding fragments thereof disclosed herein comprise a human IgG1 heavy chain constant region, a human IgG2 heavy chain constant region, a human IgG4 heavy chain constant region, or an allotype thereof. The human IgG1 heavy chain constant region may be selected from among human IgG1 (SEQ ID NO: 13), human IgG1(ΔK) (SEQ ID NO: 14), human IgG1 252Y / 254T / 256E (SEQ ID NO: 15), human IgG1 252Y / 254T / 256E(ΔK) (SEQ ID NO: 16), human IgG1 L234A / L235A / G237A (SEQ ID NO: 16), human IgG1 L234A / L235A / G237A (SEQ ID NO: 17), human IgG1 L234A / L235A / G237A(ΔK) (SEQ ID NO: 18), human IgG1 L235A / G237A (SEQ ID NO: 19), and human IgG1 L235A / G237A(ΔK) (SEQ ID NO: 20). The human IgG2 heavy chain constant region may be selected from among human IgG2 with or without ΔK (SEQ ID NO: 21 and SEQ ID NO: 22) and human IgG2 A330S / P331S with or without (ΔK) (SEQ ID NO: 23 and SEQ ID NO: 24). The human IgG4 heavy chain constant region may be selected from among human IgG4 S228P (SEQ ID NO: 25), human IgG4 S228P(ΔK) (SEQ ID NO: 26), human IgG4 228P / 252Y / 254T / 256E (SEQ ID NO: 27) and human IgG4 228P / 252Y / 254T / 256E(ΔK) (SEQ ID NO: 28). It will be understood that the IgG4 heavy chain may be used without the stabilizing substitution S228P (e.g., IgG4 with YTE alone, IgG4 with YTE and ΔK, or IgG4 with ΔK alone).

[0098] In some embodiments, the antibodies or antigen-binding fragments thereof disclosed herein comprise a human lambda light chain constant region or an allotype thereof. The human light chain lambda constant region may comprise SEQ ID NO:29.

[0099] In some embodiments, the antibodies or antigen-binding fragments thereof disclosed herein bind to human TL1A and can bind to one or more of cynomolgus monkey TL1A, mouse TL1A, rat TL1A, guinea pig TL1A, feline TL1A, dog TL1A, pig TL1A, or rabbit TL1A. In some embodiments, the antibodies or antigen-binding fragments thereof can bind to TL1A from multiple different species, such as when a shared epitope is present. In some embodiments, human TL1A comprises the amino acid sequence of SEQ ID NO: 34, SEQ ID NO: 35, or SEQ ID NO: 36. In some embodiments, cynomolgus monkey TL1A comprises the amino acid sequence of SEQ ID NO: 37. In some embodiments, mouse TL1A comprises the amino acid sequence of SEQ ID NO: 38. In some embodiments, rat TL1A comprises the amino acid sequence of SEQ ID NO: 39. In some embodiments, guinea pig TL1A comprises the amino acid sequence of SEQ ID NO: 40. In some embodiments, cat TL1A comprises the amino acid sequence of SEQ ID NO: 41. In some embodiments, pig TL1A comprises the amino acid sequence of SEQ ID NO: 42. In some embodiments, rabbit TL1A comprises the amino acid sequence of SEQ ID NO: 43. In some embodiments, canine TL1A comprises the amino acid sequence of SEQ ID NO: 44.

[0100] The antibodies or antigen-binding fragments thereof disclosed herein have an equilibrium dissociation constant (K D ) which can be measured according to an equilibrium exclusion method such as the KINEXA® assay (Sapidyne Instruments Inc., Boise, ID). D In some embodiments, the K of TL1A binding measured by equilibrium exclusion method is less than about 1000 pM. D In some embodiments, the K of TL1A binding measured by equilibrium exclusion method is less than about 500 pM, or less than about 400 pM, or less than about 300 pM, or less than about 200 pM. D is less than about 100 pM.

[0101] K of TL1A binding measured by equilibrium exclusion methodD The K of TL1A binding measured by the equilibrium exclusion method can be about 10 pM to about 100 pM. D The K of TL1A binding measured by the equilibrium exclusion method can be about 25 pM to about 75 pM. D The K of TL1A binding measured by the equilibrium exclusion method can be about 30 pM to about 60 pM. D The K of TL1A binding measured by the equilibrium exclusion method can be about 30 pM to about 50 pM. D The K of TL1A binding measured by the equilibrium exclusion method can be about 35 pM to about 50 pM. D The K of TL1A binding measured by the equilibrium exclusion method can be about 36 pM to about 46 pM. D The K of TL1A binding measured by the equilibrium exclusion method can be about 38 pM to about 44 pM. D The K of TL1A binding measured by the equilibrium exclusion method can be about 39 pM to about 43 pM. D The K of TL1A binding measured by the equilibrium exclusion method can be about 40 pM to about 45 pM. D The K of TL1A binding measured by the equilibrium exclusion method can be about 35 pM to about 42 pM. D The K of TL1A binding measured by the equilibrium exclusion method can be approximately 40 pM. D The K of TL1A binding measured by the equilibrium exclusion method can be approximately 41 pM. D can be about 42 pM. Equilibrium exclusion methods can use antibody molecules or TL1A molecules as constant binding partners and other molecules as titrants.

[0102] The antibodies or antigen-binding fragments thereof disclosed herein can bind to TL1A-positive cells. The antibodies or antigen-binding fragments thereof disclosed herein have an EC of less than about 100 nM, less than about 75 nM, less than about 50 nM, less than about 30 nM, less than about 25 nM, less than about 20 nM, less than about 18 nM, less than about 15 nM, less than about 13 nM, or less than about 10 nM. 50 It can bind to TL1A-positive cells at low levels.

[0103] The antibodies or antigen-binding fragments thereof disclosed herein may be monoclonal. In some embodiments, the antibodies or antigen-binding fragments thereof disclosed herein are full-length antibodies comprising two heavy chains and two light chains. In some embodiments, the antibodies or antigen-binding fragments thereof disclosed herein include derivatives or fragments or portions of antibodies that retain the antigen-binding specificity and most or all of the affinity of the full-length antibody. For example, the derivative may comprise at least one variable region (either a heavy chain variable region or a light chain variable region). The derivative may comprise at least two variable regions, for example, at least one heavy chain variable region and at least one light chain variable region. Other examples of suitable antibody derivatives and fragments include, but are not limited to, antibodies with multiepitopic specificity, bispecific antibodies, multispecific antibodies, diabodies, single-chain molecules, FAb, F(Ab'), Fd, Fabc, Fv molecules, single-chain (Sc) antibodies, single-chain Fv antibodies (scFv), individual antibody light chains, individual antibody heavy chains, fusions of antibody chains with other molecules, heavy chain monomers or dimers, light chain monomers or dimers, dimers consisting of one heavy chain and one light chain, and other multimers. Single-chain Fv antibodies may be multivalent. All antibody isotypes can be used to generate antibody derivatives, fragments, and portions thereof. Antibody derivatives, fragments, and / or portions thereof can be recombinantly produced and expressed by any cell type, prokaryotic or eukaryotic.

[0104] In a full-length antibody, each heavy chain is composed of a heavy chain variable region (abbreviated herein as HCVR or VH) and a heavy chain constant region. The heavy chain constant region is composed of three domains: CH1, CH2, and CH3. Each light chain is composed of a light chain variable region (abbreviated herein as LCVR or VL) and a light chain constant region. The light chain constant region is composed of one domain, CL. The VH and VL regions are further subdivided into regions of hypervariability termed complementarity-determining regions (CDRs), which may be interspersed with more conserved regions termed framework regions (FRs). Each VH and VL is composed of three CDRs and four FRs arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. Typically, the antigen-binding properties of an antibody are less affected by changes in the FR sequences than by changes in the CDR sequences. Immunoglobulin molecules can be of any type (e.g., IgG, IgE, IgM, IgD, IgA, and IgY), class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2), or subclass.

[0105] In some aspects, the antibodies or antigen-binding fragments thereof disclosed herein are fully human. A fully human antibody is one in which the entire molecule is of human or otherwise human origin or contains an amino acid sequence identical to a human form of an antibody. Fully human antibodies include, for example, antibodies obtained from human V gene libraries in which human genes encoding antibody variable regions are recombinantly expressed. Fully human antibodies can be expressed in cells from other organisms (e.g., mice and xenomouse technology) or transformed with genes encoding human antibodies. Nevertheless, fully human antibodies may contain amino acid residues not encoded by human sequences, e.g., mutations introduced by random or site-directed mutagenesis.

[0106] In some embodiments, the antibodies or antigen-binding fragments thereof disclosed herein comprise a protein framework of non-immunoglobulin origin. For example, Ku & Schutz, Proc. Natl. Acad. Sci. USA, 92:6552-6 (1995), describe the four-helix bundle protein cytochrome b562, which has two loops randomized to create CDRs selected for antigen binding.

[0107] In some aspects, the antibodies or antigen-binding fragments thereof disclosed herein comprise post-translational modifications or moieties that can affect antibody activity or stability. These modifications or moieties include, but are not limited to, methylated, acetylated, glycosylated, sulfated, phosphorylated, carboxylated, amidated, and other moieties known in the art. Each moiety includes any chemical group or combination of groups typically found on immunoglobulin molecules in nature or otherwise added to antibodies by recombinant expression systems, including prokaryotic and eukaryotic expression systems.

[0108] Examples of side chain modifications contemplated by the present disclosure include modification of amino groups by reductive alkylation by reaction with an aldehyde followed by reduction with NaBH4; amidation with methyl acetimidate; acylation with acetic anhydride; carbamoylation of amino groups with cyanate; trinitrobenzylation of amino groups with 2,4,6-trinitrobenzenesulfonic acid (TNBS); acylation of amino groups with succinic anhydride and tetrahydrophthalic anhydride; pyridoxylation of lysine with pyridoxal-5-phosphate followed by reduction with NaBH4, and the like.

[0109] The guanidine group of arginine residues can be modified by the formation of heterocyclic condensation products with reagents such as 2,3-butanedione, phenylglyoxal, and glyoxal. Carboxyl groups can be modified by carbodiimide activation via O-acylisourea formation, followed by derivatization to the corresponding amide, for example. Sulfhydryl groups can be modified by carboxymethylation with iodoacetic acid or iodoacetamide; performic acid oxidation to cysteic acid; mixed disulfide formation with other thiol compounds; reaction with maleimide, maleic anhydride, or other substituted maleimides; formation of mercury derivatives with 4-chloromercuribenzoate, 4-chloromercuriphenylsulfonic acid, phenylmercuric chloride, 2-chloromercuri-4-nitrophenol, and other mercuric acids; and carbamoylation with cyanate at alkaline pH. Tryptophan residues may be modified by, for example, oxidation with N-bromosuccinimide or alkylation of the indole ring with 2-hydroxy-5-nitrobenzyl bromide or sulfenyl halides, while tyrosine residues can be nitrated with tetranitromethane to form 3-nitrotyrosine derivatives. Modification of the imidazole ring of a histidine residue can be accomplished by alkylation with iodoacetic acid derivatives or N-carbethoxylation with diethylpyrocarbonate.

[0110] In some aspects, the antibodies or antigen-binding fragments thereof disclosed herein comprise one or more modifications that modify serum half-life and biodistribution, including, but not limited to, modifications that modify the antibody's interaction with the neonatal Fc receptor (FcRn), a receptor that protects IgG from catabolism and plays an important role in maintaining high serum antibody concentrations. Modifications that modulate serum half-life include triple substitutions of M252Y / S254T / T256E (EU numbering system (Edelman, G. et al., J. Immunol. 2009; 2010; 2011; 2012; 2013; 2014; 2015; 2016; 2017; 2019; 2020; 2021; 2022; 2023; 2024; 2025; 2026; 2027; 203; 203; 203; 204; 204; 205; 206; 207; 208; 210; 211; 212; 213; 214; 215; 216; 217; 218; 219; 220; 221; 222; 223; 224; 225; 226; 227; 228; 229; 230; 231; 232; 233; 234; 235; 236; 237; 238; 239; 240; 241; 242; 243; 244; 245; 246; 247; 248; 249; 250; 251; 252; 253; Substitutions can occur in the Fc region of IgG1, IgG2, or IgG4, including substitutions of the "YTE" substitution according to the numbering system of [their] and [their] numbers (see, e.g., U.S. Pat. No. 7,217,797), as well as at positions 307, 380, and 434 (see, e.g., WO 00 / 042072). Substitutions can affect Fc receptor binding and subsequent function by these receptors, including FcRn binding and serum half-life. Examples of amino acid substitutions in the constant domain that modulate functions mediated by the constant domain are described in U.S. Patent Application Publication Nos. 2009 / 0142340, 2009 / 0068175, and 2009 / 0092599. In either class of antibody, the C-terminal lysine of the heavy chain can be omitted or removed to reduce heterogeneity (ΔK). The S228P (EU numbering) substitution in human IgG4 can stabilize antibody Fab-arm exchange in vivo (Labrin et al., Nature Biotechnology 27(8):767-73 (2009)), and this substitution can occur simultaneously with YTE and / or ΔK modifications.

[0111] The antibodies or antigen-binding fragments thereof disclosed herein can be labeled, coupled, or conjugated to any chemical or biomolecular moiety. Labeled antibodies can be used for therapeutic, diagnostic, and basic research applications. Such labels / conjugates can be detectable, for example, fluorescent dyes, electrochemiluminescent probes, quantum dots, radioactive labels, enzymes, fluorescent proteins, luminescent proteins, biotin, and the like.

[0112] The antibodies or antigen-binding fragments thereof disclosed herein can be derivatized with known protecting / blocking groups to prevent proteolytic cleavage or to enhance activity or stability.

[0113] Administration of the antibodies or antigen-binding fragments thereof disclosed herein can include administering the antibodies or antigen-binding fragments thereof subcutaneously. Thus, in some aspects, the formulations provided herein are formulated for subcutaneous administration. Administration can include administering the antibodies or antigen-binding fragments thereof intravenously. Thus, in some aspects, the formulations provided herein are formulated for intravenous administration.

[0114] The present disclosure further provides compositions for use according to any of the methods disclosed herein.

[0115] Polynucleotides and Vectors Polynucleotide sequences encoding recombinant antibodies or antigen-binding fragments thereof, as well as subdomains thereof (e.g., FRs and CDRs), are disclosed herein. Polynucleotides include, but are not limited to, RNA, DNA, cDNA, hybrids of RNA and DNA, and single-, double-, or triple-stranded RNA, DNA, or hybrids thereof. Polynucleotides may include nucleic acid sequences encoding the heavy chain variable region and / or light chain variable region of an antibody described or exemplified herein. Complements of the polynucleotide sequences are also within the scope of this disclosure.

[0116] In some embodiments, the polynucleotide may comprise a nucleic acid sequence encoding an antibody heavy chain variable region consisting of the amino acid sequence of SEQ ID NO: 7. The polynucleotide encoding the amino acid sequence of SEQ ID NO: 7 may comprise the nucleic acid sequence of SEQ ID NO: 30 or SEQ ID NO: 31.

[0117] In some embodiments, the polynucleotide may comprise a nucleic acid sequence encoding an antibody light chain variable region comprising the amino acid sequence of SEQ ID NO: 8. The polynucleotide encoding the amino acid sequence of SEQ ID NO: 8 may comprise the nucleic acid sequence of SEQ ID NO: 32 or SEQ ID NO: 33.

[0118] In some embodiments, the polynucleotide may comprise a first nucleic acid sequence encoding an antibody heavy chain variable region and a second nucleic acid sequence encoding an antibody light chain variable region. The first nucleic acid sequence may encode an antibody heavy chain variable region consisting of the amino acid sequence of SEQ ID NO: 7, and the second nucleic acid sequence may encode an antibody light chain variable region consisting of the amino acid sequence of SEQ ID NO: 8. The first nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 7 may comprise the nucleic acid sequence of SEQ ID NO: 30 or SEQ ID NO: 31, and the second nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 8 may comprise the nucleic acid sequence of SEQ ID NO: 32 or SEQ ID NO: 33.

[0119] In some embodiments, the polynucleotide may comprise a first nucleic acid sequence encoding an antibody heavy chain variable region and a second nucleic acid sequence encoding a heavy chain constant region. In some embodiments, the polynucleotide comprises a first nucleic acid sequence encoding an antibody heavy chain variable region comprising the amino acid sequence of SEQ ID NO:7 and a second nucleic acid sequence encoding an IgG1(ΔK) heavy chain constant region of SEQ ID NO:14, e.g., a polynucleotide comprising the nucleic acid sequence of SEQ ID NO:31.

[0120] In some embodiments, the polynucleotide may comprise a first nucleic acid sequence encoding an antibody light chain variable region and a second nucleic acid sequence encoding a light chain constant region. In some embodiments, the polynucleotide comprises a first nucleic acid sequence encoding an antibody light chain variable region comprising the amino acid sequence of SEQ ID NO:8 and a second nucleic acid sequence encoding the lambda light chain constant region of SEQ ID NO:29, e.g., a polynucleotide comprising the nucleic acid sequence of SEQ ID NO:33.

[0121] Any of the polynucleotides described or exemplified herein can be incorporated into a vector. Thus, vectors containing polynucleotides are provided as part of the present disclosure. The vector can be an expression vector. Thus, a recombinant expression vector containing a sequence encoding a polypeptide of interest is provided. The expression vector can contain one or more additional sequences, such as, but not limited to, a regulatory sequence, a selection marker, a purification tag, or a polyadenylation signal. Such regulatory elements include a transcription promoter, an enhancer, an mRNA ribosomal binding site, or a sequence controlling the termination of transcription and translation.

[0122] Expression vectors, particularly mammalian expression vectors, may contain one or more nontranscribed elements such as an origin of replication, a suitable promoter and enhancer linked to the gene to be expressed, other 5' or 3' flanking nontranscribed sequences, 5' or 3' nontranslated sequences (such as necessary ribosome binding sites), a polyadenylation site, splice donor and acceptor sites, or a transcription termination sequence. An origin of replication that confers the ability to replicate in a particular host may also be incorporated.

[0123] Vectors can be used to transform any of a wide range of host cells known to those skilled in the art and capable of expressing antibodies. Vectors include, but are not limited to, plasmids, phagemids, cosmids, bacmids, bacterial artificial chromosomes (BACs), yeast artificial chromosomes (YACs), baculoviruses, and other bacterial, eukaryotic, yeast, and viral vectors. Suitable host cells include, but are not limited to, CHO cells, NS0 cells, HEK293 cells, or any known or developed eukaryotic stable cell line, including bacterial, yeast, and insect cells.

[0124] In some aspects, the antibodies or antigen-binding fragments thereof provided herein were produced in human embryonic kidney cells. In some aspects, the antibodies or antigen-binding fragments thereof provided herein were produced in HEK293 cells.

[0125] Antibodies or antigen-binding fragments thereof may also be produced by hybridoma cells. Methods for producing hybridomas are well known and established in the art.

[0126] Kits and containers The present disclosure also features a container containing any of the pharmaceutical formulations described and exemplified herein. In some embodiments, the container is a glass vial. In some embodiments, the container is a glass vial with a fill volume of 3 mL. In some embodiments, the container is a glass vial with a fill volume of 3 mL or less. In some embodiments, the container is a glass vial with a fill volume of 2 mL or less.

[0127] The present disclosure also features kits including any of the pharmaceutical formulations or antibodies or antigen-binding fragments thereof described and exemplified herein. The kits can be used to provide, among other things, pharmaceutical formulations, antibodies, antigen-binding fragments thereof, and other agents for use in diagnostic, basic research, or therapeutic methods. In some aspects, the kits include one or more of the pharmaceutical formulations, antibodies, or antigen-binding fragments thereof described or exemplified herein, and instructions for using the one or more pharmaceutical formulations, antibodies, or antigen-binding fragments thereof in a method of treating a disease in a subject in need thereof. The method can include administering to a subject any of the pharmaceutical formulations provided herein or any of the kits or containers provided herein. In some aspects, the disease is a respiratory disease, a gastrointestinal disease, a skin disease, or arthritis.

[0128] In some embodiments, the antibody or antigen-binding fragment thereof is formulated in a volume of 3 mL or less. In some embodiments, the antibody or antigen-binding fragment thereof is formulated in a volume of 2 mL or less.

[0129] In some embodiments, the airway disease is asthma, chronic obstructive pulmonary disease (COPD), pulmonary fibrosis, pulmonary sarcoidosis, allergic rhinitis, or cystic fibrosis.

[0130] In some aspects, the gastrointestinal condition is inflammatory bowel disease, Crohn's disease, colitis, ulcerative colitis, eosinophilic esophagitis, or irritable bowel syndrome.

[0131] In some embodiments, the arthritis is rheumatoid arthritis.

[0132] In some embodiments, the skin disorder is atopic dermatitis, eczema, or scleroderma.

[0133] Exemplary embodiments provided herein

[0013] In one embodiment (Embodiment 1, A1) herein, a pharmaceutical formulation is provided, comprising: (a) 150 mg / mL of an antibody or antigen-binding fragment thereof that specifically binds to TNF-like ligand 1A (TL1A), the antibody or antigen-binding fragment thereof comprising: (i) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 7 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 8; (ii) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 2, a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 3, and a light chain variable region CDR4 comprising the amino acid sequence of SEQ ID NO: 4. a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 5, a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 5, and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 6; or (iii) an antibody or antigen-binding fragment thereof comprising 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; (b) 10 mM histidine; (c) 100 mM arginine hydrochloride (Arg-HCl); (d) 5% (w / v) sucrose; and (e) 0.02% (w / v) polysorbate-80.

[0134] In one embodiment of A1, namely A2, the antibody or antigen-binding fragment of (i) or (ii) comprises an IgG1 constant region.

[0135] In one embodiment of A1 or A2, namely A3, the pharmaceutical formulation is lyophilized.

[0136] In one embodiment of A1 or A2, namely A4, the pharmaceutical formulation is a liquid.

[0137] In any one embodiment of A1-A4, ie, A5, the pharmaceutical composition has a pH of 6.0±0.5 after storage at room temperature for 24 hours or at 2-8° C. for 24 hours, 72 hours, or 10 days.

[0138] In any one of A1 to A5, i.e., A6, the pharmaceutical composition has an osmolality of 200 mOsm / kg to 500 mOsm / kg after storage at room temperature for 24 hours or at 2 to 8°C for 24 hours, 72 hours, or 10 days.

[0139] In any one of A1 to A6, ie, A7, the pharmaceutical composition has an antibody monomer content of at least 99% after storage at 2 to 8°C for 24 hours, 72 hours, or 10 days.

[0140] In any one embodiment of A1-A7, ie, A8, the pharmaceutical composition has no significant change in charge heterogeneity profile after storage at 2-8° C. for 24 hours, 72 hours, or 10 days.

[0141] In any one embodiment of A1-A8, ie, A9, the pharmaceutical composition has no significant change in purity after storage at room temperature for 24 hours or at 2-8° C. for 24 hours, 72 hours, or 10 days.

[0142] In any one embodiment of A1-A8, ie, A10, the pharmaceutical composition has a purity of at least 90% after storage at room temperature for 24 hours or at 2-8° C. for 24 hours, 72 hours, or 10 days.

[0143] In any one embodiment of A1-A10, ie, A11, the pharmaceutical composition has no significant change in particle concentration after storage at room temperature for 24 hours or at 2-8°C for 24 hours, 72 hours, or 10 days.

[0144] In any one embodiment of A1-A11, ie, A12, the pharmaceutical composition has no significant difference in visual appearance after storage at 2-8° C. for up to 36 months.

[0145] In any one of A1-A12, i.e., A13, the pharmaceutical composition has no significant difference in protein concentration, osmolality, or viscosity after storage at 2-8°C, 25°C, or 40°C for up to 36 months.

[0146] In any one of A1-A13, i.e., A14, the pharmaceutical composition has a monomer content of 95% or more, a dimer content of 5.0% or less, or no significant difference in low molecular weight species content after storage at 2-8°C for up to 36 months.

[0147] In any one of A1-A14, ie, A15, the pharmaceutical composition has a purity of 90% or greater after storage at 2-8° C. for up to 36 months.

[0148] In any one of A1-A15, i.e., A16, the pharmaceutical composition has a major species content of 50%-90%, an acidic species content of 10%-40%, and / or a basic species content of 0%-20% after storage at 2-8°C for up to 36 months.

[0149] In any one of A1-A16, ie, A17, the pharmaceutical composition has no significant difference in subvisible particle content after storage at 2-8°C, 25°C, or 40°C for up to 36 months.

[0150] In any one of A1-A17, i.e., A18, the pharmaceutical composition has no significant difference in oxidation of methionine 81 and / or methionine 254 of TEV-48574 and / or deamidation of asparagine 317 of TEV-48574 after storage at 2-8°C, 25°C, or 40°C for up to 36 months.

[0151] In any one embodiment of A1-A18, i.e., A19, the pharmaceutical composition has a relative potency of 70% to 135% as measured by enzyme-linked immunosorbent assay (ELISA) after storage at 2-8°C for up to 36 months.

[0152] In any one of A1-A19, ie, A20, the pharmaceutical composition has no significant difference in thermal stability after storage at 2-8°C, 25°C, or 40°C for up to 6 months.

[0153] In any one embodiment of A1-A20, ie, A21, the pharmaceutical composition has no significant difference in thermal stability after storage at 2-8°C for up to 36 months.

[0154] In any one of A1-A21, i.e., A22, the pharmaceutical composition has no significant difference in secondary and / or tertiary protein structure after storage at 2-8°C, 25°C, or 40°C for up to 3 months.

[0155] In any one of A1-A22, ie, A23, the pharmaceutical composition has no significant difference in secondary protein structure after storage at 2-8° C. for up to 36 months.

[0156] In any one embodiment of A1-A23, ie, A24, the pharmaceutical composition has no significant difference in concentration of polysorbate-80 after storage at 2-8° C. for up to 24 months.

[0157] In one embodiment of any one of A1 to A24, ie, A25, the antibody or antigen-binding fragment thereof is produced in Chinese hamster ovary cells.

[0158] In one embodiment, namely A26, there is provided a container comprising the pharmaceutical formulation of any one of A1-A25.

[0159] In one embodiment of A26, ie, A27, the container is a glass vial.

[0160] In one embodiment of A27, ie, A28, the container is a glass vial having a fill volume of 3 mL.

[0161] In one aspect, namely A29, provided herein is a method of treating a disease in a subject in need thereof, the method comprising administering to the subject a pharmaceutical formulation of any one of A1-A25 or a container of any one of A26-A28, optionally wherein the disease is a respiratory disease, a gastrointestinal disease, a skin disease, or arthritis.

[0162] In one embodiment of A29, ie, A30, the airway disease is asthma, chronic obstructive pulmonary disease (COPD), pulmonary fibrosis, pulmonary sarcoidosis, allergic rhinitis, or cystic fibrosis.

[0163] In one embodiment of A29, ie, A31, the gastrointestinal disease is inflammatory bowel disease, Crohn's disease, colitis, ulcerative colitis, eosinophilic esophagitis, or irritable bowel syndrome.

[0164] In one embodiment of A29, ie, A32, the arthritis is rheumatoid arthritis.

[0165] In one embodiment of A29, ie, A33, the skin disease is atopic dermatitis, eczema, or scleroderma.

[0166] In one embodiment of any one of A1 to A25, ie A34, the formulation is for use according to one of the methods of A29 to A33.

[0167] In one embodiment, i.e., B1, a pharmaceutical formulation includes: (a) about 100 mg / mL to about 250 mg / mL of an antibody or antigen-binding fragment thereof that specifically binds to TNF-like ligand 1A (TL1A), wherein the antibody or antigen-binding fragment thereof comprises a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 2, a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 3, a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 4, and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 5. and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 6; (b) about 5 mM to about 15 mM histidine; (c) about 50 mM to about 150 mM arginine hydrochloride (Arg-HCl); (d) about 2.5% (w / v) to about 7.5% (w / v) sucrose; and (e) about 0.01% (w / v) to about 0.03% (w / v) polysorbate-80.

[0168] In one embodiment of B1, namely B2, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:7 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:8.

[0169] In one embodiment of either B1 or B2, i.e., B3, the antibody or antigen-binding fragment comprises an IgG1 constant region.

[0170] In any one embodiment of B1 to B3, ie, B4, the antibody or antigen-binding fragment thereof 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.

[0171] In any one embodiment of B1-B4, ie, B5, the pharmaceutical formulation comprises about 100, about 150, about 200, about 225, or about 250 mg / mL of the antibody or antigen-binding fragment thereof.

[0172] In any one embodiment of B1-B5, ie, B6, the pharmaceutical formulation comprises about 5 mM, about 10 mM, or about 15 mM histidine.

[0173] In any one embodiment of B1-B6, ie, B7, the pharmaceutical formulation comprises about 50 mM, about 100 mM, or about 150 mM arginine hydrochloride (Arg-HCl).

[0174] In any one embodiment of B1-B7, ie, B8, the pharmaceutical formulation comprises about 2.5%, about 5%, or about 7.5% (w / v) sucrose.

[0175] In any one embodiment of B1-B8, ie, B9, the pharmaceutical formulation comprises about 0.01%, about 0.02%, or about 0.03% (w / v) polysorbate-80.

[0176] In any one of B1-B9, ie, B10, the pharmaceutical formulation is lyophilized.

[0177] In one embodiment of any one of B1-B9, ie, B11, the pharmaceutical formulation is a liquid.

[0178] In any one embodiment of B1-B11, ie, B12, the pharmaceutical composition has a pH of 6.0±0.5 after storage at room temperature for 24 hours or at 2-8° C. for 24 hours, 72 hours, or 10 days.

[0179] In any one embodiment of B1 to B12, i.e., B13, the pharmaceutical composition has an osmolality of 200 mOsm / kg to 500 mOsm / kg after storage at room temperature for 24 hours or at 2 to 8°C for 24 hours, 72 hours, or 10 days.

[0180] In any one of B1 to B13, ie, B14, the pharmaceutical composition has an antibody monomer content of at least 99% after storage at 2-8°C for 24 hours, 72 hours, or 10 days.

[0181] In any one embodiment of B1-B14, ie, B15, the pharmaceutical composition has no significant change in charge heterogeneity profile after storage at 2-8° C. for 24 hours, 72 hours, or 10 days.

[0182] In any one embodiment of B1-B15, ie, B16, the pharmaceutical composition has no significant change in purity after storage at room temperature for 24 hours or at 2-8° C. for 24 hours, 72 hours, or 10 days.

[0183] In any one embodiment of B1-B15, ie, B17, the pharmaceutical composition has a purity of at least 90% after storage at room temperature for 24 hours or at 2-8° C. for 24 hours, 72 hours, or 10 days.

[0184] In any one embodiment of B1-B17, ie, B18, the pharmaceutical composition has no significant change in particle concentration after storage at room temperature for 24 hours or at 2-8° C. for 24 hours, 72 hours, or 10 days.

[0185] In any one embodiment of B1-B18, ie, B19, the pharmaceutical composition has no significant difference in visual appearance after storage at 2-8° C. for up to 36 months.

[0186] In any one embodiment of B1-B19, i.e., B20, the pharmaceutical composition has no significant difference in protein concentration, osmolality, or viscosity after storage at 2-8°C, 25°C, or 40°C for up to 36 months.

[0187] In any one embodiment of B1-B20, i.e., B21, the pharmaceutical composition has a monomer content of 95% or more, a dimer content of 5.0% or less, or no significant difference in low molecular weight species content after storage at 2-8°C for up to 36 months.

[0188] In any one of B1 to B21, ie, B22, the pharmaceutical composition has a purity of 90% or greater after storage at 2-8° C. for up to 36 months.

[0189] In any one of embodiments B1 to B22, i.e., B23, the pharmaceutical composition has a major species content of 50% to 90%, an acidic species content of 10% to 40%, and / or a basic species content of 0% to 20%, after storage at 2 to 8°C for up to 36 months.

[0190] In any one embodiment of B1-B23, ie, B24, the pharmaceutical composition has no significant difference in subvisible particle content after storage at 2-8°C, 25°C, or 40°C for up to 36 months.

[0191] In any one of embodiments B1-B24, i.e., B25, the pharmaceutical composition has no significant difference in oxidation of methionine 81 and / or methionine 254 of TEV-48574 and / or deamidation of asparagine 317 of TEV-48574 after storage at 2-8°C, 25°C, or 40°C for up to 36 months.

[0192] In any one embodiment of B1 to B25, i.e., B26, the pharmaceutical composition has a relative potency of 70% to 135% as measured by enzyme-linked immunosorbent assay (ELISA) after storage at 2 to 8°C for up to 36 months.

[0193] In any one embodiment of B1 to B26, ie, B27, the pharmaceutical composition has no significant difference in thermal stability after storage at 2-8°C, 25°C, or 40°C for up to 6 months.

[0194] In any one embodiment of B1-B27, ie, B28, the pharmaceutical composition has no significant difference in thermal stability after storage at 2-8° C. for up to 36 months.

[0195] In any one embodiment of B1-B28, i.e., B29, the pharmaceutical composition has no significant difference in secondary and / or tertiary protein structure after storage at 2-8°C, 25°C, or 40°C for up to 3 months.

[0196] In any one embodiment of B1-B29, ie, B30, the pharmaceutical composition has no significant difference in secondary protein structure after storage at 2-8° C. for up to 36 months.

[0197] In any one embodiment of B1 to B30, ie, B31, the pharmaceutical composition has no significant difference in concentration of polysorbate-80 after storage at 2 to 8° C. for up to 24 months.

[0198] In any one embodiment of B1 to B31, ie, B32, the antibody or antigen-binding fragment thereof is produced in Chinese hamster ovary cells.

[0199] In one embodiment, namely B33, there is provided a container comprising the pharmaceutical formulation of any one of B1 to B32.

[0200] In one embodiment of B33, ie, B34, the container is a glass vial.

[0201] In one embodiment of B34, ie, B35, the container is a glass vial having a fill volume of 3 mL.

[0202] In one embodiment of B33, ie, B36, the container is a syringe, and optionally the syringe is a pre-filled syringe.

[0203] In one aspect, namely B37, provided herein is a method of treating a disease in a subject in need thereof, the method comprising administering to the subject a pharmaceutical formulation of any one of B1-B32 or a container of any one of B33-B36, optionally wherein the disease is a respiratory disease, a gastrointestinal disease, a skin disease, or arthritis.

[0204] In one embodiment of B37, ie, B38, the airway disease is asthma, chronic obstructive pulmonary disease (COPD), pulmonary fibrosis, pulmonary sarcoidosis, allergic rhinitis, or cystic fibrosis.

[0205] In one embodiment of B37, ie, B39, the gastrointestinal disease is inflammatory bowel disease, Crohn's disease, colitis, ulcerative colitis, eosinophilic esophagitis, or irritable bowel syndrome.

[0206] In one embodiment of B37, ie, B40, the arthritis is rheumatoid arthritis.

[0207] In one embodiment of B37, ie, B41, the skin disease is atopic dermatitis, eczema, or scleroderma.

[0208] In one embodiment of B37-B41, ie, B42, the pharmaceutical preparation is administered intravenously. In one embodiment of B37-B41, ie, B43, the pharmaceutical preparation is administered subcutaneously.

[0209] In an embodiment of any one of B1-B32 (ie, B44), the formulation is for use according to any one of the methods of B37-B43.

[0210] The following examples are offered by way of illustration and not by way of limitation. [Example]

[0211] Example 1: Stability of reconstituted TEV-48574 drug product solution The in-use stability of the TEV-48574 reconstituted drug product was evaluated.

[0212] In this study, TEV-48574 was lyophilized to a protein concentration of 150 mg / mL in 10 mM histidine, 5% (w / v) sucrose, 100 mM arginine hydrochloride (Arg-HCl), 0.02% (w / v) polysorbate-80 (PS-80), pH 6.0. The lyophilized formulation was packaged in 5 cc vials with a 20 mm neck. The vials were stored at 2-8 °C and allowed to warm to room temperature before reconstitution. Approximately seven vials were reconstituted with 2.0 mL of sterile water for injection (WFI) per vial to form a stock solution. This stock solution was then diluted in different Falcon tubes to concentrations of 50, 20, and 5 mg / mL with the formulation buffer: 10 mM histidine, 5% (w / v) sucrose, 100 mM Arg-HCl, 0.02% (w / v) PS-80, pH 6.0. After dilution, 4 mL samples were incubated at 2-8°C under light protection for 24 hours, 72 hours, 10 days, and under normal light conditions at room temperature for 24 hours. Table 1 shows the specific time points and conditions tested.

[0213] [Table 2]

[0214] At the end of each time point and condition, samples were analyzed for visual appearance, pH, osmolality, protein concentration, and subvisible particles using size exclusion chromatography (SEC), capillary sodium dodecyl sulfate gel electrophoresis (cSDS), capillary isoelectric focusing (cIEF), and microflow imaging (MFI).

[0215] As shown in Table 2, no significant differences were observed in visual appearance, pH, osmolality, or protein concentration in the reconstituted samples after dilution. The pH and osmolality of the samples were 6.0 ± 0.5 pH, respectively, and were also within the range of 200–500 mOsm / kg.

[0216] [Table 3]

[0217] SEC analysis showed that the antibody monomer concentration in the samples did not change significantly over the stability period (see Table 3). These results indicate that storage of the diluted reconstituted drug product at 2-8°C for up to 10 days does not affect the percentage of monomer at the time of incubation.

[0218] [Table 4]

[0219] cIEF analysis showed that dilution of the reconstituted drug product and incubation at 2-8°C for up to 10 days did not significantly affect the charge heterogeneity profile of TEV-48574. See Table 4.

[0220] [Table 5]

[0221] As shown in Table 5, cSDS analysis revealed that the purity of the reconstituted drug product was greater than 90%, and dilution of the reconstituted drug product and incubation under various conditions did not significantly affect the purity.

[0222] [Table 6]

[0223] The 50 mg / mL sample contained slightly more subvisible particles according to the MFI analysis (see Table 6). However, when comparing the first time point sample with the later time point samples, no significant increase in particle concentration was observed overall for the different concentration samples.

[0224] [Table 7]

[0225] Taken together, these results indicate that the reconstituted TEV-48574 drug product is stable for up to 10 days when diluted and stored at 2-8°C, protected from light, and under normal light conditions for 24 hours at room temperature.

[0226] Example 2: Stability of TEV-48574 Liquid Formulations at 100 mg / ml and 150 mg / ml The objective of this study was to evaluate the long-term stability of liquid and lyophilized TEV-48574 formulations. Three formulations were tested: a lyophilized drug product containing 100 mg / mL of drug product in 10 mM histidine, 5% (w / v) sucrose, 100 mM Arg-HCl, 0.02% (w / v) PS-80, pH 6.0 (Formulation 1), and two liquid formulations containing either 100 mg / mL or 150 mg / mL of drug product in 10 mM histidine, 5% (w / v) sucrose, 100 mM Arg-HCl, 0.02% (w / v) PS-80, pH 6.0 (Formulations 2 and 3, respectively). The stability of these formulations was evaluated in 5 cc vials (Type I glass) under the following conditions: "standard" conditions (2-8°C), "accelerated" conditions (25±2°C / 60±5% relative humidity (RH)), and "stress" conditions (40±2°C / 75±5% RH). Additionally, the stability of Formulation 3 was evaluated under the same conditions, except the formulation was filled into 2.25 mL Nipro prefilled syringes (Nipro PFS) fitted with waist plunger stoppers. The effect of these conditions on several product quality attributes was tested for each formulation.

[0227] Visual appearance, protein concentration, osmolality and viscosity The results of the visual appearance, protein concentration, and osmolality analyses of Formulations 1–3 are shown in Tables 7–10. No significant differences were observed between the formulations at standard storage conditions of 2–8°C. Few visible particles were observed at intermittent time points. However, this may be related to the developmental testing in which the drug product was manually filled into vials. Furthermore, these particles were not considered product-related because they did not increase over time and were not consistently detected in all samples at different time points. Regarding visual appearance under stress conditions at 40°C, the solution appeared slightly yellow at later time points. No significant differences were observed in visual appearance, protein concentration, osmolality, or viscosity for Formulation 3 filled in Nipro PFS compared to Formulation 3 stored in glass vials for up to 24 months (data not shown), indicating that these attributes of the formulations are not affected by contact with Nipro PFS.

[0228] Protein concentrations measured under standard (2-8°C), accelerated (25°C), and stressed (40°C) conditions were close to the expected nominal concentrations for all three formulations and did not change over time. Osmolality and viscosity measured under standard storage conditions at time zero (T0), 24 months (24M), and 36 months (36M) showed similar trends and no significant differences over time.

[0229] [Table 8]

[0230] [Table 9]

[0231] [Table 10]

[0232] [Table 11]

[0233] Size Exclusion Chromatography (SEC) Tables 11-13 and Figures 1A-1C, 2A-2C, and 3A-3C show the percentages (%) of monomer, dimer, and low molecular weight species for formulations 1, 2, and 3, as measured by SEC. All three formulations stored at standard storage conditions of 2-8°C met the acceptance criteria for monomer and dimer content for up to 24 and 36 months, respectively. Compared to formulation 1, formulations 2 and 3 exhibited an increase in the percentage of low molecular weight species, while a slight decrease in the percentage of monomer was observed. However, this is an expected observation given that lyophilized formulations are typically more stable than liquid formulations. Under accelerated and stress conditions, protein fragmentation was more pronounced, indicating that the primary degradation pathway for liquid formulations is the formation of low molecular weight species. The protein degradation rates for formulations 2 and 3 were similar, indicating that protein concentration does not significantly affect degradation. No significant differences were observed in the proportions of monomer, dimer, and low molecular weight species for up to 24 months in formulation 3 filled in Nipro PFS compared to formulation 3 stored in glass vials (data not shown), indicating that the stability profile of the formulation was not affected by contact with Nipro PFS.

[0234] [Table 12]

[0235] [Table 13]

[0236] [Table 14]

[0237] Capillary gel electrophoresis (reducing and non-reducing) For formulations 1-3, the percent (%) of immunoglobulin G (IgG) + 125 kDa peak was measured using non-reducing capillary gel electrophoresis (CGE). Additionally, for formulations 1-3, the heavy chain + light chain ratio was measured using reducing CGE. These results are shown in Tables 14-15, Figures 4A-4C, and Figures 5A-5C, respectively. For all three formulations stored at standard storage conditions of 2-8°C, the % purity met acceptance criteria for up to 24 months and up to 36 months. Under accelerated and stressed conditions, protein fragmentation was observed in formulations 2 and 3 compared to formulation 1. However, this is an expected observation given that lyophilized formulations are typically more stable than liquid formulations. Furthermore, the proteolytic degradation trends of formulations 2 and 3 were similar, indicating no significant effect of protein concentration. No significant differences were observed in the percentage of the IgG+125 kDa peak for up to 24 months for formulation 3 filled in Nipro PFS compared to formulation 3 stored in glass vials (data not shown), indicating that the stability profile of the formulation was not affected by contact with Nipro PFS.

[0238] [Table 15]

[0239] [Table 16]

[0240] Capillary isoelectric focusing (icIEF) The percentages of major species, acidic species, and basic species for formulations 1–3 (F1–F3) were measured using capillary isoelectric focusing (icIEF). The results are shown in Tables 16–18. Figures 6A–6C, 7A–7C, and 8A–8C show the stability trends of charge heterogeneity overlaid. Under standard storage conditions at 2–8°C (Figures 6A, 7A, and 8A), all three formulations (F1–F3) met the acceptance criteria for up to 24 and 36 months, respectively. Under accelerated storage conditions (25°C; Figures 6B, 7B, and 8B) and stressed storage conditions (40°C; Figures 6C, 7C, and 8C), the percentage of the main peak decreased for formulations 2 and 3, while the percentage of acidic species increased. No significant changes were observed for formulation 1. However, this is an expected observation, given that lyophilized formulations are typically more stable than liquid formulations. No significant differences were observed in the proportions of the main peak, acidic species, and basic species for up to 24 months in Formulation 3 filled in Nipro PFS compared to Formulation 3 stored in glass vials (data not shown), indicating that the stability profile of the formulation was not affected by contact with Nipro PFS.

[0241] [Table 17]

[0242] [Table 18]

[0243] [Table 19]

[0244] Subvisible Particles Using Microflow Imaging (MFI) Subvisible particles in Formulations 1-3 were measured at different time points under standard conditions of 2-8°C, accelerated conditions of 25°C, and stress conditions of 40°C. The results are shown in Tables 19-21, respectively. No significant changes in subvisible particles were observed. More subvisible particles were observed at intermittent time points than at other time points. This may be related to the method, where greater variability and sensitivity was observed for subvisible particles measured using MFI. Overall, subvisible particles ≥10 μm (and above) were less than 6,000 particles / mL. In the ≥25 μm particle size range, subvisible particles were less than 600 particles / mL, which is below the USP 1000 standard, even considering the increased sensitivity of using MFI to detect subvisible particles. <788> The stability of Formulation 3 filled in Nipro PFS was within the acceptable range. No significant differences were observed in the percentage of subvisible particles up to 24 months compared with Formulation 3 stored in glass vials (data not shown), indicating that the stability profile of the formulation was not affected by contact with Nipro PFS.

[0245] [Table 20]

[0246] [Table 21]

[0247] [Table 22]

[0248] Chemical modification of primary structure by peptide mapping Formulations 1-3 were monitored for amino acids that may undergo chemical modification over time and affect protein structure. Specifically, the amino acid residues methionine 81 and methionine 254 of TEV-48574 were monitored because they may undergo oxidation, potentially affecting the primary structure. Similarly, asparagine 317 of TEV-48574 was monitored because it may undergo deamidation to succinimide. However, as shown in Tables 22-24, no significant changes in these modifications were observed over time. Results showed that the oxidation rate of Met81, deamidation rate of Asn317, and oxidation rate of Met254 were comparable between Formulation 3 filled in glass vials and the equivalent formulation filled in Nipro PFS at long-term storage conditions (2-8°C) for up to 24 months (data not shown), indicating that the stability profile of the formulations is not affected by contact with Nipro PFS.

[0249] [Table 23]

[0250] [Table 24]

[0251] [Table 25]

[0252] Titer evaluation by ELISA The percent potency (%) of Formulations 1-3 was measured by enzyme-linked immunosorbent assay (ELISA). The results are shown in Table 25. Under standard storage conditions (2-8°C), no significant changes were observed between the formulations, and the acceptance criteria of 70%-135% potency were met for up to 24 months and up to 36 months. No significant differences were observed in percent potency for up to 24 months for Formulation 3 filled in Nipro PFS compared to Formulation 3 stored in glass vials (data not shown), indicating that the stability profile of the formulations is not affected by contact with Nipro PFS.

[0253] [Table 26]

[0254] Differential scanning calorimetry (DSC) analysis for thermal stability DSC was used to evaluate the thermal stability of Formulations 1-3. DSC analysis was performed on time zero (T0) samples and 3-, 6-, and 36-month samples of Formulations 1-3 at 2-8°C, 25°C, and 40°C. Figure 9A shows the thermal stability of Formulations 1-3 at time zero (T0) and 3 months at 2-8°C, 25°C, and 40°C. Figure 9B shows the thermal stability of Formulations 1-3 at time zero (T0) and 6 months at 2-8°C, 25°C, and 40°C. Figure 9C shows the thermal stability of Formulations 1-3 at time zero (T0) and 36 months at 2-8°C. The transition temperatures (Tm) of all formulations overlapped for thermal stability at T0, 3 months, 6 months, and 36 months, and no significant differences were observed among the three formulations. A slight decrease in enthalpy was observed for formulations stored under accelerated and stressed conditions. This may be due to the enthalpy drop caused by protein fragmentation observed in the samples under these conditions.No significant differences in thermal stability were observed for Formulation 3 filled in Nipro PFS compared to Formulation 3 stored in glass vials for up to 6 months, indicating that the stability profile of the formulation is not affected by contact with Nipro PFS.

[0255] Protein secondary and tertiary structure analysis using circular dichroism (CD) spectroscopy The secondary protein structures of Formulations 1-3 were analyzed using far-UV circular dichroism (CD), and the tertiary protein structures of Formulations 1-3 were analyzed using near-UV CD. Secondary and tertiary structure analyses were performed on time zero (T0) samples of Formulations 1, 2, and 3, 3-month samples at 2-8°C, 25°C, and 40°C, 24-month samples at 2-8°C, and 36-month samples at 2-8°C. The results are shown in Figures 10A-10F (Figures 10A-10F). Specifically, Figure 10A shows the secondary structures of Formulations 1-3 at T0 and after 3 months at 2-8°C, 25°C, and 40°C using far-UV CD. Figure 10B shows the secondary structures of Formulations 1-3 at T0 and after 24 months at 2-8°C using far-UV CD. Figure 10C shows the secondary structures of Formulations 1-3 at T0 and after 36 months at 2-8°C using far-UV CD. Figure 10D shows the tertiary structures of Formulations 1-3 at T0 and after 3 months at 2-8°C, 25°C, and 40°C using UV CD. Figure 10E shows the tertiary structures of Formulations 1-3 at T0 and after 24 months at 2-8°C using near-UV CD. Figure 10F shows the tertiary structures of Formulations 1-3 at T0 and after 36 months at 2-8°C using near-UV CD.

[0256] The far-UV CD spectra showed a negative maximum near 217 nm, and for the three formulations, the TO sample exhibited the beta-sheet structure expected for a monoclonal antibody. The near-UV CD spectra showed a positive maximum near 292 nm, indicating absorption due to tryptophan residues, and a negative maximum near 276 nm, indicating absorption due to tyrosine residues. No significant changes in the secondary or tertiary structure of the three formulations were observed up to 3 months. Small changes in the CD spectra near 200 nm were observed at 24 and 36 months. This is likely due to protein fragmentation, as absorption in this region is primarily due to absorption of peptide bonds. However, no significant changes in the protein's secondary structure (absorption at 217 nm) were observed.

[0257] Figure 11A shows the secondary structure of Formulation 3 stored in Nipro prefilled syringes (PFS) at time zero (T0) and after 3 months (3M), 6 months (6M), and 24 months (24M) storage at 2-8°C and after 3 months (3M) and 6 months (6M) storage at 25°C and 40°C using far-UVCD. Figure 11B shows the tertiary structure of Formulation 3 stored in Nipro prefilled syringes (PFS) at time zero (T0) and after 3 months (3M), 6 months (6M), and 24 months (24M) storage at 2-8°C and after 3 months (3M) and 6 months (6M) storage at 25°C and 40°C using near-UVCD. The spectra obtained for the Nipro PFS samples were comparable to those obtained for Formulation 3 stored in glass vials (Figures 10A-10B and 10D-10E).

[0258] Polysorbate 80 Analysis Polysorbate 80 (PS80), a surfactant / excipient in formulations 1-3, can undergo degradation, resulting in reactive peroxides that may affect protein stability during product shelf life. Formulations 1-3 were analyzed for PS80 levels after 24 and 36 months of storage at 2-8°C. As shown in Table 26, the measured PS80 concentration was 0.02% (w / v) for the 24-month samples of all formulations, which correlated with the expected PS80 concentration. After 36 months, the PS80 values of liquid formulations 2 and 3 showed a decrease. No significant differences in PS80 levels were observed after 24 months for formulation 3 filled in Nipro PFS compared to formulation 3 stored in glass vials, indicating that the stability profile of the formulations is not affected by contact with Nipro PFS.

[0259] [Table 27]

[0260] conclusion The stability of TEV-48574 in the lyophilized drug product (Formulation 1, 100 mg / mL) was comparable to that of the 100 mg / mL (Formulation 2) and 150 mg / mL (Formulation 3) liquid drug products stored at 2-8°C, the standard storage conditions for drug products. Under accelerated (25°C) and stressed (40°C) conditions, both liquid formulations exhibited protein fragmentation as the primary degradation pathway, but this was not observed in lyophilized Formulation 1. However, this is an expected observation given the greater stability of the lyophilized formulations compared to the liquid formulations.

[0261] In liquid formulations, the formation of acidic species was observed compared to lyophilized formulations, as measured by icIEF, concomitant with the protein fragmentation observed using SEC and CGE. This indicates that specific amino acid residues are deamidated, potentially leading to fragmentation. See Wang, W. et al., J. Pharm. Sci., 96:1-26 (2007).

[0262] Based on these data, lyophilized Formulation 1 can be converted to 100 mg / mL and 150 mg / mL liquid formulations with comparable stability profiles, meeting acceptance criteria of up to 24 months and up to 36 months under storage conditions at 2-8°C.

[0263] Furthermore, the stability of 150 mg / mL TEV-48574 (Formulation 3) stored in Nipro PFS was found to be comparable to that of Formulation 3 samples stored in glass vials. This data indicates that Nipro PFS is compatible with storage at 2-8°C, the standard storage conditions for drug products, for up to 24 months.

[0264] Example 3: Stability of TEV-48574 liquid formulation at 200 mg / ml The objective of this study was to evaluate the long-term stability of high-concentration liquid TEV-48574 formulations compared with lyophilized drug product. Two formulations were tested: a lyophilized drug product containing 100 mg / mL of drug product in 10 mM histidine, 5% (w / v) sucrose, 100 mM Arg-HCl, 0.02% (w / v) PS-80, pH 6.0 (Formulation 4 or F4) and a liquid drug product containing 200 mg / mL of drug product in 10 mM histidine, 5% (w / v) sucrose, 100 mM Arg-HCl, 0.02% (w / v) PS-80, pH 6.0 (Formulation 5 or F5). The stability of these formulations was evaluated under long-term storage conditions at 2-8°C in 20 mm stoppered 5 cc vials. The effect of these conditions on several quality attributes of the formulations was examined.

[0265] Visual appearance, protein concentration, osmolality and viscosity The results of the visual appearance, protein concentration, and osmolality analysis of Formulations 4 and 5 are shown in Tables 27-28. No significant differences were observed between the formulations at long-term storage conditions of 2-8°C.

[0266] The protein concentrations of both formulations measured under long-term storage conditions at 2-8°C were close to the expected nominal concentrations and did not vary over time. The osmolality and viscosity of the drug products were evaluated at time zero (T0). The osmolality and viscosity of the 200 mg / mL liquid formulation (F5) were observed to be higher than those observed for the 100 mg / mL lyophilized formulation (F4).

[0267] [Table 28]

[0268] [Table 29]

[0269] Size Exclusion Chromatography (SEC) Table 29 and Figures 12A-12C show the percent (%) of monomer, dimer, and low molecular weight species measured by SEC for Formulations 4 and 5. Both formulations stored under long-term storage conditions at 2-8°C met the acceptance criteria for monomer and dimer content for up to 24 months. For Formulation 5, there was a slight decrease in the percentage of monomer compared to Formulation 4, with a concomitant increase in the percentage of low molecular weight species.

[0270] [Table 30]

[0271] Capillary gel electrophoresis (reducing and non-reducing) For formulations 4 and 5, the percent (%) of immunoglobulin G (IgG) + 125 kDa peak was measured using non-reducing capillary gel electrophoresis (CGE). Additionally, for formulations 4 and 5, the heavy chain + light chain ratio was measured using reducing CGE. These results are shown in Tables 30-31 and Figures 13A-13B, respectively. For both formulations stored under long-term storage conditions at 2-8°C, the % purity met acceptance criteria for up to 24 months.

[0272] [Table 31]

[0273] [Table 32]

[0274] Capillary isoelectric focusing (icIEF) The percentage of major species, percentage of acidic species, and percentage of basic species for formulations 4 and 5 (F4 and F5) were measured using capillary isoelectric focusing (icIEF). The results are shown in Table 32. An overlay of the stability trends in charge heterogeneity is shown in Figures 14A-14C. Under long-term storage conditions of 2-8°C (Figures 14A, 14B, 14C), both formulations (F4 and F5) met the acceptance criteria over a 24-month period.

[0275] [Table 33]

[0276] Subvisible Particles Using Microflow Imaging (MFI) Subvisible particles in Formulations 4 and 5 were measured at different time points under long-term storage conditions at 2-8°C. The results are shown in Table 33. No significant changes in subvisible particles were observed. Overall, subvisible particles ≥10 μm (or larger) were less than 6,000 particles / mL. In the ≥25 μm particle size range, subvisible particles were less than 600 particles / mL, exceeding the USP 10 ... <788> The data indicated that the 200 mg / mL formulation (F5) did not significantly affect the number of subvisible particles under long-term storage conditions.

[0277] [Table 34]

[0278] Chemical modification of primary structure by peptide mapping Formulations 4 and 5 were monitored for amino acids that may undergo chemical modification over time and affect protein structure. Specifically, amino acid residues methionine 81 and methionine 254 of TEV-48574 were monitored because they may undergo oxidation, potentially affecting the primary structure. Similarly, asparagine 317 of TEV-48574 was monitored because it may undergo deamidation to succinimide. As shown in Table 34, no significant changes were observed in the oxidation rates of Met81 and Met254 over time under long-term storage conditions of 2-8°C for 24 months. At 24 months, the liquid drug product (F5) showed a higher level of Asn317 deamidation compared to the lyophilized drug product (F4). This difference in the extent of Asn deamidation is likely a result of the lower stability of the liquid drug product compared to the lyophilized drug product and is not due to the higher nominal protein concentration of F5.

[0279] [Table 35]

[0280] Titer evaluation by ELISA The percent (%) potency of formulations 4 and 5 was measured by enzyme-linked immunosorbent assay (ELISA). The results are shown in Table 35. Under long-term storage conditions (2-8°C), no significant changes were observed between the formulations, and the acceptance criteria of 70%-135% potency were met over 24 months.

[0281] [Table 36]

[0282] Protein secondary and tertiary structure analysis using circular dichroism (CD) spectroscopy The secondary protein structure of Formulations 4 and 5 was analyzed using far-UV circular dichroism (CD), and the tertiary protein structure of Formulations 4 and 5 was analyzed using near-UV CD. Secondary structure analysis was performed on the time zero (T0) sample and 3-, 6-, and 12-month samples of Formulation 4 at 25°C, and on the T0 sample and 3- and 12-month samples of Formulation 5 at 25°C. Tertiary structure analysis was performed on the time zero (T0) sample and 3- and 12-month samples of Formulation 4 at 25°C, and on the T0 sample and 3-, 6-, and 12-month samples of Formulation 5 at 25°C. The results are shown in Figures 15A-15B (Figures 15A-15B). Specifically, Figure 15A shows the secondary structure of Formulation 4 (F4) at time zero (T0) and after 3 months (3M), 6 months (6M), and 12 months (12M) of storage at 25°C, and Formulation 5 (F5) after storage at T0 and 3M and 12M at 25°C, using far-UV CD. Figure 15B shows the tertiary structure of F4 after storage at T0, 3M, and 12M at 25°C, and F5 after storage at T0, 3M, 6M, and 12M at 25°C, using near-UV CD. No significant changes in secondary or tertiary structure were observed in either the lyophilized formulation (F4) or the concentrated liquid formulation (F5).

[0283] conclusion The stability of the lyophilized formulation (Formulation 4, 100 mg / mL) of TEV-48574 was comparable to that of the liquid formulation (Formulation 5, 200 mg / mL). Under long-term storage conditions of 2-8°C, drug product stability data for both formulations were confirmed to be comparable.

[0284] At 24M, the liquid drug product showed a higher level of Asn317 deamidation compared to the lyophilized drug product. This difference in the extent of Asn deamidation is likely a result of the lower stability of the liquid drug product compared to the lyophilized drug product and is not due to the higher nominal protein concentration of F2. A slight decrease in the proportion of monomer, along with an increase in the proportion of low molecular weight species, was also observed in F5 compared to F4. However, lyophilized formulations are typically more stable than liquid formulations.

[0285] These data confirm that the 200 mg / mL high-concentration liquid drug product is stable at 2-8°C without significant impact on the critical quality attributes of TEV-48574.

[0286] Example 4: Evaluation of Arginine Hydrochloride as an Excipient in TEV-48574 Liquid Formulations The objective of this study was to evaluate the effectiveness of arginine hydrochloride (Arg-HCl) as an excipient and its effect on the stability of the TEV-48574 drug product. Four liquid formulations were tested. Two formulations were prepared in 10 mM histidine, 5% (w / v) sucrose, 100 mM Arg-HCl, 0.02% (w / v) PS-80, pH 6.0, containing either 150 mg / mL (Formulation 1A) or 100 mg / mL (Formulation 2A) of drug product. Two formulations were prepared in 10 mM histidine, 5% (w / v) sucrose, 0.02% (w / v) PS-80, pH 6.0, containing either 150 mg / mL (Formulation 1B) or 100 mg / mL (Formulation 2B) of drug product. The stability of these formulations was evaluated under stressful storage conditions at 40°C. The effect of these conditions on product quality attributes was evaluated.

[0287] Visual appearance, protein concentration, osmolality and viscosity The results of the visual appearance, protein concentration, and osmolality analyses of Formulations 1A-2A and 1B-2B are shown in Tables 36-38. At 40°C storage conditions, a slight yellow color was observed in the visual appearance of Formulations 1A and 1B, which may be related to the deterioration of these formulations over time at elevated temperatures. The measured protein concentrations of the four formulations at 40°C were close to the expected nominal concentrations and did not change over time. The osmolality and viscosity of the drug products were evaluated and no significant differences were observed over time.

[0288] [Table 37]

[0289] [Table 38]

[0290] [Table 39]

[0291] The osmolality of Formulations 1A and 2A containing Arg-HCl was higher than that of Formulations 1B and 2B without Arg-HCl. The viscosity of Formulation 1B without Arg-HCl was observed to be higher than that of Formulation 1A with Arg-HCl at 150 mg / mL, as shown in Table 39 and Figure 16.

[0292] [Table 40]

[0293] Size Exclusion Chromatography (SEC) Tables 40-41 and Figures 17A-17C show the percent (%) of monomer, dimer, and fragment species measured by SEC for Formulations 1A-1B and 2A-2B after storage at 40°C for up to 8 weeks. The reported values are within the variability of the assay, so no significant differences in monomer or fragment percentages were observed among the four formulations. Formulations 1B and 2B without Arg-HCl showed an increased rate of dimer formation compared to Formulations 1A and 2A with Arg-HCl, indicating the stabilizing effect of Arg-HCl on the drug product.

[0294] [Table 41]

[0295] [Table 42]

[0296] Capillary gel electrophoresis (reducing and non-reducing) For Formulations 1A-1B and 2A-2B, the percent (%) of immunoglobulin G (IgG) + 125 kDa peak was measured using non-reducing capillary gel electrophoresis (CGE) under stress conditions at 40°C for up to 8 weeks. Additionally, for Formulations 1A-1B and 2A-2B, the percentage of heavy chains plus light chains was measured using reducing CGE under stress conditions at 40°C for up to 8 weeks. These results are shown in Tables 42-43 and Figures 18A-18B, respectively. Formulations 1B and 2B without Arg-HCl were observed to have increased rates of fragmentation compared to Formulations 1A and 2A with Arg-HCl, demonstrating the stabilizing effect of Arg-HCl on the fragmentation pattern of the drug product.

[0297] [Table 43]

[0298] [Table 44]

[0299] Capillary isoelectric focusing (icIEF) The percentages of major, acidic, and basic species for Formulations 1A-1B and 2A-2B were measured using capillary isoelectric focusing (icIEF) under stress conditions at 40°C for up to 8 weeks. The results are shown in Tables 44-45. An overlay of the stability trends in charge heterogeneity is shown in Figures 19A-19C. Slightly higher levels of acidic species were observed at the end of 8 weeks for Formulations 1B and 2B without Arg-HCl. This may be related to the higher fragmentation rates of these formulations compared to Formulations 1A and 2A, as reported in Table 43.

[0300] [Table 45]

[0301] [Table 46]

[0302] Chemical modification of primary structure by peptide mapping For Formulations 1A-1B and 2A-2B, amino acids that may undergo chemical modification over time and affect protein structure were monitored under stress conditions at 40°C for up to 8 weeks. Specifically, the amino acid residues methionine 81 and methionine 254 of TEV-48574 were monitored because they may undergo oxidation, potentially affecting the primary structure. Similarly, asparagine 317 of TEV-48574 was monitored because it may undergo deamidation to succinimide. The oxidation rate of Met81, deamidation rate of Asn317, and oxidation rate of Met254 did not change significantly over time under storage conditions at 40°C for up to 8 weeks (data not shown), suggesting that the presence of Arg-HCl does not affect these attributes.

[0303] Titer evaluation by ELISA The percent potency of Formulations 1A-1B and 2A-2B was measured by enzyme-linked immunosorbent assay (ELISA) under stress conditions at 40°C for up to 8 weeks. The results are shown in Table 46. The percent potency trends were similar for all formulations, regardless of drug product concentration or Arg-HCl content.

[0304] [Table 47]

[0305] Dynamic Light Scattering (DLS) for Particle Size Characterization DLS was employed to measure the polydispersity (%PD) and hydrodynamic radius of the potential drug product and nanoparticles contained in Formulations 1A-1B and 2A-2B. DLS analysis was performed on time zero (T0) samples of Formulations 1A-1B and 2A-2B. Table 47 shows the results of the DLS analysis. The apparent hydrodynamic radius of Formulations 1B and 2B (without Arg-HCl) was observed to be smaller compared to Formulations 1A and 2A (with Arg-HCl).

[0306] [Table 48]

[0307] Differential scanning calorimetry (DSC) analysis for thermal stability DSC was used to evaluate the thermal stability of Formulations 1A-1B and 2A-2B. DSC analysis was performed on time zero (T0) samples of Formulations 1A-1B and 2A-2B. Figure 20 shows the thermal stability of Formulations 1A-1B and 2A-2B at time zero (T0), and Table 48 shows the DSC thermograms of all four formulations. onset The transition temperatures (Tm) were approximately 78°C for all formulations.

[0308] [Table 49]

[0309] The Tm1 of formulations 1A and 2A was slightly shifted to lower temperatures, indicating that Arg-HCl may affect the thermal stability of the molecule. This may be due to the enthalpy reduction caused by protein fragmentation observed in the samples under these conditions. However, this temperature is above the recommended storage range of 2-8°C for this drug product.

[0310] conclusion The effectiveness of arginine hydrochloride (Arg-HCl) as an excipient and its effect on the stability of TEV-48574 drug product was evaluated at two different drug product concentrations (100 mg / mL and 150 mg / mL). For all four formulations tested, the quality attributes of dimer percentage and purity, measured using reduced CGE, demonstrated the stabilizing effect of 100 mM Arg-HCl compared to formulations without Arg-HCl. The viscosity of the 150 mg / mL drug product with Arg-HCl was lower than that of the formulation without Arg-HCl. A small shift of approximately 3°C in Tm1, as analyzed by DSC, was observed for the formulation containing Arg-HCl. However, because the recommended storage conditions for the drug product are 2-8°C, this Tm1 shift is not expected to significantly affect stability under typical storage and handling conditions. All other attributes showed no significant changes over 8 weeks at 40°C for any of the four formulations.

[0311] These data indicated that 100 mM Arg-HCl in the TEV-48574 formulation may stabilize the formulation and minimize viscosity even as the drug product concentration increases.

[0312] It is understood that for interpreting the claims, reference is intended to be made to the Detailed Description section, and not to the Summary and Abstract sections. The Summary and Abstract sections may describe one or more, but not all, exemplary aspects of the invention contemplated by the inventors, and are therefore not intended to limit the scope of the invention and the appended claims in any way.

[0313] The foregoing description of specific embodiments fully reveals the general nature of the present invention so that others, by applying knowledge within the skill of those skilled in the art, can readily modify and / or adapt such specific embodiments for various uses without departing from the general concept of the present invention and without undue experimentation. Such adaptations and modifications are therefore intended to be within the meaning and range of equivalents of the disclosed embodiments, based on the teaching and guidance presented herein. It is to be understood that the phrases or terms used herein are intended to be descriptive rather than limiting, as the terms or terms used herein would be interpreted by one of ordinary skill in the art in light of the teaching and guidance.

[0314] The breadth and scope of the present invention should not be limited by any of the above-described exemplary embodiments, but should be defined only in accordance with the following claims and their equivalents.

[0315] Various publications, including patents, published applications, accession numbers, technical papers and scholarly articles, are cited throughout this specification, each of which is incorporated herein by reference in its entirety and for all purposes.

[0316] [Table 50]

[0317] [Table 51]

[0318] [Table 52]

[0319] [Table 53]

[0320] [Table 54]

[0321] Table 55

[0322] Table 56

[0323] Table 57

Claims

1. 1. A pharmaceutical formulation comprising: (a) about 100 mg / mL to about 250 mg / mL of an antibody or antigen-binding fragment thereof that specifically binds to TNF-like ligand 1A (TL1A), the antibody or antigen-binding fragment thereof comprising a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 2, a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 3, a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 5, and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 6; (b) about 5 mM to about 15 mM histidine; and (c) about 50 mM to about 150 mM arginine hydrochloride (Arg-HCl); (d) about 2.5% (w / v) to about 7.5% (w / v) sucrose; and (e) about 0.01% (w / v) to about 0.03% (w / v) polysorbate-80; and 10. A pharmaceutical formulation comprising:

2. 2. The pharmaceutical formulation of claim 1, wherein the antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:7 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:

8.

3. 3. The pharmaceutical formulation of claim 1 or 2, wherein the antibody or antigen-binding fragment comprises an IgG1 constant region.

4. 4. The pharmaceutical formulation of claim 1, wherein the antibody or antigen-binding fragment thereof 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.

5. 5. The pharmaceutical formulation of claim 1, comprising about 100 mg / mL of the antibody or antigen-binding fragment thereof.

6. 5. The pharmaceutical formulation of claim 1, comprising about 150 mg / mL of the antibody or antigen-binding fragment thereof.

7. 5. The pharmaceutical formulation of claim 1, comprising about 200 mg / mL of the antibody or antigen-binding fragment thereof.

8. 5. The pharmaceutical formulation of claim 1, comprising about 225 mg / mL of the antibody or antigen-binding fragment thereof.

9. 5. The pharmaceutical formulation of claim 1, comprising about 250 mg / mL of the antibody or antigen-binding fragment thereof.

10. 10. The pharmaceutical formulation of any one of claims 1 to 9, comprising about 5 mM histidine.

11. 10. The pharmaceutical formulation of any one of claims 1 to 9, comprising about 10 mM histidine.

12. 10. The pharmaceutical formulation of any one of claims 1 to 9, comprising about 15 mM histidine.

13. 13. The pharmaceutical formulation of any one of claims 1 to 12, comprising about 50 mM arginine hydrochloride (Arg-HCl).

14. 13. The pharmaceutical formulation of any one of claims 1 to 12, comprising about 100 mM arginine hydrochloride (Arg-HCl).

15. 13. The pharmaceutical formulation of any one of claims 1 to 12, comprising about 150 mM arginine hydrochloride (Arg-HCl).

16. 16. The pharmaceutical formulation of any one of claims 1 to 15, comprising about 2.5% (w / v) sucrose.

17. 16. The pharmaceutical formulation of any one of claims 1 to 15, comprising about 5% (w / v) sucrose.

18. 16. The pharmaceutical formulation of any one of claims 1 to 15, comprising about 7.5% (w / v) sucrose.

19. 19. The pharmaceutical formulation of any one of claims 1 to 18, comprising about 0.01% (w / v) polysorbate-80.

20. 19. The pharmaceutical formulation of any one of claims 1 to 18, comprising about 0.02% (w / v) polysorbate-80.

21. 19. The pharmaceutical formulation of any one of claims 1 to 18, comprising about 0.03% (w / v) polysorbate-80.

22. 5. The pharmaceutical formulation of claim 1, comprising about 250 mg / mL of the antibody or antigen-binding fragment thereof, about 10 mM histidine, about 100 mM arginine hydrochloride (Arg-HCl), about 5% (w / v) sucrose, and about 0.02% (w / v) polysorbate-80.

23. 5. The pharmaceutical formulation of claim 1, comprising about 200 mg / mL of the antibody or antigen-binding fragment thereof, about 10 mM histidine, about 100 mM arginine hydrochloride (Arg-HCl), about 5% (w / v) sucrose, and about 0.02% (w / v) polysorbate-80.

24. 5. The pharmaceutical formulation of claim 1, comprising about 150 mg / mL of the antibody or antigen-binding fragment thereof, about 10 mM histidine, about 100 mM arginine hydrochloride (Arg-HCl), about 5% (w / v) sucrose, and about 0.02% (w / v) polysorbate-80.

25. The pharmaceutical formulation of any one of claims 1 to 24, which is freeze-dried.

26. The pharmaceutical formulation according to any one of claims 1 to 24, which is a liquid.

27. 27. The pharmaceutical formulation of any one of claims 1 to 26, having a pH of 6.0±0.5 after storage at room temperature for 24 hours or at 2-8°C for 24 hours, 72 hours or 10 days.

28. 28. The pharmaceutical formulation of any one of claims 1 to 27, having an osmolality of 200 mOsm / kg to 500 mOsm / kg after storage at room temperature for 24 hours or at 2-8°C for 24 hours, 72 hours or 10 days.

29. 29. The pharmaceutical formulation of any one of claims 1 to 28, having an antibody monomer content of at least 99% after storage at 2-8°C for 24 hours, 72 hours or 10 days.

30. 30. The pharmaceutical formulation of any one of claims 1 to 29, having no significant change in charge heterogeneity profile after storage at 2-8°C for 24 hours, 72 hours or 10 days.

31. 31. The pharmaceutical formulation of any one of claims 1 to 30, which has no significant change in purity after storage at room temperature for 24 hours or at 2-8°C for 24 hours, 72 hours or 10 days.

32. 31. The pharmaceutical formulation of any one of claims 1 to 30, having a purity of at least 90% after storage at room temperature for 24 hours or at 2-8°C for 24 hours, 72 hours or 10 days.

33. 33. The pharmaceutical formulation of any one of claims 1 to 32, which has no significant change in particle concentration after storage at room temperature for 24 hours or at 2-8°C for 24 hours, 72 hours or 10 days.

34. 34. A pharmaceutical formulation according to any one of claims 1 to 33, which has no significant difference in visual appearance after storage at 2-8°C for up to 36 months.

35. 35. The pharmaceutical formulation of any one of claims 1 to 34, which has no significant difference in protein concentration, osmolality or viscosity after storage at 2-8°C, 25°C or 40°C for up to 36 months.

36. 36. The pharmaceutical formulation of any one of claims 1 to 35, having a monomer content of 95% or more, a dimer content of 5.0% or less, or no significant difference in low molecular weight species content after storage at 2-8°C for up to 36 months.

37. 37. The pharmaceutical formulation of any one of claims 1 to 36, having a purity of 90% or greater after storage at 2-8°C for up to 36 months.

38. 38. A pharmaceutical formulation according to any one of claims 1 to 37, having a major species content of 50% to 90%, an acidic species content of 10% to 40% and / or a basic species content of 0% to 20% after storage at 2 to 8°C for up to 36 months.

39. 39. The pharmaceutical formulation of any one of claims 1 to 38, which has no significant difference in subvisible particle content after storage at 2-8°C, 25°C or 40°C for up to 36 months.

40. 40. The pharmaceutical formulation of any one of claims 1 to 39, having no significant difference in oxidation of methionine 81 and / or methionine 254 of TEV-48574 and / or deamidation of asparagine 317 of TEV-48574 after storage at 2-8°C, 25°C or 40°C for up to 36 months.

41. 41. The pharmaceutical formulation of any one of claims 1 to 40, having a relative potency of 70% to 135% as measured by enzyme-linked immunosorbent assay (ELISA) after storage at 2-8°C for up to 36 months.

42. 42. The pharmaceutical formulation of any one of claims 1 to 41, which has no significant difference in thermal stability after storage at 2-8°C, 25°C or 40°C for up to 6 months.

43. 43. The pharmaceutical formulation of any one of claims 1 to 42, which has no significant difference in thermal stability after storage at 2-8°C for up to 36 months.

44. 44. A pharmaceutical formulation according to any one of claims 1 to 43, which has no significant changes in secondary and / or tertiary protein structure after storage at 2-8°C, 25°C or 40°C for up to 3 months.

45. 45. The pharmaceutical formulation of any one of claims 1 to 44, which has no significant difference in secondary protein structure after storage at 2-8°C for up to 36 months.

46. 46. The pharmaceutical formulation of any one of claims 1 to 45, having no significant difference in polysorbate-80 concentration after storage at 2-8°C for up to 24 months.

47. 47. The pharmaceutical formulation of any one of claims 1 to 46, wherein the antibody or antigen-binding fragment thereof is produced in Chinese hamster ovary cells.

48. A container comprising the pharmaceutical formulation of any one of claims 1 to 47.

49. 49. The container of claim 48, which is a glass vial.

50. 50. The container of claim 49, which is a glass vial having a fill volume of 3 mL.

51. 49. The container of claim 48, which is a syringe, optionally wherein the syringe is a pre-filled syringe.

52. 52. A method of treating a disease in a subject in need thereof, comprising administering to said subject a pharmaceutical formulation according to any one of claims 1 to 47 or a container according to any one of claims 48 to 51, optionally wherein said disease is a respiratory tract disease, a gastrointestinal disease, a skin disease or arthritis.

53. 53. The method of claim 52, wherein the airway disease is asthma, chronic obstructive pulmonary disease (COPD), pulmonary fibrosis, pulmonary sarcoidosis, allergic rhinitis, or cystic fibrosis.

54. 53. The method of claim 52, wherein the gastrointestinal disease is inflammatory bowel disease, Crohn's disease, colitis, ulcerative colitis, eosinophilic esophagitis, or irritable bowel syndrome.

55. 53. The method of claim 52, wherein the arthritis is rheumatoid arthritis.

56. 53. The method of claim 52, wherein the skin disease is atopic dermatitis, eczema, or scleroderma.

57. 57. The method of any one of claims 52 to 56, wherein the pharmaceutical preparation is administered intravenously.

58. 57. The method of any one of claims 52 to 56, wherein the pharmaceutical formulation is administered subcutaneously.

59. A formulation according to any one of claims 1 to 47 for use according to a method according to any one of claims 52 to 58.

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