Anti-TL1A antibody formulations

Abstract

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

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This international application claims the benefit of priority to U.S. Provisional Application No. 63 / 369,638, filed on July 27, 2022, which is incorporated herein by reference in its entirety. References to electronically submitted sequence listings

[0002] 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 its entirety. Background Art

[0003] TNF-like ligand 1A (TL1A, synonymous TNF superfamily member 15 (TNFSF15); TL1 and VEGI) is a member of the tumor necrosis factor superfamily that is expressed by antigen-presenting cells (including dendritic cells, B cells and macrophages), CD4+ and CD8+ T cells and endothelial cells. TL1A can be 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 and FOXP3+ regulatory T (Treg) cells and type 2 and 3 innate lymphoid cells (ILC2 and ILC3).

[0004] TL1A can also bind to a decoy receptor (DcR3), which is a competitive inhibitor of DR3. DcR3 also acts as a decoy receptor for Fas-ligand (Fas-L) and lymphotoxin-like inducible protein, which competes with glycoprotein D for binding to herpes virus invasion mediator (LIGHT) on T cells. Therefore, DcR3 is an important regulator of several signal transduction pathways.

[0005] The TL1A / DR3 signaling pathway has been implicated in several biological systems that are associated with human disease. For example, TL1A has been shown to play a role in immunity of barrier tissues, angiogenesis, and homeostasis. Inhibition of the interaction of TL1A with DR3 has also been shown to promote therapeutic benefit in several immune-mediated disorders, such as 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 treatments for these diseases.

[0006] However, in order to use such antibodies and provide treatments for these diseases, stable antibody formulations are required. Summary of the invention

[0007] Provided herein is a pharmaceutical formulation comprising: (a) about 100 mg / mL to about 250 mg / mL of an antibody or an antigen-binding fragment thereof that specifically binds to TNF-like ligand 1A (TL1A); wherein the antibody or the 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, 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. NO:6; (b) about 5mM to about 15mM histidine; (c) about 50mM to about 150mM 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 aspects, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising an amino acid sequence of SEQ ID NO:7 and a light chain variable region comprising an amino acid sequence of SEQ ID NO:8. In some aspects, the antibody or antigen-binding fragment thereof comprises an IgG1 constant region. In some aspects, the antibody or antigen-binding fragment thereof comprises a heavy chain comprising an amino acid sequence of SEQ ID NO:9 and a light chain comprising an amino acid sequence of SEQ ID NO:10.

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

[0009] In some aspects, the pharmaceutical formulation disclosed herein comprises 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 formulation disclosed herein comprises 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 formulation disclosed herein comprises 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 aspects, the pharmaceutical formulation is lyophilized. In some aspects, the pharmaceutical formulation is liquid. In some aspects, the pharmaceutical formulation has a pH of 6.0 ± 0.5 after storage at room temperature for 24 hours or at 2°C-8°C for 24 hours, 72 hours or 10 days.

[0013] In some aspects, the drug formulation is stored at room temperature for 24 hours or at 2 ° C-8 ° C for 24 hours, 72 hours or 10 days after having an osmotic pressure of from 200mOsm / kg to 500mOsm / kg. In some aspects, the drug formulation has at least 99% antibody monomer content after being stored at 2 ° C-8 ° C for 24 hours, 72 hours or 10 days. In some aspects, the drug formulation has no significant change in charge heterogeneity characteristics after being stored at 2 ° C-8 ° C for 24 hours, 72 hours or 10 days. In some aspects, the drug formulation has no significant change in purity after being stored at room temperature for 24 hours or at 2 ° C-8 ° C for 24 hours, 72 hours or 10 days. In some aspects, the drug formulation has at least 90% purity after being stored at room temperature for 24 hours or at 2 ° C-8 ° C for 24 hours, 72 hours or 10 days. In some aspects, the pharmaceutical formulation is stored at room temperature for 24 hours or at 2 ° C-8 ° C for 24 hours, 72 hours or 10 days after there is no significant change in particle concentration. In some aspects, the pharmaceutical formulation is stored at 2 ° C-8 ° C for up to 36 months and there is no significant difference in visual appearance. In some aspects, the pharmaceutical formulation disclosed herein is stored at 2 ° C-8 ° C, 25 ° C or 40 ° C for up to 36 months and there is no significant difference in protein concentration, osmotic pressure or viscosity. In some aspects, the pharmaceutical formulation disclosed herein has ≥ (greater than or equal to) 95% monomer content, ≤ (less than or equal to) 5.0% dimer content or low molecular weight material content after being stored at 2 ° C-8 ° C for up to 36 months. There is no significant difference. In some aspects, the pharmaceutical formulation disclosed herein has ≥ (greater than or equal to) 90% purity after being stored at 2 ° C-8 ° C for up to 36 months. In some aspects, the pharmaceutical formulation disclosed herein has a main substance content of 50% to 90%, an acidic substance content of 10% to 40%, and / or a basic substance content of 0% to 20% after storage for up to 36 months at 2°C-8°C. In some aspects, the pharmaceutical formulation disclosed herein has no significant difference in sub-visible particle content after storage for up to 36 months at 2°C-8°C, 25°C or 40°C. In some aspects, the pharmaceutical formulation disclosed herein has no significant difference in the oxidation of methionine 81 and / or methionine 254 of TEV-48574 and / or the deamidation of asparagine 317 of TEV-48574 after storage for up to 36 months at 2°C-8°C, 25°C or 40°C. In some aspects, the pharmaceutical formulation disclosed herein has a relative potency of 70% to 135% measured by enzyme-linked immunosorbent assay (ELISA) after storage for up to 36 months at 2°C-8°C. In some aspects, the pharmaceutical formulations disclosed herein have no significant differences in thermal stability after storage at 2°C-8°C, 25°C, or 40°C for up to 6 months.In some aspects, the pharmaceutical formulation disclosed herein has no significant difference in thermal stability after being stored at 2°C-8°C for up to 36 months. In some aspects, the pharmaceutical formulation disclosed herein has no significant difference in secondary and / or tertiary protein structure after being stored at 2°C-8°C, 25°C or 40°C for up to 3 months. In some aspects, the pharmaceutical formulation disclosed herein has no significant difference in secondary protein structure after being stored at 2°C-8°C for up to 36 months. In some aspects, the pharmaceutical formulation disclosed herein has no significant difference in the concentration of polysorbate-80 after being stored at 2°C-8°C for up to 24 months.

[0014] Also provided herein is a container comprising a pharmaceutical formulation disclosed herein. In some aspects, the container is a glass vial. In some aspects, the container is a glass vial with a 3 mL fill volume. In some aspects, the container is a glass vial with a 3 mL or less fill volume. In some aspects, the container is a syringe, such as a pre-filled syringe. In some aspects, the pre-filled syringe has a volume of 2 mL or less.

[0015] In some aspects, the antibody or its Fab is present in a pharmaceutical formulation or container disclosed herein. In some aspects, the antibody or its Fab is prepared in a volume of 3 mL or less. In some aspects, the antibody or its Fab is prepared in a volume of 2 mL or less.

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

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

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

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

[0020] In some aspects, the skin disease is atopic dermatitis, eczema, or scleroderma.

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

[0022] Also provided herein is a composition for use according to the methods disclosed herein.

[0023] Provided herein is a pharmaceutical formulation comprising: (a) 150 mg / mL of an antibody or an antigen-binding fragment thereof that specifically binds to TNF-like ligand 1A (TL1A); wherein the antibody or the antigen-binding fragment thereof comprises: (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, 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; or (iii) a heavy chain comprising the amino acid sequence of SEQ ID NO: 9 and a light chain variable region CDR4 comprising the amino acid sequence of SEQ ID NO: 10. NO: light chain of the amino acid sequence of 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 aspects, the antibody or antigen-binding fragment of (i) or (ii) comprises an IgG1 constant region.

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

[0025] In some aspects, the drug formulation is stored at room temperature for 24 hours or at 2 ° C-8 ° C for 24 hours, 72 hours or 10 days after having an osmotic pressure of from 200mOsm / kg to 500mOsm / kg. In some aspects, the drug formulation has at least 99% antibody monomer content after being stored at 2 ° C-8 ° C for 24 hours, 72 hours or 10 days. In some aspects, the drug formulation has no significant change in charge heterogeneity characteristics after being stored at 2 ° C-8 ° C for 24 hours, 72 hours or 10 days. In some aspects, the drug formulation has no significant change in purity after being stored at room temperature for 24 hours or at 2 ° C-8 ° C for 24 hours, 72 hours or 10 days. In some aspects, the drug formulation has at least 90% purity after being stored at room temperature for 24 hours or at 2 ° C-8 ° C for 24 hours, 72 hours or 10 days. In some aspects, the pharmaceutical formulation is stored at room temperature for 24 hours or at 2 ° C-8 ° C for 24 hours, 72 hours or 10 days after there is no significant change in particle concentration. In some aspects, the pharmaceutical formulation is stored at 2 ° C-8 ° C for up to 36 months and there is no significant difference in visual appearance. In some aspects, the pharmaceutical formulation disclosed herein is stored at 2 ° C-8 ° C, 25 ° C or 40 ° C for up to 36 months and there is no significant difference in protein concentration, osmotic pressure or viscosity. In some aspects, the pharmaceutical formulation disclosed herein has ≥ (greater than or equal to) 95% monomer content, ≤ (less than or equal to) 5.0% dimer content or low molecular weight material content after being stored at 2 ° C-8 ° C for up to 36 months. There is no significant difference. In some aspects, the pharmaceutical formulation disclosed herein has ≥ (greater than or equal to) 90% purity after being stored at 2 ° C-8 ° C for up to 36 months. In some aspects, the pharmaceutical formulation disclosed herein has a main substance content of 50% to 90%, an acidic substance content of 10% to 40%, and / or a basic substance content of 0% to 20% after storage for up to 36 months at 2°C-8°C. In some aspects, the pharmaceutical formulation disclosed herein has no significant difference in sub-visible particle content after storage for up to 36 months at 2°C-8°C, 25°C or 40°C. In some aspects, the pharmaceutical formulation disclosed herein has no significant difference in the oxidation of methionine 81 and / or methionine 254 of TEV-48574 and / or the deamidation of asparagine 317 of TEV-48574 after storage for up to 36 months at 2°C-8°C, 25°C or 40°C. In some aspects, the pharmaceutical formulation disclosed herein has a relative potency of 70% to 135% measured by enzyme-linked immunosorbent assay (ELISA) after storage for up to 36 months at 2°C-8°C. In some aspects, the pharmaceutical formulations disclosed herein have no significant differences in thermal stability after storage at 2°C-8°C, 25°C, or 40°C for up to 6 months.In some aspects, the pharmaceutical formulation disclosed herein has no significant difference in thermal stability after being stored at 2°C-8°C for up to 36 months. In some aspects, the pharmaceutical formulation disclosed herein has no significant difference in secondary and / or tertiary protein structure after being stored at 2°C-8°C, 25°C or 40°C for up to 3 months. In some aspects, the pharmaceutical formulation disclosed herein has no significant difference in secondary protein structure after being stored at 2°C-8°C for up to 36 months. In some aspects, the pharmaceutical formulation disclosed herein has no significant difference in the concentration of polysorbate-80 after being stored at 2°C-8°C for up to 24 months.

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

[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, there is provided herein a method for treating a disease in a subject in need thereof, the method comprising administering to the subject any pharmaceutical formulation provided herein, including a pharmaceutical formulation in any container provided herein. In some aspects, the disease is a respiratory disease, a gastrointestinal disease, a skin disease, or arthritis.

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

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

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

[0032] In some aspects, the skin disease is atopic dermatitis, eczema, or scleroderma.

[0033] Also provided herein is a composition for use according to the methods disclosed herein. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figures 1A-1C Three TEV-48574 formulations (F1-F3) are shown at 2°C-8°C ( Figure 1A )、25℃( Figure 1B ) and 40℃( Figure 1C ) after storage under .

[0035] Figures 2A-2CThree TEV-48574 formulations (F1-F3) are shown at 2°C-8°C ( Figure 2A )、25℃( Figure 2B ) and 40℃( Figure 2C ) after storage under .

[0036] Figures 3A-3C Three TEV-48574 formulations (F1-F3) are shown at 2°C-8°C ( Figure 3A )、25℃( Figure 3B ) and 40℃( Figure 3C ) after storage under low molecular weight (LWM) protein fragment composition percentage (%).

[0037] Figures 4A-4C Three TEV-48574 formulations (F1-F3) are shown at 2°C-8°C ( Figure 4A )、25℃( Figure 4B ) and 40℃( Figure 4C ) after storage under 4% purity, determined using CGE non-reducing conditions (IgG+125 kDa peak %).

[0038] Figures 5A-5C Three TEV-48574 formulations (F1-F3) are shown at 2°C-8°C ( Figure 5A )、25℃( Figure 5B ) and 40℃( Figure 5C ) after storage under reduced pressure, determined using CGE reducing conditions (heavy chain + light chain %).

[0039] Figures 6A-6C It is shown that the formulations 1-3 (F1-F3) at 2°C-8°C ( Fig. 6A )、25℃( Figure 6B ) and 40℃( Figure 6C ) after storage under , as the percentage (%) of the main species (main peak).

[0040] Figures 7A-7C It is shown that the formulations 1-3 (F1-F3) at 2°C-8°C ( Fig. 7A )、25℃( Figure 7B ) and 40℃( Figure 7C ) after storage under , expressed as the percentage (%) of acidic substances (acidic peak).

[0041] Figures 8A-8C It is shown that the formulations 1-3 (F1-F3) at 2°C-8°C ( Fig. 8A )、25℃( Figure 8B ) and 40℃( Figure 8C) after storage under , expressed as the percentage (%) of basic substances (basic peak).

[0042] Fig. 9A Shown are the thermal stabilities of Formulations 1-3 (F1-F3) at time zero (TO) and after 3 months storage at 2-8°C, 25°C and 40°C.

[0043] Fig. 9B The thermal stability of Formulations 1-3 (F1-F3) at time zero (TO) and after 6 months storage at 2-8°C, 25°C and 40°C is shown.

[0044] Fig. 9C Shown are the thermal stabilities of Formulations 1-3 (F1-F3) at time zero (TO) and after storage for 36 months at 2-8°C.

[0045] Fig. 10A Shown are the secondary structures of Formulations 1-3 at time zero (T0) and after 3 months storage at 2-8°C, 25°C, and 40°C, as determined using far ultraviolet (UV) circular dichroism (CD).

[0046] Fig. 10B Shown are the secondary structures of Formulations 1-3 at time zero (TO) and after storage for 24 months at 2-8°C, as determined using far UV CD.

[0047] Fig. 10C Shown are the secondary structures of Formulations 1-3 at time zero (TO) and after storage for 36 months at 2-8°C, as determined using far UV CD.

[0048] Fig. 10D Shown are the tertiary structures of Formulations 1-3 at time zero (TO) and after storage for 6 months at 2-8°C, 25°C and 40°C, as determined using near UV CD.

[0049] Fig.10E Shown are the tertiary structures of Formulations 1-3 at time zero (TO) and after storage for 24 months at 2-8°C, as determined using near UV CD.

[0050] Fig.10F Shown are the tertiary structures of Formulations 1-3 at time zero (TO) and after storage for 36 months at 2-8°C, as determined using near UV CD.

[0051] Fig.11AShown are the secondary structures 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) at 2°C-8°C, and after 3 months (3M) and 6 months (6M) at 25°C and 40°C, as determined using far UV CD.

[0052] Fig. 11B Shown is the tertiary structure of Formulation 3 stored in a Nipro prefilled syringe (PFS) at time zero (T0) and after 3 months (3M), 6 months (6M) and 24 months (24M) storage at 2°C-8°C, and after 3 months (3M) and 6 months (6M) storage at 25°C and 40°C, as determined using near UV CD.

[0053] Fig. 12A Shown are the monomer composition percentages (%) of a 100 mg / mL lyophilized TEV-48574 formulation (F4) and a 200 mg / mL liquid TEV-48574 formulation (F5) after storage at 2°C-8°C for up to 24 months.

[0054] Fig. 12B Shown are the dimer composition percentage (%) of a 100 mg / mL lyophilized TEV-48574 formulation (F4) and a 200 mg / mL liquid TEV-48574 formulation (F5) after storage at 2°C-8°C for up to 24 months.

[0055] Fig. 12C Shown are the percentage (%) of 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 at 2°C-8°C for up to 24 months.

[0056] Fig.13A Shown are the percent (%) purity of two TEV-48574 formulations (F4 and F5) after storage at 2°C-8°C for up to 24 months, as determined using CGE non-reducing conditions (IgG+125 kDa peak %).

[0057] Fig. 13B Shown are the percent (%) purity of two TEV-48574 formulations (F4 and F5) after storage at 2°C-8°C for up to 24 months, as determined using CGE reducing conditions (heavy chain (HC) + light chain (LC) %).

[0058] Fig.14A The charge heterogeneity characteristics of Formulations 4 and 5 (F4 and F5) after storage at 2°C-8°C for up to 24 months are shown as the percentage (%) of the major species (main peak).

[0059] Fig. 14B Shown are the charge heterogeneity characteristics of Formulations 4 and 5 (F4 and F5) after storage at 2°C-8°C for up to 24 months, expressed as the percentage (%) of acidic species (acidic peak).

[0060] Fig. 14C Shown are the charge heterogeneity characteristics of Formulations 4 and 5 (F4 and F5) after storage at 2°C-8°C for up to 24 months, expressed as the percentage (%) of basic material (basic peak).

[0061] Fig.15A Shown are the secondary structures 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, as determined using far UV CD.

[0062] Fig. 15B Shown are the tertiary structures 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, as determined using near UV CD.

[0063] Fig.16 Shown are the viscosities of liquid formulations at 20°C with increasing concentrations of TEV-48574 (0 to 150 mg / mL), with the addition of 100 mM arginine-HCl as an excipient (1A) or without the addition of arginine-HCl (1B).

[0064] Fig.17A Shown are the monomer composition percentages (%) of two liquid TEV-48574 formulations of 150 mg / mL (1A) and 100 mg / mL (2A) with the addition of 100 mM arginine-HCl and two liquid TEV-48574 formulations of 150 mg / mL (1B) and 100 mg / mL (2B) without arginine-HCl after storage at 40°C for up to 8 weeks.

[0065] Fig. 17B Shown are the dimer composition percentage (%) of two liquid TEV-48574 formulations of 150 mg / mL (1A) and 100 mg / mL (2A) with the addition of 100 mM arginine-HCl and two liquid TEV-48574 formulations of 150 mg / mL (1B) and 100 mg / mL (2B) without arginine-HCl after storage at 40°C for up to 8 weeks.

[0066] Fig. 17CShown are the percentage (%) of fragments after storage at 40°C for up to 8 weeks for two liquid TEV-48574 formulations of 150 mg / mL (1A) and 100 mg / mL (2A) with the addition of 100 mM arginine-HCl and two liquid TEV-48574 formulations of 150 mg / mL (1B) and 100 mg / mL (2B) without arginine-HCl.

[0067] Fig.18A Shown are the percent (%) purity of two liquid TEV-48574 formulations at 150 mg / mL (1A) and 100 mg / mL (2A) with the addition of 100 mM arginine-HCl and two liquid TEV-48574 formulations at 150 mg / mL (1B) and 100 mg / mL (2B) without arginine-HCl after storage at 40°C for up to 8 weeks, as determined using CGE non-reducing conditions (IgG+125 kDa peak %).

[0068] Fig.18B Shown are the percent purity (%) of two liquid TEV-48574 formulations at 150 mg / mL (1A) and 100 mg / mL (2A) with the addition of 100 mM arginine-HCl and two liquid TEV-48574 formulations at 150 mg / mL (1B) and 100 mg / mL (2B) without arginine-HCl after storage at 40°C for up to 8 weeks, as determined using CGE reducing conditions (heavy chain (HC) + light chain (LC) %).

[0069] Fig.19A Shown are the charge heterogeneity characteristics of two liquid TEV-48574 formulations of 150 mg / mL (1A) and 100 mg / mL (2A) with the addition of 100 mM arginine-HCl and two liquid TEV-48574 formulations of 150 mg / mL (1B) and 100 mg / mL (2B) without arginine-HCl after storage at 40°C for up to 8 weeks as the percentage (%) of the main species (main peak).

[0070] Fig.19B Shown are the charge heterogeneity characteristics of two liquid TEV-48574 formulations of 150 mg / mL (1A) and 100 mg / mL (2A) with the addition of 100 mM arginine-HCl and two liquid TEV-48574 formulations of 150 mg / mL (1B) and 100 mg / mL (2B) without arginine-HCl after storage at 40°C for up to 8 weeks, expressed as the percentage (%) of acidic species (acidic peak).

[0071] Fig.19CShown are the charge heterogeneity characteristics of two liquid TEV-48574 formulations of 150 mg / mL (1A) and 100 mg / mL (2A) with the addition of 100 mM arginine-HCl and two liquid TEV-48574 formulations of 150 mg / mL (1B) and 100 mg / mL (2B) without arginine-HCl after storage at 40°C for up to 8 weeks, expressed as the percentage (%) of basic species (basic peak).

[0072] Fig. 20 Shown are the thermal stabilities at time zero (T0) of two liquid TEV-48574 formulations of 150 mg / mL (1A) and 100 mg / mL (2A) with the addition of 100 mM arginine-HCl and two liquid TEV-48574 formulations of 150 mg / mL (1B) and 100 mg / mL (2B) without arginine-HCl. DETAILED DESCRIPTION

[0073] In order to more easily understand the present disclosure, some terms are first defined. As used in this application, unless otherwise expressly provided herein, each of the following terms should have the meaning described below. Additional definitions are set forth throughout the application. definition

[0074] Various terms related to various aspects of the present disclosure are used throughout this specification and claims. Unless otherwise specified, such terms will be given their ordinary meanings in the art. Other specifically defined terms should be interpreted in a manner consistent with the definitions provided herein.

[0075] The term "antibody" means an immunoglobulin molecule that is recognized by at least one antigen recognition site in the variable region of an immunoglobulin molecule and specifically binds to a target (such as a protein, polypeptide, peptide, carbohydrate, polynucleotide, lipid, or a combination thereof). As used herein, the term "antibody" encompasses complete polyclonal antibodies, complete monoclonal antibodies, chimeric antibodies, humanized antibodies, human antibodies, fusion proteins comprising antibodies, and any other modified immunoglobulin molecules, as long as these antibodies exhibit the desired biological activity. The antibody can be any of the five major classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM (referred to as α, δ, ε, γ, and μ, respectively, based on the identity of their heavy chain constant domains), or its subclass (isotype) (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2). Different classes of immunoglobulins have different and well-known subunit structures and three-dimensional configurations. The antibody can be a naked antibody or conjugated with other molecules (such as radioisotopes).

[0076] In the absence of explicit indication, and unless the context indicates otherwise, the term "antibody" includes monospecific antibodies, bispecific antibodies, or multispecific antibodies. In some aspects, the antibody is a bispecific antibody. The term "bispecific antibody" refers to an antibody that binds to two different epitopes.

[0077] The term "antibody fragment" refers to a part of a complete antibody. "Antigen binding fragment", "antigen binding domain" or "antigen binding region" refers to a part of a complete antibody that binds to an antigen. In the context of bispecific antibodies, an "antigen binding fragment" binds two antigens. An antigen binding fragment may contain the antigen recognition site of a complete antibody (e.g., a complementary determining region (CDR) sufficient to specifically bind to an 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, such as rodents (e.g., mice, rats or hamsters) and humans, or may be artificially produced.

[0078] "Monoclonal" antibody or its antigen binding fragment refers to a homogeneous antibody or antigen binding fragment population that participates in a highly specific binding of a single antigenic determinant or epitope. This is in contrast to polyclonal antibodies that typically include different antibodies for different antigenic determinants. The term "monoclonal" antibody or its antigen binding fragment encompasses complete and full-length monoclonal antibodies as well as antibody fragments (such as Fab, Fab', F(ab')2, Fv), single-chain (scFv) mutants, fusion proteins comprising antibody portions, and any other modified immunoglobulin molecules comprising antigen recognition sites. In addition, "monoclonal" antibody or its antigen binding fragment refers to such antibodies and their antigen binding fragments prepared in any number of ways, including but not limited to by hybridoma, phage selection, recombinant expression, and transgenic animals.

[0079] As used herein, the terms "variable region" or "variable region domain" are used interchangeably and are common in the art. A variable region typically refers to a part of an antibody, usually a part of a light chain or a heavy chain, typically around the amino terminal 110 to 120 amino acids or 110 to 125 amino acids in a mature heavy chain and about 90 to 115 amino acids in a mature light chain, whose sequence is different between antibodies and is used for the binding and specificity of a specific antibody to its specific antigen. The variability of the sequence is concentrated in those regions called complementary determining regions (CDRs), while the more highly conserved regions in the variable domains are referred to as framework regions (FRs). Without wishing to be constrained by any particular mechanism or theory, it is believed that the CDRs of light and heavy chains are primarily responsible for the interaction and specificity of antibodies to antigens. In some aspects of the present disclosure, the variable region is a human variable region. In some aspects of the present disclosure, the variable region comprises rodent or mouse CDRs and human framework regions (FRs). In a particular aspect of the present disclosure, the variable region is a primate (e.g., non-human primate) variable region. In some aspects of the disclosure, the variable regions comprise rodent or murine CDRs and primate (eg, non-human primate) framework regions (FRs).

[0080] The terms "VL" and "VL domain" are used interchangeably to refer to the light chain variable region of an antibody.

[0081] The terms "VH" and "VH domain" are used interchangeably to refer to the heavy chain variable region of an antibody.

[0082] The term "Kabat numbering" and similar terms are well-known in the art, and refer to a system in which the amino acid residues in the heavy chain and light chain variable region of an antibody or its antigen-binding fragment are numbered. In some aspects, CDR can be determined according to the Kabat numbering system (see, for example, Kabat EA and Wu TT (1971) Ann NY Acad Sci [Annuals of the New York Academy of Sciences] 190: 382-391 and Kabat EA et al., (1991) Sequences of Proteins of Immunological Interest [sequences of immunology-related proteins], the 5th edition, U.S. Department of Health and Human Services, NIH Publication No. 91-3242). Using the Kabat numbering system, the CDR in the antibody heavy chain molecule is typically present in amino acid positions 31 to 35 (which optionally can include one or two additional amino acids (referred to as 35A and 35B in the Kabat numbering scheme) after 35) (CDR1), amino acid positions 50 to 65 (CDR2) and amino acid positions 95 to 102 (CDR3). Using the Kabat numbering system, CDRs within an antibody light chain molecule typically occur at amino acid positions 24 to 34 (CDR1), amino acid positions 50 to 56 (CDR2), and amino acid positions 89 to 97 (CDR3).

[0083] Chothia refers to the location of the structural loops (Chothia and Lesk, J. Mol. Biol. 196:901-917 (1987)). When numbered using the Kabat numbering convention, the ends of the Chothia CDR-H1 loop vary between H32 and H34, depending on the length of the loop (this is because the Kabat numbering scheme places the insertion 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; 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.

[0084] As used herein, the terms "constant region" or "constant domain" are interchangeable and have common meanings in the art. The constant region is an antibody portion, such as the carboxyl terminal portion of a light chain and / or a heavy chain, which is not directly involved in the binding of the antibody to an antigen, but can exhibit various effector functions, such as interactions with Fc receptors. The constant region of an immunoglobulin molecule typically has an amino acid sequence that is more conservative relative to an immunoglobulin variable domain. In some aspects, an antibody or antigen-binding fragment comprises a constant region or portion thereof sufficient for antibody-dependent cell-mediated cytotoxicity (ADCC).

[0085] As used herein, the term "heavy chain" when used with respect to an antibody can refer to any of the different types, such as alpha (α), delta (δ), epsilon (ε), gamma (γ), and mu (μ), based on the amino acid sequence of the constant domain, which respectively produce the IgA, IgD, IgE, IgG, and IgM classes of antibodies, including subclasses of IgG, such as 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.

[0086] As used herein, the term "light chain" when used with respect to an antibody can refer to any of the different types, such as kappa (κ) or lambda (λ), 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.

[0087] The term "chimeric" antibody or antigen-binding fragment thereof refers to an antibody or antigen-binding fragment thereof in which the 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 derived from one mammalian species (e.g., mouse, rat, rabbit, etc.) with the desired specificity, affinity, and capacity, while the constant regions are homologous to sequences in an antibody or antigen-binding fragment thereof derived from another species (usually human) to avoid eliciting an immune response in this species.

[0088] The term "humanized" antibody or antigen-binding fragment thereof refers to a form of a non-human (e.g., murine) antibody or antigen-binding fragment that is a specific immunoglobulin chain, chimeric immunoglobulin, or fragment thereof containing minimal non-human (e.g., murine) sequence. Typically, a humanized antibody or antigen-binding fragment thereof is a human immunoglobulin in which residues from a complementary determining region (CDR) are replaced with residues from a CDR of a non-human species (e.g., mouse, rat, rabbit, hamster) with the desired specificity, affinity, and capacity ("CDR-grafted") (Jones et al., Nature [Nature] 321: 522-525 (1986); Riechmann et al., Nature [Nature] 332: 323-327 (1988); Verhoeyen et al., Science [Nature] 239: 1534-1536 (1988)). In some cases, some Fv framework region (FR) residues of human immunoglobulin are replaced by corresponding residues with desired specificity, affinity and ability in antibodies or fragments from non-human species. Humanized antibody or its Fab can be further modified by replacing other residues in Fv framework region and / or in non-human CDR residues, to improve and optimize the specificity, affinity and / or ability of antibody or its Fab. Usually, humanized antibody or its Fab will comprise variable domains, and these variable domains contain all or substantially all CDR regions corresponding to non-human immunoglobulin, and all or substantially all FR regions are those FR regions of human immunoglobulin consensus sequence. Humanized antibody or its Fab can also comprise at least a portion of immunoglobulin constant region or domain (Fc), typically at least a portion of constant region or domain of human immunoglobulin. Examples of methods for generating humanized antibodies are described in: U.S. Pat. No. 5,225,539; Roguska et al., Proc. Natl. Acad. Sci., USA [Proceedings of the National Academy of Sciences of the United States of America], 91(3):969-73 (1994), and Roguska et al., Protein Eng. [Protein Engineering] 9(10):895-904 (1996). In some aspects of the present disclosure, a "humanized antibody" is a resurfaced antibody.

[0089] The term "human" antibody or antigen-binding fragment thereof refers to an antibody or antibody-binding fragment thereof having an amino acid sequence derived from a human immunoglobulin locus, wherein such antibody or antigen-binding fragment is prepared using any technique known in the art. This definition of human antibodies or antigen-binding fragments thereof includes complete or full-length antibodies and fragments thereof.

[0090] "Binding affinity" generally refers to the strength of the sum 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 indicated, as used herein, "binding affinity" refers to the intrinsic binding affinity that reflects 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 as a dissociation constant (KD). Affinity can be measured and / or expressed in a variety of ways known in the art, including, but not limited to, the equilibrium dissociation constant (KD) and the equilibrium association constant (KA). KD is given by k off / k on , and KA is calculated by k off / k on The quotient of K is calculated. on refers to, for example, the association rate constant of an antibody or antigen-binding fragment thereof with an antigen, and k off Refers to, for example, the dissociation of an antibody or its antigen-binding fragment from an antigen. on and k off This can be done by techniques known to those of ordinary skill in the art (such as or KinExA) to determine.

[0091] As used herein, "epitope" is a term in the art, and refers to a local region of an antigen to which an antibody or its Fab can specifically bind. An epitope can be, for example, a continuous amino acid (linear or continuous epitope) of a polypeptide, or an epitope can be, for example, from two or more non-continuous regions (conformational, non-linear, discontinuous or non-continuous epitopes) of one or more polypeptides. In some aspects of the present disclosure, the epitope to which an antibody or its Fab specifically binds can be determined by, for example, NMR spectroscopy, X-ray diffraction crystallography, ELISA assay, hydrogen / deuterium exchange coupled to 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 method 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 comprising an antigen in complex with an antibody or antigen-binding fragment can be studied using well-known X-ray diffraction techniques and can be improved using computer software such as X-PLOR (Yale University, 1992, published by Molecular Simulations, Inc.; see, e.g., Meth Enzymol (1985) Vols. 114 and 115, Wyckoff HW et al., eds.; US2004 / 0014194) and BUSTER (Bricogne G (1993) Acta Crystallogr D Biol Crystallogr 49(Pt 1):37-60; Bricogne G (1997) Meth Enzymol 276A:361-423, Carter CW ed.; Roversi P et al., (2000) Acta Crystallogr D Biol Crystallogr 50(Pt 2):54-57. Crystallogr [Acta Crystallogrica Sinica D - Structural Biology] 56 (Pt 10): 1316-1323). Mutagenesis mapping studies can be performed using any method known to those skilled in the art.For a description of mutagenesis techniques, including alanine scanning mutagenesis, see, for example, Champe M et al., (1995) J Biol Chem 270: 1388-1394 and Cunningham BC & Wells JA (1989) Science 244: 1081-1085.

[0092] "Separated" polypeptides, antibodies, polynucleotides, vectors, cells or compositions are polypeptides, antibodies, polynucleotides, vectors, cells or compositions that are in a form that does not exist in nature. Separated polypeptides, antibodies, polynucleotides, vectors, cells or compositions include those that have been purified to the extent that they are no longer in the form in which they are found in nature. In some aspects of the present disclosure, separated antibodies, polynucleotides, vectors, cells or compositions are substantially pure. As used herein, "substantially pure" refers to at least 50% pure (i.e., free of contaminants), at least 90% pure, at least 95% pure, at least 98% pure or at least 99% pure material.

[0093] The terms "polypeptide," "peptide," and "protein" are used interchangeably herein to refer to polymers of amino acids of any length. A polymer may be linear or branched, it may contain modified amino acids, and it may be interrupted by non-amino acids. The term also encompasses amino acid polymers that have been modified naturally or by intervention; for example, disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or any other manipulation or modification, such as conjugation with a labeling component. Also included in the definition are, for example, polypeptides containing one or more analogs of amino acids (including, for example, non-natural amino acids, etc.), as well as other modifications known in the art. It should be understood that because the polypeptides of the present disclosure are antibody-based, in some aspects of the present disclosure, the polypeptides may exist as single chains or associated chains.

[0094] As used herein, the term "pharmaceutical formulation" refers to a preparation that is in a form that permits the biological activity of the active ingredient to be effective and does not contain additional components that are unacceptably toxic to the subject to which the formulation is to be administered. The formulation can be sterile. In some aspects, the formulation is suitable for therapeutic use in human subjects.

[0095] As used herein, the terms "administer / administering / administration" and the like refer to methods that can be used to enable delivery of a drug (e.g., an anti-TL1A antibody or 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 found, 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).

[0096] As used herein, the term "combination" or "administered in combination" means that the antibodies or antigen-binding fragments thereof described herein can be administered together with one or more additional therapeutic agents. In some aspects of the combinations provided herein, the antibodies or antigen-binding fragments thereof can be administered simultaneously or sequentially with one or more additional therapeutic agents. In some aspects, the antibodies or antigen-binding fragments thereof described herein can be administered together with one or more additional therapeutic agents in the same composition or in different compositions.

[0097] The terms "subject" and "patient" are used interchangeably and include any animal. In some aspects, the subject is a mammal, including companions (e.g., cats, dogs) and farm mammals (e.g., pigs, horses, cattle) and rodents, including mice, rabbits and rats, guinea pigs and other rodents. In some aspects, the subject is a non-human primate, such as a cynomolgus monkey. In some aspects, the subject is a human.

[0098] The term "therapeutically effective amount" refers to an amount of a drug (e.g., an anti-TL1A antibody (e.g., TEV-48574) or an antigen-binding fragment thereof) that is effective to treat a disease or disorder in a subject. Terms such as "treating / treatment / to treat" and "alleviating / to alleviate" refer to utilizing a method to obtain a beneficial or desired clinical result, including but not limited to methods for achieving such a beneficial or desired clinical result, wherein the clinical result may include a therapeutic measure that improves, cures, alleviates, attenuates the symptoms of a pathological condition or disorder, and / or stops the progression of a pathological condition or disorder. Those in need of treatment may include those who have been diagnosed with a disorder or are suspected of having a disorder.

[0099] In the context of antibody-antigen interactions, "specificity" is not necessarily an absolute term, but can constitute a relative term, indicating the degree of selectivity of the antibody for antigen-positive cells compared to antigen-negative cells. The specificity of the antibody for antigen-positive cells is mediated by the variable region of the antibody, and usually by the complementarity determining region (CDR) of the antibody. The construct can have from about 100-fold to about 1000-fold specificity for antigen-positive cells compared to antigen-negative cells.

[0100] As used herein, the term "recombinant" includes the expression of genes produced by genetic engineering or otherwise by laboratory manipulation.

[0101] As used herein, the term "TEV 48574" or "TEV-48574" refers to a highly potent, fully human immunoglobulin G (IgG) subclass 1 (IgG1) (λ) monoclonal antibody (mAb) that binds to human, cynomolgus monkey, and rat TL1A with high affinity. 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, the antibody prevents activation of the DR3 signaling pathway. TEV-48574 also inhibits the binding of TL1A to DcR3, although TEV-48574 inhibits the TL1A-DR3 interaction more than the TL1A-DcR3 interaction. TEV-48574 exhibits 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 as "320-587" in US Patent No. 10,138,296, which is incorporated herein by reference in its entirety.

[0102] 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 inhibitor (VEGI), a member of the TNF superfamily, which is expressed by antigen presenting cells (including dendritic cells, B cells and macrophages), CD4+ and CD8+ T cells and endothelial cells. TL1A can be expressed on the cell surface or secreted as a soluble cytokine. TL1A's cognate signaling receptor death receptor 3 (DR3) is expressed by a variety of cells, including CD4+ and CD8+ T cells, NK cells, NKT cells and FOXP3+ regulatory T (Treg) cells and type 2 and type 3 innate lymphoid cells (ILC2 and ILC3). TL1A can also bind to a decoy receptor (DcR3), which is a competitive inhibitor of DR3. DcR3 also acts as a decoy receptor for Fas-ligand (Fas-L) and lymphotoxin-like inducible protein, which competes with glycoprotein D for binding to herpes virus entry mediator (LIGHT) on T cells. Thus, DcR3 is an important regulator of several signal transduction pathways.

[0103] As used in the disclosure and claims, the singular forms "a," "an," and "the" include plural forms unless the context clearly dictates otherwise.

[0104] It should be understood that wherever aspects of the present disclosure are described herein with the language "comprising," otherwise similar aspects described with the phrases "consisting of" and / or "consisting essentially of" are also provided.

[0105] Unless expressly stated or obvious from the context, as used herein, the term "or" is to be understood to be inclusive. The term "and / or" as used in phrases such as "A and / or B" herein is intended to include both "A and B", "A or B", "A" and "B". Similarly, the term "and / or" as used in phrases such as "A, B and / or C" is intended to cover each of the following: 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).

[0106] 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 value or range remains within the intended meaning of the listed value or range. As will be appreciated by those skilled in the art, references herein to "about" values ​​or ranges include (and describe) references to the value or range itself. For example, a description referring to "about X" includes a description of "X".

[0107] Any composition or method provided herein can be combined with one or more of any other compositions and methods provided herein.

[0108] Units, prefixes and symbols are all expressed in the form accepted by the International System of Units (SI). Numerical ranges include numbers that limit the range. The titles provided herein are not limitations on the various aspects of the present disclosure, which can be obtained by reference to the entire specification. Therefore, the terms of the definitions immediately below are more fully defined by reference to the specification (in its entirety). Pharmaceutical composition

[0109] The present disclosure provides pharmaceutical compositions comprising antibodies or antigen-binding fragments that bind to TL1A. The pharmaceutical compositions may comprise any antibody or antigen-binding fragment thereof described and / or exemplified herein and an acceptable carrier, such as a pharmaceutically acceptable carrier. Suitable carriers include any culture medium that does not interfere with the biological activity of the antibody or antigen-binding fragment thereof and is nontoxic to the host to which it is administered. The pharmaceutical composition may be formulated for administration to a subject in any suitable dosage form.

[0110] Pharmaceutical compositions suitable for administration to human patients are typically formulated for parenteral administration, for example in a liquid vehicle, or suitable for reconstitution into a liquid solution or suspension for intravenous or subcutaneous administration.

[0111] Usually, such compositions typically include a pharmaceutically acceptable carrier. As used herein, the term "pharmaceutically acceptable" means approved by a government regulatory agency or listed in the U.S. Pharmacopeia or another generally recognized pharmacopoeia for use in animals, particularly in humans. The term "carrier" refers to a diluent, adjuvant, excipient or vehicle used together with the compound. Such a pharmaceutical carrier can be a sterile liquid. Water or saline solutions and aqueous glucose and glycerol solutions can also be used as carriers, particularly for injection solutions. Liquid compositions for parenteral administration can be formulated for administration by injection or continuous infusion. The route of administration by injection or infusion includes intravenous and subcutaneous.

[0112] In some aspects, 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); wherein the antibody or antigen-binding fragment thereof comprises: (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, 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; or (iii) a heavy chain comprising the amino acid sequence of SEQ ID NO:9 and a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO:10. NO:10 light chain of the amino acid sequence; (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 aspects, the pharmaceutical formulation is lyophilized. In some aspects, the pharmaceutical formulation is liquid.

[0113] In some aspects, 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); wherein the antibody or antigen-binding fragment thereof comprises: (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, 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; or (iii) a heavy chain comprising the amino acid sequence of SEQ ID NO:9 and a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO:10. NO:10 light chain of the amino acid sequence; (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 aspects, the pharmaceutical formulation is lyophilized. In some aspects, the pharmaceutical formulation is liquid.

[0114] In some aspects, 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); wherein the antibody or antigen-binding fragment thereof comprises: (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, 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; or (iii) a heavy chain comprising the amino acid sequence of SEQ ID NO:9 and a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO:10. NO: light chain of the amino acid sequence of 10; (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.

[0115] In some aspects, 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); wherein the antibody or antigen-binding fragment thereof comprises: (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, 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; or (iii) a heavy chain comprising the amino acid sequence of SEQ ID NO:9 and a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO:10. NO: light chain of the amino acid sequence of 10; (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.

[0116] In some aspects, the pharmaceutical formulations provided herein are stored at room temperature for 24 hours or at 2 ° C-8 ° C for 24 hours, 72 hours or 10 days after having a pH of 6.0 ± 0.5. In some aspects, the pharmaceutical formulations provided herein are stored at room temperature for 24 hours or at 2 ° C-8 ° C for 24 hours, 72 hours or 10 days after having an osmotic pressure from 200mOsm / kg to 500mOsm / kg. In some aspects, the pharmaceutical formulations provided herein are stored at 2 ° C-8 ° C for 24 hours, 72 hours or 10 days after having at least 99% antibody monomer content. In some aspects, the pharmaceutical formulations provided herein are stored at 2 ° C-8 ° C for 24 hours, 72 hours or 10 days after having no significant changes in charge heterogeneity characteristics. In some aspects, the pharmaceutical formulations provided herein are stored at room temperature for 24 hours or at 2 ° C-8 ° C for 24 hours, 72 hours or 10 days after having no significant changes in purity. In some aspects, the pharmaceutical formulations provided herein are stored at room temperature for 24 hours or stored at 2 ° C-8 ° C for 24 hours, 72 hours or 10 days with at least 90% purity. In some aspects, the pharmaceutical formulations provided herein are stored at room temperature for 24 hours or stored at 2 ° C-8 ° C for 24 hours, 72 hours or 10 days after there is no significant change in particle concentration. In some aspects, the pharmaceutical formulations provided herein are stored at 2 ° C-8 ° C for up to 24 months and there is no significant difference in visual appearance. In some aspects, the pharmaceutical formulations provided herein are stored at 2 ° C-8 ° C for up to 36 months and there is no significant difference in visual appearance. In some aspects, the pharmaceutical formulations provided herein are stored at 2 ° C-8 ° C, 25 ° C or 40 ° C for up to 24 months and there is no significant difference in protein concentration, osmotic pressure or viscosity. In some aspects, the pharmaceutical formulations provided herein are stored at 2 ° C-8 ° C, 25 ° C or 40 ° C for up to 36 months and there is no significant difference in protein concentration, osmotic pressure or viscosity. In some respects, the pharmaceutical formulations provided herein are stored at 2 ℃-8 ℃ for up to 24 months and have ≥ (greater than or equal to) 95% monomer content, ≤ (less than or equal to) 5.0% dimer content or no significant difference in low molecular weight content. In some respects, the pharmaceutical formulations provided herein are stored at 2 ℃-8 ℃ for up to 36 months and have ≥ (greater than or equal to) 95% monomer content, ≤ (less than or equal to) 5.0% dimer content or no significant difference in low molecular weight content. In some respects, the pharmaceutical formulations provided herein are stored at 2 ℃-8 ℃ for up to 24 months and have ≥ (greater than or equal to) 90% purity. In some respects, the pharmaceutical formulations provided herein are stored at 2 ℃-8 ℃ for up to 36 months and have ≥ (greater than or equal to) 90% purity.In some aspects, the pharmaceutical formulations provided herein have a main substance content of from 50% to 90%, an acidic substance content of from 10% to 40%, and / or a basic substance content of from 0% to 20% after being stored at 2°C-8°C for up to 24 months. In some aspects, the pharmaceutical formulations provided herein have a main substance content of from 50% to 90%, an acidic substance content of from 10% to 40%, and / or a basic substance content of from 0% to 20% after being stored at 2°C-8°C for up to 36 months. In some aspects, the pharmaceutical formulations provided herein have no significant differences in sub-visible particle content after being stored at 2°C-8°C, 25°C or 40°C for up to 24 months. In some aspects, the pharmaceutical formulations provided herein have no significant differences in sub-visible particle content after being stored at 2°C-8°C, 25°C or 40°C for up to 36 months. In some aspects, the pharmaceutical formulations provided herein are stored at 2°C-8°C, 25°C or 40°C for up to 24 months after the oxidation of methionine 81 and / or methionine 254 of TEV-48574 and / or the deamidation of asparagine 317 of TEV-48574. In some aspects, the pharmaceutical formulations provided herein are stored at 2°C-8°C, 25°C or 40°C for up to 36 months after the oxidation of methionine 81 and / or methionine 254 of TEV-48574 and / or the deamidation of asparagine 317 of TEV-48574. In some aspects, the pharmaceutical formulations provided herein have a relative potency of from 70% to 135% measured by enzyme-linked immunosorbent assay (ELISA) after storage at 2°C-8°C for up to 24 months. In some aspects, the pharmaceutical formulations provided herein are stored at 2 ℃-8 ℃ for up to 36 months and have a relative efficacy of 70% to 135% measured by enzyme-linked immunosorbent assay (ELISA). In some aspects, the pharmaceutical formulations provided herein are stored at 2 ℃-8 ℃, 25 ℃ or 40 ℃ for up to 6 months and have no significant difference in thermal stability. In some aspects, the pharmaceutical formulations provided herein are stored at 2 ℃-8 ℃ for up to 36 months and have no significant difference in thermal stability. In some aspects, the pharmaceutical formulations provided herein are stored at 2 ℃-8 ℃, 25 ℃ or 40 ℃ for up to 3 months and have no significant difference in secondary and / or tertiary protein structure. In some aspects, the pharmaceutical formulations provided herein are stored at 2 ℃-8 ℃ for up to 36 months and have no significant difference in secondary protein structure. In some aspects, the pharmaceutical formulations provided herein are stored at 2 ℃-8 ℃ for up to 24 months and have no significant difference in the concentration of polysorbate-80. method

[0117] The present disclosure provides a method for treating a disease in a subject in need thereof, the method comprising administering to the subject any pharmaceutical formulation provided herein or any container provided herein. In some aspects, the disease is a respiratory disease, a gastrointestinal disease, a skin disease, or arthritis.

[0118] In some aspects, the respiratory disease is asthma, chronic obstructive pulmonary disease (COPD), pulmonary fibrosis, pulmonary sarcoidosis, allergic rhinitis, or cystic fibrosis.

[0119] In some aspects, the gastrointestinal disease is inflammatory bowel disease, Crohn's disease, colitis, ulcerative colitis, eosinophilic esophagitis, or irritable bowel syndrome.

[0120] In some aspects, the arthritis is rheumatoid arthritis.

[0121] In some aspects, the skin disease is atopic dermatitis, eczema, or scleroderma.

[0122] In some aspects, the subject is a human subject. In some aspects, the subject is a non-human primate, such as a cynomolgus monkey. In some aspects, the subject is a non-human mammal, such as a mouse, rat, guinea pig, cat, pig, rabbit or dog. Anti-TL1A antibody

[0123] In some aspects, the formulations provided herein comprise an anti-TL1A antibody or antigen binding fragment thereof. In some aspects, the antibody or antigen binding fragment thereof inhibits the ability of TL1A to interact with DR3, and in some aspects, also inhibits the ability of TL1A to interact with DcR3 and further inhibits signal transduction induced by the interaction of TL1A with DR3. In some aspects, the antibody or antigen binding fragment thereof has enhanced potency relative to antibody 320-179. In some aspects, the antibody or antigen binding fragment thereof has enhanced affinity for TL1A relative to antibody 320-179.

[0124] In some aspects, an antibody or antigen-binding fragment thereof as 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 is capable of inhibiting the interaction between TL1A and DR3.

[0125] In some aspects, an antibody or antigen-binding fragment thereof as disclosed herein comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 7. In some aspects, the antibody or antigen-binding fragment thereof comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 8. In some aspects, 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.

[0126] In some aspects, the heavy chain variable region is connected to a heavy chain constant region. In some aspects, the heavy chain constant region is an IgG1, IgG2 or IgG4 heavy chain constant region. In some aspects, the heavy chain variable region of SEQ ID NO:7 is connected 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 aspects, the heavy chain constant region provided herein does not comprise a C-terminal lysine. In some aspects, the light chain variable region of SEQ ID NO:8 is connected to a lambda human light chain constant region (e.g., SEQ ID NO:29) such that the light chain comprises SEQ ID NO:10.

[0127] In some aspects, the antibodies or antigen-binding fragments thereof as disclosed herein are monoclonal antibodies or antigen-binding fragments thereof.

[0128] In some aspects, the antibody or antigen-binding fragment thereof disclosed herein is a recombinant antibody or antigen-binding fragment thereof. In some aspects, the recombinant antibody is a full-length antibody. In some aspects, the recombinant antibody is a monoclonal antibody.

[0129] In some aspects, an antibody or antigen-binding fragment thereof as disclosed herein binds to TL1A with enhanced affinity relative to the 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 aspects, an antibody or antigen-binding fragment thereof as disclosed herein has enhanced potency relative to the 320-179 anti-TL1A antibody. The enhanced potency can be at least about 10 times, at least about 12 times, at least about 13 times, at least about 15 times, at least about 20 times, at least about 25 times, at least about 27 times, at least about 40 times, about 10 times to about 40 times, about 12 times to about 40 times, about 13 times to about 40 times, about 15 times to about 40 times, about 20 times to about 40 times, about 25 times to about 40 times, or about 27 times to about 40 times greater potency relative to the 320-179 anti-TL1A antibody. The potency enhancement factor can be determined in TF-1 cells according to a TL1A-induced caspase potency assay. See, e.g., U.S. Patent No. 10,138,296. The 320-179 antibody has favorable biophysical properties, is an effective inhibitor of TL1A, and has a low predicted immunogenicity profile. U.S. Patent No. 10,138,296 and U.S. Publication No. 2014 / 0255302 are incorporated herein by reference in their entirety.

[0130] In some aspects, an antibody as disclosed herein is TEV-48574, also referred to as the 320-587 antibody in US Pat. No. 10,138,296 (in this disclosure, VH is SEQ ID NO: 3 and VL is SEQ ID NO: 4).

[0131] In some aspects, an antibody or antigen-binding fragment thereof as disclosed herein 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, and binds to TL1A with enhanced affinity relative to an 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. Anti-TL1A 320-179 antibodies have been previously described in U.S. Pat. No. 10,138,296 (in this publication, VH is SEQ ID NO: 1 and VL is SEQ ID NO: 2) and U.S. Publication No. 2014 / 0255302 (in this publication, VH is SEQ ID NO: 186 and VL is SEQ ID NO: 199). The 320-179 antibody has favorable biophysical properties, is a potent inhibitor of TL1A, and has a low predicted immunogenicity profile. US Patent No. 10,138,296 and US Publication No. 2014 / 0255302 are incorporated herein by reference in their entirety.

[0132] In some aspects, an antibody or antigen-binding fragment thereof as disclosed herein comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 7 and a light chain variable region anti-TL1A antibody comprising the amino acid sequence of SEQ ID NO: 8, and has enhanced potency relative to the 320-179 anti-TL1A antibody. The enhanced potency can be at least about 10 times, at least about 12 times, at least about 13 times, at least about 15 times, at least about 20 times, at least about 25 times, at least about 27 times, or at least about 40 times greater potency relative to the 320-179 anti-TL1A antibody. The potency enhancement factor can be determined in TF-1 human erythroleukemia antibody cells (ATCC: CRL-2003) according to the TL1A-induced caspase potency assay. See, e.g., U.S. Pat. No. 10,138,296.

[0133] In some aspects, the antibodies or antigen-binding fragments thereof as disclosed herein are full-length antibodies. In some aspects, the antibodies as disclosed herein are monoclonal antibodies.

[0134] In some aspects, the antibodies or antigen-binding fragments thereof disclosed herein comprise a human IgG1 heavy chain constant region, a human IgG2 heavy chain constant region, or a human IgG4 heavy chain constant region, or any allotype thereof. The human IgG1 heavy chain constant region may be selected from 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: 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 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 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 should be understood that the IgG4 heavy chain can be used without the stabilizing substitution S228P (e.g., IgG4 and YTE alone, IgG4 and YTE and ΔK, or IgG4 and ΔK alone).

[0135] In some aspects, the antibodies or antigen-binding fragments thereof as 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.

[0136] In some aspects, an antibody or antigen-binding fragment thereof as disclosed herein binds to human TL1A, and may bind to one or more of cynomolgus monkey TL1A, mouse TL1A, rat TL1A, guinea pig TL1A, cat TL1A, dog TL1A, pig TL1A, or rabbit TL1A. In some aspects, for example, if the epitope is shared, the antibody or antigen-binding fragment thereof may bind to TL1A of multiple different species. In some aspects, human TL1A comprises the amino acid sequence of SEQ ID NO:34, SEQ ID NO:35, or SEQ ID NO:36. In some aspects, cynomolgus monkey TL1A comprises the amino acid sequence of SEQ ID NO:37. In some aspects, mouse TL1A comprises the amino acid sequence of SEQ ID NO:38. In some aspects, rat TL1A comprises the amino acid sequence of SEQ ID NO:39. In some aspects, guinea pig TL1A comprises the amino acid sequence of SEQ ID NO:40. In some aspects, cat TL1A comprises the amino acid sequence of SEQ ID NO:41. In some aspects, the pig TL1A comprises the amino acid sequence of SEQ ID NO: 42. In some aspects, the rabbit TL1A comprises the amino acid sequence of SEQ ID NO: 43. In some aspects, the dog TL1A comprises the amino acid sequence of SEQ ID NO: 44.

[0137] The antibodies or antigen-binding fragments thereof disclosed herein have a binding affinity for an epitope on TL1A, the binding affinity comprising an equilibrium dissociation constant (K D ), the equilibrium dissociation constant can be determined by kinetic exclusion (such as The K of TL1A binding determined by kinetic exclusion assay was measured using the kinetic exclusion assay (Sapidyne Instruments, Boise, ID). D Less than about 1000 pM. In some aspects, the K of TL1A binding determined by kinetic exclusion assay is D In some aspects, the K of TL1A binding determined by kinetic exclusion assay 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 Less than about 100 pM.

[0138] K of TL1A binding determined by kinetic exclusion assay D The K of TL1A binding determined by kinetic exclusion assay is from about 10 pM to about 100 pM. D The K of TL1A binding determined by kinetic exclusion assay is from about 25 pM to about 75 pM. D The K of TL1A binding determined by kinetic exclusion assay is from about 30 pM to about 60 pM.D The K of TL1A binding determined by kinetic exclusion assay is from about 30 pM to about 50 pM. D The K of TL1A binding determined by kinetic exclusion assay is from about 35 pM to about 50 pM. D The K of TL1A binding determined by kinetic exclusion assay is from about 36 pM to about 46 pM. D The K of TL1A binding determined by kinetic exclusion assay is from about 38 pM to about 44 pM. D The K of TL1A binding determined by kinetic exclusion assay is from about 39 pM to about 43 pM. D The K of TL1A binding determined by kinetic exclusion assay is from about 40 pM to about 45 pM. D The K of TL1A binding determined by kinetic exclusion assay is from about 35 pM to about 42 pM. D The K of TL1A binding determined by kinetic exclusion assay is about 40 pM. D The K of TL1A binding determined by kinetic exclusion assay is about 41 pM. D It may be about 42 pM. Kinetic exclusion assays may use either the antibody molecule or the TL1A molecule as a constant binding partner and another molecule as a titrant.

[0139] The antibodies or antigen-binding fragments thereof disclosed herein can bind to TL1A positive cells. The antibodies or antigen-binding fragments thereof disclosed herein can bind to TL1A positive cells with an EC 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 Values ​​were combined with TL1A-positive cells.

[0140] As disclosed herein, the antibody or its antigen-binding fragment can be monoclonal. In some aspects, as disclosed herein, the antibody or its antigen-binding fragment is a full-length antibody comprising two heavy chains and two light chains. In some aspects, as disclosed herein, the antibody or its antigen-binding fragment comprises a derivative or fragment or part of the antibody that retains the antigen-binding specificity of the full-length antibody and also retains most or all of the affinity. For example, the derivative can include at least one variable region (heavy chain or light chain variable region). The derivative can include at least two variable regions, such as 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 multi-epitope specificity, bispecific antibodies, multispecific antibodies, double antibodies, single-chain molecules, and FAb, F(Ab')2, Fd, Fabc and Fv molecules, single-chain (Sc) antibodies, single-chain Fv antibodies (scFv), single antibody light chains, single antibody heavy chains, fusions between antibody chains and other molecules, heavy chain monomers or dimers, light chain monomers or dimers, dimers consisting of one heavy chain and one light chain, and other polymers. Single-chain Fv antibodies can be multivalent. All antibody isotypes can be used to produce antibody derivatives, fragments and parts. Antibody derivatives, fragments and / or parts can be recombinantly produced and expressed by any cell type of prokaryotic or eukaryotic.

[0141] In a full-length antibody, each heavy chain comprises a heavy chain variable region (abbreviated herein as HCVR or VH) and a heavy chain constant region. The heavy chain constant region comprises three domains, CH1, CH2 and CH3. Each light chain comprises a light chain variable region (abbreviated herein as LCVR or VL) and a light chain constant region. The light chain constant region comprises one domain, CL. The VH and VL regions can be further subdivided into regions of high variability, called complementarity determining regions (CDRs), interspersed with more conserved regions, called framework regions (FRs). Each VH and VL consists of three CDRs and four FRs arranged in the following order from amino terminus to carboxyl terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. Typically, the antigen binding properties of an antibody are less likely to be disturbed by changes in the FR sequences than by changes in the CDR sequences. The immunoglobulin molecule 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 of immunoglobulin molecule.

[0142] In some aspects, antibodies or their antigen-binding fragments as disclosed herein are fully human. Fully human antibodies are those antibodies wherein the entire molecule is human or otherwise human-derived, or comprising the same amino acid sequence as the human form of the antibody. Fully human antibodies include those obtained from human V gene libraries, for example, wherein the human genes encoding the variable regions of antibodies are recombinantly expressed. Fully human antibodies can be expressed in cells of other organisms (e.g., mice and xenomouse technology) or from other organisms transformed with genes encoding human antibodies. However, fully human antibodies may include amino acid residues not encoded by human sequences, such as mutations introduced by random or site-directed mutagenesis.

[0143] In some aspects, antibodies or antigen-binding fragments thereof as disclosed herein comprise a non-immunoglobulin derived protein framework. For example, reference can be made to Ku and Schutz, Proc. Natl. Acad. Sci. USA, 92:6552-6 (1995), which describes a four-helix bundle protein cytochrome b562 having two randomized loops to generate CDRs that have been selected for antigen binding.

[0144] In some aspects, antibodies or antigen-binding fragments thereof as disclosed herein comprise post-translational modifications or moieties that may affect antibody activity or stability. These modifications or moieties include, but are not limited to, methylated, acetylated, glycosylated, sulfated, phosphorylated, carboxylated and amidated moieties and other moieties well known in the art. Moieties include any chemical group or combination of groups that are often found on immunoglobulin molecules in nature or added to antibodies by recombinant expression systems (including prokaryotic and eukaryotic expression systems).

[0145] Examples of side chain modifications contemplated by the present disclosure include modifications of amino groups, such as reductive alkylation by reaction with an aldehyde followed by reduction with NaBH4; amidination 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; and pyridoxylation of lysine with pyridoxal-5-phosphate followed by reduction with NaBH4.

[0146] The guanidinium group of arginine residues can be modified by forming heterocyclic condensation products with reagents such as 2,3-butanedione, phenylglyoxal and glyoxal. Carboxyl groups can be formed via O-acylisourea followed by derivatization to, for example, the corresponding amide for modification by carbodiimide activation. Sulfonyl groups can be modified by methods such as carboxymethylation with iodoacetic acid or iodoacetamide; oxidation with performic acid to cysteine; formation of mixed disulfides with other thiol compounds; reaction with maleimide, maleic anhydride or other substituted maleimides; formation of mercury derivatives using 4-chloromercurybenzoate, 4-chloromercurybenzenesulfonic acid, phenylmercuric chloride, 2-chloromercury-4-nitrophenol and other mercury compounds; carbamoylation with cyanate at alkaline pH. Tryptophan residues can be modified by, for example, oxidation with N-bromosuccinimide or alkylation of the indole ring with 2-hydroxy-5-nitrobenzyl bromide or sulfinyl halides. On the other hand, tyrosine residues can be modified by nitration with tetranitromethane to form 3-nitrotyrosine derivatives. Modification of the imidazole ring of histidine residues can be achieved by alkylation with iodoacetic acid derivatives or N-ethoxylation with diethylpyrocarbonate.

[0147] In some aspects, antibodies or their antigen-binding fragments as disclosed herein comprise one or more modifications that regulate serum half-life and biodistribution, including but not limited to modifications that regulate the interaction of antibodies with neonatal Fc receptors (FcRn), which are receptors that play a key role in protecting IgG from catabolism and maintaining high serum antibody concentrations. Serum half-life regulation modifications can occur in the Fc region of IgG1, IgG2, or IgG4, including triple substitutions of M252Y / S254T / T256E ("YTE" substitutions, numbered according to the EU numbering system (Edelman, GM et al., Proc. Natl. Acad. USA [Proceedings of the National Academy of Sciences of the United States], 63:78-85 (1969)), as described in U.S. Patent No. 7,083,784. Other substitutions can occur at positions 250 and 428 (see, e.g., U.S. Patent No. 7,217,797) and positions 307, 380, and 434 (see, e.g., PCT Publication No. WO 00 / 042072). Examples of constant domain amino acid substitutions that modulate binding to Fc receptors and subsequent functions mediated by these receptors (including FcRn binding and serum half-life) are described in U.S. Publication Nos. 2009 / 0142340, 2009 / 0068175, and 2009 / 0092599. Antibodies of any class may omit or remove the heavy chain C-terminal lysine to reduce heterogeneity (ΔK). Substitution of S228P (EU numbering) 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 be present simultaneously with YTE and / or ΔK modifications.

[0148] Antibodies or their antigen-binding fragments as disclosed herein may be labeled, bound or conjugated to any chemical or biomolecular moiety. Labeled antibodies may be used for treatment, diagnosis or basic research applications. Such labels / conjugates may be detectable, such as fluorescent dyes, electrochemiluminescent probes, quantum dots, radioactive labels, enzymes, fluorescent proteins, luminescent proteins and biotin.

[0149] The antibodies or antigen-binding fragments thereof as disclosed herein can be derivatized with known protecting / blocking groups to prevent proteolytic cleavage or to enhance activity or stability.

[0150] Administration of an antibody or antigen binding fragment thereof as disclosed herein may include subcutaneous administration of the antibody or antigen binding fragment thereof. Thus, in some aspects, the formulations provided herein are formulated for subcutaneous administration. Administration may include intravenous administration of the antibody or antigen binding fragment thereof. Thus, in some aspects, the formulations provided herein are formulated for intravenous administration.

[0151] The disclosure further provides compositions for use according to any of the methods disclosed herein. Polynucleotides and vectors

[0152] Disclosed herein are polynucleotide sequences encoding recombinant antibodies or their antigen-binding fragments and their subdomains (e.g., FR and CDR). Polynucleotides include, but are not limited to, RNA, DNA, cDNA, hybrids of RNA and DNA, and single-stranded, double-stranded or triple-stranded chains of RNA, DNA or their hybrids. Polynucleotides may include nucleic acid sequences encoding the heavy chain variable region and / or light chain variable region of an antibody as described or exemplified herein. The complement sequence of a polynucleotide sequence is also within the scope of this disclosure.

[0153] In some aspects, the polynucleotide may comprise a nucleic acid sequence encoding an antibody heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 7. The polynucleotide encoding the amino acid sequence of SEQ ID NO: 7 may comprise a nucleic acid sequence of SEQ ID NO: 30 or SEQ ID NO: 31.

[0154] In some aspects, 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 a nucleic acid sequence of SEQ ID NO: 32 or SEQ ID NO: 33.

[0155] In some aspects, the polynucleotide may include 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 comprising an amino acid sequence of SEQ ID NO: 7, and the second nucleic acid sequence may encode an antibody light chain variable region comprising an amino acid sequence of SEQ ID NO: 8. The first nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 7 may include a 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 include a nucleic acid sequence of SEQ ID NO: 32 or SEQ ID NO: 33.

[0156] In some aspects, the polynucleotide may include 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 aspects, the polynucleotide includes a first nucleic acid sequence encoding an antibody heavy chain variable region comprising an 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, for example, a polynucleotide comprising a nucleic acid sequence of SEQ ID NO: 31.

[0157] In some aspects, the polynucleotide may include 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 aspects, the polynucleotide includes a first nucleic acid sequence encoding an antibody light chain variable region comprising an amino acid sequence of SEQ ID NO: 8 and a second nucleic acid sequence encoding a lambda light chain constant region of SEQ ID NO: 29, for example, a polynucleotide comprising a nucleic acid sequence of SEQ ID NO: 33.

[0158] Any polynucleotide described or exemplified herein may be contained in a vector. Therefore, vectors containing polynucleotides are provided as part of the present disclosure. These vectors may be expression vectors. Therefore, a recombinant expression vector containing a sequence encoding a polypeptide of interest is provided. The expression vector may contain one or more additional sequences, such as but not limited to regulatory sequences, selection markers, purification tags, or polyadenylation signals. Such regulatory elements may include transcriptional promoters, enhancers, mRNA ribosome binding sites, or sequences that control transcription and translation termination.

[0159] Expression vectors, especially mammalian expression vectors, can include one or more non-transcribed elements, such as a replication origin, a suitable promoter and enhancer linked to the gene to be expressed, other 5' or 3' flanking non-transcribed sequences, 5' or 3' non-translated sequences (such as essential ribosome binding sites), polyadenylation sites, splice donor and acceptor sites, or transcription termination sequences. A replication origin that confers replication ability in a specific host can also be incorporated.

[0160] These vectors can be used to transform any of a variety of host cells well known to those skilled in the art and host cells capable of expressing antibodies. Vectors include, but are not limited to, plasmids, phagemids, cosmids, baculovirus shuttle vectors, bacterial artificial chromosomes (BAC), yeast artificial chromosomes (YAC) and baculovirus, as well as other bacteria, eukaryotic, yeast and viral vectors. Suitable host cells include, but are not limited to, CHO cells, NSO cells, HEK293 cells, or any eukaryotic stable cell lines known or produced, and also include bacteria, yeast and insect cells.

[0161] In some aspects, the antibodies or antigen-binding fragments thereof provided herein are produced in human embryonic kidney cells. In some aspects, the antibodies or antigen-binding fragments thereof provided herein are produced in HEK293 cells.

[0162] The 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. Kits and Containers

[0163] The present disclosure is also characterized by a container comprising any pharmaceutical formulation described and exemplified herein. In some aspects, the container is a glass vial. In some aspects, the container is a glass vial with a 3 mL fill volume. In some aspects, the container is a glass vial with a 3 mL or less fill volume. In some aspects, the container is a glass vial with a 2 mL or less fill volume.

[0164] The present disclosure is also characterized by comprising a kit comprising any pharmaceutical formulation described and exemplified herein, or an antibody or its antigen binding fragment. These kits can be used to provide pharmaceutical formulations, antibodies, their antigen binding fragments, and other agents for diagnosis, basic research or methods of treatment, etc. In some aspects, these kits include any one or more of the pharmaceutical formulations, antibodies or their antigen binding fragments described or exemplified herein, and instructions for using the one or more pharmaceutical formulations, antibodies or their antigen binding fragments in a method for treating a disease in a subject in need. The method may include administering any pharmaceutical formulation provided herein or any kit or container provided herein to the subject. In some aspects, the disease is a respiratory disease, a gastrointestinal disease, a skin disease, or an arthritis.

[0165] In some aspects, the antibody or antigen-binding fragment thereof is formulated in a volume of 3 mL or less. In some aspects, the antibody or antigen-binding fragment thereof is formulated in a volume of 2 mL or less.

[0166] In some aspects, the respiratory disease is asthma, chronic obstructive pulmonary disease (COPD), pulmonary fibrosis, pulmonary sarcoidosis, allergic rhinitis, or cystic fibrosis.

[0167] In some aspects, the gastrointestinal disease is inflammatory bowel disease, Crohn's disease, colitis, ulcerative colitis, eosinophilic esophagitis, or irritable bowel syndrome.

[0168] In some aspects, the arthritis is rheumatoid arthritis.

[0169] In some aspects, the skin disease is atopic dermatitis, eczema, or scleroderma. Exemplary aspects provided herein

[0170] In one aspect (Aspect 1; A1), provided herein is 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); wherein the antibody or antigen-binding fragment thereof comprises: (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, 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; or (iii) a heavy chain comprising the amino acid sequence of SEQ ID NO:9 and a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO:10. Light chain of the amino acid sequence of 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.

[0171] In one aspect of A1 , ie A2, the antibody or antigen-binding fragment of (i) or (ii) comprises an IgG1 constant region.

[0172] In one aspect of A1 or A2, ie A3, the pharmaceutical formulation is lyophilized.

[0173] In one aspect of A1 or A2, ie A4, the pharmaceutical formulation is liquid.

[0174] In one aspect of any one 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°C-8°C for 24 hours, 72 hours or 10 days.

[0175] In one aspect of any one of A1-A5, ie A6, the pharmaceutical composition has an osmotic pressure from 200 to 500 mOsm / kg after storage at room temperature for 24 hours or at 2°C-8°C for 24 hours, 72 hours or 10 days.

[0176] In one aspect of any one of A1-A6, ie A7, the pharmaceutical composition has an antibody monomer content of at least 99% after storage at 2°C-8°C for 24 hours, 72 hours or 10 days.

[0177] In one aspect of any one of A1-A7, ie A8, the pharmaceutical composition has no significant change in charge heterogeneity characteristics after storage at 2°C-8°C for 24 hours, 72 hours or 10 days.

[0178] In one aspect of any one 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°C-8°C for 24 hours, 72 hours or 10 days.

[0179] In one aspect of any one of A1-A8, ie A10, the pharmaceutical composition has at least 90% purity after storage at room temperature for 24 hours or at 2°C-8°C for 24 hours, 72 hours or 10 days.

[0180] In one aspect of any one 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°C-8°C for 24 hours, 72 hours or 10 days.

[0181] In an aspect of any one of A1-A11, ie A12, the pharmaceutical composition has no significant difference in visual appearance after storage at 2°C-8°C for up to 36 months.

[0182] In one aspect of any one of A1-A12, ie A13, the pharmaceutical composition has no significant difference in protein concentration, osmotic pressure or viscosity after storage at 2°C-8°C, 25°C or 40°C for up to 36 months.

[0183] In one aspect of any one of A1-A13, ie A14, the pharmaceutical composition has ≥95% monomer content, ≤5.0% dimer content, or no significant difference in low molecular weight material content after storage at 2°C-8°C for up to 36 months.

[0184] In an aspect of any one of A1-A14, ie A15, the pharmaceutical composition has a purity of ≥ 90% after storage at 2°C-8°C for up to 36 months.

[0185] In one aspect of any one of A1-A15, i.e. A16, the pharmaceutical composition has a main substance content from 50% to 90%, an acidic substance content from 10% to 40%, and / or a basic substance content from 0% to 20% after storage at 2°C-8°C for up to 36 months.

[0186] In an aspect of any one of A1-A16, ie A17, the pharmaceutical composition has no significant difference in subvisible particulate content after storage at 2°C-8°C, 25°C or 40°C for up to 36 months.

[0187] In one aspect of 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°C-8°C, 25°C or 40°C for up to 36 months.

[0188] In one aspect of any one of A1-A18, ie A19, the pharmaceutical composition has a relative potency of from 70% to 135% after storage at 2°C-8°C for up to 36 months as measured by enzyme-linked immunosorbent assay (ELISA).

[0189] In an aspect of any one of A1-A19, ie A20, the pharmaceutical composition has no significant difference in thermal stability after storage at 2°C-8°C, 25°C or 40°C for up to 6 months.

[0190] In an aspect of any one of A1-A20, ie A21, the pharmaceutical composition has no significant difference in thermal stability after storage at 2°C-8°C for up to 36 months.

[0191] In one aspect of any one of A1-A21, ie A22, the pharmaceutical composition has no significant differences in secondary and / or tertiary protein structure after storage at 2°C-8°C, 25°C or 40°C for up to 3 months.

[0192] In an aspect of any one of A1-A22, ie A23, the pharmaceutical composition has no significant difference in secondary protein structure after storage at 2°C-8°C for up to 36 months.

[0193] In an aspect of any one of A1-A23, ie A24, the pharmaceutical composition has no significant difference in the concentration of polysorbate-80 after storage at 2°C-8°C for up to 24 months.

[0194] In one aspect of any one of A1-A24, i.e. A25, the antibody or antigen-binding fragment thereof is produced in Chinese hamster ovary cells.

[0195] In one aspect, A26, a container is provided, comprising a pharmaceutical formulation as described in any one of A1-A25.

[0196] In one aspect of A26, namely A27, the container is a glass vial.

[0197] In one aspect of A27, namely A28, the container is a glass vial having a fill volume of 3 mL.

[0198] In one aspect, i.e. 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 as described in any one of A1-A25 or a container as described in any one of A26-A28, optionally wherein the disease is a respiratory disease, a gastrointestinal disease, a skin disease, or arthritis.

[0199] In one aspect of A29, i.e. A30, the respiratory disease is asthma, chronic obstructive pulmonary disease (COPD), pulmonary fibrosis, pulmonary sarcoidosis, allergic rhinitis, or cystic fibrosis.

[0200] In one aspect of A29, i.e. A31, the gastrointestinal disease is inflammatory bowel disease, Crohn's disease, colitis, ulcerative colitis, eosinophilic esophagitis, or irritable bowel syndrome.

[0201] In one aspect of A29, i.e. A32, the arthritis is rheumatoid arthritis.

[0202] In one aspect of A29, i.e. A33, the skin disease is atopic dermatitis, eczema or scleroderma.

[0203] In an aspect of any one of A1-A25, ie A34, the formulation is for use according to a method as described in any one of A29-A33.

[0204] In one aspect, i.e., B1, a pharmaceutical formulation is provided herein, comprising: (a) about 100 mg / mL to about 250 mg / mL of an antibody or an antigen-binding fragment thereof that specifically binds to TNF-like ligand 1A (TL1A); wherein the antibody or the 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, 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. NO:6 amino acid sequence of light chain variable region CDR3; (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.

[0205] In one aspect 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.

[0206] In one aspect of B1 or B2, ie, B3, the antibody or antigen-binding fragment comprises an IgG1 constant region.

[0207] In one aspect of any one of B1-B3, i.e. 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.

[0208] In one aspect of any one of B1-B4, ie, B5, the pharmaceutical formulation comprises about 100 mg / mL, about 150 mg / mL, about 200 mg / mL, about 225 mg / mL, or about 250 mg / mL of the antibody or antigen-binding fragment thereof.

[0209] In one aspect of any one of B1-B5, ie, B6, the pharmaceutical formulation comprises about 5 mM, about 10 mM, or about 15 mM histidine.

[0210] In one aspect of any one of B1-B6, ie, B7, the pharmaceutical formulation comprises about 50 mM, about 100 mM, or about 150 mM arginine-hydrochloride (Arg-HCl).

[0211] In one aspect of any one of B1-B7, ie, B8, the pharmaceutical formulation comprises about 2.5%, about 5%, or about 7.5% (w / v) sucrose.

[0212] In one aspect of any one of B1-B8, ie, B9, the pharmaceutical formulation comprises about 0.01%, about 0.02%, or about 0.03% (w / v) polysorbate-80.

[0213] In one aspect of any one of B1-B9, ie B10, the pharmaceutical formulation is lyophilized.

[0214] In one aspect of any one of B1-B9, ie B11, the pharmaceutical formulation is liquid.

[0215] In one aspect of any one 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°C-8°C for 24 hours, 72 hours or 10 days.

[0216] In one aspect of any one of B1-B12, ie B13, the pharmaceutical composition has an osmotic pressure from 200 to 500 mOsm / kg after storage at room temperature for 24 hours or at 2°C-8°C for 24 hours, 72 hours or 10 days.

[0217] In one aspect of any one of B1-B13, ie B14, the pharmaceutical composition has an antibody monomer content of at least 99% after storage at 2°C-8°C for 24 hours, 72 hours or 10 days.

[0218] In one aspect of any one of B1-B14, ie B15, the pharmaceutical composition has no significant change in charge heterogeneity characteristics after storage at 2°C-8°C for 24 hours, 72 hours or 10 days.

[0219] In one aspect of any one 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°C-8°C for 24 hours, 72 hours or 10 days.

[0220] In one aspect of any one of B1-B15, ie B17, the pharmaceutical composition has at least 90% purity after storage at room temperature for 24 hours or at 2°C-8°C for 24 hours, 72 hours or 10 days.

[0221] In one aspect of any one 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°C-8°C for 24 hours, 72 hours or 10 days.

[0222] In an aspect of any one of B1-B18, ie B19, the pharmaceutical composition has no significant difference in visual appearance after storage at 2°C-8°C for up to 36 months.

[0223] In one aspect of any one of B1-B19, ie B20, the pharmaceutical composition has no significant difference in protein concentration, osmotic pressure or viscosity after storage at 2°C-8°C, 25°C or 40°C for up to 36 months.

[0224] In one aspect of any one of B1-B20, ie B21, the pharmaceutical composition has ≥95% monomer content, ≤5.0% dimer content, or no significant difference in low molecular weight material content after storage at 2°C-8°C for up to 36 months.

[0225] In one aspect of any one of B1-B21, ie B22, the pharmaceutical composition has a purity of ≥90% after storage at 2°C-8°C for up to 36 months.

[0226] In one aspect of any one of B1-B22, i.e. B23, the pharmaceutical composition has a main substance content from 50% to 90%, an acidic substance content from 10% to 40%, and / or a basic substance content from 0% to 20% after storage at 2°C-8°C for up to 36 months.

[0227] In an aspect of any one of B1-B23, ie B24, the pharmaceutical composition has no significant difference in subvisible particulate content after storage at 2°C-8°C, 25°C or 40°C for up to 36 months.

[0228] In one aspect of any one of 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°C-8°C, 25°C or 40°C for up to 36 months.

[0229] In one aspect of any one of B1-B25, ie B26, the pharmaceutical composition has a relative potency of from 70% to 135% after storage at 2°C-8°C for up to 36 months as measured by enzyme-linked immunosorbent assay (ELISA).

[0230] In one aspect of any one of B1-B26, ie B27, the pharmaceutical composition has no significant difference in thermal stability after storage at 2°C-8°C, 25°C or 40°C for up to 6 months.

[0231] In one aspect of any one of B1-B27, ie B28, the pharmaceutical composition has no significant difference in thermal stability after storage at 2°C-8°C for up to 36 months.

[0232] In one aspect of any one of B1-B28, ie B29, the pharmaceutical composition has no significant differences in secondary and / or tertiary protein structure after storage at 2°C-8°C, 25°C or 40°C for up to 3 months.

[0233] In an aspect of any one of B1-B29, ie B30, the pharmaceutical composition has no significant difference in secondary protein structure after storage at 2°C-8°C for up to 36 months.

[0234] In an aspect of any one of B1-B30, ie B31, the pharmaceutical composition has no significant difference in the concentration of polysorbate-80 after storage at 2°C-8°C for up to 24 months.

[0235] In one aspect of any one of B1-B31, i.e., B32, the antibody or antigen-binding fragment thereof is produced in Chinese hamster ovary cells.

[0236] In one aspect, B33, a container is provided, comprising a pharmaceutical formulation as described in any one of B1-B32.

[0237] In one aspect of B33, namely B34, the container is a glass vial.

[0238] In one aspect of B34, namely B35, the container is a glass vial having a fill volume of 3 mL.

[0239] In one aspect of B33, i.e. B36, the container is a syringe, optionally wherein the syringe is a pre-filled syringe.

[0240] In one aspect, i.e., 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 as described in any one of B1-B32 or a container as described in any one of B33-B36, optionally wherein the disease is a respiratory disease, a gastrointestinal disease, a skin disease, or arthritis.

[0241] In one aspect of B37, i.e. B38, the respiratory disease is asthma, chronic obstructive pulmonary disease (COPD), pulmonary fibrosis, pulmonary sarcoidosis, allergic rhinitis, or cystic fibrosis.

[0242] In one aspect of B37, i.e. B39, the gastrointestinal disease is inflammatory bowel disease, Crohn's disease, colitis, ulcerative colitis, eosinophilic esophagitis, or irritable bowel syndrome.

[0243] In one aspect of B37, i.e. B40, the arthritis is rheumatoid arthritis.

[0244] In one aspect of B37, i.e. B41, the skin disease is atopic dermatitis, eczema or scleroderma.

[0245] In one aspect of any one of B37-B41, i.e. B42, the pharmaceutical formulation is administered intravenously. In one aspect of any one of B37-B41, i.e. B43, the pharmaceutical formulation is administered subcutaneously.

[0246] In one aspect of any one of B1-B32 (ie, B44), the formulation is for use according to a method as described in any one of B37-B43.

[0247] The following examples are offered by way of illustration and not by way of limitation. Examples Example 1: Stability of Reconstituted TEV-48574 Drug Product Solution

[0248] To evaluate the in-use stability of the TEV-48574 reconstituted drug product.

[0249] For this study, TEV-48574 is presented in lyophilized form, at pH 6.0, in 10mM histidine, 5% (w / v) sucrose, 100mM arginine-hydrochloride (Arg-HCl), 0.02% (w / v) polysorbate-80 (PS-80), the protein concentration is 150mg / mL. The lyophilized formulation is contained in a 5cc vial with a bottleneck size of 20mm. The vial is stored at 2°C-8°C and allowed to reach room temperature before reconstitution. About 7 vials are reconstituted in each vial using 2.0mL sterile water for injection (WFI) to prepare a stock solution. The stock solution is then diluted into different falcon tubes using a formulation buffer (pH 6.0) of 10mM histidine, 5% (w / v) sucrose, 100mM Arg-HCl and 0.02% (w / v) PS80 to a concentration of 50, 20 and 5mg / mL. After dilution, 4 mL samples were incubated at 2°C-8°C in the dark for 24 hours, 72 hours and 10 days, and at room temperature under normal light conditions for 24 hours. Table 1 shows the specific time points and conditions tested. Table 1: Conditions and time points tested for stability study of reconstituted TEV-48574 Note: T0 = time zero

[0250] At each time point and at the end of the condition, samples were analyzed by visual appearance, pH, osmolarity, protein concentration, size exclusion chromatography (SEC), sodium dodecyl sulfate capillary gel electrophoresis (cSDS), capillary isoelectric focusing (cIEF), and subvisible particle analysis using microfluidics imaging (MFI).

[0251] No significant differences were observed in visual appearance, pH, osmolarity, and protein concentration among the reconstituted samples after dilution as shown in Table 2. In addition, the pH and osmolarity values ​​of the samples were within the limits of 6.0 ± 0.5 pH and 200-500 mOsm / kg, respectively. Table 2: Visual appearance, pH, osmolarity and protein concentration of the reconstituted product Note: T0 = time zero; RT = room temperature; C = clear solution, L = colorless, F = no visible particles

[0252] SEC analysis showed that the % monomer level of the antibody in the samples did not change significantly over the duration of the stability evaluation. See Table 3. These results indicate that storage of the diluted reconstituted drug product at 2°C-8°C for up to 10 days did not have any effect on the % monomer level after incubation. Table 3: Percentage levels of monomers in reconstituted products Conditions and timing HMW% Dimer % Main peak% 5 mg / mL, T0 0.7 0.1 99.3 5 mg / mL, RT, 24 hours 0.7 0.1 99.3 5 mg / mL, 2°C-8°C, 24 hours 0.6 0.1 99.3 5 mg / mL, 2°C-8°C, 72 hours 0.7 0.1 99.3 5 mg / mL, 2°C-8°C, 10 days 0.6 0.1 99.3 20 mg / mL, T0 0.7 0.1 99.3 20 mg / mL, RT, 24 hours 0.6 0.1 99.3 20 mg / mL, 2°C-8°C, 24 hours 0.7 0.1 99.2 20 mg / mL, 2°C-8°C, 72 hours 0.7 0.1 99.3 20 mg / mL, 2°C-8°C, 10 days 0.7 0.1 99.2 50 mg / mL, T0 0.7 0.1 99.3 50 mg / mL, RT, 24 hours 0.7 0.1 99.2 50 mg / mL, 2°C-8°C, 24 hours 0.7 0.1 99.2 50 mg / mL, 2°C-8°C, 72 hours 0.7 0.1 99.2 50 mg / mL, 2°C-8°C, 10 days 0.7 0.1 99.2 Note: T0 = time zero; RT = room temperature; HMW = high molecular weight species

[0253] cIEF analysis showed that dilution of the reconstituted drug product and its incubation at 2°C-8°C for up to 10 days did not have any significant effect on the charge heterogeneity profile of TEV-48574. See Table 4. Table 4: cIEF analysis of reconstituted products Note: T0 = time zero

[0254] As shown in Table 5, cSDS analysis revealed that the reconstituted drug product had a purity of >90%, and dilution and incubation of the reconstituted drug product at different conditions and time points had no significant effect on the purity. Table 5: cSDS analysis of reconstituted products Note: NR = non-reducing; R = reducing; HC = heavy chain; LC = light chain; HMW = high molecular weight species; T0 = time zero; RT = room temperature

[0255] MFI analysis showed that the 50 mg / mL sample contained a slightly higher number of subvisible particles. See Table 6. However, when comparing the initial time point samples to the later time point samples, no significant increase in particle concentration was observed overall for the different concentration samples. Table 6: Particle size concentration of reconstituted products Note: T0 = time zero; RT = room temperature

[0256] In summary, these results show that the reconstituted TEV-48574 drug product is stable upon dilution after storage at 2°C-8°C protected from light for 10 days and at room temperature under normal light conditions for 24 hours. Example 2: Stability of TEV-48574 liquid formulations at 100 mg / ml and 150 mg / ml

[0257] The purpose of this study was to evaluate the long-term stability of liquid and lyophilized TEV-48574 formulations. Three formulations were tested: a lyophilized form 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 at pH 6.0 (Formulation 1); and two liquid forms 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 at pH 6.0 (Formulations 2 and 3, respectively). The stability of these formulations was evaluated in 5cc vials (Type I glass) under the following conditions: "standard" conditions (2°C-8°C), "accelerated" conditions (25°C±2°C / 60%±5% relative humidity (RH)), and "stressed" conditions (40°C±2°C / 75%±5% RH). In addition, the stability of Formulation 3 was evaluated under the same conditions, but the formulation was filled into a 2.25mL Nipro prefilled syringe (Nipro PFS) equipped with a West plunger stopper. The effects of these conditions on several product quality attributes were tested for the formulations. Visual appearance, protein concentration, osmolarity, and viscosity

[0258] The results of the visual appearance, protein concentration and osmotic pressure analysis of formulations 1-3 are shown in Tables 7-10. Under standard storage conditions of 2°C-8°C, no significant differences across formulations were observed. At intermittent time points, very few visible particles were observed. However, this may be related to the development nature of the study, in which the drug product was manually filled into vials. In addition, these particles are unlikely to be product-related because they do not increase over time and are not consistently detected at different time points in all samples. For the visual appearance under stress conditions of 40°C, the solution looks slightly yellow at a later time point. Compared to formulation 3 stored in glass vials for up to 24 months, no significant differences were observed between formulation 3 filled into Nipro PFS in terms of visual appearance, protein concentration, osmotic pressure and viscosity (data not shown), indicating that these attributes of the formulation are not affected by contact with Nipro PFS.

[0259] Under standard (2°C-8°C), accelerated (25°C) and stressed (40°C) conditions, the measured protein concentrations of all three formulations were close to the expected nominal concentrations and did not change over time. Similar trends were observed for osmotic pressure and viscosity measured under standard storage conditions at time zero (T0), 24 months (24M) and 36 months (36M), with no significant differences observed over time. Table 7: Visual appearance and protein concentration results at 2°C-8°C T0: time zero; M: month; F1: formulation 1; F2: formulation 2; F3: formulation 3; S: slightly milky; L: colorless; FFVP: contains no visible particles; SY: slightly yellow; NT: not tested Table 8: Osmotic pressure and viscosity results at 2°C-8°C T0: time zero; M: month; F1: formulation 1; F2: formulation 2; F3: formulation 3; NT: not tested Table 9: Visual appearance and protein concentration results at 25°C T0: time zero; WK: week; M: month; F1: formulation 1; F2: formulation 2; F3: formulation 3; S: slightly milky; L: colorless; FFVP: no visible particles; SY: slightly yellow; NT: not tested Table 10: Visual appearance and protein concentration results at 40°C T0: time zero; WK: week; M: month; F1: formulation 1; F2: formulation 2; F3: formulation 3; S: slightly milky; L: colorless; FFVP: no visible particles; SY: slightly yellow; NT: not tested Size Exclusion Chromatography (SEC)

[0260] Table 11-13 and Figures 1A-1C, 2A-2C and 3A-3C show the monomer percentage (%), dimer % and low molecular weight substances % of formulations 1, 2 and 3 measured by SEC. All three formulations stored under standard storage conditions of 2°C-8°C meet the acceptance criteria for monomer % and dimer % levels for up to 24 months and up to 36 months. Compared with formulation 1, a slight decrease in monomer % and a simultaneous increase in low molecular weight substances % were observed for formulations 2 and 3. However, considering that lyophilized formulations are typically more stable than liquid formulations, this is an expected observation. Under accelerated and stressed conditions, protein fragmentation is more common, indicating the formation of low molecular weight substances as the main degradation pathway for liquid formulations. The protein degradation rates of formulations 2 and 3 are similar, indicating that protein concentration has no significant effect on degradation. No significant differences were observed in the % monomer, % dimer, and % low molecular weight species up to 24 months between Formulation 3 filled into Nipro PFS compared to Formulation 3 stored in glass vials (data not shown), indicating that the stability characteristics of the formulations were not affected by contact with Nipro PFS. Table 11: Monomer %, dimer % and low molecular weight species % at 2°C-8°C LMW: low molecular weight substances Table 12: Monomer %, dimer % and low molecular weight species % at 25°C LMW: low molecular weight substances Table 13: Monomer %, dimer % and low molecular weight % at 40°C LMW: low molecular weight substances Capillary gel electrophoresis (reducing and non-reducing)

[0261] The percentage (%) of the immunoglobulin G (IgG) + 125 kDa peak of formulations 1-3 was determined using non-reducing capillary gel electrophoresis (CGE). In addition, the heavy chain + light chain % of formulations 1-3 was determined using reducing CGE. These results are shown in Tables 14-15 and Figures 4A-4Cand 5A-5C. In all three formulations stored under standard storage conditions of 2°C-8°C, the purity % met the acceptance criteria for up to 24 months and up to 36 months. Under accelerated and stressed conditions, protein fragmentation was observed for Formulations 2 and 3 compared to Formulation 1. However, this was an expected observation, considering that lyophilized formulations are typically more stable than liquid formulations. In addition, the protein degradation trends for Formulations 2 and 3 were similar, indicating that protein concentration had no significant effect. No significant differences were observed between Formulation 3 filled into Nipro PFS in terms of IgG+125kDa peak % for up to 24 months compared to Formulation 3 stored in glass vials (data not shown), indicating that the stability characteristics of the formulations were not affected by contact with Nipro PFS. Table 14: % IgG + 125 kDa peak measured using non-reducing CGE at 2°C-8°C, 25°C and 40°C for Formulations 1, 2 and 3 NT: Not tested Table 15: % Heavy and Light Chains measured using reduced CGE at 2°C-8°C, 25°C and 40°C for Formulations 1, 2 and 3 NT: Not tested Capillary isoelectric focusing (icIEF)

[0262] Capillary isoelectric focusing (icIEF) was used to measure the content percentage (%) of the main substance (main peak), acidic substance (acidic peak) % and basic substance (basic peak) % of Formulations 1-3 (F1-F3). The results are shown in Tables 16-18. Figures 6A-6C , 7A-7C and 8A-8C. Under standard storage conditions of 2°C-8°C ( Fig. 6A , 7A and 8A), all three formulations (F1-F3) met the acceptance criteria for up to 24 months and up to 36 months. At accelerated (25°C; Figure 6B , 7B and 8B) and stress (40°C; Figure 6C , 7CUnder the storage conditions of 8C and 8C, a decrease in the main peak % and a simultaneous increase in the acidic substance % were observed for Formulations 2 and 3. No significant changes were observed for Formulation 1. However, considering that lyophilized formulations are typically more stable than liquid formulations, this is an expected observation. Compared to Formulation 3 stored in glass vials, no significant differences were observed between Formulation 3 filled into Nipro PFS in terms of the main peak %, acidic substance % and basic substance % for up to 24 months (data not shown), indicating that the stability characteristics of the formulations were not affected by contact with Nipro PFS. Table 16: Main peak %, acidic % and basic % determined using icIEF at 2°C-8°C for Formulations 1, 2 and 3 Table 17: Main peak %, acidic species % and basic species % determined using icIEF at 25°C for Formulations 1, 2 and 3 Table 18: Main peak %, acidic species % and basic species % determined using icIEF at 40°C for Formulations 1, 2 and 3 Subvisible particles using microfluidics imaging (MFI)

[0263] The subvisible particles in Formulations 1-3 were measured at different time points under standard conditions of 2°C-8°C, accelerated conditions of 25°C, and stressed conditions of 40°C. The results are shown in Tables 19-21, respectively. No significant changes in subvisible particles were observed. At intermittent time points, higher subvisible particles were observed compared to other time points. This may be related to the method in which greater variability and sensitivity of subvisible particles measured using MFI have been observed. Overall, the subvisible particles in the size range of ≥ (greater than or equal to) 10 μm were less than 6,000 particles / mL. In the size range of ≥ (greater than or equal to) 25 μm, even considering the increased sensitivity of using MFI for subvisible particle detection, the subvisible particles were less than 600 particles / mL and were well within the USP <788> No significant differences were observed in subvisible particles up to 24 months between Formulation 3 filled into Nipro PFS compared to Formulation 3 stored in glass vials (data not shown), indicating that the stability profile of the formulations was not affected by contact with Nipro PFS. Table 19: Subvisible particles of Formulations 1-3 at 2°C-8°C SbVP: Subvisible particles Table 20: Subvisible particles of Formulations 1-3 at 25°C SbVP: Subvisible particles Table 21: Subvisible particles of Formulations 1-3 at 40°C Chemical modification of primary structure using peptide mapping

[0264] The amino acids that may potentially undergo chemical modification and affect protein structure over time in formulations 1-3 were monitored. In particular, the amino acid residues methionine 81 and methionine 254 of TEV-48574 may potentially undergo oxidation, thereby affecting the primary structure, and were monitored. Similarly, asparagine 317 of TEV-48574 may potentially undergo deamidation, thereby producing succinimide, and were monitored. However, as shown in Tables 22-24, no significant changes in these modifications over time were observed. Under long-term storage conditions (2°C-8°C) for up to 24 months, compared to formulation 3 stored in a glass vial, the results in terms of Met81 oxidation %, Asn317 deamidation % and Met254 oxidation % between formulation 3 filled into Nipro PFS were comparable (data not shown), indicating that the stability characteristics of the formulation were not affected by contact with Nipro PFS. Table 22: % Oxidation of Met81 in Formulations 1-3 NT: not tested; F1: Formulation 1; F2: Formulation 2; F3: Formulation 3; Met81: Methionine residue 81 of TEV-48574 Table 23: % Deamidation of Asn317 in Formulations 1-3 NT: not tested; F1: Formulation 1; F2: Formulation 2; F3: Formulation 3; Asn317: Asparagine residue 317 of TEV-48574 Table 24: % Oxidation of Met254 in Formulations 1-3 NT: not tested; F1: Formulation 1; F2: Formulation 2; F3: Formulation 3; Met254: methionine residue 254 of TEV-48574 Potency determined by ELISA

[0265] The percent potency (%) of Formulations 1-3 was determined by enzyme-linked immunosorbent assay (ELISA). The results are shown in Table 25. Under standard storage conditions (2°C-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 between Formulation 3 filled into Nipro PFS in terms of potency % for up to 24 months compared to Formulation 3 stored in glass vials (data not shown), indicating that the stability characteristics of the formulations were not affected by contact with Nipro PFS. Table 25: Relative % potency of Formulations 1-3 as determined by ELISA NT: Not tested Differential Scanning Calorimetry (DSC) Analysis of Thermal Stability

[0266] DSC was used to evaluate the thermal stability of Formulations 1 to 3. DSC analysis was performed on time zero (T0) samples and 3-month, 6-month, and 36-month samples of Formulations 1 to 3 at 2°C-8°C, 25°C, and 40°C. Fig. 9A ( Fig. 9A ) shows the thermal stability of Formulations 1-3 at time zero (T0) and after 3 months at 2°C-8°C, 25°C and 40°C. Fig. 9B ( Fig. 9B ) shows the thermal stability of Formulations 1-3 at time zero (T0) and after 6 months at 2°C-8°C, 25°C and 40°C. Fig. 9C ( Fig. 9C ) shows the thermal stability of formulations 1-3 at time zero (T0) and after 36 months at 2°C-8°C. For the three formulations, no significant differences in thermal stability were observed at T0, 3 months, 6 months and 36 months because the transition temperatures (Tm) of all formulations overlap. A slight decrease in enthalpy was observed for formulations stored under accelerated and stressed conditions. This may be due to protein fragmentation observed in the samples under these conditions, resulting in a decrease in enthalpy. Compared to formulation 3 stored in glass vials, no significant differences were observed between formulation 3 filled into Nipro PFS in terms of thermal stability for up to 6 months, indicating that the stability characteristics of the formulations were not affected by contact with Nipro PFS. Secondary and tertiary protein structure analysis using circular dichroism (CD) spectroscopy

[0267] The secondary protein structure of Formulations 1-3 was analyzed using far ultraviolet (far UV) circular dichroism (CD), and the tertiary protein structure of Formulations 1-3 was analyzed using near UV CD. Secondary and tertiary structure analysis was performed on: time zero (T0) samples of Formulations 1, 2, and 3; 3 month samples at 2°C-8°C, 25°C, and 40°C; 24 month samples at 2°C-8°C; and 36 month samples at 2°C-8°C. The results are in Figures 10A-10F ( Figures 10A-10F ). In particular, Fig. 10A Shown are the secondary structures of Formulations 1-3 at T0 and after 3 months at 2-8°C, 25°C and 40°C, determined using far UV CD. Fig. 10B Shown are the secondary structures of Formulations 1-3 at T0 and after 24 months at 2°C-8°C, determined using far UV CD. Fig. 10C Shown are the secondary structures of Formulations 1-3 at T0 and after 36 months at 2°C-8°C, determined using far UV CD. Fig. 10D Shown are the tertiary structures of Formulations 1-3 at T0 and after 3 months at 2-8°C, 25°C and 40°C, determined using near UV CD. Fig.10E The tertiary structures of Formulations 1-3 at T0 and after 24 months at 2°C-8°C, determined using near UVCD, are shown. Fig.10F Shown are the tertiary structures of Formulations 1-3 at T0 and after 36 months at 2°C-8°C, determined using near UVCD.

[0268] The far UV CD spectra show a negative maximum at about 217 nm, indicating a beta-folded structure of the T0 sample in all three formulations, which is expected for a monoclonal antibody. The near UV CD spectra show a positive maximum at about 292 nm, indicating the absorption of tryptophan residues, and a negative maximum at 276 nm, indicating the absorption of tyrosine residues. For the three formulations, no significant changes in secondary or tertiary structure were observed for up to 3 months. At 24 and 36 months, slight changes in the CD spectra were observed at about 200 nm. This may be due to protein fragmentation, because the absorption in this region is mainly due to peptide bond absorption. However, no significant changes in the secondary structure of the protein (absorption at 217 nm) were observed.

[0269] Fig.11A Shown are the secondary structures 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) at 2°C-8°C, and after 3 months (3M) and 6 months (6M) at 25°C and 40°C, as determined using far UV CD. Fig. 11BThe tertiary structure of Formulation 3 stored in a Nipro prefilled syringe (PFS) at time zero (T0) and after 3 months (3M), 6 months (6M), and 24 months (24M) storage at 2°C-8°C and after 3 months (3M) and 6 months (6M) storage at 25°C and 40°C, as determined using near-UV CD, is shown. The spectra obtained for the Nipro PFS sample are comparable to those obtained for Formulation 3 stored in glass vials ( Figures 10A-10B and Figures 10D-10E ). Polysorbate 80 Analysis

[0270] The polysorbate 80 (PS80) present as surfactant / excipient in formulations 1-3 may potentially undergo degradation, which produces reactive peroxides that may affect protein stability during the shelf life of the product. After storage at 2°C-8°C for 24 months and 36 months, formulations 1-3 were subjected to PS80 analysis by testing PS80 levels. As shown in Table 26, in the 24-month samples in all formulations, the PS80 level measured was 0.02% (w / v), which is related to the expected PS80 concentration. At 36 months, the PS80 levels in formulations 2 and 3 as liquid formulations showed a reduction. Compared with formulation 3 stored in glass vials, no significant differences were observed in the PS80 levels at 24 months between formulation 3 filled in Nipro PFS, indicating that the stability characteristics of the formulations were not affected by contact with Nipro PFS. Table 26: Polysorbate 80 (PS80) levels detected in Formulations 1-3 after 24 and 36 months storage at 2°C-8°C Preparations at 2°C-8°C PS80 level at 24M (w / v) PS80 level at 36M (w / v) Preparation 1 0.02％ 0.02％ Preparation 2 0.02％ 0.01％ Preparation 3 0.02％ 0.01％ in conclusion

[0271] The stability of TEV-48574 in lyophilized form (Formulation 1 at 100 mg / mL) was comparable to that of the liquid formulations at 100 mg / mL (Formulation 2) and 150 mg / mL (Formulation 3) stored at 2°C-8°C (standard storage conditions for drug products). Under accelerated (25°C) and stressed (40°C) conditions, both liquid formulations showed protein fragmentation as the major degradation pathway, which was not observed for lyophilized Formulation 1. However, this was an expected observation given that the lyophilized formulations were more stable compared to the liquid formulations.

[0272] Compared to the lyophilized formulation, the formation of acidic species was observed in the liquid formulation (measured using icIEF), and concomitant protein fragmentation was observed using SEC and CGE. This indicates deamidation of certain amino acid residues, potentially leading to fragmentation. See Wang, W. et al., J. Pharm. Sci., 96: 1-26 (2007).

[0273] Based on these data, lyophilized Formulation 1 can be bridged to 100 mg / mL and 150 mg / mL liquid formulations that have comparable stability characteristics and meet acceptance criteria for up to 24 months and up to 36 months at 2°C-8°C storage conditions.

[0274] Furthermore, the stability of 150 mg / mL TEV-48574 (Formulation 3) stored in Nipro PFS was found to be comparable to Formulation 3 samples stored in glass vials. Based on this data, storage for 24 months at 2°C-8°C (standard storage conditions for drug products) is compatible with Nipro PFS. Example 3: Stability of 200 mg / ml TEV-48574 liquid formulation

[0275] The purpose of this study was to evaluate the long-term stability of a high concentration liquid TEV-48574 formulation compared to a lyophilized drug product. Two formulations were tested: a lyophilized form 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 at pH 6.0 (Formulation 4 or F4); and a liquid form 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 at pH 6.0 (Formulation 5 or F5). The stability of these formulations was evaluated in 5 cc vials with 20 mm sized stops under long-term storage conditions of 2°C-8°C. The effects of these conditions on several product quality attributes were tested for the formulations. Visual appearance, protein concentration, osmolarity, and viscosity

[0276] The results of the visual appearance, protein concentration and osmotic pressure analysis of Formulations 4 and 5 are shown in Tables 27-28. No significant differences across formulations were observed under long-term storage conditions at 2°C-8°C.

[0277] Under long-term storage conditions of 2°C-8°C, the measured protein concentrations of both formulations were close to the expected nominal concentrations and did not change over time. The osmotic pressure and viscosity of the drug product were evaluated at time zero (T0). The osmotic pressure 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). Table 27: Visual appearance and protein concentration results at 2°C-8°C T0: time zero; M: month; F4: formulation 4; F5: formulation 5; S: slightly milky; O: milky; V: distinctly milky; L: colorless; FFVP: contains no visible particles; C: clear; VSY: distinctly slightly yellow; SP: slightly pink; NT: not tested Table 28: Osmotic pressure and viscosity results at 2°C-8°C T0: time zero; F4: preparation 4; F5: preparation 5 Size Exclusion Chromatography (SEC)

[0278] Table 29 and Figures 12A-12C The monomer percentage (%), dimer % and low molecular weight species % of Formulations 4 and 5 measured by SEC are shown. Both formulations stored under long-term storage conditions of 2°C-8°C met the acceptance criteria for monomer % and dimer % levels for up to 24 months. A slight decrease in monomer % and a concurrent increase in low molecular weight species % were observed for Formulation 5 compared to Formulation 4. Table 29: Monomer %, dimer % and low molecular weight species % at 2°C-8°C LMW: low molecular weight substances; NT: not tested Capillary gel electrophoresis (reducing and non-reducing)

[0279] Non-reducing capillary gel electrophoresis (CGE) was used to determine the percentage (%) of the immunoglobulin G (IgG) + 125 kDa peak for Formulations 4 and 5. Additionally, reducing CGE was used to determine the heavy chain + light chain % for Formulations 4 and 5. These results are shown in Tables 30-31 and Figures 13A-13B In both formulations stored at long term storage conditions of 2°C-8°C, the % purity met the acceptance criteria for up to 24 months. Table 30: % IgG + 125 kDa peak measured using non-reducing CGE at 2°C-8°C for Formulations 4 and 5 Table 31: Heavy and light chain % measured using reduced CGE at 2°C-8°C for Formulations 4 and 5 Capillary isoelectric focusing (icIEF)

[0280] Capillary isoelectric focusing (icIEF) was used to measure the content percentage (%) of the main substance (main peak), acidic substance (acidic peak) % and basic substance (basic peak) % of Formulations 4 and 5 (F4 and F5). The results are shown in Table 32. The stability trend of charge heterogeneity is superimposed on Figures 14A-14C Under long-term storage conditions of 2°C-8°C ( Fig.14A , 14B and 14C), both formulations (F4 and F5) met the acceptance criteria for 24 months. Table 32: Main peak %, acidic % and basic % determined using icIEF at 2°C-8°C for Formulations 1 and 2 Subvisible particles using microfluidics imaging (MFI)

[0281] Subvisible particles in Formulations 4 and 5 were measured at different time points under long-term storage conditions at 2°C-8°C. The results are shown in Table 33. No significant changes in subvisible particles were observed. Overall, subvisible particles in the size range of ≥ (greater than or equal to) 10 μm were less than 6,000 particles / mL. In the size range of ≥ (greater than or equal to) 25 μm, even considering the increased sensitivity of using MFI for subvisible particle detection, subvisible particles were less than 600 particles / mL and well within USP <788> This data indicates that the 200 mg / mL formulation (F5) has no significant effect on subvisible particle counts under long-term storage conditions. Table 33: Subvisible particles of Formulations 4 and 5 at 2°C-8°C SbVP: subvisible particles; NT: not tested Chemical modification of primary structure using peptide mapping

[0282] The amino acids that may potentially undergo chemical modification and affect the protein structure over time in formulations 4 and 5 were monitored. In particular, the amino acid residues methionine 81 and methionine 254 of TEV-48574 may potentially undergo oxidation, thereby affecting the primary structure, and are monitored. Similarly, asparagine 317 of TEV-48574 may potentially undergo deamidation, thereby producing succinimide, and is monitored. As shown in Table 34, under long-term storage conditions of 2°C-8°C, no significant changes in Met81 oxidation % and Met254 oxidation % over time were observed within 24 months. At the 24M time point, the liquid drug product (F5) showed a higher level of Asn317 deamidation compared to the lyophilized drug product (F4). This difference in the degree of Asn deamidation may be a result of the lower stability of the liquid drug product compared to the lyophilized drug product, and may not be attributed to the high nominal protein concentration in F5. Table 34: Met81 oxidation %, Asn317 deamidation % and Met254 oxidation % in Formulations 4 and 5 F4: Formulation 4; F5: Formulation 5; Met81: methionine residue 81 of TEV-48574; Asn317: asparagine residue 317 of TEV-48574; Met254: methionine residue 254 of TEV-48574 Potency determined by ELISA

[0283] The percent (%) potency of Formulations 4 and 5 was determined by enzyme-linked immunosorbent assay (ELISA). The results are shown in Table 35. Under long-term storage conditions (2°C-8°C), no significant changes were observed between the formulations, and the acceptance criteria of 70%-135% potency were met within 24 months. Table 35: Relative % potency of Formulations 4 and 5 as determined by ELISA Secondary and tertiary protein structure analysis using circular dichroism (CD) spectroscopy

[0284] The secondary protein structure of Formulations 4 and 5 was analyzed using far ultraviolet (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-month, 6-month, and 12-month samples of Formulation 4 at 25°C; and secondary structure analysis was performed on the T0 sample and 3-month and 12-month samples of Formulation 5 at 25°C. Tertiary structure analysis was performed on the time zero (T0) sample and 3-month and 12-month samples of Formulation 4 at 25°C; and tertiary structure analysis was performed on the T0 sample and 3-month, 6-month, and 12-month samples of Formulation 5 at 25°C. The results were Figures 15A-15B ( Figures 15A-15B ). In particular, Fig.15A Shown are the secondary structures of Formulation 4 (F4) at time zero (T0) and after 3 months (3M), 6 months (6M) and 12 months (12M) storage at 25°C, and Formulation 5 (F5) at T0 and after 3M and 12M storage at 25°C, as determined using far UV CD. Fig. 15B The tertiary structure of F4 at T0 and after storage at 25°C for 3M and 12M and F5 at T0 and after storage at 25°C for 3M, 6M and 12M, determined using near UV CD, is shown. No significant changes in secondary or tertiary structure were observed for the lyophilized formulation (F4) or the high concentration liquid formulation (F5). in conclusion

[0285] The stability of TEV-48574 in lyophilized form (100 mg / mL Formulation 4) was comparable to the 200 mg / mL liquid formulation (Formulation 5). Under long-term storage conditions of 2°C-8°C, drug product stability data was observed to be comparable for both formulations.

[0286] At the 24M time point, 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 may be a result of the lower stability of the liquid drug product compared to the lyophilized drug product and may not be due to the high nominal protein concentration in F2. A slight decrease in the % monomer and a concurrent increase in the % low molecular weight species were also observed for F5 compared to F4. However, lyophilized formulations are typically more stable than liquid formulations.

[0287] Based on these data, it was observed that the 200 mg / mL high concentration liquid drug product was stable at 2°C-8°C with no significant impact on the critical quality attributes of TEV-48574. Example 4: Evaluation of Arginine-HCl as an Excipient in TEV-48574 Liquid Formulations

[0288] The objective of this study was to evaluate the effectiveness of arginine-HCl (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 at pH 6.0 with either 150 mg / mL (Formulation 1A) or 100 mg / mL (Formulation 2A) of the drug product. Two additional formulations were prepared in 10 mM histidine, 5% (w / v) sucrose, 0.02% (w / v) PS-80 at pH 6.0 with either 150 mg / mL (Formulation 1B) or 100 mg / mL (Formulation 2B) of the drug product. The stability of these formulations was evaluated under stress storage conditions at 40°C. The effect of these conditions on product quality attributes was evaluated. Visual appearance, protein concentration, osmolarity, and viscosity

[0289] The results of the visual appearance, protein concentration and osmotic pressure analysis of formulations 1A-2A and 1B-2B are shown in Tables 36-38. Under stress storage conditions at 40°C, the visual appearance of formulations 1A and 1B was observed to be slightly yellow, potentially related to the degradation of these formulations over time at elevated temperatures. At 40°C, the measured protein concentrations of the four formulations were close to the expected nominal concentrations and did not change over time. The osmotic pressure and viscosity of the drug product were evaluated, and no significant differences were observed over time. Table 36: Visual appearance results at 40°C T0: time zero; WK: week; 1A: formulation 1A; 1B: formulation 1B; 2A: formulation 2A; 2B: formulation 2B; L: colorless; FFVP: contains no visible particles; O: milky white; SY: slightly yellow; VSY: clearly slightly yellow; S: slightly milky white Table 37: Protein concentration results at 40°C T0: time zero; WK: week; 1A: preparation 1A; 1B: preparation 1B; 2A: preparation 2A; 2B: preparation 2B Table 38: Osmotic pressure results at 40°C T0: time zero; WK: week; 1A: preparation 1A; 1B: preparation 1B; 2A: preparation 2A; 2B: preparation 2B

[0290] The osmotic pressure of Formulations 1A and 2A containing Arg-HCl was higher than that of Formulations 1B and 2B without Arg-HCl. It was observed that at 150 mg / mL, the viscosity of the Formulation without Arg-HCl (1B) was higher than that of the Formulation with Arg-HCl (1A), as shown in Tables 39 and Fig.16 as shown in . Table 39: Viscosity of Formulations 1A and 1B at 20°C T0: time zero; 1A: preparation 1A; 1B: preparation 1B Size Exclusion Chromatography (SEC)

[0291] Table 40-41 and Figures 17A-17C The monomer percentage (%), dimer % and fragment species % of formulations 1A-1B and 2A-2B measured by SEC after storage at 40°C for up to 8 weeks are shown. No significant differences in monomer % or fragment % were observed among the 4 formulations, as the reported values ​​were within the method variability. Formulations 1B and 2B without Arg-HCl showed an increased rate of dimer formation compared to formulations 1A and 2A containing Arg-HCl, indicating the stabilizing effect of Arg-HCl on the drug product. Table 40: % Monomer, % Dimer and % Fragment Species of Formulations 1A-1B at 40°C T0: time zero; WK: week Table 41: % Monomer, % Dimer and % Fragment Species of Formulations 2A-2B at 40°C T0: time zero; WK: week Capillary gel electrophoresis (reducing and non-reducing)

[0292] The percentage (%) of immunoglobulin G (IgG) +125 kDa peak was determined for Formulations 1A-1B and 2A-2B using non-reducing capillary gel electrophoresis (CGE) under stress conditions at 40°C for up to 8 weeks. Additionally, the heavy chain + light chain % was determined for Formulations 1A-1B and 2A-2B 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 An increased fragmentation rate was observed for Formulations 1B and 2B lacking Arg-HCl compared to Formulations 1A and 2A containing Arg-HCl, demonstrating the stabilizing effect of Arg-HCl on the fragmentation pattern of the drug product. Table 42: % IgG + 125 kDa peak measured using non-reducing CGE at 40°C for Formulations 1A-1B and 2A-2B Time Preparation 1A Preparation 1B Preparation 2A Preparation 2B T0 98.5 98.4 98.4 98.4 1WK 98.0 98.1 98.1 98.2 2WK 97.7 97.8 97.8 97.9 3WK 97.5 97.5 97.3 97.4 4WK 96.9 97.0 96.9 97.1 6WK 96.2 96.1 96.2 96.5 8WK 95.5 95.3 95.4 95.6 T0: time zero; WK: week Table 43: % Heavy and Light Chains Measured Using Reduced CGE at 40°C for Formulations 1A-1B and 2A-2B Time Preparation 1A Preparation 1B Preparation 2A Preparation 2B T0 97.9 98.0 98.0 98.0 1WK 97.8 97.4 97.7 97.6 2WK 97.6 97.3 97.4 97.3 3WK 97.1 96.7 97.0 96.7 4WK 96.8 96.3 96.8 96.4 6WK 96.1 95.3 95.9 95.4 8WK 95.3 94.4 95.1 94.6 T0: time zero; WK: week Capillary isoelectric focusing (icIEF)

[0293] The content percentage (%) of the main species (main peak), acidic species (acidic peak) % and basic species (basic peak) % of 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. Figures 19A-19C For Formulations 1B and 2B lacking Arg-HCl, slightly higher levels of acidic species were observed at the end of 8 weeks. This may be related to the significantly higher fragmentation rates of these formulations compared to Formulations 1A and 2A, as reported in Table 43. Table 44: Main Peak %, Acidic %, and Basic % Determined Using icIEF at 40°C for Formulations 1A and 1B T0: time zero; WK: week Table 45: Main peak %, acidic species %, and basic species % determined using icIEF at 40°C for Formulations 2A and 2B T0: time zero; WK: week Chemical modification of primary structure using peptide mapping

[0294] Formulations 1A-1B and 2A-2B were monitored for amino acids that could potentially undergo chemical modification over time and affect protein structure under stress conditions at 40°C for up to 8 weeks. In particular, amino acid residues methionine 81 and methionine 254 of TEV-48574 could potentially undergo oxidation, thereby affecting primary structure, and were monitored. Similarly, asparagine 317 of TEV-48574 could potentially undergo deamidation, thereby producing succinimide, and were monitored. No significant changes were observed over time in Met81 oxidation %, Asn317 deamidation %, or Met254 oxidation % under stress storage conditions at 40°C for up to 8 weeks (data not shown), indicating that the presence of Arg-HCl did not appear to have an effect on this property. Potency determined by ELISA

[0295] The percent potency (%) of Formulations 1A-1B and 2A-2B was determined 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 trend of the potency % was comparable among the formulations regardless of drug product concentration or the inclusion of Arg-HCl. Table 46: Relative % potency of Formulations 1A-1B and 2A-2B as determined by ELISA T0: time zero; WK: week; 1A: preparation 1A; 1B: preparation 1B; 2A: preparation 2A; 2B: preparation 2B Dynamic Light Scattering (DLS) for Particle Size Characterization

[0296] DLS was used to measure the polydispersity % (PD%) and hydrodynamic radius of the drug product and nanoparticles that may be present 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. It was observed that the apparent hydrodynamic radius of formulations 1B and 2B (without Arg-HCl) was smaller than that of formulations 1A and 2A (with Arg-HCl). Table 47: Polydispersity % (PD%) and Radius of Particles of Formulations 1A-1B and 2A-2B T0: Time Zero Differential Scanning Calorimetry (DSC) Analysis of Thermal Stability

[0297] 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. Fig. 20 The thermal stabilities of Formulations 1A-1B and 2A-2B at time zero (T0) are shown, and the DSC thermograms for all four formulations are shown in Table 48. onset is about 60°C, and the T of Formulations 1B and 2B onset was about 63° C. For all formulations, the transition temperature (Tm) was about 78° C. Table 48: DSC Thermograms of Formulations 1A-1B and 2A-2B Preparations Tm1(℃) Tm2(℃) 1A 67.9 78.8 1B 71.2 79.8 2A 67.5 78.6 2B 70.7 79.5

[0298] The Tm1 of Formulations 1A and 2A was found to be slightly shifted to lower temperatures, demonstrating that Arg-HCl may affect the thermal stability of the molecule. This may be due to protein fragmentation observed in the samples under these conditions, resulting in a decrease in enthalpy. However, this temperature is higher than the recommended storage condition of 2°C-8°C for this drug product. in conclusion

[0299] The effectiveness of arginine-HCl (Arg-HCl) as an excipient and its effect on the stability of the TEV-48574 drug product were evaluated in two different drug product concentrations (100 mg / mL and 150 mg / mL). For all 4 formulations tested, the quality attributes of dimer% and purity% measured using reduced CGE indicated the stabilizing effect of 100 mM Arg-HCl compared to formulations without Arg-HCl. The viscosity of the drug product with Arg-HCl at 150 mg / mL was lower than that of the formulation without Arg-HCl. A slight shift of about 3 ° C of Tm1 analyzed by DSC was observed for the formulation containing Arg-HCl. However, the recommended storage conditions for drug products are 2 ° C-8 ° C, so it is expected that this observed shift of Tm1 will not significantly affect the stability during typical storage and handling conditions. At 40 ° C, all other attributes did not show significant changes in 8 weeks in all 4 formulations.

[0300] Based on these data, 100 mM Arg-HCl in the TEV-48574 formulation has shown the potential to stabilize the formulation and minimize viscosity as the concentration of drug product increases.

[0301] It should be understood that the detailed description section, rather than the summary and abstract sections, is intended to be used to interpret the claims. The summary and abstract sections may set forth one or more but not all exemplary aspects of the invention as contemplated by the inventors, and therefore are not intended to limit the invention and the appended claims in any way.

[0302] The foregoing description of particular aspects will so fully disclose the general nature of the invention that others can easily modify and / or adapt such particular aspects for a variety of applications by applying knowledge within the skill of the art without undue experimentation and without departing from the overall concept of the invention. Therefore, based on the teachings and guidance set forth herein, such adaptations and modifications are intended to be within the meaning and range of equivalents of the disclosed aspects. It should be understood that the terms or terminology herein are for purposes of description and not limitation, and therefore the terms or terminology of this specification will be interpreted by those skilled in the art based on the teachings and guidance.

[0303] The breadth and scope of the present invention should not be limited by any of the above-described exemplary aspects, but should be defined only in accordance with the following claims and their equivalents.

[0304] Various publications, including patents, published applications, accession numbers, technical articles, and academic articles, are cited throughout this specification. Each of these cited publications is incorporated into this document in its entirety and for all purposes by reference. Sequence Listing

Claims

1. A pharmaceutical preparation 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); 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, 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; (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.

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

4. The pharmaceutical formulation of any one of claims 1-3, 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. The pharmaceutical formulation of any one of claims 1-4, comprising about 100 mg / mL of the antibody or antigen-binding fragment thereof.

6. The pharmaceutical formulation of any one of claims 1-4, comprising about 150 mg / mL of the antibody or antigen-binding fragment thereof.

7. The pharmaceutical formulation of any one of claims 1-4, comprising about 200 mg / mL of the antibody or antigen-binding fragment thereof.

8. The pharmaceutical formulation of any one of claims 1-4, comprising about 225 mg / mL of the antibody or antigen-binding fragment thereof.

9. The pharmaceutical formulation of any one of claims 1-4, comprising about 250 mg / mL of the antibody or antigen-binding fragment thereof.

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

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

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

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

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

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

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

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

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

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

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

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

22. The pharmaceutical formulation of any one of claims 1-4, 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. The pharmaceutical formulation of any one of claims 1-4, 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. The pharmaceutical formulation of any one of claims 1-4, 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 1-24, wherein the pharmaceutical formulation is lyophilized.

26. The pharmaceutical formulation of any one of claims 1-24, wherein the pharmaceutical formulation is liquid.

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

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

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

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

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

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

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

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

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

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

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

38. The pharmaceutical formulation of any one of claims 1-37, having a main substance content of from 50% to 90%, an acid substance content of from 10% to 40%, and / or a basic substance content of from 0% to 20% after storage at 2°C-8°C for up to 36 months.

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

40. The pharmaceutical formulation of any one of claims 1-39, which 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°C-8°C, 25°C or 40°C for up to 36 months.

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

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

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

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

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

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

47. The pharmaceutical formulation of any one of claims 1-46, 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 1-47.

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

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

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

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

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

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

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

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

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

59. The formulation of any one of claims 1-47, for use according to the method of any one of claims 52-58.

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