Preparations containing anti-TIGIT antibodies and methods of using the same

Abstract

Provided are pharmaceutical formulations of antibodies, or antigen-binding fragments thereof, against the T cell immunoreceptor having Ig and ITIM domains (TIGIT). The pharmaceutical formulations exhibit a significant degree of anti-TIGIT antibody stability after being subjected to stress conditions, accelerated storage, and long-term storage. Also provided are methods of making and using such antibody formulations.

Description

[Technical Field]

[0001] Disclosed herein are stable formulations comprising an antibody or antigen-binding fragment thereof that binds to a T cell immunoreceptor having Ig and ITIM domains (TIGIT). Also disclosed herein are methods for preparing the disclosed formulations and methods for treating cancer with the formulations. [Background technology]

[0002] TIGIT is a newly discovered immunosuppressive costimulatory molecule. It is primarily expressed in T cells and NK cells and directly inhibits the killing effect of T cells and NK cells against tumor cells via its ITIM domain. Similar to the inhibitory receptors CTLA4 and PD-1, TIGIT also plays an important role in autoimmunity, making it another promising immunotherapy target.

[0003] Monoclonal antibody drugs are injected intravenously or subcutaneously. Antibodies can degrade and / or aggregate due to physical and chemical instability during storage, transportation, and use, resulting in decreased biological activity and increased immunogenicity. Physical instability includes antibody denaturation, aggregation, and / or precipitation. Chemical instability is caused by deamidation, isomerization, oxidation, or hydrolysis. To address these issues, some antibody formulations contain additional stabilizers, antioxidants, preservatives, etc. to improve antibody stability. However, the introduction of additional excipients increases the chance of infusion reactions in patients. Therefore, appropriate formulations of antibodies can maintain their physical and chemical stability. Summary of the Invention

[0004] The present invention provides an anti-TIGIT antibody pharmaceutical formulation.

[0005] Pharmaceutical preparations including: about 5 mg / mL to about 200 mg / mL of an anti-TIGIT antibody or an antigen-binding fragment thereof; about 5 mM to about 50 mM of a formulation buffer solution that provides a pH of about 5.0 to about 7.0; Stabilizer: about 30 mM to about 300 mM Nonionic surfactant: about 0.01 mg / ml to about 1 mg / ml.

[0006] The formulation, wherein the anti-TIGIT antibody or its antigen-binding fragment comprises a heavy chain variable region comprising HCDR1 (heavy chain complementarity determining region 1) of SEQ ID NO: 1, HCDR2 of SEQ ID NO: 2, and HCDR3 of SEQ ID NO: 3, and a light chain variable region comprising LCDR1 (light chain complementarity determining region 1) of SEQ ID NO: 4, LCDR2 of SEQ ID NO: 5, and LCDR3 of SEQ ID NO: 6.

[0007] The formulation, wherein the anti-TIGIT antibody or antigen-binding fragment thereof comprises SEQ ID NO:7 and SEQ ID NO:8.

[0008] The formulation, wherein the formulation buffer is selected from the group consisting of histidine, acetate, citrate, succinate, phosphate, a mixture of histidine and acetic acid, or a mixture of histidine and citric acid.

[0009] The formulation, wherein the formulation buffer is histidine.

[0010] The preparation, wherein the concentration of the histidine buffer is 10 mM to 30 mM.

[0011] The formulation comprises a 20 mM histidine buffer.

[0012] The formulation, wherein the pH is in the range of pH 5.2 to 6.2.

[0013] The formulation, wherein the stabilizer is selected from the group consisting of trehalose, sucrose, sorbitol, mannitol, maltose, dextran, (2-hydroxypropyl)-β-cyclodextrin, sodium chloride, magnesium chloride, calcium chloride, sodium sulfate, sodium dihydrogen phosphate, or disodium hydrogen phosphate.

[0014] The formulation, wherein the stabilizer is trehalose.

[0015] The preparation, wherein the trehalose concentration is 50 mM to 280 mM.

[0016] The preparation, wherein the trehalose concentration is 150 mM to 250 mM.

[0017] The formulation, wherein the stabilizer is sucrose.

[0018] The preparation, wherein the sucrose concentration is 50 mM to 280 mM.

[0019] The preparation, wherein the sucrose concentration is 150 mM to 250 mM.

[0020] The formulation, wherein the non-ionic surfactant is selected from the group consisting of polysorbate 20, polysorbate 80, or poloxamer 188.

[0021] The preparation, wherein the concentration of the polysorbate 20 is 0.1 mg / ml to 0.8 mg / ml.

[0022] The preparation, wherein the concentration of the polysorbate 20 is 0.2 mg / ml to 0.6 mg / ml.

[0023] The preparation, wherein the concentration of the polysorbate 80 is 0.1 mg / ml to 0.8 mg / ml.

[0024] The preparation, wherein the concentration of the polysorbate 80 is 0.2 mg / ml to 0.6 mg / ml.

[0025] The preparation, wherein the concentration of the poloxamer 188 is 0.1 mg / ml to 0.8 mg / ml.

[0026] The formulation, wherein the concentration of the poloxamer 188 is 0.2 mg / ml to 0.6 mg / ml.

[0027] The formulation comprises 30 mM acetic acid-sodium acetate, 240 mM sucrose, and 0.2 mg / ml polysorbate 80, and has a pH of pH 5.5.

[0028] The formulation comprises 20 mM histidine-histidine HCl, 240 mM trehalose, and 0.2 mg / ml polysorbate 20, and has a pH of pH 5.8.

[0029] The formulation comprises 20 mM histidine-histidine HCl, 70 mM NaCl, 80 mM trehalose, and 0.8 mg / ml polysorbate 20, and has a pH of pH 6.0.

[0030] The formulation, wherein the concentration of the anti-TIGIT antibody or antigen-binding fragment thereof is about 10 mg / mL to 150 mg / mL.

[0031] A method for producing an antibody formulation, comprising: a. exchanging the anti-TIGIT antibody for about 5 mM to about 50 mM of a buffer solution that provides a pH of about 5.0 to about 7.0; b. concentrating the antibody formulation of (a) to an antibody concentration of about 5 to 200 mg / mL; c. Adding a nonionic surfactant to the antibody formulation of (c) to obtain an antibody formulation having a surfactant concentration of 0.01 mg / ml or more; d. adding a stabilizer to the antibody to obtain an antibody formulation having a stabilizer concentration of 30 mM or more, wherein the anti-TIGIT antibody comprises a heavy chain variable region comprising HCDR1 (heavy chain complementarity determining region 1) of SEQ ID NO: 1, HCDR2 of SEQ ID NO: 2, and HCDR3 of SEQ ID NO: 3, and a light chain variable region comprising LCDR1 (light chain complementarity determining region 1) of SEQ ID NO: 4, LCDR2 of SEQ ID NO: 5, and LCDR3 of SEQ ID NO: 6.

[0032] The formulation, wherein the anti-TIGIT antibody or antigen-binding fragment thereof comprises SEQ ID NO:7 and SEQ ID NO:8.

[0033] A method for treating cancer in a human patient in need thereof, comprising administering an effective amount of the anti-TIGIT antibody formulation of claim 1.

[0034] The method, wherein the anti-TIGIT antibody formulation is administered at a dose of about 200 mg to about 2400 mg.

[0035] The method, wherein the anti-TIGIT antibody formulation is administered once every three weeks.

[0036] The method, wherein the human patient is administered at least one other therapeutic agent selected from the group consisting of zanubrutinib, pamiparib, tislelizumab, an anti-LAG3 antibody, a second anti-TIGIT antibody, an anti-4-1BB antibody, an anti-OX40 antibody, an anti-TIM-3 antibody, a CD40 agonist, a TLR agonist, a CAR-T cell, or a chemotherapeutic agent.

[0037] In some embodiments, the antibody formulation comprises an anti-TIGIT antibody or antigen-binding fragment thereof, a formulation buffer, a stabilizer, and a non-ionic surfactant. In some embodiments, the formulation buffer provides a pH range of 5.0 to 7.0. In some embodiments, the antibody formulation is stable to freeze-thawing and heat stress.

[0038] In some embodiments, the antibody formulation can comprise or consist essentially of about 10 mg / mL to about 40 mg / mL of an anti-TIGIT antibody or antigen-binding fragment thereof, a formulation buffer, a stabilizer, and a non-ionic surfactant, and has a pH range of 5.2 to 6.2. In some embodiments, the antibody formulation is stable under stress conditions, accelerated storage, and long-term storage.

[0039] In some embodiments, the formulation buffer is selected from the group consisting of histidine, acetate, citrate, succinate, phosphate, a mixture of histidine and acetate, a mixture of histidine and citric acid, or any combination thereof. In some embodiments, the formulation buffer can be a histidine buffer. In some embodiments, the histidine buffer has a concentration of about 10 mM to about 30 mM. In some embodiments, the histidine buffer has a concentration of about 20 mM histidine.

[0040] In some embodiments, the stabilizer is selected from the group consisting of trehalose, sucrose, sorbitol, mannitol, maltose, dextran, (2-hydroxypropyl)-β-cyclodextrin, sodium chloride, magnesium chloride, calcium chloride, sodium sulfate, sodium dihydrogen phosphate, or disodium hydrogen phosphate. In some embodiments, the stabilizer may be trehalose. In some embodiments, the trehalose is a,α-trehalose dihydrate. In other embodiments, the stabilizer may be sucrose. In some embodiments, the concentration of the stabilizer may be about 30 mM to about 300 mM. In some embodiments, the concentration of the stabilizer may be about 50 mM to about 280 mM, preferably about 150 mM, about 170 mM, about 190 mM, about 210 mM, about 230 mM, or about 250 mM.

[0041] In some embodiments, the nonionic surfactant is selected from the group consisting of polysorbate 80 (PS80), polysorbate 20 (PS20), or poloxamer 188 (P188). In some embodiments, the concentration of the nonionic surfactant can be about 0.1 mg / ml to about 0.8 mg / ml. In some embodiments, the concentration of the nonionic surfactant is about 0.2 mg / ml, about 0.3 mg / ml, about 0.4 mg / ml, about 0.5 mg / ml, or about 0.6 mg / ml. In some embodiments, the nonionic surfactant is polysorbate 20. In some embodiments, the nonionic surfactant is polysorbate 80. In some embodiments, the nonionic surfactant is poloxamer 188.

[0042] In some embodiments, the antibody formulation consists essentially of about 10 mg / mL, about 20 mg / mL, about 30 mg / mL, about 40 mg / mL, about 50 mg / mL, about 60 mg / mL, about 70 mg / mL, about 80 mg / mL, about 90 mg / mL, about 100 mg / mL, about 110 mg / mL, about 120 mg / mL, about 130 mg / mL, about 140 mg / mL, or about 150 mg / mL of an anti-TIGIT antibody or antigen-binding fragment thereof, about 20 mM histidine buffer, about 240 mM a,a-trehalose dihydrate or sucrose, and about 0.2 mg / mL to about 0.6 mg / mL of polysorbate 20 or polysorbate 80 or poloxamer 188, and the antibody formulation has a pH of 5.8±0.4.

[0043] Also provided herein is a method for producing a stable anti-TIGIT antibody formulation, the method comprising: exchanging the anti-TIGIT antibody into about 5 mM to about 50 mM of a buffer solution that provides a pH of about 5.0 to about 7.0; concentrating the antibody formulation to 5 to 200 mg / mL; adding a stabilizer to the antibody to obtain an antibody formulation having a stabilizer concentration of 30 mM or greater; and adding a non-ionic surfactant to obtain an antibody formulation having a surfactant concentration of 0.001% (w / v) or greater.

[0044] Also provided herein is a method of inhibiting cancer growth in a human patient with cancer, the method comprising administering to the patient an effective amount of an antibody formulation described herein.

[0045] Provided herein are methods of inhibiting cancer growth in a human patient, the methods comprising administering to the patient an effective amount of an antibody formulation described herein.

[0046] In some embodiments, the antibody formulation has an antibody concentration of about 200 mg to 2400 mg. In other embodiments, the antibody formulation has an antibody concentration of about 200 mg, about 300 mg, about 400 mg, about 500 mg, about 600 mg, about 700 mg, about 800 mg, about 900 mg, about 1000 mg, about 1100 mg, about 1200 mg, about 1300 mg, about 1400 mg, about 1500 mg, about 1600 mg, about 1700 mg, about 1800 mg, about 1900 mg, about 2000 mg, about 2100 mg, about 2200 mg, about 2300 mg, or about 2400 mg. In some embodiments, the antibody formulation is administered once every three weeks. In some embodiments, the antibody formulation is about 400 mg to 1200 mg and is administered once every three weeks.

[0047] In some embodiments, the present disclosure provides methods of treating cancer with a combination of an anti-TIGIT subcutaneous antibody formulation and another therapeutic agent, such as zanubrutinib, pamiparib, tislelizumab (BGB-A317), an anti-LAG3 antibody, a second anti-TIGIT antibody, an anti-4-1BB antibody, an anti-OX40 antibody, an anti-TIM-3 antibody, a CD40 agonist, a TLR agonist, a CAR-T cell, or a chemotherapeutic agent. [Brief explanation of the drawings]

[0048] [Figure 1A] 1 shows the results of different pH and buffer solutions for stability from 5 cycles of freeze-thaw conditions (labeled "5FT" on the graph), showing data for particles that are not visible to the naked eye. [Figure 1B]Figure 1 shows the results of different pH and buffer solutions for stability from 5 cycles of freeze-thaw conditions (labeled "5FT" on the graph). Turbidity data. [Figure 1C] 1 shows the results of different pH and buffer solutions on stability from 5 cycles of freeze-thaw conditions (labeled "5FT" on the graph). SEC % monomer data. [Figure 2A] The results of different protein concentrations on stability from 5 cycles of freeze-thaw conditions (labeled "5FT" on the graph) are shown. Data are for particles that are not visible to the naked eye. [Figure 2B] Figure 1 shows the results of different protein concentrations on stability from 5 cycles of freeze-thaw conditions (labeled "5FT" on the graph). Turbidity data. [Figure 2C] Figure 1 shows the results of different protein concentrations on stability from 5 cycles of freeze-thaw conditions (labeled "5FT" on the graph). SEC % monomer data. [Figure 3A] The results of different surfactants for stability under different stress conditions are shown: three cycles of freeze-thaw (labeled "3FT" on the graph), two days of shaking (labeled "Shake 2D" on the graph), two weeks of photostability (labeled "Photo 2W" on the graph), and heat stress of a formulation kept at 37°C for four weeks (labeled "37C4W" on the graph). Data are for particles not visible to the naked eye. [Figure 3B] The results of different surfactants for stability under different stress conditions, namely, three cycles of freeze-thaw (labeled "3FT" on the graph), two days of shaking (labeled "Shake 2D" on the graph), two weeks of photostability (labeled "Photo 2W" on the graph), and heat stress of a formulation kept at 37°C for four weeks (labeled "37C4W" on the graph), are shown. Turbidity data. [Figure 3C]The results of different surfactants for stability under different stress conditions, namely, three cycles of freeze-thaw (labeled "3FT" on the graph), two days of shaking (labeled "Shake 2D" on the graph), two weeks of photostability (labeled "Photo 2W" on the graph), and heat stress of a formulation kept at 37°C for four weeks (labeled "37C4W" on the graph), are shown. SEC % monomer data. [Figure 3D] 1 shows the results of different surfactants for stability under different stress conditions, namely, 3 cycles of freeze-thaw (labeled "3FT" on the graph), 2 days of shaking (labeled "Shake 2D" on the graph), 2 weeks of photostability (labeled "Photo 2W" on the graph), and heat stress of the formulation kept at 37°C for 4 weeks (labeled "37C4W" on the graph). % of IEC main peak data. [Figure 3E] The results of different surfactants for stability under different stress conditions are shown: three cycles of freeze-thaw (labeled "3FT" on the graph), two days of shaking (labeled "Shake 2D" on the graph), two weeks of photostability (labeled "Photo 2W" on the graph), and heat stress of the formulation maintained at 37°C for four weeks (labeled "37C4W" on the graph). CE-SDS(NR) % intact peak data. [Figure 4A] The graph shows the results of stability under stress conditions, namely, heat stress in which the antibody formulation was maintained at 40°C for two weeks (labeled "40C2W" on the graph), light stability of the formulation over two weeks (labeled "Photo 2W" on the graph), and freeze-thawing (labeled "6FT" on the graph). The data shows particles that cannot be seen with the naked eye. [Figure 4B] The graph shows the stability results under stress conditions, namely, heat stress in which the antibody formulation was maintained at 40°C for two weeks (labeled "40C2W" on the graph), light stability of the formulation over two weeks (labeled "Photo 2W" on the graph), and freeze-thaw (labeled "6FT" on the graph). The data show the % monomer data from SEC. [Figure 4C]The graph shows the stability results under stress conditions, namely, heat stress in which the antibody formulation was maintained at 40°C for two weeks (labeled "40C2W" on the graph), light stability of the formulation over two weeks (labeled "Photo 2W" on the graph), and freeze-thaw (labeled "6FT" on the graph). The data are the percentage of the main IEC peak. [Figure 5A] Stability data at 25°C is shown, showing particles that are not visible to the naked eye. [Figure 5B] The stability data at 25° C. are shown. SEC % monomer data. [Figure 5C] The stability data at 25°C is shown. The data is the percentage of the IEC main peak. [Figure 5D] The stability data at 25°C is shown. The data shows the % of CE-SDS(NR) intact peak. [Figure 6A] Stability data at 5±3°C are shown. Data are for particles not visible to the naked eye. [Figure 6B] The stability data at 5±3° C. are shown. SEC data of % monomer. [Figure 6C] The stability data at 5±3°C is shown. The data shows the percentage of the IEC main peak. [Figure 6D] The stability data at 5±3°C is shown. The data shows the % of CE-SDS(NR) intact peaks. DETAILED DESCRIPTION OF THE INVENTION

[0049] definition Unless specifically defined elsewhere in this document, all other technical and scientific terms used herein have the meaning commonly understood by those of ordinary skill in the art.

[0050] As used in this specification, including the appended claims, singular words such as "a," "an," and "the" include their corresponding plural referents unless the context clearly indicates otherwise.

[0051] The term "or" is used to mean, and is used interchangeably with, the term "and / or," unless context clearly indicates otherwise.

[0052] Throughout this specification and the claims that follow, unless the context requires otherwise, the word "comprise" and variations such as "comprises" and "comprising" will be understood to mean the inclusion of a stated amino acid sequence, DNA sequence, step, or group thereof, but not the exclusion of any other amino acid sequences, DNA sequences, or steps. As used herein, the term "comprising" can be interchanged with the terms "containing," "including," or, where appropriate, "having."

[0053] As used herein, the terms "administration," "administering," "treating," and "treatment," when applied to an animal, human, experimental subject, cell, tissue, organ, or biological fluid, refer to contacting an exogenous pharmaceutical, therapeutic, diagnostic, or antibody preparation with the animal, human, subject, cell, tissue, organ, or biological fluid. Treatment of a cell encompasses contacting a reagent with the cell and contacting a reagent with a fluid (wherein the fluid is in contact with the cell). The terms "administration" and "treatment" also refer to in vitro and ex vivo treatments, e.g., treatment of a cell with a reagent, diagnostic, binding compound, or another cell. As used herein, the term "subject" includes any organism, preferably an animal, more preferably a mammal (e.g., a rat, mouse, dog, cat, rabbit), and most preferably a human. In one aspect, treating any disease or disorder refers to ameliorating the disease or disorder (i.e., delaying, preventing, or reducing the onset of the disease or at least one of its clinical symptoms). In another embodiment, "treat," "treating," or "treatment" refers to alleviating or improving at least one physical parameter, including those that may not be discernible by the patient. In yet another embodiment, "treat," "treating," or "treatment" refers to modulating a disease or disorder physically (e.g., stabilization of discernible symptoms), physiologically (e.g., stabilization of physical parameters), or both.

[0054] As used herein, a "subject" is a mammal, e.g., a rodent or a primate, preferably a higher primate, e.g., a human (e.g., a patient having or at risk of having a disorder described herein).

[0055] As used herein, the term "therapeutically effective amount" refers to the amount of an anti-TIGIT antibody that, when administered to a subject to treat a disease or at least one clinical symptom of a disease or disorder, is sufficient to affect treatment for the disease, disorder, or condition. A "therapeutically effective amount" may vary depending on the drug, disease, disorder, and / or symptoms of the disease or disorder, the severity of the disease, disorder, and / or symptoms of the disease or disorder, the age of the subject being treated, and / or the weight of the subject being treated. The appropriate amount in any given case will be apparent to one skilled in the art or can be determined by routine experimentation. In the case of combination therapy, a "therapeutically effective amount" refers to the total amount of the combined components for effective treatment of a disease, disorder, or condition. In some embodiments of the present disclosure, the subject is a human.

[0056] The term "antibody" as used herein is used in the broadest sense and specifically encompasses antibodies (including full-length monoclonal antibodies) and antibody fragments, so long as they recognize an antigen, e.g., TIGIT. Antibodies are usually monospecific but may be described as idiospecific, heterospecific, or multispecific. An antibody molecule binds to a specific antigenic determinant or epitope on an antigen via a specific binding site.

[0057] As used herein, the term "monoclonal antibody" or "mAb" or "Mab" refers to a population of substantially homogeneous antibodies, i.e., the antibody molecules within the population are identical in amino acid sequence except for possible naturally occurring mutations that may be present in minor amounts. In contrast, conventional (polyclonal) antibody preparations typically contain a large number of different antibodies with different amino acid sequences within the variable domains, particularly the complementarity-determining regions (CDRs), which are often specific for different epitopes. The modifier "monoclonal" indicates the character of the antibody as being obtained from a population of substantially homogeneous antibodies and is not to be construed as requiring production of the antibody by any particular method. Monoclonal antibodies (mAbs) may be obtained by methods known to those of skill in the art. See, for example, Kohler G et al., Nature 1975 256:495-497; U.S. Patent No. 4,376,110; Ausubel FM et al., CURRENT PROTOCOLS IN MOLECULAR BIOLOGY 1992; Harlow E et al., ANTIBODIES: A LABORATORY MANUAL, Cold Spring Harbor Laboratory 1988; and Colligan JE et al., CURRENT PROTOCOLS IN IMMUNOLOGY 1993. The monoclonal antibodies disclosed herein may be of any immunoglobulin class, including IgG, IgM, IgD, IgE, IgA, and any subclass thereof. Hybridomas producing monoclonal antibodies can be cultured in vitro or in vivo. High-titer monoclonal antibodies can be obtained by in vivo production. Here, cells from individual hybridomas are injected intraperitoneally into mice, such as pristine-primed Balb / c mice, to produce ascites fluid containing high concentrations of the desired monoclonal antibodies. Monoclonal antibodies of the IgM or IgG isotype can be purified from such ascites fluid or from the culture supernatant using column chromatography methods well known to those skilled in the art.

[0058] Generally, the basic structural unit of an antibody comprises a tetramer. Each tetramer contains two identical pairs of polypeptide chains, each pair having one "light chain" (approximately 25 kDa) and one "heavy chain" (approximately 50-70 kDa). The amino-terminal portion of each chain contains a variable region of approximately 100-110 amino acids in length, primarily responsible for antigen recognition. The carboxy-terminal portion of the heavy chain may define a constant region primarily responsible for effector function. Human light chains are typically classified as kappa and lambda light chains. Human heavy chains are further typically classified as α, δ, ε, γ, or μ, and the antibody isotype is defined as IgA, IgD, IgE, IgG, and IgM, respectively. Within the light and heavy chains, the variable and constant regions are connected by a "J" region of approximately 12 or more amino acids, and heavy chains also contain a "D" region of approximately 10 amino acids.

[0059] The variable regions of each light chain / heavy chain (VL / VH) pair form the antibody binding site. Therefore, an intact antibody generally has two binding sites. Except for bifunctional or bispecific antibodies, the two binding sites are generally the same. Typically, both the heavy and light chain variable domains contain three hypervariable regions, also called "complementarity-determining regions (CDRs)," located between relatively conserved framework regions (FRs). The CDRs are usually aligned by the framework regions, enabling binding to a specific epitope. Generally, both the light and heavy chain variable domains contain, from the N-terminus to the C-terminus, FR-1 (or FR1), CDR-1 (or CDR1), FR-2 (FR2), CDR-2 (CDR2), FR-3 (or FR3), CDR-3 (CDR3), and FR-4 (or FR4), in that order. The assignment of amino acids to each domain generally follows the definitions in Sequences of Proteins of Immunological Interest, Kabat, et al., National Institutes of Health, Bethesda, Md.; 5th ed.; NIH Publ. No. 91-3242 (1991), Kabat (1978) Adv. Prot. Chem. 32:1-75, Kabat, et al., (1977) J. Biol. Chem. 252:6609-6616, Chothia, et al., (1987) J Mol. Biol. 196:901-917 or Chothia, et al., (1989) Nature 342:878-883.

[0060] The term "hypervariable region" refers to the amino acid residues of an antibody that are involved in antigen binding. The hypervariable region comprises the amino acid residues of the "CDRs" (i.e., VL-CDR1, VL-CDR2, and VL-CDR3 of the light chain variable domain and VH-CDR1, VH-CDR2, and VH-CDR3 of the heavy chain variable domain). See Kabat et al. (1991) Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (where antibody CDR regions are defined by sequence); see also Chothia and Lesk (1987) J. Mol. Biol. 196:901-917 (where antibody CDR regions are defined by structure). The term "framework" or "FR" residues refers to variable domain residues other than the hypervariable region residues defined herein as CDR residues.

[0061] Unless otherwise indicated, "antibody fragment" or "antigen-binding fragment" refers to an antigen-binding fragment of an antibody, i.e., an antibody fragment that retains the ability to specifically bind to the antigen bound by the full-length antibody, e.g., a fragment that retains one or more CDR regions. Examples of antigen-binding fragments include, but are not limited to, Fab, Fab', F(ab'), and Fv fragments, diabodies, linear antibodies, single-chain antibody molecules such as single-chain Fvs (ScFvs), nanobodies formed from antibody fragments, and multispecific antibodies.

[0062] An antibody that specifically binds to a particular target protein is also described as specifically binding to the particular target protein. This means that the antibody exhibits preferential binding to that target compared to other proteins, but this specificity does not require absolute binding specificity. An antibody is considered "specific" for an intended target if its binding determines the presence of the target protein in a sample and does not produce undesirable results, such as false positives. Antibodies or binding fragments thereof useful in the present invention bind to the target protein with an affinity that is at least 2-fold, preferably at least 10-fold, more preferably at least 20-fold, and most preferably at least 100-fold greater than the affinity for a non-target protein. Antibodies herein are said to specifically bind to a polypeptide comprising a given amino acid sequence.

[0063] The term "human antibody" herein refers to an antibody that contains only human immunoglobulin protein sequences. A human antibody may contain murine carbohydrate chains if produced in a mouse, a mouse cell, or a hybridoma derived from a mouse cell. Similarly, a "mouse antibody" or a "rat antibody" refers to an antibody that contains only mouse or rat immunoglobulin protein sequences, respectively.

[0064] The term "humanized antibody" refers to forms of antibodies that contain both non-human (e.g., murine) and human antibody sequences. Such antibodies contain minimal sequence derived from non-human immunoglobulins. Generally, humanized antibodies contain substantially all of at least one, and typically two, variable domains, in which all or substantially all hypervariable loops correspond to those of a non-human immunoglobulin and all or substantially all FR regions are those of a human immunoglobulin. A humanized antibody optionally also contains at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. When it is necessary to distinguish a humanized antibody from its parent rodent antibody, the prefix "hum," "hu," "Hu," or "h" is added to the name of the antibody clone. Humanized forms of rodent antibodies generally contain the same CDR sequences as the parent rodent antibody but may contain certain amino acid substitutions for improved affinity, increased stability of the humanized antibody, or other reasons.

[0065] The antibodies of the present application have potential therapeutic applications in the treatment of cancer. As used herein, the term "cancer" or "tumor" refers to or describes a physiological condition in mammals that is typically characterized by unregulated cell growth. Examples of cancer include, but are not limited to, lung cancer (including small cell lung cancer or non-small cell lung cancer), adrenal cancer, liver cancer, gastric cancer, cervical cancer, melanoma, kidney cancer, breast cancer, colorectal cancer, leukemia, bladder cancer, bone cancer, brain tumor, endometrial cancer, head and neck cancer, lymphoma, ovarian cancer, skin cancer, thyroid tumor, or metastatic lesions of cancer.

[0066] Furthermore, the antibodies of the present application have potential therapeutic applications in the control of viral infections and other human diseases that mechanistically involve immune tolerance or "exhaustion." In the context of the present application, the term "exhaustion" refers to a process that leads to the exhaustion of immune cells' ability to respond to cancer or chronic viral infection.

[0067] Anti-TIGIT antibody The present disclosure provides anti-TIGIT antibodies and formulations thereof. For example, osipellimab (BGB-A1217) is an anti-TIGIT antibody disclosed in PCT Patent No. WO2019 / 129261, and has the sequence shown in Table 1 below. [Table 1]

[0068] Treatment method The antibodies or antigen-binding fragments of the present disclosure are useful for various applications, including, but not limited to, methods for treating TIGIT-related disorders or diseases. In one embodiment, the TIGIT-related disorder or disease is cancer.

[0069] In one embodiment, the present disclosure provides a method for treating cancer.In certain embodiments, the method comprises administering an effective amount of anti-TIGIT antibody or antigen-binding fragment to a patient in need thereof.The cancer can include but is not limited to lung cancer (including small cell lung cancer or non-small cell lung cancer), adrenal cancer, liver cancer, gastric cancer, cervical cancer, melanoma, kidney cancer, breast cancer, colorectal cancer, leukemia, bladder cancer, bone cancer, brain tumor, endometrial cancer, head and neck cancer, lymphoma, ovarian cancer, skin cancer, thyroid tumor or metastatic lesion of cancer.

[0070] The antibodies or antigen-binding fragments of the present invention can be administered by any suitable means, including parenteral, intrapulmonary, and intranasal administration, as well as intralesional administration if desired for localized treatment. Parenteral infusions include intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration. Dosing can be by any suitable route, for example, injection, such as intravenous or subcutaneous injection, depending in part on whether administration is brief or chronic. Various dosing schedules are contemplated herein, including, but not limited to, single or multiple doses over various time periods, bolus administration, and pulse infusion.

[0071] Anti-TIGIT antibodies or antigen-binding fragments can be formulated, dispensed, and administered in a manner consistent with the principles of good medical practice. Factors to consider in this regard include the particular disorder being treated, the particular mammal being treated, the clinical condition of the individual patient, the cause of the disorder, the site of drug delivery, the method of administration, the schedule of administration, and other factors known to medical professionals. The antibody is optionally, but need not be, formulated with one or more agents currently used to prevent or treat the disorder in question. The effective amount of such other agents will vary depending on the amount of antibody present in the formulation, the type of disorder or treatment, and the other factors discussed above. These will generally be used in the same dosages and by the same routes of administration as those described herein, or about 1-99% of the dosages described herein, or at any dosage and by any route empirically / clinically determined to be appropriate.

[0072] The appropriate dosage of the antibody or antigen-binding fragment of the present invention for the prevention or treatment of disease will vary depending on the type of disease being treated, the type of antibody, the severity and course of the disease, whether the antibody is administered for prophylactic or therapeutic purposes, previous therapy, the patient's clinical history and response to the antibody, and the judgment of the attending physician. The antibody is appropriately administered to the patient at one time or over a series of treatments. Depending on the type and severity of the disease, an initial dosage of about 200 mg to 2400 mg of antibody can be administered to the patient, for example, in one or more individual administrations or by continuous infusion. A typical daily dosage can range from about 200 mg to 2400 mg of antibody or more, depending on the factors mentioned above. For repeated administration over several or longer days, depending on the condition, the treatment is generally sustained until a desired suppression of disease symptoms occurs. Such doses can be administered intermittently, for example, weekly, every two weeks, or every three weeks. An initial higher loading dose can be administered, followed by one or more lower doses. However, other dosage regimens may be useful. The progress of this therapy is easily monitored by conventional techniques and assays.

[0073] Pharmaceutical Compositions and Formulations Also provided is a composition, including pharmaceutical preparations, comprising anti-TIGIT antibody or its antigen-binding fragment, or a polynucleotide comprising a sequence encoding anti-TIGIT antibody or antigen-binding fragment.In certain embodiments, the composition comprises one or more antibodies or antigen-binding fragments that bind to TIGIT, or one or more polynucleotides comprising a sequence encoding one or more antibodies or antigen-binding fragments that bind to TIGIT.These compositions can further comprise suitable carriers, for example, pharmaceutically acceptable excipients such as buffers well known in the art.

[0074] Pharmaceutical formulations of the anti-TIGIT antibodies or antigen-binding fragments described herein are prepared in the form of lyophilized formulations or aqueous solutions by mixing such antibodies or antigen-binding fragments having the desired purity with one or more pharmaceutically acceptable carriers. Pharmaceutically acceptable carriers are generally non-toxic to recipients at the dosages and concentrations used and may contain buffers, e.g., phosphate, citrate, and other organic acids, antioxidants including ascorbic acid and methionine, preservatives (such as octadecyldimethylbenzylammonium chloride, hexamethonium chloride, benzalkonium chloride, benzethonium chloride, phenol, butyl or benzyl alcohol, alkyl parabens such as methyl or propyl paraben, catechol, resorcinol, cyclohexanol, 3-pentanol, and m-cresol), low molecular weight (less than about 10 residues) polypeptides, proteins, etc. Examples of suitable surfactants include, but are not limited to, surfactants such as serum albumin, gelatin, or immunoglobulins, hydrophilic polymers such as polyvinylpyrrolidone, amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine, monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins, chelating agents such as EDTA, sugars such as sucrose, mannitol, trehalose, or sorbitol, salt-forming counterions such as sodium, metal complexes (e.g., Zn-protein complexes), and / or non-ionic surfactants such as polyethylene glycol (PEG).

[0075] Sustained-release preparations can be prepared. Suitable examples of sustained-release preparations include semipermeable matrices of solid hydrophobic polymers containing the antibody, which matrices are in the form of shaped articles, e.g., films, or microcapsules.

[0076] Formulations to be used for in vivo administration are generally sterile. Sterility may be readily accomplished, for example, by filtration through sterile filtration membranes. [Example]

[0077] The examples and descriptions of certain embodiments should be considered illustrative rather than limiting of the invention as defined by the claims. As will be readily appreciated, numerous variations and combinations of the features described above can be utilized without departing from the invention as set forth in the claims. All such variations are intended to be within the scope of the present invention. All references are incorporated herein by reference in their entirety.

[0078] Analysis method The Methods section outlines the methods used in Examples 1-4 below.

[0079] SEC-HPLC The formation of soluble aggregates and fragments is analyzed by size exclusion chromatography (SEC) on a Waters™ HPLC system. Using an isocratic gradient, proteins are separated based on molecular size on a TSK gel Super SW™ mAb HR, 7.8 x 300 mm column maintained at 37 ± 5°C. Molecular weight species are eluted and detected by UV absorbance at 215 nm. The distribution of aggregates, monomers, and fragments is quantified by peak areas of standards and samples.

[0080] IEC-HPLC Ion Exchange Chromatography (IEC) IEC-HPLC method was used to evaluate charge variants by cation exchange chromatography. The HPLC system (Waters) was equipped with a Thermo MabPac SCX-10™ analytical column (4 x 250 mm) at 37 ± 5°C. Gradient NaCl elution was performed at a constant flow rate of 1.0 mL / min, and UV signals were acquired at 280 nm. Peaks in the IEC-HPLC chromatogram were integrated, and the peak area percentage of each peak was calculated.

[0081] CE-SDS(NR). Sample purity is determined by capillary gel electrophoresis (CE) using a PA800 Plus™ (Beckman). Samples are denatured with sodium dodecyl sulfate (SDS) and separated based on size in a capillary filled with gel, which acts as a sieving medium. For non-reduced (NR) samples, the alkylating agent N-ethylmaleimide (NEM) is added to avoid any fragmentation caused by the sample preparation and ensure that the major IgG peak remains intact. Samples are injected electrokinetically, and a UV detector is used to detect the migrated proteins by UV absorbance at 200 nm. The reportable value for non-reduced samples is the time-corrected area percent (TCA)% of the major IgG peak.

[0082] Protein concentration Protein concentration is determined by UV 280 nm.

[0083] Tm and Tag Stability determinations were performed using Uncle™ (Unchained Labs), which combines three different measurement modes: fluorescence, static light scattering (SLS), and dynamic light scattering (DLS). SLS and intrinsic fluorescence methods were performed to determine the aggregation onset temperature (Tagg) and melting temperature (Tm) of the formulations, respectively.

[0084] Turbidity The turbidity test was performed using a NEPHELOstar plus™ with a laser light of 635 nm wavelength. Samples were placed in a 96-well plate and the turbidity was read by the NEPHELOstar plus™.

[0085] Visible particles Visible particles were inspected against black and white backgrounds using approximately 2000 lux white fluorescent lighting. The vial under inspection was gently rotated and inspected for at least 10 seconds against each background. Total inspection time was approximately 20 seconds. Particles that cannot be seen with the naked eye Micro-Flow Imaging (MFI, Micro-Flow Imaging™ 5200, ProteinSimple) was used to analyze particles that are not visible to the naked eye. A water flush was performed before each sample analysis to ensure background counts were adequate for testing. The average cumulative counts per mL were reported.

[0086] Example 1: Effect of pH and buffer on the stability of anti-TIGIT antibody formulations To determine the antibody stability at different pHs and buffers, osipellimab was dialyzed into different buffers using a 10 kDa MWCO dialysis cassette and prepared into the formulations listed in Table 2. These formulations were filtered through Millex™ GP 0.22 mm PES 33 mm filters and filled into 2 mL ready-to-use glass vials. To assess the impact on antibody stability, the Tm and Tag of these antibody samples were determined using Uncle™ (Unchained Labs), and the data are shown in Table 2. Additionally, these antibody samples were subjected to five freeze-thaw cycles (denoted "5FT" in Figure 2). As a control, these antibody samples were analyzed at the starting time point, designated TO in Figures 1A-C. Subvisible particles, turbidity, and purity by SEC-HPLC were measured for all samples. The results are shown graphically in Figures 1A-C. [Table 2]

[0087] Table 2 shows that all formulations had Tm1 above 65°C, Tm2 above 75°C, and Tg above 70°C, which are good indicators of stability. Subvisible particles and turbidity increased in formulations without stabilizers or surfactants after five freeze-thaw cycles, whereas no significant increase was observed in antibody formulations containing stabilizers and surfactants. All of these formulations showed no significant changes after five freeze-thaw cycles and demonstrated 95% or greater monomer content in SEC. Collectively, the subvisible particle data (Figure 1A), turbidity data (Figure 1B), and SEC-HPLC data (Figure 1C) demonstrated that the osipellimab antibody was stable in formulations containing stabilizers and surfactants in different buffers at pH 5.0-7.0.

[0088] Example 2: Effect of protein concentration on the stability of anti-TIGIT antibody formulations To investigate the effect of osipellimab antibody protein concentration on the stability of osipellimab antibody formulations, stress tests under freeze-thaw conditions were performed. For F11-F13, a 30 kDa Amicon Ultra™ centrifugal filter was used to generate a concentrated stock solution of osipellimab. This stock solution was dialyzed into 30 mM acetic acid-sodium acetate buffer and then used to produce the formulations listed in Table 3. For F14, a 30 kDa Amicon Ultra™ centrifugal filter was used to generate a concentrated stock solution of osipellimab, which was then dialyzed into 20 mM histidine-histidine HCl buffer and then adjusted to the F14 formulation shown in Table 3. Each of the formulated solutions was filtered using a 0.22 mm PES syringe filter and subjected to freeze-thaw stress conditions. To assess the impact on antibody stability, the Tm and Tagg of these antibody samples were examined using Uncle™ from Unchained Labs and the data are shown in Table 3. [Table 3]

[0089] Table 3 shows that all formulations had Tm1 above 65°C, Tm2 above 75°C, and Tg above 70°C, which are good indicators of stability. In this freeze-thaw stability test, subvisible particles increased slightly compared to the time point zero (T0) samples, particularly subvisible particles smaller than 10 μm. However, the total number of subvisible particles, especially those larger than 10 μm, remained low. Turbidity data showed an increase in turbidity with increasing protein concentration, but there was no apparent change after five freeze-thaw cycles, indicating that the osipellimab antibody was stable for these formulations. SEC-HPLC data showed no apparent change in SEC monomer after five freeze-thaw cycles. All samples had greater than 95% SEC monomer, indicating that these formulations containing osipellimab antibody at concentrations of 10-150 mg / ml were less prone to aggregation and particle formation. Combining the data shown in Figures 3A-C, all of the formulations tested were stable over a wide range of osipellimab antibody protein concentrations, ranging from 10 mg / ml to 150 mg / ml.

[0090] Example 3: Effect of surfactants on the stability of anti-TIGIT antibody formulations This experiment investigated the effect of various surfactants on the stability of osipellimab antibody formulations. In this study, osipellimab was buffer exchanged into 20 mM histidine-histidine HCl buffer (pH 5.8) by dialysis and then adjusted with different surfactants to form the formulations listed in Table 4. Each of the formulated solutions was filtered using a 0.22 mm PES syringe filter and subjected to stress conditions. [Table 4]

[0091] Freeze-thaw tests were performed on these formulations by subjecting them to three cycles of freezing at -70°C and then thawing at ambient temperature (denoted "3FT" in Figures 3A-E). To test the formulations' high-temperature and photostability, the osipellimab antibody formulations were stored in a stability chamber at 37°C for four weeks (denoted "37C4W" in tables and graphs) or placed in a photostability chamber with an illumination level of 1.2 million lux-hours or greater for two weeks (denoted "photo 2W" in figures). Mechanical stress tests were performed by subjecting the formulations to shaking conditions using 800 rpm for two days (denoted "shake 2D" in figures). These formulations were evaluated for subvisible particles, turbidity, SEC (purity), IEC (charge profile), and CE-SDS (NR) (purity). As controls, these samples were analyzed at the starting time point (denoted as TO in figures).

[0092] As shown in Figures 3A-B, osipellimab formulations F15, F16, and F17 showed no significant increase in subvisible particles or turbidity after three freeze-thaw cycles, two days of shaking, two weeks of photostability, and four weeks of storage at 37°C. These data indicate that osipellimab is less likely to aggregate and form particles in these formulations. Note that F17, an antibody formulation containing poloxamer 188, showed fewer subvisible particles and turbidity than formulations containing polysorbate 20 and polysorbate 80. As shown in Figures 3C-E, formulations F15, F16, and F17 showed no significant changes in SEC, IEC, or CE-SDS(NR) after three freeze-thaw cycles, two days of shaking, and four weeks of storage at 37°C. Under the two-week stress test conditions under photostability conditions, formulations F15, F16, and F17 showed no obvious changes in SEC, IEC, or CE-SDS (NR) purity when compared to the starting point (T0). Considering these stability data, polysorbate 20, polysorbate 80, and poloxamer 188 contributed to the stability of the osipellimab antibody in the formulation.

[0093] Example 4. Stability of anti-TIGIT antibody formulations The stability of osipellimab antibody was evaluated in various formulations listed in Table 5. All formulations were prepared in 20 mM histidine-histidine HCl buffers with a pH range of 5.2 to 6.2. Trehalose at a concentration of 240 mM was used as a stabilizer and osmolality adjuster. The surfactant was polysorbate 20 at a concentration of 0.2 mg / ml. The osipellimab antibody was buffer exchanged by dialysis into 20 mM histidine-histidine HCl buffers of different pHs (pH 5.2, 5.5, 5.8, 6.0, and 6.2) to generate osipellimab stock solutions of the intended pH and used an osipellimab antibody concentration of 20 mg / ml. Each formulated solution was filtered using a 0.22 mm PES syringe filter and subjected to the stability studies shown in Table 6. [Table 5] [Table 6]

[0094] Freeze-thaw testing was performed on these samples by subjecting them to six cycles of freezing at -70°C and thawing at ambient temperature (denoted "6FT" in Table 7). To determine the effect of high-temperature and light stability on the formulations, samples were stored in a stability chamber at 40°C for two weeks (denoted "40C 2W" in tables and graphs) or placed in a light stability chamber with an illumination level of 1.2 million lux-hours or greater for two weeks (denoted "photo 2W" in Table 7). Accelerated stability testing at 25°C and long-term stability testing at 5±3°C were also investigated (denoted "25C" and "5C" in Tables 8 and 9). These formulations were evaluated by visible particles, subvisible particles, SEC (purity), IEC (charge profile), and CE-SDS (NR) (purity). As a control, these samples were analyzed at the starting time point (denoted as TO). The results are shown in Tables 7-9 and Figures 4-6.

[0095] From the visible particle data in Table 7, all formulations were clear after six freeze-thaw cycles, and no visible particles were observed for the formulations tested at 40°C for two weeks and for light stability. From the subvisible particle data in Figure 4A, all formulations were stable after six freeze-thaw cycles, two weeks at 40°C, and light stability, with no apparent increase in subvisible particles. From the SEC data in Figure 4B, all formulations were stable after six freeze-thaw cycles, with a slight decrease in monomer purity and a slight decrease in light stability after two weeks at 40°C. From the IEC data in Figure 4C, all formulations were stable after six freeze-thaw cycles, with a slight decrease in the main peak and a slight decrease in light stability after two weeks at 40°C. [Table 7]

[0096] Stability data for osipellimab formulations stored at 25°C are shown in Table 8 and Figure 5. When stored at 25°C for 6 months, all formulations were clear, with no visible particles observed. From the subvisible particle data in Figure 5A, all formulations were stable at 25°C for 6 months, with no increase in subvisible particles. From the SEC data in Figure 5B, the % monomer decreased slightly over time but remained above 95% at the longest endpoint of 6 months at 25°C. This indicates that the formulations pass the 95% SEC standard (a general specification for monoclonal antibodies). From the IEC data in Figure 5C, the % main peak decreased over time but remained above 40% at 25°C for 6 months. From the CE-SDS(NR) data in Figure 5D, the % intact peak decreased over time but remained above 94% when measured at 25°C for 6 months. [Table 8]

[0097] The long-term stability of osipellimab antibody formulations stored at 5±3°C at various time points is shown in Table 9 and Figure 6. All formulations remained clear at 5±3°C for 24 months, with no visible particles observed. From the subvisible particle data in Figure 6A, all formulations were stable at 5±3°C for 24 months, with no increase in subvisible particles. From the SEC data shown in Figure 6B, the % monomer decreased slightly over time but remained at 97% at 5±3°C for 24 months. Again, this indicates that the formulations disclosed herein pass the 95% SEC standard (a common specification for monoclonal antibodies). From the IEC data in Figure 6C, the % main peak did not change over time at 5±3°C for 24 months. From the CE-SDS(NR) data in Figure 6D, the % intact peak decreased slightly over time but remained above 96% at 5±3°C for 24 months. [Table 9]

[0098] In conclusion, these results demonstrated that osipellimab antibody formulations F18, F19, F20, F21, and F22 were stable for 24 months at 5±3°C and 25°C, within acceptable parameters for clinical use.

Claims

1. A pharmaceutical formulation comprising: i. about 5 mg / mL to about 200 mg / mL of an anti-TIGIT antibody, or antigen-binding fragment thereof; ii. A formulation buffer of about 5 mM to about 50 mM to provide a pH of about 5.0 to about 7.0 iii. about 30 mM to about 300 mM of a stabilizer; iv. about 0.01 mg / ml to about 1 mg / ml of a non-ionic surfactant.

2. The formulation of claim 1, wherein the anti-TIGIT antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising HCDR1 (heavy chain complementarity determining region 1) of SEQ ID NO: 1, HCDR2 of SEQ ID NO: 2, and HCDR3 of SEQ ID NO: 3, and a light chain variable region comprising LCDR1 (light chain complementarity determining region 1) of SEQ ID NO: 4, LCDR2 of SEQ ID NO: 5, and LCDR3 of SEQ ID NO:

6.

3. The formulation of claim 1 , wherein the anti-TIGIT antibody or antigen-binding fragment thereof comprises SEQ ID NO: 7 and SEQ ID NO:

8.

4. 10. The formulation of claim 1, wherein the formulation buffer is selected from the group consisting of histidine, acetate, citrate, succinate, phosphate, a mixture of histidine and acetic acid, or a mixture of histidine and citric acid.

5. 5. The formulation of claim 4, wherein the formulation buffer is histidine.

6. 6. The formulation of claim 5, wherein the concentration of the histidine buffer is 10 mM to 30 mM.

7. 7. The formulation of claim 6, comprising 20 mM of said histidine buffer.

8. 8. The formulation of claim 6 or 7, wherein the pH is in the range of 5.2 to 6.

2.

9. 9. The formulation of any one of claims 1 to 8, wherein the stabilizer is selected from the group consisting of trehalose, sucrose, sorbitol, mannitol, maltose, dextran, (2-hydroxypropyl)-b-cyclodextrin, sodium chloride, magnesium chloride, calcium chloride, sodium sulfate, sodium dihydrogen phosphate, or disodium hydrogen phosphate.

10. 10. The formulation of claim 9, wherein the stabilizer is trehalose.

11. The formulation according to any one of claims 1 to 10, wherein the concentration of the trehalose is from 50 mM to 280 mM.

12. 12. The formulation of claim 11, wherein the concentration of trehalose is from 150 mM to 250 mM.

13. 10. The formulation of claim 9, wherein the stabilizer is sucrose.

14. 14. The formulation of claim 13, wherein the concentration of sucrose is from 50 mM to 280 mM.

15. 15. The formulation of claim 14, wherein the concentration of sucrose is from 150 mM to 250 mM.

16. 16. The formulation of any one of claims 1 to 15, wherein the non-ionic surfactant is selected from the group consisting of polysorbate 20, polysorbate 80, or poloxamer 188.

17. 17. The formulation of claim 16, wherein the concentration of polysorbate 20 is from 0.1 mg / ml to 0.8 mg / ml.

18. 18. The formulation of claim 17, wherein the concentration of polysorbate 20 is from 0.2 mg / ml to 0.6 mg / ml.

19. 17. The formulation of claim 16, wherein the concentration of polysorbate 80 is from 0.1 mg / ml to 0.8 mg / ml.

20. 20. The formulation of claim 19, wherein the concentration of polysorbate 80 is from 0.2 mg / ml to 0.6 mg / ml.

21. 17. The formulation of claim 16, wherein the concentration of the poloxamer 188 is from 0.1 mg / ml to 0.8 mg / ml.

22. 22. The formulation of claim 21, wherein the concentration of the poloxamer 188 is from 0.2 mg / ml to 0.6 mg / ml.

23. 10. The formulation of claim 1 comprising 30 mM acetic acid-sodium acetate, 240 mM sucrose, and 0.2 mg / ml polysorbate 80, and having a pH of pH 5.

5.

24. 10. The formulation of claim 1 comprising 20 mM histidine-histidine HCl, 240 mM trehalose, and 0.2 mg / ml polysorbate 20, and having a pH of pH 5.

8.

25. 10. The formulation of claim 1 comprising 20 mM histidine-histidine HCl, 70 mM NaCl, 80 mM trehalose, and 0.8 mg / ml polysorbate 20, and having a pH of pH 6.

0.

26. 26. The formulation of any one of claims 1 to 25, wherein the concentration of the anti-TIGIT antibody, or antigen-binding fragment thereof, is about 10 mg / mL to 150 mg / mL.

27. A method for producing an antibody formulation, comprising: a. exchanging the anti-TIGIT antibody into about 5 mM to about 50 mM of a buffer providing a pH of about 5.0 to about 7.0; b. concentrating the antibody formulation of (a) to an antibody concentration of about 5 to 200 mg / mL; c. adding a nonionic surfactant to the antibody formulation of (c) to obtain an antibody formulation having a surfactant concentration of 0.01 mg / ml or more; d. adding a stabilizer to the antibody to obtain an antibody formulation having a stabilizer concentration of 30 mM or more, wherein the anti-TIGIT antibody is The method comprises a heavy chain variable region comprising HCDR1 (heavy chain complementarity determining region 1) of SEQ ID NO: 1, HCDR2 of SEQ ID NO: 2, and HCDR3 of SEQ ID NO: 3, and a light chain variable region comprising LCDR1 (light chain complementarity determining region 1) of SEQ ID NO: 4, LCDR2 of SEQ ID NO: 5, and LCDR3 of SEQ ID NO:

6.

28. 28. The formulation of claim 27, wherein the anti-TIGIT antibody or antigen-binding fragment thereof comprises SEQ ID NO:7 and SEQ ID NO:

8.

29. A method for treating cancer in a human patient in need thereof, comprising administering an effective amount of the anti-TIGIT antibody formulation of claim 1.

30. 30. The method of claim 29, wherein the anti-TIGIT antibody formulation is administered at a dose of about 200 mg to about 2400 mg.

31. 31. The method of claim 30, wherein the anti-TIGIT antibody formulation is administered once every three weeks.

32. 30. The method of claim 29, wherein the human patient is administered at least one other therapeutic agent selected from the group consisting of zanubrutinib, pamiparib, tislelizumab, an anti-LAG3 antibody, a second anti-TIGIT antibody, an anti-4-1BB antibody, an anti-OX40 antibody, an anti-TIM-3 antibody, a CD40 agonist, a TLR agonist, a CAR-T cell, or a chemotherapeutic agent.

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