Pharmaceutical composition of Anti-tim-3 antibody and use thereof

Abstract

Provided are a pharmaceutical composition comprising an anti-TIM-3 antibody and a use thereof. The pharmaceutical composition comprises an anti-TIM-3 antibody and a buffer, and the pharmaceutical composition may further comprise a surfactant and a stabilizer. The pharmaceutical composition exhibits good stability.

Description

Pharmaceutical compositions and uses of anti-TIM-3 antibodies Technical Field

[0001] The present disclosure belongs to the field of preparations. Specifically, the present disclosure relates to a pharmaceutical composition comprising an anti-TIM-3 antibody, and the use of the pharmaceutical composition for treating tumors. Background Art

[0002] T cell immunoglobulin and mucin domain-containing protein 3 (TIM-3), also known as hepatitis A virus cellular receptor 2 (HAVCR2), is a member of the TIM family of immunoregulatory proteins (the human TIM family includes TIM-1, 3, and 4). TIM-3 is selectively expressed on the surface of activated Th1 cells and is also expressed on myeloid cells, dendritic cells, natural killer (NK) cells, macrophages, and various tumor cells. TIM-3 has multiple ligands, including galectin-9, phosphatidylserine (PtdSer), high mobility group protein B1 (HMGB1), and carcinoembryonic antigen cell adhesion molecule 1 (CEACAM1). As an immune checkpoint, TIM-3 negatively regulates the body's immune response, preventing damage from excessive immune or autoimmune responses. However, increasing evidence shows that TIM-3 protein and / or mRNA are upregulated in a variety of tumor tissues and tumor-associated immune cells, participating in tumor immune escape and immune response, and promoting tumor development.

[0003] There is a need in the art to develop pharmaceutical compositions comprising anti-TIM-3 antibodies that are suitable for production and administration to patients and that maintain biological activity and stability during storage and subsequent use.

[0004] Summary of the Invention

[0005] Pharmaceutical composition

[0006] The present disclosure provides a stable pharmaceutical composition comprising an anti-TIM-3 antibody or an antigen-binding fragment thereof.

[0007] The present disclosure provides a pharmaceutical composition comprising: (a) an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) a buffer, (c) a surfactant, and (d) a stabilizer.

[0008] In some embodiments, the anti-TIM-3 antibody or antigen-binding fragment thereof comprises: a heavy chain CDR1 (HCDR1) of the amino acid sequence of SEQ ID NO: 1 or 11, a HCDR2 of the amino acid sequence of SEQ ID NO: 2 or 12, a HCDR3 of the amino acid sequence of SEQ ID NO: 3 or 13, a light chain CDR1 (LCDR1) of the amino acid sequence of SEQ ID NO: 4 or 14, a LCDR2 of the amino acid sequence of SEQ ID NO: 5 or 15, and a LCDR3 of the amino acid sequence of SEQ ID NO: 6 or 16. In some embodiments, the anti-TIM-3 antibody or antigen-binding fragment thereof comprises: a HCDR1 of the amino acid sequence of SEQ ID NO: 1, a HCDR2 of the amino acid sequence of SEQ ID NO: 2, a HCDR3 of the amino acid sequence of SEQ ID NO: 3, a LCDR1 of the amino acid sequence of SEQ ID NO: 4, a LCDR2 of the amino acid sequence of SEQ ID NO: 5, and a LCDR3 of the amino acid sequence of SEQ ID NO: 6. In some embodiments, the anti-TIM-3 antibody or antigen-binding fragment thereof comprises: a HCDR1 of the amino acid sequence set forth in SEQ ID NO: 11, a HCDR2 of the amino acid sequence set forth in SEQ ID NO: 12, a HCDR3 of the amino acid sequence set forth in SEQ ID NO: 13, a LCDR1 of the amino acid sequence set forth in SEQ ID NO: 14, a LCDR2 of the amino acid sequence set forth in SEQ ID NO: 15, and a LCDR3 of the amino acid sequence set forth in SEQ ID NO: 16.

[0009] In some embodiments, the anti-TIM-3 antibody or antigen-binding fragment thereof comprises: HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as set forth in SEQ ID NO: 7 or 17, and LCDR1, LCDR2, and LCDR3 in the light chain variable region as set forth in SEQ ID NO: 8 or 18. In some embodiments, the anti-TIM-3 antibody or antigen-binding fragment thereof comprises: HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as set forth in SEQ ID NO: 7, and LCDR1, LCDR2, and LCDR3 in the light chain variable region as set forth in SEQ ID NO: 8. In some embodiments, the anti-TIM-3 antibody or antigen-binding fragment thereof comprises: HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as set forth in SEQ ID NO: 17, and LCDR1, LCDR2, and LCDR3 in the light chain variable region as set forth in SEQ ID NO: 18.

[0010] The CDR sequences of exemplary anti-TIM-3 antibodies or antigen-binding fragments thereof are shown in Table 1.

[0011] Table 1. CDR sequences of anti-TIM-3 antibodies or antigen-binding fragments thereof

[0012] It will be understood by those skilled in the art that, unless otherwise specified, the term "CDR" or "complementarity determining region" of a given antibody or region thereof (e.g., variable region) should be understood to encompass complementarity determining regions defined by any known scheme. Although Table 1 shows CDR sequences (wherein the CDR regions shown in SEQ ID NOs: 1-6 are defined by the AbM numbering system), when referring to antibodies defined by the specific CDR sequences of the present disclosure, the scope of the antibodies also encompasses antibodies defined by CDR sequences converted to any other numbering system definition (e.g., one or more combinations of the definitions of Kabat, Chothia, CCG, IMGT, or Contact, etc., which are well known in the art). For example, an anti-TIM-3 antibody or antigen-binding fragment thereof comprising the following amino acid sequence: HCDR1 of the amino acid sequence set forth in SEQ ID NO: 1, HCDR2 of the amino acid sequence set forth in SEQ ID NO: 2, HCDR3 of the amino acid sequence set forth in ARRYYGYDAMDY (SEQ ID NO: 21), LCDR1 of the amino acid sequence set forth in SEQ ID NO: 4, LCDR2 of the amino acid sequence set forth in SEQ ID NO: 5, and LCDR3 of the amino acid sequence set forth in SEQ ID NO: 6 is also encompassed within the scope of the anti-TIM-3 antibodies or antigen-binding fragments thereof of the present disclosure.

[0013] In some embodiments, the anti-TIM-3 antibody or antigen-binding fragment thereof comprises a heavy chain variable region having an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 7 or 17. In some embodiments, the anti-TIM-3 antibody or antigen-binding fragment thereof comprises a light chain variable region having an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 8 or 18. In some embodiments, the anti-TIM-3 antibody or antigen-binding fragment thereof comprises a heavy chain variable region set forth in SEQ ID NO: 7. In some embodiments, the anti-TIM-3 antibody or antigen-binding fragment thereof comprises a light chain variable region set forth in SEQ ID NO: 8. In some embodiments, the anti-TIM-3 antibody or antigen-binding fragment thereof comprises a heavy chain variable region having an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 7 or 17, and a light chain variable region having an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 8 or 18. In some embodiments, the anti-TIM-3 antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the amino acid sequence of SEQ ID NO: 7 and a light chain variable region having the amino acid sequence of SEQ ID NO: 8. In some embodiments, the anti-TIM-3 antibody or antigen-binding fragment thereof comprises a heavy chain variable region having the amino acid sequence of SEQ ID NO: 17 and a light chain variable region having the amino acid sequence of SEQ ID NO: 18. In some specific embodiments, the amino acid sequence of the heavy chain variable region of the anti-TIM-3 antibody or antigen-binding fragment thereof is as set forth in SEQ ID NO: 7, and the amino acid sequence of the light chain variable region is as set forth in SEQ ID NO: 8.In some specific embodiments, the amino acid sequence of the heavy chain variable region of the anti-TIM-3 antibody or antigen-binding fragment thereof is shown in SEQ ID NO: 17, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 18.

[0014] In some embodiments, the anti-TIM-3 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HCDR1, HCDR2, and HCDR3, and the light chain variable region comprises LCDR1, LCDR2, and LCDR3, wherein HCDR1 comprises the amino acid sequence of SEQ ID NO:1, HCDR2 comprises the amino acid sequence of SEQ ID NO:2, HCDR3 comprises the amino acid sequence of SEQ ID NO:3, LCDR1 comprises the amino acid sequence of SEQ ID NO:4, LCDR2 comprises the amino acid sequence of SEQ ID NO:5, and LCDR3 comprises the amino acid sequence of SEQ ID NO:6; and the heavy chain variable region comprises an amino acid sequence that has at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 110%, 111%, 112%, 113%, 114%, 115%, 116%, 117%, 118%, 119%, 120%, 121%, 122%, 123%, 124%, 125%, 126%, 127%, 128%, 129%, 130%, 131%, 132%, 133%, 134%, 135%, 136%, 137%, 138%, 139%, 200%, 201 The light chain variable region comprises an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:8.

[0015] In some embodiments, the anti-TIM-3 antibody or antigen-binding fragment thereof may further comprise an immunoglobulin constant region, or a fragment, analog, variant, or derivative of the constant region. In some embodiments, the constant region comprises a heavy chain constant region and a light chain constant region. In some embodiments, the heavy chain constant region is derived from a human immunoglobulin heavy chain, such as IgG1, IgG2, IgG3, and IgG4 or other classes of immunoglobulin heavy chains, preferably IgG4 heavy chains. In some embodiments, the light chain constant region is derived from a human immunoglobulin light chain, such as a kappa light chain or a lambda light chain of a human immunoglobulin. In some embodiments, the constant region may comprise any modification described herein, such as insertion, deletion, substitution, or chemical modification of amino acids. In some embodiments, the constant region comprises a mutation that alters effector function. In some embodiments, any amino acid residue in the constant region may be substituted with an amino acid residue of any allotype.

[0016] In some embodiments, the anti-TIM-3 antibody or antigen-binding fragment thereof comprises a heavy chain having an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 9 or 19. In some embodiments, the anti-TIM-3 antibody or antigen-binding fragment thereof comprises a light chain having an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 10 or 20. In some embodiments, the anti-TIM-3 antibody or antigen-binding fragment thereof comprises the heavy chain set forth in SEQ ID NO: 9. In some embodiments, the anti-TIM-3 antibody or antigen-binding fragment thereof comprises the light chain set forth in SEQ ID NO: 10. In some embodiments, the anti-TIM-3 antibody or antigen-binding fragment thereof comprises a heavy chain having an amino acid sequence at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 9 or 19, and a light chain having an amino acid sequence at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 10 or 20. In some embodiments, the anti-TIM-3 antibody comprises a heavy chain having an amino acid sequence as set forth in SEQ ID NO:9, and a light chain having an amino acid sequence as set forth in SEQ ID NO:10. In some embodiments, the anti-TIM-3 antibody comprises a heavy chain having an amino acid sequence as set forth in SEQ ID NO:19, and a light chain having an amino acid sequence as set forth in SEQ ID NO:20. In some specific embodiments, the amino acid sequence of the heavy chain of the anti-TIM-3 antibody is as set forth in SEQ ID NO:9, and the amino acid sequence of the light chain is as set forth in SEQ ID NO:10. In some specific embodiments, the amino acid sequence of the heavy chain of the anti-TIM-3 antibody is as set forth in SEQ ID NO:19, and the amino acid sequence of the light chain is as set forth in SEQ ID NO:20. The terminal amino acid K in SEQ ID NOs:9 and 19 may or may not be present.

[0017] In some embodiments, the anti-TIM-3 antibody or antigen-binding fragment thereof of the present disclosure is mAb 50B5 or an antigen-binding fragment thereof described in the patent application document with publication number CN112566936A, or a chimeric antibody or an antigen-binding fragment thereof of mAb 50B5, or a humanized antibody or an antigen-binding fragment thereof of mAb 50B5.

[0018] In other embodiments, the anti-TIM-3 antibody or antigen-binding fragment thereof disclosed herein is mAb 15B4 or an antigen-binding fragment thereof described in patent application publication number CN112566936A, or a chimeric antibody or an antigen-binding fragment thereof of mAb 15B4, or a humanized antibody or an antigen-binding fragment thereof of mAb 15B4.

[0019] The entire contents of the CN112566936A patent application document are incorporated herein by reference.

[0020] In other embodiments, the anti-TIM-3 antibody or antigen-binding fragment thereof of the present disclosure is selected from Sabatolimab, Cobolimab, Surzebiclimab, Lomvastomig, SHR-1702 of Hengrui Medicine, Verzistobart (INCAGN-2390) of Agenus, BC-3402 of WuXi Biologics, LBL-003 of Weilizhibo, LNL-005 of Jianxin Bio, or BMS-986258 of BMS.

[0021] In some embodiments, the pharmaceutical composition is an injection. In some embodiments, the pharmaceutical composition of the present disclosure is a lyophilized formulation; preferably, the lyophilized formulation is a lyophilized injection. In some embodiments, the pharmaceutical composition of the present disclosure is an aqueous formulation; preferably, the aqueous formulation is an aqueous injection.

[0022] In some embodiments, the amount of the anti-TIM-3 antibody or antigen-binding fragment thereof in the pharmaceutical composition is 1 mg to 120 mg, 10 mg to 100 mg, 15 mg to 90 mg, 20 mg to 80 mg, 25 mg to 70 mg, 30 mg to 60 mg, 55 mg to 65 mg, or 59 mg to 61 mg, or multiples thereof. In some embodiments, the amount of the anti-TIM-3 antibody or antigen-binding fragment thereof is 30 mg to 60 mg, or multiples thereof. In some embodiments, examples of the amount of the anti-TIM-3 antibody or antigen-binding fragment thereof include, but are not limited to, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 59 mg, 60 mg, 61 mg, 65 mg, 70 mg, 75 mg, 80 mg, 85 mg, 90 mg, 95 mg, or 100 mg, or multiples thereof. In some embodiments, the amount of the anti-TIM-3 antibody or antigen-binding fragment thereof is 30 mg or a multiple thereof. In some embodiments, the amount of the anti-TIM-3 antibody or antigen-binding fragment thereof is 60 mg or a multiple thereof (e.g., 240 mg, 360 mg, 480 mg, 600 mg, or 1200 mg).

[0023] In some embodiments, the concentration of the anti-TIM-3 antibody or antigen-binding fragment thereof in the pharmaceutical composition is 1 mg / mL to 120 mg / mL, 10 mg / mL to 100 mg / mL, 15 mg / mL to 90 mg / mL, 20 mg / mL to 80 mg / mL, 25 mg / mL to 70 mg / mL, 30 mg / mL to 60 mg / mL, 55 mg / mL to 65 mg / mL, or 59 mg / mL to 61 mg / mL. In some embodiments, the concentration of the anti-TIM-3 antibody or antigen-binding fragment thereof is 30 mg / mL to 60 mg / mL. In some embodiments, examples of concentrations of the anti-TIM-3 antibody or antigen-binding fragment thereof include, but are not limited to, 10 mg / mL, 15 mg / mL, 20 mg / mL, 25 mg / mL, 30 mg / mL, 35 mg / mL, 40 mg / mL, 45 mg / mL, 50 mg / mL, 55 mg / mL, 59 mg / mL, 60 mg / mL, 61 mg / mL, 65 mg / mL, 70 mg / mL, 75 mg / mL, 80 mg / mL, 85 mg / mL, 90 mg / mL, 95 mg / mL, or 100 mg / mL, or any range thereof. In some embodiments, the concentration of the anti-TIM-3 antibody or antigen-binding fragment thereof is 30 mg / mL. In some embodiments, the concentration of the anti-TIM-3 antibody or antigen-binding fragment thereof is 60 mg / mL.

[0024] In some embodiments, in the pharmaceutical composition, the buffer is a histidine buffer, a phosphate buffer, a succinate buffer, a citrate buffer and / or an acetate buffer. In some embodiments, the buffer is a histidine buffer, a phosphate buffer or a citrate buffer.

[0025] In some embodiments, the histidine buffer includes but is not limited to histidine-acetate buffer, histidine-hydrochloric acid buffer, histidine-phosphate buffer or histidine-histidine hydrochloride buffer. In some embodiments, the phosphate buffer includes but is not limited to sodium phosphate buffer or potassium phosphate buffer. In some embodiments, the succinate buffer includes but is not limited to sodium succinate buffer or potassium succinate buffer. In some embodiments, the citrate buffer includes but is not limited to sodium citrate buffer or potassium citrate buffer. In some embodiments, the acetate buffer includes but is not limited to potassium acetate buffer, ammonium acetate buffer or sodium acetate buffer.

[0026] In some embodiments, in the pharmaceutical composition, the buffer is a histidine buffer. In some embodiments, the buffer is a histidine-acetate buffer.

[0027] In some embodiments, the amount of the buffer in the pharmaceutical composition is 1 μmol to 100 μmol, 2 μmol to 80 μmol, 5 μmol to 60 μmol, 8 μmol to 40 μmol, 10 μmol to 30 μmol, 10 μmol to 20 μmol, 20 μmol to 30 μmol, 15 μmol to 25 μmol, or 19 μmol to 21 μmol per 60 mg of the anti-TIM-3 antibody or antigen-binding fragment thereof. In some embodiments, the amount of the buffer is 10 μmol to 30 μmol per 60 mg of the anti-TIM-3 antibody or antigen-binding fragment thereof. In some embodiments, examples of the amount of the buffer include, but are not limited to, 8 μmol, 9 μmol, 10 μmol, 11 μmol, 12 μmol, 13 μmol, 14 μmol, 15 μmol, 16 μmol, 17 μmol, 18 μmol, 19 μmol, 20 μmol, 21 μmol, 22 μmol, 23 μmol, 24 μmol, 25 μmol, 26 μmol, 27 μmol, 28 μmol, 30 μmol, 32 μmol, 34 μmol, 36 μmol, 38 μmol, 40 μmol, 50 μmol, 60 μmol, 70 μmol, 80 μmol, 90 μmol, or 100 μmol, or a range of any values ​​per 60 mg of the anti-TIM-3 antibody or antigen-binding fragment thereof. In some embodiments, the amount of the buffer is 10 μmol per 60 mg of the anti-TIM-3 antibody or antigen-binding fragment thereof. In some embodiments, the amount of the buffer is 20 μmol per 60 mg of the anti-TIM-3 antibody or antigen-binding fragment thereof. In some embodiments, the amount of the buffer is 30 μmol per 60 mg of the anti-TIM-3 antibody or antigen-binding fragment thereof.

[0028] In some embodiments, the amount of the histidine buffer in the pharmaceutical composition is 1 μmol to 100 μmol, 2 μmol to 80 μmol, 5 μmol to 60 μmol, 8 μmol to 40 μmol, 10 μmol to 30 μmol, 10 μmol to 20 μmol, 20 μmol to 30 μmol, 15 μmol to 25 μmol, or 19 μmol to 21 μmol per 60 mg of the anti-TIM-3 antibody or antigen-binding fragment thereof. In some embodiments, the amount of the histidine buffer is 10 μmol to 30 μmol per 60 mg of the anti-TIM-3 antibody or antigen-binding fragment thereof. In some embodiments, examples of the amount of the histidine buffer include, but are not limited to, 8 μmol, 9 μmol, 10 μmol, 11 μmol, 12 μmol, 13 μmol, 14 μmol, 15 μmol, 16 μmol, 17 μmol, 18 μmol, 19 μmol, 20 μmol, 21 μmol, 22 μmol, 23 μmol, 24 μmol, 25 μmol, 26 μmol, 27 μmol, 28 μmol, 30 μmol, 32 μmol, 34 μmol, 36 μmol, 38 μmol, 40 μmol, 50 μmol, 60 μmol, 70 μmol, 80 μmol, 90 μmol, or 100 μmol, or a range of any values ​​per 60 mg of the anti-TIM-3 antibody or antigen-binding fragment thereof. In some embodiments, the amount of the histidine buffer is 10 μmol per 60 mg of the anti-TIM-3 antibody or antigen-binding fragment thereof. In some embodiments, the amount of the histidine buffer is 20 μmol per 60 mg of the anti-TIM-3 antibody or antigen-binding fragment thereof. In some embodiments, the amount of the histidine buffer is 30 μmol per 60 mg of the anti-TIM-3 antibody or antigen-binding fragment thereof. In some embodiments, the histidine buffer is a histidine-acetate buffer.

[0029] In some embodiments, the amount of the histidine-acetate buffer in the pharmaceutical composition is 1 μmol to 100 μmol, 2 μmol to 80 μmol, 5 μmol to 60 μmol, 8 μmol to 40 μmol, 10 μmol to 30 μmol, 10 μmol to 20 μmol, 20 μmol to 30 μmol, 15 μmol to 25 μmol, or 19 μmol to 21 μmol per 60 mg of the anti-TIM-3 antibody or antigen-binding fragment thereof. In some embodiments, the amount of the histidine-acetate buffer is 10 μmol to 30 μmol per 60 mg of the anti-TIM-3 antibody or antigen-binding fragment thereof. In some embodiments, examples of the amount of the histidine-acetate buffer include, but are not limited to, 8 μmol, 9 μmol, 10 μmol, 11 μmol, 12 μmol, 13 μmol, 14 μmol, 15 μmol, 16 μmol, 17 μmol, 18 μmol, 19 μmol, 20 μmol, 21 μmol, 22 μmol, 23 μmol, 24 μmol, 25 μmol, 26 μmol, 27 μmol, 28 μmol, 30 μmol, 32 μmol, 34 μmol, 36 μmol, 38 μmol, 40 μmol, 50 μmol, 60 μmol, 70 μmol, 80 μmol, 90 μmol, or 100 μmol, or a range thereof, per 60 mg of the anti-TIM-3 antibody or antigen-binding fragment thereof. In some embodiments, the amount of the histidine-acetate buffer is 10 μmol per 60 mg of the anti-TIM-3 antibody or antigen-binding fragment thereof. In some embodiments, the amount of the histidine-acetate buffer is 20 μmol per 60 mg of the anti-TIM-3 antibody or antigen-binding fragment thereof. In some embodiments, the amount of the histidine-acetate buffer is 30 μmol per 60 mg of the anti-TIM-3 antibody or antigen-binding fragment thereof.

[0030] In some embodiments, in the pharmaceutical composition, the concentration of the buffer is 1 mM to 100 mM, 2 mM to 80 mM, 5 mM to 60 mM, 8 mM to 40 mM, 10 mM to 30 mM, 10 mM to 20 mM, 20 mM to 30 mM, 15 mM to 25 mM, or 19 mM to 21 mM. In some embodiments, the concentration of the buffer is 10 mM to 30 mM. In some embodiments, examples of the concentration of the buffer include, but are not limited to, 8mM, 9mM, 10mM, 11mM, 12mM, 13mM, 14mM, 15mM, 16mM, 17mM, 18mM, 19mM, 20mM, 21mM, 22mM, 23mM, 24mM, 25mM, 26mM, 27mM, 28mM, 30mM, 32mM, 34mM, 36mM, 38mM, 40mM, 50mM, 60mM, 70mM, 80mM, 90mM, or 100mM, or a range formed by any value. In some embodiments, the concentration of the buffer is 10mM. In some embodiments, the concentration of the buffer is 20mM. In some embodiments, the concentration of the buffer is 30mM.

[0031] In some embodiments, in the pharmaceutical composition, the concentration of the histidine buffer is 1 mM to 100 mM, 2 mM to 80 mM, 5 mM to 60 mM, 8 mM to 40 mM, 10 mM to 30 mM, 10 mM to 20 mM, 20 mM to 30 mM, 15 mM to 25 mM, or 19 mM to 21 mM. In some embodiments, the concentration of the histidine buffer is 10 mM to 30 mM. In some embodiments, examples of the concentration of the histidine buffer include, but are not limited to, 8 mM, 9 mM, 10 mM, 11 mM, 12 mM, 13 mM, 14 mM, 15 mM, 16 mM, 17 mM, 18 mM, 19 mM, 20 mM, 21 mM, 22 mM, 23 mM, 24 mM, 25 mM, 26 mM, 27 mM, 28 mM, 30 mM, 32 mM, 34 mM, 36 mM, 38 mM, 40 mM, 50 mM, 60 mM, 70 mM, 80 mM, 90 mM, or 100 mM, or a range of any value. In some embodiments, the concentration of the histidine buffer is 10 mM. In some embodiments, the concentration of the histidine buffer is 20 mM. In some embodiments, the concentration of the histidine buffer is 30 mM. In some embodiments, the histidine buffer is a histidine-acetate buffer.

[0032] In some embodiments, in the pharmaceutical composition, the concentration of the histidine-acetate buffer is 1 mM to 100 mM, 2 mM to 80 mM, 5 mM to 60 mM, 8 mM to 40 mM, 10 mM to 30 mM, 10 mM to 20 mM, 20 mM to 30 mM, 15 mM to 25 mM, or 19 mM to 21 mM. In some embodiments, the concentration of the histidine-acetate buffer is 10 mM to 30 mM. In some embodiments, examples of the concentration of the histidine-acetate buffer include, but are not limited to, 8 mM, 9 mM, 10 mM, 11 mM, 12 mM, 13 mM, 14 mM, 15 mM, 16 mM, 17 mM, 18 mM, 19 mM, 20 mM, 21 mM, 22 mM, 23 mM, 24 mM, 25 mM, 26 mM, 27 mM, 28 mM, 30 mM, 32 mM, 34 mM, 36 mM, 38 mM, 40 mM, 50 mM, 60 mM, 70 mM, 80 mM, 90 mM, or 100 mM, or a range of any value. In some embodiments, the concentration of the histidine-acetate buffer is 10 mM. In some embodiments, the concentration of the histidine-acetate buffer is 20 mM. In some embodiments, the concentration of the histidine-acetate buffer is 30 mM.

[0033] In some embodiments, the pH of the pharmaceutical composition of the present disclosure is 4 to 7, 5 to 7, 5.5 to 7, 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5. In some embodiments, examples of the pH of the pharmaceutical composition include, but are not limited to, 5, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, or 7, or a range formed by any value. In some embodiments, the pH of the pharmaceutical composition is 5. In some embodiments, the pH of the pharmaceutical composition is 6. In some embodiments, the pH of the pharmaceutical composition is 6.5. In some embodiments, the pH of the pharmaceutical composition is 7.

[0034] In some embodiments, in the pharmaceutical composition, the surfactant is a polysorbate (e.g., polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 65, polysorbate 80, polysorbate 81, or polysorbate 85), a poloxamer (e.g., poloxamer 181, poloxamer 188, or poloxamer 407), or polyethylene glycol. In some embodiments, in the pharmaceutical composition, the surfactant is polysorbate 80 or polysorbate 20. In some embodiments, in the pharmaceutical composition, the surfactant is polysorbate 80.

[0035] In some embodiments, the amount of the surfactant in the pharmaceutical composition is 0.01 mg to 2 mg, 0.05 mg to 1.5 mg, 0.1 mg to 1 mg, 0.2 mg to 0.8 mg, 0.4 mg to 0.8 mg, 0.3 mg to 0.5 mg, or 0.7 mg to 0.9 mg per 60 mg of the anti-TIM-3 antibody or antigen-binding fragment thereof. In some embodiments, the amount of the surfactant is 0.2 mg to 0.8 mg, or 0.4 mg to 0.8 mg per 60 mg of the anti-TIM-3 antibody or antigen-binding fragment thereof. In some embodiments, the amount of the surfactant is 0.4 mg to 1.2 mg, or 0.6 mg to 1 mg per 60 mg of the anti-TIM-3 antibody or antigen-binding fragment thereof. In some embodiments, examples of the amount of the surfactant include, but are not limited to, 0.05 mg, 0.06 mg, 0.07 mg, 0.08 mg, 0.09 mg, 0.1 mg, 0.2 mg, 0.3 mg, 0.4 mg, 0.5 mg, 0.6 mg, 0.7 mg, 0.8 mg, 0.9 mg, 1 mg, 1.1 mg, 1.2 mg, 1.3 mg, 1.4 mg, or 1.5 mg per 60 mg of anti-TIM-3 antibody or antigen-binding fragment thereof, or a range thereof. In some embodiments, the amount of the surfactant is 0.2 mg per 60 mg of anti-TIM-3 antibody or antigen-binding fragment thereof. In some embodiments, the amount of the surfactant is 0.4 mg per 60 mg of anti-TIM-3 antibody or antigen-binding fragment thereof. In some embodiments, the amount of the surfactant is 0.8 mg per 60 mg of anti-TIM-3 antibody or antigen-binding fragment thereof. In some embodiments, the surfactant in the pharmaceutical composition of the present disclosure is polysorbate 20 or polysorbate 80. In some embodiments, the surfactant in the pharmaceutical composition of the present disclosure is preferably polysorbate 80.

[0036] In some embodiments, in the pharmaceutical composition, the concentration of the surfactant is 0.01 mg / mL to 2 mg / mL, 0.05 mg / mL to 1.5 mg / mL, 0.1 mg / mL to 1 mg / mL, 0.2 mg / mL to 0.8 mg / mL, 0.4 mg / mL to 0.8 mg / mL, 0.3 mg / mL to 0.5 mg / mL, or 0.7 mg / mL to 0.9 mg / mL. In some embodiments, the concentration of the surfactant is 0.2 mg / mL to 0.8 mg / mL, or 0.4 mg / mL to 0.8 mg / mL. In some embodiments, the concentration of the surfactant is 0.4 mg / mL to 1.2 mg / mL, or 0.6 mg / mL to 1 mg / mL. In some embodiments, the example of the concentration of the surfactant includes but is not limited to: 0.05mg / mL, 0.06mg / mL, 0.07mg / mL, 0.08mg / mL, 0.09mg / mL, 0.1mg / mL, 0.2mg / mL, 0.3mg / mL, 0.4mg / mL, 0.5mg / mL, 0.6mg / mL, 0.7mg / mL, 0.8mg / mL, 0.9mg / mL, 1mg / mL, 1.1mg / mL, 1.2mg / mL, 1.3mg / mL, 1.4mg / mL or 1.5mg / mL or the range formed by any value. In some embodiments, the concentration of the surfactant is 0.2mg / mL. In some embodiments, the concentration of the surfactant is 0.4mg / mL. In some embodiments, the concentration of the surfactant is 0.8mg / mL. In some embodiments, the surfactant in the pharmaceutical composition of the present disclosure is polysorbate 20 or polysorbate 80. In some embodiments, the surfactant in the pharmaceutical composition of the present disclosure is preferably polysorbate 80.

[0037] In some embodiments, in the pharmaceutical composition, the concentration of the surfactant is 0.001% (w / v) to 0.2% (w / v), 0.005% (w / v) to 0.15% (w / v), 0.01% (w / v) to 0.1% (w / v), 0.02% (w / v) to 0.08% (w / v), 0.04% (w / v) to 0.08% (w / v), 0.03% (w / v) to 0.05% (w / v), or 0.07% (w / v) to 0.09% (w / v). In some embodiments, the concentration of the surfactant is 0.02% (w / v) to 0.08% (w / v), or 0.04% (w / v) to 0.08% (w / v). In some embodiments, the concentration of the surfactant is 0.04% (w / v) to 0.12% (w / v), or 0.06% (w / v) to 0.1% (w / v). In some embodiments, the concentration of the surfactant includes, but is not limited to, 0.005% (w / v), 0.006% (w / v), 0.007% (w / v), 0.008% (w / v), 0.009% (w / v), 0.01% (w / v), 0.02% (w / v), 0.03% (w / v), 0.04% (w / v), 0.05% (w / v), 0.06% (w / v), 0.07% (w / v), 0.08% (w / v), 0.09% (w / v), 0.1% (w / v), 0.11% (w / v), 0.12% (w / v), 0.13% (w / v), 0.14% (w / v), or 0.15% (w / v), or any range thereof. In some embodiments, the concentration of the surfactant is 0.02% (w / v). In some embodiments, the concentration of the surfactant is 0.04% (w / v). In some embodiments, the concentration of the surfactant is 0.08% (w / v). In some embodiments, the surfactant in the pharmaceutical composition of the present disclosure is polysorbate 20 or polysorbate 80. In some embodiments, the surfactant in the pharmaceutical composition of the present disclosure is preferably polysorbate 80.

[0038] In some embodiments, the amount of polysorbate 80 in the pharmaceutical composition is 0.01 mg to 2 mg, 0.05 mg to 1.5 mg, 0.1 mg to 1 mg, 0.2 mg to 0.8 mg, 0.4 mg to 0.8 mg, 0.3 mg to 0.5 mg, or 0.7 mg to 0.9 mg per 60 mg of anti-TIM-3 antibody or antigen-binding fragment thereof. In some embodiments, the amount of polysorbate 80 is 0.2 mg to 0.8 mg, or 0.4 mg to 0.8 mg per 60 mg of anti-TIM-3 antibody or antigen-binding fragment thereof. In some embodiments, the amount of polysorbate 80 is 0.4 mg to 1.2 mg, or 0.6 mg to 1 mg per 60 mg of anti-TIM-3 antibody or antigen-binding fragment thereof. In some embodiments, examples of amounts of polysorbate 80 include, but are not limited to, 0.05 mg, 0.06 mg, 0.07 mg, 0.08 mg, 0.09 mg, 0.1 mg, 0.2 mg, 0.3 mg, 0.4 mg, 0.5 mg, 0.6 mg, 0.7 mg, 0.8 mg, 0.9 mg, 1 mg, 1.1 mg, 1.2 mg, 1.3 mg, 1.4 mg, or 1.5 mg per 60 mg of anti-TIM-3 antibody or antigen-binding fragment thereof, or any range thereof. In some embodiments, the amount of polysorbate 80 is 0.2 mg per 60 mg of anti-TIM-3 antibody or antigen-binding fragment thereof. In some embodiments, the amount of polysorbate 80 is 0.4 mg per 60 mg of anti-TIM-3 antibody or antigen-binding fragment thereof. In some embodiments, the amount of polysorbate 80 is 0.8 mg per 60 mg of anti-TIM-3 antibody or antigen-binding fragment thereof.

[0039] In some embodiments, in the pharmaceutical composition, the concentration of the polysorbate 80 is 0.01 mg / mL to 2 mg / mL, 0.05 mg / mL to 1.5 mg / mL, 0.1 mg / mL to 1 mg / mL, 0.2 mg / mL to 0.8 mg / mL, 0.4 mg / mL to 0.8 mg / mL, 0.3 mg / mL to 0.5 mg / mL, or 0.7 mg / mL to 0.9 mg / mL. In some embodiments, the concentration of the polysorbate 80 is 0.2 mg / mL to 0.8 mg / mL, or 0.4 mg / mL to 0.8 mg / mL. In some embodiments, the concentration of the polysorbate 80 is 0.4 mg / mL to 1.2 mg / mL, or 0.6 mg / mL to 1 mg / mL. In some embodiments, the example of the concentration of the polysorbate 80 includes but is not limited to: 0.05mg / mL, 0.06mg / mL, 0.07mg / mL, 0.08mg / mL, 0.09mg / mL, 0.1mg / mL, 0.2mg / mL, 0.3mg / mL, 0.4mg / mL, 0.5mg / mL, 0.6mg / mL, 0.7mg / mL, 0.8mg / mL, 0.9mg / mL, 1mg / mL, 1.1mg / mL, 1.2mg / mL, 1.3mg / mL, 1.4mg / mL or 1.5mg / mL or the range formed by any value. In some embodiments, the concentration of the polysorbate 80 is 0.2mg / mL. In some embodiments, the concentration of the polysorbate 80 is 0.4mg / mL. In some embodiments, the concentration of the polysorbate 80 is 0.8mg / mL.

[0040] In some embodiments, in the pharmaceutical composition, the concentration of polysorbate 80 is 0.001% (w / v) to 0.2% (w / v), 0.005% (w / v) to 0.15% (w / v), 0.01% (w / v) to 0.1% (w / v), 0.02% (w / v) to 0.08% (w / v), 0.04% (w / v) to 0.08% (w / v), 0.03% (w / v) to 0.05% (w / v), or 0.07% (w / v) to 0.09% (w / v). In some embodiments, the concentration of polysorbate 80 is 0.02% (w / v) to 0.08% (w / v), or 0.04% (w / v) to 0.08% (w / v). In some embodiments, the concentration of polysorbate 80 is 0.04% (w / v) to 0.12% (w / v), or 0.06% (w / v) to 0.1% (w / v). In some embodiments, examples of the concentration of polysorbate 80 include, but are not limited to, 0.005% (w / v), 0.006% (w / v), 0.007% (w / v), 0.008% (w / v), 0.009% (w / v), 0.01% (w / v), 0.02% (w / v), 0.03% (w / v), 0.04% (w / v), 0.05% (w / v), 0.06% (w / v), 0.07% (w / v), 0.08% (w / v), 0.09% (w / v), 0.1% (w / v), 0.11% (w / v), 0.12% (w / v), 0.13% (w / v), 0.14% (w / v), or 0.15% (w / v), or a range of any value. In some embodiments, the concentration of polysorbate 80 is 0.02% (w / v). In some embodiments, the concentration of polysorbate 80 is 0.04% (w / v). In some embodiments, the concentration of polysorbate 80 is 0.08% (w / v).

[0041] In some embodiments, in the pharmaceutical composition, the stabilizer includes but is not limited to sugar alcohols (e.g., mannitol, sorbitol), disaccharides (e.g., trehalose, sucrose, maltose, lactose), monosaccharides (e.g., dextrose (D-glucose)), amino acids (e.g., lysine, glycine, proline, arginine, or pharmaceutically acceptable salts thereof), and / or salts (e.g., sodium chloride). In some embodiments, the stabilizer comprises mannitol, sorbitol, trehalose, sucrose, lysine, glycine, proline, arginine, or pharmaceutically acceptable salts thereof, and / or sodium chloride. In some embodiments, the stabilizer comprises a disaccharide (e.g., sucrose or trehalose). In some embodiments, the stabilizer comprises sucrose. In some embodiments, the stabilizer comprises sucrose and sodium chloride.

[0042] In some embodiments, the amount of sucrose in the pharmaceutical composition is 30 to 960 μmol, 60 to 720 μmol, 120 to 480 μmol, 240 to 480 μmol, 210 to 270 μmol, 230 to 250 μmol, 450 to 510 μmol, or 470 to 490 μmol per 60 mg of the anti-TIM-3 antibody or antigen-binding fragment thereof. In some embodiments, the amount of sucrose is 120 to 480 μmol, or 240 to 480 μmol per 60 mg of the anti-TIM-3 antibody or antigen-binding fragment thereof. In some embodiments, examples of the amount of sucrose include, but are not limited to, 30 μmol, 60 μmol, 90 μmol, 120 μmol, 150 μmol, 180 μmol, 200 μmol, 210 μmol, 230 μmol, 240 μmol, 250 μmol, 270 μmol, 300 μmol, 330 μmol, 360 μmol, 390 μmol, 400 μmol, 420 μmol, 450 μmol per 60 mg of anti-TIM-3 antibody or antigen-binding fragment thereof. In some embodiments, the amount of sucrose is 120 μmol per 60 mg of anti-TIM-3 antibody or antigen-binding fragment thereof. In some embodiments, the amount of sucrose is 240 μmol per 60 mg of anti-TIM-3 antibody or antigen-binding fragment thereof. In some embodiments, the amount of sucrose is 480 μmol per 60 mg of anti-TIM-3 antibody or antigen-binding fragment thereof.

[0043] In some embodiments, in the pharmaceutical composition, the concentration of sucrose is 30mM to 960mM, 60mM to 720mM, 120mM to 480mM, 240mM to 480mM, 210mM ​​to 270mM, 230mM to 250mM, 450mM to 510mM, or 470mM to 490mM. In some embodiments, the concentration of sucrose is 120mM to 480mM, or 240mM to 480mM. In some embodiments, examples of the concentration of sucrose include, but are not limited to, 30 mM, 60 mM, 90 mM, 120 mM, 150 mM, 180 mM, 200 mM, 210 mM, 230 mM, 240 mM, 250 mM, 270 mM, 300 mM, 330 mM, 360 mM, 390 mM, 400 mM, 420 mM, 450 mM, 470 mM, In some embodiments, the concentration of sucrose is 100mM, 200mM, 300mM, 400mM, 500mM, 510mM, 540mM, 570mM, 600mM, 630mM, 660mM, 690mM, 700mM, 720mM, 750mM, 780mM, 800mM, 810mM, 840mM, 870mM, 900mM, 930mM or 960mM or the range formed by any value. In some embodiments, the concentration of the sucrose is 120mM. In some embodiments, the concentration of the sucrose is 240mM. In some embodiments, the concentration of the sucrose is 480mM.

[0044] In some embodiments, in the pharmaceutical composition, the amount of sodium chloride is 5 μmol to 200 μmol, 10 μmol to 100 μmol, 20 μmol to 100 μmol, or 50 μmol to 100 μmol per 60 mg of the anti-TIM-3 antibody or antigen-binding fragment thereof. In some embodiments, the amount of sodium chloride is 50 μmol to 100 μmol per 60 mg of the anti-TIM-3 antibody or antigen-binding fragment thereof. In some embodiments, examples of the amount of sodium chloride include, but are not limited to, 5 μmol, 10 μmol, 15 μmol, 20 μmol, 25 μmol, 30 μmol, 35 μmol, 40 μmol, 45 μmol, 50 μmol, 55 μmol, 60 μmol, 65 μmol, 70 μmol, 75 μmol, 80 μmol, 85 μmol, 90 μmol, 95 μmol, 100 μmol, 110 μmol, 120 μmol, 130 μmol, 140 μmol, 150 μmol, 160 μmol, 170 μmol, 180 μmol, 190 μmol, or 200 μmol, or a range of any values ​​per 60 mg of the anti-TIM-3 antibody or antigen-binding fragment thereof. In some embodiments, the amount of sodium chloride is 50 μmol per 60 mg of the anti-TIM-3 antibody or antigen-binding fragment thereof. In some embodiments, the amount of sodium chloride is 100 μmol per 60 mg of the anti-TIM-3 antibody or antigen-binding fragment thereof.

[0045] In some embodiments, in the pharmaceutical composition, the concentration of sodium chloride is 5 mM to 200 mM, 10 mM to 100 mM, 20 mM to 100 mM, or 50 mM to 100 mM. In some embodiments, the concentration of sodium chloride is 50 mM to 100 mM. In some embodiments, examples of the concentration of sodium chloride include, but are not limited to, 5 mM, 10 mM, 15 mM, 20 mM, 25 mM, 30 mM, 35 mM, 40 mM, 45 mM, 50 mM, 55 mM, 60 mM, 65 mM, 70 mM, 75 mM, 80 mM, 85 mM, 90 mM, 95 mM, 100 mM, 110 mM, 120 mM, 130 mM, 140 mM, 150 mM, 160 mM, 170 mM, 180 mM, 190 mM, or 200 mM, or a range of any value. In some embodiments, the concentration of sodium chloride is 50 mM. In some embodiments, the concentration of sodium chloride is 100 mM.

[0046] In some embodiments, the pharmaceutical composition comprises: (a) an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) a histidine buffer, a phosphate buffer, a succinate buffer, a citrate buffer, and / or an acetate buffer, (c) polysorbate 80 or polysorbate 20, and (d) mannitol, sorbitol, trehalose, sucrose, lysine, glycine, proline, arginine, or a pharmaceutically acceptable salt thereof, and / or sodium chloride.

[0047] In some embodiments, the pharmaceutical composition comprises: (a) an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) a histidine buffer, a phosphate buffer, or a citrate buffer, (c) polysorbate 80 or polysorbate 20, and (d) sucrose and / or sodium chloride.

[0048] In some embodiments, the pharmaceutical composition comprises: (a) an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) a histidine buffer, (c) polysorbate 80 or polysorbate 20, and (d) sucrose.

[0049] In some embodiments, the pharmaceutical composition comprises: (a) an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) a histidine buffer, (c) polysorbate 80 or polysorbate 20, and (d) sucrose and sodium chloride.

[0050] In some embodiments, the pharmaceutical composition comprises: (a) an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) a phosphate buffer, (c) polysorbate 80 or polysorbate 20, and (d) sucrose.

[0051] In some embodiments, the pharmaceutical composition comprises: (a) an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) a citrate buffer, (c) polysorbate 80 or polysorbate 20, and (d) sucrose.

[0052] In some embodiments, the pharmaceutical composition comprises: (a) an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) a histidine buffer, a phosphate buffer, or a citrate buffer, (c) polysorbate 80, and (d) sucrose and / or sodium chloride.

[0053] In some embodiments, the pharmaceutical composition comprises: (a) an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) a histidine buffer, (c) polysorbate 80, and (d) sucrose.

[0054] In some embodiments, the pharmaceutical composition comprises: (a) an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) a histidine buffer, (c) polysorbate 80, and (d) sucrose and sodium chloride.

[0055] In some embodiments, the pharmaceutical composition comprises: (a) an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) a phosphate buffer, (c) polysorbate 80, and (d) sucrose.

[0056] In some embodiments, the pharmaceutical composition comprises: (a) an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) a citrate buffer, (c) polysorbate 80, and (d) sucrose.

[0057] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 1 μmol to 100 μmol of a buffer, (c) 0.01 mg to 2 mg of a surfactant, and (d) a stabilizer comprising sucrose and / or sodium chloride.

[0058] In some embodiments, the pharmaceutical composition comprises: (a) 1 mg / mL to 120 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 1 mM to 100 mM buffer, (c) 0.01 mg / mL to 2 mg / mL of a surfactant, and (d) a stabilizer comprising sucrose and / or sodium chloride; and the pH of the pharmaceutical composition is 4 to 7, 5 to 7, 5.5 to 7, 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5.

[0059] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 2 μmol to 80 μmol of a buffer, (c) 0.05 mg to 1.5 mg of a surfactant, and (d) a stabilizer comprising sucrose and / or sodium chloride.

[0060] In some embodiments, the pharmaceutical composition comprises: (a) 10 mg / mL to 100 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 2 mM to 80 mM buffer, (c) 0.05 mg / mL to 1.5 mg / mL of a surfactant, and (d) a stabilizer comprising sucrose and / or sodium chloride; and the pH of the pharmaceutical composition is 5 to 7, 5.5 to 7, 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5.

[0061] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 5 μmol to 60 μmol of a buffer, (c) 0.1 mg to 1 mg of a surfactant, and (d) a stabilizer comprising sucrose and / or sodium chloride.

[0062] In some embodiments, the pharmaceutical composition comprises: (a) 15 mg / mL to 90 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 5 mM to 60 mM buffer, (c) 0.1 mg / mL to 1 mg / mL of a surfactant, and (d) a stabilizer comprising sucrose and / or sodium chloride; and the pH of the pharmaceutical composition is 5 to 7, 5.5 to 7, 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5.

[0063] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 8 μmol to 40 μmol of a buffer, (c) 0.2 mg to 0.8 mg of a surfactant, and (d) a stabilizer comprising sucrose and / or sodium chloride.

[0064] In some embodiments, the pharmaceutical composition comprises: (a) 20 mg / mL to 80 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 8 mM to 40 mM buffer, (c) 0.2 mg / mL to 0.8 mg / mL of a surfactant, and (d) a stabilizer comprising sucrose and / or sodium chloride; and the pH of the pharmaceutical composition is 5 to 7, 5.5 to 7, 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5.

[0065] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 10 μmol to 30 μmol of a buffer, (c) 0.2 mg to 0.8 mg of a surfactant, and (d) a stabilizer comprising sucrose and / or sodium chloride.

[0066] In some embodiments, the pharmaceutical composition comprises: (a) 30 mg / mL to 60 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 10 mM to 30 mM of a buffer, (c) 0.2 mg / mL to 0.8 mg / mL of a surfactant, and (d) a stabilizer comprising sucrose and / or sodium chloride; and the pH of the pharmaceutical composition is 5 to 7, 5.5 to 7, 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5. In some embodiments, the pH of the pharmaceutical composition is preferably 6 to 7.

[0067] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 10 μmol to 30 μmol of a buffer, (c) 0.4 mg to 1.2 mg of a surfactant, and (d) a stabilizer comprising sucrose and / or sodium chloride.

[0068] In some embodiments, the pharmaceutical composition comprises: (a) 30 mg / mL to 60 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 10 mM to 30 mM of a buffer, (c) 0.4 mg / mL to 1.2 mg / mL of a surfactant, and (d) a stabilizer comprising sucrose and / or sodium chloride; and the pH of the pharmaceutical composition is 5 to 7, 5.5 to 7, 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5. In some embodiments, the pH of the pharmaceutical composition is preferably 6 to 7.

[0069] In some embodiments, the pharmaceutical composition comprises: (a) per 60 mg of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 10 μmol to 30 μmol of a buffer, (c) 0.4 mg to 0.8 mg of a surfactant, and (d) a stabilizer comprising sucrose and / or sodium chloride.

[0070] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 10 mM to 30 mM of a buffer, (c) 0.4 mg / mL to 0.8 mg / mL of a surfactant, and (d) a stabilizer comprising sucrose and / or sodium chloride; and the pH of the pharmaceutical composition is 5 to 7, 5.5 to 7, 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5. In some embodiments, the pH of the pharmaceutical composition is preferably 6 to 7.

[0071] In some embodiments, the pharmaceutical composition comprises: (a) per 60 mg of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 10 μmol to 30 μmol of a buffer, (c) 0.6 mg to 1 mg of a surfactant, and (d) a stabilizer comprising sucrose and / or sodium chloride.

[0072] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 10 mM to 30 mM buffer, (c) 0.6 mg / mL to 1 mg / mL of a surfactant, and (d) a stabilizer comprising sucrose and / or sodium chloride; and the pH of the pharmaceutical composition is 5 to 7, 5.5 to 7, 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5. In some embodiments, the pH of the pharmaceutical composition is preferably 6 to 7.

[0073] In some embodiments, the pharmaceutical composition comprises: (a) per 60 mg of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 15 μmol to 25 μmol of a buffer, (c) 0.7 mg to 0.9 mg of a surfactant, and (d) a stabilizer comprising sucrose and / or sodium chloride.

[0074] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 15 mM to 25 mM buffer, (c) 0.7 mg / mL to 0.9 mg / mL of a surfactant, and (d) a stabilizer comprising sucrose and / or sodium chloride; and the pH of the pharmaceutical composition is 5 to 7, 5.5 to 7, 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5. In some embodiments, the pH of the pharmaceutical composition is preferably 6 to 7.

[0075] In some embodiments, the pharmaceutical composition comprises: (a) per 60 mg of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 15 to 25 μmol of a buffer, (c) 0.4 to 0.8 mg of a surfactant, and (d) a stabilizer, wherein the stabilizer comprises sucrose.

[0076] In some embodiments, the pharmaceutical composition comprises: (a) 55 mg / mL to 65 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 15 mM to 25 mM of a buffer, (c) 0.4 mg / mL to 0.8 mg / mL of a surfactant, and (d) a stabilizer comprising sucrose; and the pH of the pharmaceutical composition is 5 to 7, 5.5 to 7, 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5. In some embodiments, the pH of the pharmaceutical composition is preferably 6 to 7.

[0077] In some embodiments, the pharmaceutical composition comprises: (a) per 60 mg of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 20 μmol of a buffer, (c) 0.4 mg to 0.8 mg of a surfactant, and (d) a stabilizer, wherein the stabilizer comprises sucrose.

[0078] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg / mL anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 20 mM buffer, (c) 0.4 mg / mL to 0.8 mg / mL surfactant, and (d) a stabilizer comprising sucrose; and the pH of the pharmaceutical composition is 5 to 7, 5.5 to 7, 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5. In some embodiments, the pH of the pharmaceutical composition is preferably 6 to 7.

[0079] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 20 μmol of a buffer, (c) 0.4 mg of a surfactant, and (d) a stabilizer, wherein the stabilizer comprises sucrose.

[0080] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 20 mM of a buffer, (c) 0.4 mg / mL of a surfactant, and (d) a stabilizer comprising sucrose; and the pH of the pharmaceutical composition is 5 to 7, 5.5 to 7, 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5. In some embodiments, the pH of the pharmaceutical composition is preferably 6 to 7.

[0081] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 20 μmol of a buffer, (c) 0.8 mg of a surfactant, and (d) a stabilizer, wherein the stabilizer comprises sucrose.

[0082] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 20 mM of a buffer, (c) 0.8 mg / mL of a surfactant, and (d) a stabilizer comprising sucrose; and the pH of the pharmaceutical composition is 5 to 7, 5.5 to 7, 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5. In some embodiments, the pH of the pharmaceutical composition is preferably 6 to 7.

[0083] In some embodiments, the pharmaceutical composition comprises: (a) per 60 mg of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 1 μmol to 100 μmol of a histidine buffer, a phosphate buffer, or a citrate buffer, (c) 0.01 mg to 2 mg of polysorbate 80, and (d) 30 μmol to 960 μmol of sucrose and / or 5 μmol to 200 μmol of sodium chloride.

[0084] In some embodiments, the pharmaceutical composition comprises: (a) 1 mg / mL to 120 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 1 mM to 100 mM histidine buffer, phosphate buffer, or citrate buffer, (c) 0.01 mg / mL to 2 mg / mL polysorbate 80, and (d) 30 mM to 960 mM sucrose and / or 5 mM to 200 mM sodium chloride; and the pH of the pharmaceutical composition is 4 to 7, 5 to 7, 5.5 to 7, 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5.

[0085] In some embodiments, the pharmaceutical composition comprises: (a) per 60 mg of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 2 μmol to 80 μmol of a histidine buffer, a phosphate buffer, or a citrate buffer, (c) 0.05 mg to 1.5 mg of polysorbate 80, and (d) 60 μmol to 720 μmol of sucrose and / or 10 μmol to 100 μmol of sodium chloride.

[0086] In some embodiments, the pharmaceutical composition comprises: (a) 10 mg / mL to 100 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 2 mM to 80 mM histidine buffer, phosphate buffer, or citrate buffer, (c) 0.05 mg / mL to 1.5 mg / mL polysorbate 80, and (d) 60 mM to 720 mM sucrose and / or 10 mM to 100 mM sodium chloride; and the pH of the pharmaceutical composition is 4 to 7, 5 to 7, 5.5 to 7, 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5.

[0087] In some embodiments, the pharmaceutical composition comprises: (a) per 60 mg of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 5 to 60 μmol of a histidine buffer, a phosphate buffer, or a citrate buffer, (c) 0.1 to 1 mg of polysorbate 80, and (d) 120 to 480 μmol of sucrose and / or 20 to 100 μmol of sodium chloride.

[0088] In some embodiments, the pharmaceutical composition comprises: (a) 15 mg / mL to 90 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 5 mM to 60 mM histidine buffer, phosphate buffer, or citrate buffer, (c) 0.1 mg / mL to 1 mg / mL polysorbate 80, and (d) 120 mM to 480 mM sucrose and / or 20 mM to 100 mM sodium chloride; and the pH of the pharmaceutical composition is 5 to 7, 5.5 to 7, 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5.

[0089] In some embodiments, the pharmaceutical composition comprises: (a) per 60 mg of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 8 to 40 μmol of a histidine buffer, a phosphate buffer, or a citrate buffer, (c) 0.2 to 0.8 mg of polysorbate 80, and (d) 120 to 480 μmol of sucrose and / or 20 to 100 μmol of sodium chloride.

[0090] In some embodiments, the pharmaceutical composition comprises: (a) 20 mg / mL to 80 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 8 mM to 40 mM histidine buffer, phosphate buffer, or citrate buffer, (c) 0.2 mg / mL to 0.8 mg / mL polysorbate 80, and (d) 120 mM to 480 mM sucrose and / or 20 mM to 100 mM sodium chloride; and the pH of the pharmaceutical composition is 5 to 7, 5.5 to 7, 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5.

[0091] In some embodiments, the pharmaceutical composition comprises: (a) per 60 mg of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 10 μmol to 30 μmol of a histidine buffer, a phosphate buffer, or a citrate buffer, (c) 0.2 mg to 0.8 mg of polysorbate 80, and (d) 120 μmol to 480 μmol of sucrose and / or 50 μmol to 100 μmol of sodium chloride.

[0092] In some embodiments, the pharmaceutical composition comprises: (a) 30 mg / mL to 60 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 10 mM to 30 mM histidine buffer, phosphate buffer, or citrate buffer, (c) 0.2 mg / mL to 0.8 mg / mL polysorbate 80, and (d) 120 mM to 480 mM sucrose and / or 50 mM to 100 mM sodium chloride; and the pH of the pharmaceutical composition is 5 to 7, 5.5 to 7, 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5. In some embodiments, the pH of the pharmaceutical composition is preferably 6 to 7.

[0093] In some embodiments, the pharmaceutical composition comprises: (a) per 60 mg of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 10 μmol to 30 μmol of a histidine buffer, a phosphate buffer, or a citrate buffer, (c) 0.2 mg to 0.8 mg of polysorbate 80, and (d) 120 μmol to 480 μmol of sucrose.

[0094] In some embodiments, the pharmaceutical composition comprises: (a) 30 mg / mL to 60 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 10 mM to 30 mM histidine buffer, phosphate buffer, or citrate buffer, (c) 0.2 mg / mL to 0.8 mg / mL of polysorbate 80, and (d) 120 mM to 480 mM sucrose; and the pH of the pharmaceutical composition is 5 to 7, 5.5 to 7, 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5. In some embodiments, the pH of the pharmaceutical composition is preferably 6 to 7.

[0095] In some embodiments, the pharmaceutical composition comprises: (a) per 60 mg of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 10 μmol to 30 μmol of a histidine buffer, a phosphate buffer, or a citrate buffer, (c) 0.4 mg to 1.2 mg, or 0.6 mg to 1 mg of polysorbate 80, and (d) 120 μmol to 480 μmol of sucrose.

[0096] In some embodiments, the pharmaceutical composition comprises: (a) 30 mg / mL to 60 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 10 mM to 30 mM histidine buffer, phosphate buffer, or citrate buffer, (c) 0.4 mg / mL to 1.2 mg / mL, or 0.6 mg / mL to 1 mg / mL of polysorbate 80, and (d) 120 mM to 480 mM sucrose; and the pH of the pharmaceutical composition is 5 to 7, 5.5 to 7, 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5. In some embodiments, the pH of the pharmaceutical composition is preferably 6 to 7.

[0097] In some embodiments, the pharmaceutical composition comprises: (a) per 60 mg of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 10 μmol to 30 μmol of a histidine buffer, a phosphate buffer, or a citrate buffer, (c) 0.4 mg to 0.8 mg of polysorbate 80, and (d) 240 μmol to 480 μmol of sucrose.

[0098] In some embodiments, the pharmaceutical composition comprises: (a) 30 mg / mL to 60 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 10 mM to 30 mM histidine buffer, phosphate buffer, or citrate buffer, (c) 0.4 mg / mL to 0.8 mg / mL of polysorbate 80, and (d) 240 mM to 480 mM sucrose; and the pH of the pharmaceutical composition is 5 to 7, 5.5 to 7, 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5. In some embodiments, the pH of the pharmaceutical composition is preferably 6 to 7.

[0099] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 10 μmol to 30 μmol of a histidine buffer, (c) 0.2 mg to 0.8 mg of polysorbate 80, and (d) 120 μmol to 480 μmol of sucrose and 50 μmol to 100 μmol of sodium chloride.

[0100] In some embodiments, the pharmaceutical composition comprises: (a) 30 mg / mL to 60 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 10 mM to 30 mM histidine buffer, (c) 0.2 mg / mL to 0.8 mg / mL polysorbate 80, and (d) 120 mM to 480 mM sucrose and 50 mM to 100 mM sodium chloride; and the pH of the pharmaceutical composition is 5 to 7, 5.5 to 7, 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5.

[0101] In some embodiments, the pharmaceutical composition comprises: (a) per 60 mg of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 15 μmol to 25 μmol of a histidine buffer, a phosphate buffer, or a citrate buffer, (c) 0.4 mg to 0.8 mg of polysorbate 80, and (d) 240 μmol to 480 μmol of sucrose.

[0102] In some embodiments, the pharmaceutical composition comprises: (a) 55 mg / mL to 65 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 15 mM to 25 mM of a histidine buffer, a phosphate buffer, or a citrate buffer, (c) 0.4 mg / mL to 0.8 mg / mL of polysorbate 80, and (d) 240 mM to 480 mM of sucrose; and the pH of the pharmaceutical composition is 5 to 7, 5.5 to 7, 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5. In some embodiments, the pH of the pharmaceutical composition is preferably 6 to 7.

[0103] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 20 μmol of a histidine buffer, a phosphate buffer, or a citrate buffer, (c) 0.4 mg to 0.8 mg of polysorbate 80, and (d) 240 μmol to 480 μmol of sucrose.

[0104] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 20 mM of histidine buffer, phosphate buffer, or citrate buffer, (c) 0.4 mg / mL to 0.8 mg / mL of polysorbate 80, and (d) 240 mM to 480 mM of sucrose; and the pH of the pharmaceutical composition is 5 to 7, 5.5 to 7, 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5. In some embodiments, the pH of the pharmaceutical composition is preferably 6 to 7.

[0105] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 1 μmol to 100 μmol of a histidine buffer, (c) 0.01 mg to 2 mg of polysorbate 80, and (d) 30 μmol to 960 μmol of sucrose.

[0106] In some embodiments, the pharmaceutical composition comprises: (a) 1 mg / mL to 120 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 1 mM to 100 mM histidine buffer, (c) 0.01 mg / mL to 2 mg / mL of polysorbate 80, and (d) 30 mM to 960 mM sucrose; and the pH of the pharmaceutical composition is 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5.

[0107] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 2 μmol to 80 μmol of a histidine buffer, (c) 0.05 mg to 1.5 mg of polysorbate 80, and (d) 60 μmol to 720 μmol of sucrose.

[0108] In some embodiments, the pharmaceutical composition comprises: (a) 10 mg / mL to 100 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 2 mM to 80 mM histidine buffer, (c) 0.05 mg / mL to 1.5 mg / mL polysorbate 80, and (d) 60 mM to 720 mM sucrose; and the pH of the pharmaceutical composition is 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5.

[0109] In some embodiments, the pharmaceutical composition comprises: (a) per 60 mg of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 5 μmol to 60 μmol of a histidine buffer, (c) 0.1 mg to 1 mg of polysorbate 80, and (d) 120 μmol to 480 μmol of sucrose.

[0110] In some embodiments, the pharmaceutical composition comprises: (a) 15 mg / mL to 90 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 5 mM to 60 mM histidine buffer, (c) 0.1 mg / mL to 1 mg / mL polysorbate 80, and (d) 120 mM to 480 mM sucrose; and the pH of the pharmaceutical composition is 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5.

[0111] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 8 μmol to 40 μmol of a histidine buffer, (c) 0.2 mg to 0.8 mg of polysorbate 80, and (d) 120 μmol to 480 μmol of sucrose.

[0112] In some embodiments, the pharmaceutical composition comprises: (a) 20 mg / mL to 80 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 8 mM to 40 mM histidine buffer, (c) 0.2 mg / mL to 0.8 mg / mL polysorbate 80, and (d) 120 mM to 480 mM sucrose; and the pH of the pharmaceutical composition is 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5.

[0113] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 10 μmol to 30 μmol of a histidine buffer, (c) 0.2 mg to 0.8 mg of polysorbate 80, and (d) 120 μmol to 480 μmol of sucrose.

[0114] In some embodiments, the pharmaceutical composition comprises: (a) 30 mg / mL to 60 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 10 mM to 30 mM histidine buffer, (c) 0.2 mg / mL to 0.8 mg / mL of polysorbate 80, and (d) 120 mM to 480 mM sucrose; and the pH of the pharmaceutical composition is 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5. In some embodiments, the pH of the pharmaceutical composition is preferably 6 to 7.

[0115] In some embodiments, the pharmaceutical composition comprises: (a) per 60 mg of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 10 μmol to 30 μmol of a histidine buffer, (c) 0.4 mg to 1.2 mg, or 0.6 mg to 1 mg, of polysorbate 80, and (d) 120 μmol to 480 μmol of sucrose.

[0116] In some embodiments, the pharmaceutical composition comprises: (a) 30 mg / mL to 60 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 10 mM to 30 mM histidine buffer, (c) 0.4 mg / mL to 1.2 mg / mL, or 0.6 mg / mL to 1 mg / mL of polysorbate 80, and (d) 120 mM to 480 mM sucrose; and the pH of the pharmaceutical composition is 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5. In some embodiments, the pH of the pharmaceutical composition is preferably 6 to 7.

[0117] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 10 μmol to 30 μmol of a histidine buffer, (c) 0.4 mg to 0.8 mg of polysorbate 80, and (d) 240 μmol to 480 μmol of sucrose.

[0118] In some embodiments, the pharmaceutical composition comprises: (a) 30 mg / mL to 60 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 10 mM to 30 mM histidine buffer, (c) 0.4 mg / mL to 0.8 mg / mL of polysorbate 80, and (d) 240 mM to 480 mM sucrose; and the pH of the pharmaceutical composition is 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5. In some embodiments, the pH of the pharmaceutical composition is preferably 6 to 7.

[0119] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 15 μmol to 25 μmol of a histidine buffer, (c) 0.4 mg to 0.8 mg of polysorbate 80, and (d) 240 μmol to 480 μmol of sucrose.

[0120] In some embodiments, the pharmaceutical composition comprises: (a) 55 mg / mL to 65 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 15 mM to 25 mM histidine buffer, (c) 0.4 mg / mL to 0.8 mg / mL of polysorbate 80, and (d) 240 mM to 480 mM sucrose; and the pH of the pharmaceutical composition is 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5. In some embodiments, the pH of the pharmaceutical composition is preferably 6 to 7.

[0121] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 15 μmol to 25 μmol of a histidine buffer, (c) 0.3 mg to 0.5 mg of polysorbate 80, and (d) 210 μmol to 270 μmol of sucrose.

[0122] In some embodiments, the pharmaceutical composition comprises: (a) 55 mg / mL to 65 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 15 mM to 25 mM histidine buffer, (c) 0.3 mg / mL to 0.5 mg / mL of polysorbate 80, and (d) 210 mM to 270 mM sucrose; and the pH of the pharmaceutical composition is 5 to 7, 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5. In some embodiments, the pH of the pharmaceutical composition is preferably 6 to 7.

[0123] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 19 μmol to 21 μmol of a histidine buffer, (c) 0.3 mg to 0.5 mg of polysorbate 80, and (d) 230 μmol to 250 μmol of sucrose.

[0124] In some embodiments, the pharmaceutical composition comprises: (a) 59 mg / mL to 61 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 19 mM to 21 mM histidine buffer, (c) 0.3 mg / mL to 0.5 mg / mL of polysorbate 80, and (d) 230 mM to 250 mM sucrose; and the pH of the pharmaceutical composition is 5.5 to 6.5, or 6 to 6.5.

[0125] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 10 μmol to 30 μmol of histidine buffer, (c) 0.6 mg to 1 mg of polysorbate 80, and (d) 450 μmol to 510 μmol of sucrose.

[0126] In some embodiments, the pharmaceutical composition comprises: (a) 55 mg / mL to 65 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 10 mM to 30 mM histidine buffer, (c) 0.6 mg / mL to 1 mg / mL of polysorbate 80, and (d) 450 mM to 510 mM sucrose; and the pH of the pharmaceutical composition is 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5. In some embodiments, the pH of the pharmaceutical composition is preferably 6 to 7.

[0127] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 15 μmol to 25 μmol of histidine buffer, (c) 0.7 mg to 0.9 mg of polysorbate 80, and (d) 450 μmol to 510 μmol of sucrose.

[0128] In some embodiments, the pharmaceutical composition comprises: (a) 55 mg / mL to 65 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 15 mM to 25 mM histidine buffer, (c) 0.7 mg / mL to 0.9 mg / mL of polysorbate 80, and (d) 450 mM to 510 mM sucrose; and the pH of the pharmaceutical composition is 5 to 7, 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5. In some embodiments, the pH of the pharmaceutical composition is preferably 6 to 7.

[0129] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 19 μmol to 21 μmol of a histidine buffer, (c) 0.7 mg to 0.9 mg of polysorbate 80, and (d) 470 μmol to 490 μmol of sucrose.

[0130] In some embodiments, the pharmaceutical composition comprises: (a) 59 mg / mL to 61 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 19 mM to 21 mM histidine buffer, (c) 0.7 mg / mL to 0.9 mg / mL of polysorbate 80, and (d) 470 mM to 490 mM sucrose; and the pH of the pharmaceutical composition is 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5. In some embodiments, the pH of the pharmaceutical composition is preferably 6 to 7.

[0131] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 20 μmol of histidine buffer, (c) 0.4 mg to 0.8 mg of polysorbate 80, and (d) 240 μmol to 480 μmol of sucrose.

[0132] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 20 mM histidine buffer, (c) 0.4 mg / mL to 0.8 mg / mL of polysorbate 80, and (d) 240 mM to 480 mM sucrose; and the pH of the pharmaceutical composition is 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5. In some embodiments, the pH of the pharmaceutical composition is preferably 6 to 7.

[0133] In some embodiments, the pharmaceutical composition comprises: (a) per 60 mg of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 20 μmol of histidine buffer, (c) 0.4 mg of polysorbate 80, and (d) 240 μmol of sucrose.

[0134] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg / mL of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 20 mM histidine buffer, (c) 0.4 mg / mL polysorbate 80, and (d) 240 mM sucrose; and the pH of the pharmaceutical composition is 5.5 to 6.5, preferably 6.

[0135] In some embodiments, the pharmaceutical composition comprises: (a) per 60 mg of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 20 μmol of histidine buffer, (c) 0.8 mg of polysorbate 80, and (d) 480 μmol of sucrose.

[0136] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg / mL of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 20 mM histidine buffer, (c) 0.8 mg / mL of polysorbate 80, and (d) 480 mM sucrose; and the pH of the pharmaceutical composition is 6 to 7, preferably 6.5.

[0137] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 10 μmol to 30 μmol of histidine-acetate buffer, (c) 0.4 mg to 0.8 mg of polysorbate 80, and (d) 240 μmol to 480 μmol of sucrose.

[0138] In some embodiments, the pharmaceutical composition comprises: (a) 30 mg / mL to 60 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 10 mM to 30 mM histidine-acetate buffer, (c) 0.4 mg / mL to 0.8 mg / mL of polysorbate 80, and (d) 240 mM to 480 mM sucrose; and the pH of the pharmaceutical composition is 5 to 7, 5.5 to 7, 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5. In some embodiments, the pH of the pharmaceutical composition is preferably 6 to 7.

[0139] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 15 μmol to 25 μmol of histidine-acetate buffer, (c) 0.4 mg to 0.8 mg of polysorbate 80, and (d) 240 μmol to 480 μmol of sucrose.

[0140] In some embodiments, the pharmaceutical composition comprises: (a) 55 mg / mL to 65 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 15 mM to 25 mM histidine-acetate buffer, (c) 0.4 mg / mL to 0.8 mg / mL of polysorbate 80, and (d) 240 mM to 480 mM sucrose; and the pH of the pharmaceutical composition is 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5. In some embodiments, the pH of the pharmaceutical composition is preferably 6 to 7.

[0141] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 15 μmol to 25 μmol of histidine-acetate buffer, (c) 0.3 mg to 0.5 mg of polysorbate 80, and (d) 210 μmol to 270 μmol of sucrose.

[0142] In some embodiments, the pharmaceutical composition comprises: (a) 55 mg / mL to 65 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 15 mM to 25 mM histidine-acetate buffer, (c) 0.3 mg / mL to 0.5 mg / mL of polysorbate 80, and (d) 210 mM to 270 mM sucrose; and the pH of the pharmaceutical composition is 5 to 7, 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5. In some embodiments, the pH of the pharmaceutical composition is preferably 6 to 7.

[0143] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 19 μmol to 21 μmol of histidine-acetate buffer, (c) 0.3 mg to 0.5 mg of polysorbate 80, and (d) 230 μmol to 250 μmol of sucrose.

[0144] In some embodiments, the pharmaceutical composition comprises: (a) 59 mg / mL to 61 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 19 mM to 21 mM histidine-acetate buffer, (c) 0.3 mg / mL to 0.5 mg / mL of polysorbate 80, and (d) 230 mM to 250 mM sucrose; and the pH of the pharmaceutical composition is 5.5 to 6.5, or 6 to 6.5.

[0145] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 10 μmol to 30 μmol of histidine-acetate buffer, (c) 0.6 mg to 1 mg of polysorbate 80, and (d) 450 μmol to 510 μmol of sucrose.

[0146] In some embodiments, the pharmaceutical composition comprises: (a) 55 mg / mL to 65 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 10 mM to 30 mM histidine-acetate buffer, (c) 0.6 mg / mL to 1 mg / mL of polysorbate 80, and (d) 450 mM to 510 mM sucrose; and the pH of the pharmaceutical composition is 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5. In some embodiments, the pH of the pharmaceutical composition is preferably 6 to 7.

[0147] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 15 μmol to 25 μmol of histidine-acetate buffer, (c) 0.7 mg to 0.9 mg of polysorbate 80, and (d) 450 μmol to 510 μmol of sucrose.

[0148] In some embodiments, the pharmaceutical composition comprises: (a) 55 mg / mL to 65 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 15 mM to 25 mM histidine-acetate buffer, (c) 0.7 mg / mL to 0.9 mg / mL of polysorbate 80, and (d) 450 mM to 510 mM sucrose; and the pH of the pharmaceutical composition is 5 to 7, 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5. In some embodiments, the pH of the pharmaceutical composition is preferably 6 to 7.

[0149] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 19 μmol to 21 μmol of histidine-acetate buffer, (c) 0.7 mg to 0.9 mg of polysorbate 80, and (d) 470 μmol to 490 μmol of sucrose.

[0150] In some embodiments, the pharmaceutical composition comprises: (a) 59 mg / mL to 61 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 19 mM to 21 mM histidine-acetate buffer, (c) 0.7 mg / mL to 0.9 mg / mL of polysorbate 80, and (d) 470 mM to 490 mM sucrose; and the pH of the pharmaceutical composition is 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5. In some embodiments, the pH of the pharmaceutical composition is preferably 6 to 7.

[0151] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 20 μmol of histidine-acetate buffer, (c) 0.4 mg to 0.8 mg of polysorbate 80, and (d) 240 μmol to 480 μmol of sucrose.

[0152] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 20 mM histidine-acetate buffer, (c) 0.4 mg / mL to 0.8 mg / mL of polysorbate 80, and (d) 240 mM to 480 mM sucrose; and the pH of the pharmaceutical composition is 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5. In some embodiments, the pH of the pharmaceutical composition is preferably 6 to 7.

[0153] In some embodiments, the pharmaceutical composition comprises: (a) per 60 mg of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 20 μmol of histidine-acetate buffer, (c) 0.4 mg of polysorbate 80, and (d) 240 μmol of sucrose. Furthermore, the pharmaceutical composition has a pH of 6.

[0154] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg / mL of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 20 mM histidine-acetate buffer, (c) 0.4 mg / mL of polysorbate 80, and (d) 240 mM sucrose; and the pH of the pharmaceutical composition is 6.

[0155] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 20 μmol of histidine-acetate buffer, (c) 0.8 mg of polysorbate 80, and (d) 480 μmol of sucrose. Furthermore, the pharmaceutical composition has a pH of 6.5.

[0156] In some embodiments, the pharmaceutical composition comprises: (a) 60 mg / mL of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 20 mM histidine-acetate buffer, (c) 0.8 mg / mL polysorbate 80, and (d) 480 mM sucrose; and the pH of the pharmaceutical composition is 6.5.

[0157] In a specific embodiment, the pharmaceutical composition comprises: (a) 60 mg of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 3.1 mg of histidine, (c) 0.4 mg of polysorbate 80, and (d) 240 μmol of sucrose. Furthermore, the pharmaceutical composition further comprises acetic acid to a pH of 6.

[0158] In a specific embodiment, the pharmaceutical composition comprises: (a) 60 mg of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 3.1 mg of histidine, (c) 0.8 mg of polysorbate 80, and (d) 480 μmol of sucrose. Furthermore, the pharmaceutical composition further comprises acetic acid to a pH of 6.5.

[0159] The present disclosure also provides a method for preparing the pharmaceutical composition, which is an aqueous formulation, comprising the step of contacting the anti-TIM-3 antibody or antigen-binding fragment thereof with a buffer, for example, displacing the anti-TIM-3 antibody or antigen-binding fragment thereof into the buffer, wherein the buffer may be a histidine buffer, a phosphate buffer, a succinate buffer, a citrate buffer, and / or an acetate buffer, preferably a histidine buffer, more preferably a histidine-acetate buffer; further, the method further comprises adding a surfactant and a stabilizer, wherein the surfactant and stabilizer are added in any order, wherein the stabilizer comprises mannitol, sorbitol, trehalose, sucrose, lysine, glycine, proline, arginine, or pharmaceutically acceptable salts thereof, and / or sodium chloride, preferably sucrose and / or sodium chloride; and the surfactant is polysorbate, poloxamer, or polyethylene glycol, preferably polysorbate, more preferably polysorbate 80. In some embodiments, the method includes contacting the anti-TIM-3 antibody or antigen-binding fragment thereof with a histidine-acetate buffer, for example, by displacing the anti-TIM-3 antibody or antigen-binding fragment thereof into the histidine-acetate buffer. Furthermore, the method includes adding polysorbate 80 and sucrose, with the order of addition of polysorbate 80 and sucrose not critical. It is understood that the preparation method of the present disclosure may also include sterilizing the prepared pharmaceutical composition and packaging it (e.g., into vials). The present disclosure does not specifically limit the sterilization method, and membrane filtration may be used, but is not limited to.

[0160] The present disclosure also provides a method for preparing the pharmaceutical composition, which is a lyophilized preparation (preferably a lyophilized injection), comprising the step of freeze-drying the pharmaceutical composition in the form of an aqueous preparation. In some embodiments, the freeze-drying is carried out according to methods well known in the art, including but not limited to the steps of pre-freezing, primary drying and secondary drying. In some embodiments, the lyophilized preparation of the present disclosure can also be dried by other methods known in the art, such as spray drying and bubble drying (bubbledrying). It will be understood by those skilled in the art that any method of removing water from the pharmaceutical composition of the present disclosure is applicable to the present disclosure. It will be understood by those skilled in the art that in the pharmaceutical field, lyophilized preparations typically have a residual moisture content of about 0.1 to 5% (w / v) and are present as a powder or a physically stable cake.

[0161] The present disclosure also provides a pharmaceutical composition in the form of a lyophilized preparation, which is prepared by the method for preparing a lyophilized preparation.

[0162] The pharmaceutical composition in the form of a freeze-dried preparation disclosed herein has a good appearance and appears as a loose block.

[0163] When the pharmaceutical composition of the present disclosure is an aqueous preparation (eg, aqueous injection), the pharmaceutical composition of the present disclosure may be preferably stored under refrigeration.

[0164] When the pharmaceutical composition of the present disclosure is a lyophilized preparation (eg, lyophilized injection), the pharmaceutical composition of the present disclosure may be preferably stored in a brown vial.

[0165] The present disclosure also provides an article of manufacture comprising a container containing the pharmaceutical composition. Furthermore, the article of manufacture includes instructions for using the pharmaceutical composition. In some embodiments, the pharmaceutical composition in the container can be provided in any desired volume, such as 4 mL and 10 mL. In some embodiments, the container contains 240 mg or 600 mg of an anti-TIM-3 antibody or antigen-binding fragment thereof.

[0166] In some embodiments, the pharmaceutical compositions of the present disclosure can be stably stored at 2-8°C for at least about 1 week, at least about 2 weeks, at least about 1 month, at least about 2 months, at least about 3 months, at least about 6 months, at least about 9 months, at least about 1 year, or at least about 2 years. In some embodiments, the pharmaceutical compositions of the present disclosure can be stably stored at 25°C for at least about 1 week, at least about 2 weeks, at least about 1 month, at least about 2 months, at least about 3 months, at least about 6 months, at least about 9 months, at least about 1 year, or at least about 2 years. In some embodiments, the pharmaceutical compositions of the present disclosure can be stably stored at 40°C for at least about 1 week, at least about 2 weeks, at least about 1 month, at least about 2 months, at least about 3 months, at least about 6 months, at least about 9 months, at least about 1 year, or at least about 2 years.

[0167] Any of the pharmaceutical compositions comprising an anti-TIM-3 antibody or antigen-binding fragment thereof provided herein can achieve a stable effect and is suitable for administration to a subject. Specifically, any of the pharmaceutical compositions comprising an anti-TIM-3 antibody or antigen-binding fragment thereof provided herein has one or more of the following properties: (i) inhibition of the formation of high-molecular-weight impurities; (ii) inhibition of the formation of charge variants; (iii) maintenance of the biological activity of the antibody; and (iv) thermal stability.

[0168] Uses of the pharmaceutical composition

[0169] The pharmaceutical compositions of the present disclosure can be administered according to known methods, such as by injection or infusion over a period of time in a suitable manner, for example, by injection or infusion by subcutaneous, intravenous, intraperitoneal, intramuscular, intraarterial, intralesional or intraarticular routes, topically, by inhalation or by sustained release or delayed release.

[0170] When the pharmaceutical composition of the present disclosure is a lyophilized injection, it can be diluted to a suitable concentration with a suitable diluent before administration to provide the best desired response (e.g., therapeutic response). When the pharmaceutical composition of the present disclosure is a lyophilized injection, the lyophilized injection can be dissolved with water (preferably water for injection) and infused intravenously after dilution with a suitable diluent.

[0171] When the pharmaceutical composition of the present disclosure is in the form of an aqueous injection, it can be diluted with a suitable diluent to a suitable concentration before administration to provide the best desired response (eg, therapeutic response).

[0172] In some embodiments, examples of diluents include, but are not limited to, Water for Injection, Sodium Chloride Injection, Ringer's Solution, or Dextrose Injection.

[0173] The present disclosure provides a method for treating a disease in a subject, the method comprising administering to the subject a pharmaceutical composition of the present disclosure. The present disclosure provides a method for treating a disease in a subject, the method comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition of the present disclosure.

[0174] The present disclosure provides a method for enhancing an immune response in a subject, the method comprising administering a pharmaceutical composition of the present disclosure to the subject. The present disclosure provides a method for enhancing an immune response in a subject, the method comprising administering a therapeutically effective amount of a pharmaceutical composition of the present disclosure to the subject. The present disclosure also provides use of the pharmaceutical composition of the present disclosure in the preparation of a medicament for enhancing an immune response in a subject.

[0175] The present disclosure provides a method for treating cancer in a subject, the method comprising administering to the subject a pharmaceutical composition of the present disclosure. The present disclosure provides a method for treating cancer in a subject, the method comprising administering to the subject a therapeutically effective amount of the pharmaceutical composition of the present disclosure. The present disclosure also provides use of the pharmaceutical composition of the present disclosure in the preparation of a medicament for treating cancer in a subject.

[0176] In some embodiments, the cancer is a recurrent, refractory, metastatic, and / or advanced cancer. In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a non-solid tumor. In some embodiments, the cancer is biliary tract cancer (including bile duct cancer, gallbladder cancer), carcinosarcoma, soft tissue sarcoma, esophageal cancer (also known as esophageal cancer), gastroesophageal junction cancer, breast cancer, gastric cancer, pancreatic cancer, head and neck cancer (including head and neck squamous cell carcinoma), colorectal cancer (including colon cancer and rectal cancer), kidney cancer (including renal cell carcinoma), cervical cancer, ovarian cancer, endometrial cancer, uterine carcinosarcoma, uterine cancer, fallopian tube cancer, melanoma, pharyngeal cancer, laryngeal cancer, nasopharyngeal cancer, oral cancer, skin cancer, lung cancer (including small cell lung cancer and non-small cell lung cancer), urethral cancer, prostate cancer, bladder cancer, anal cancer, cancer, testicular cancer, vaginal cancer, gastrointestinal stromal tumor, squamous cell carcinoma, peritoneal cancer, liver cancer (including hepatocellular carcinoma), uterine cancer, salivary gland cancer, vulvar cancer, thyroid cancer, penile cancer, Merkel cell carcinoma, leukemia (including acute lymphoblastic leukemia, acute myeloid leukemia, acute promyelocytic leukemia, chronic lymphocytic leukemia, chronic myelomonocytic leukemia, and chronic myelomonocytic leukemia), malignant lymphoma (including Hodgkin lymphoma and non-Hodgkin lymphoma), glioblastoma, bone cancer, plasma cell neoplasm, myeloma, myelodysplastic syndrome, myelofibrosis, or Paget's disease.

[0177] In some embodiments, the pharmaceutical compositions of the present disclosure may be administered with one or more other cancer therapeutics known in the art.

[0178] The present disclosure provides a method for treating an infectious disease in a subject, the method comprising administering a pharmaceutical composition of the present disclosure to the subject. The present disclosure provides a method for treating an infectious disease in a subject, the method comprising administering a therapeutically effective amount of a pharmaceutical composition of the present disclosure to the subject. The present disclosure also provides the use of the pharmaceutical composition of the present disclosure in the preparation of a medicament for treating an infectious disease in a subject. In some embodiments, the infectious disease is a disease caused by an organism such as bacteria, fungi, parasites, viruses or other pathogens, including but not limited to AIDS, viral hepatitis, viral pneumonia, hand, foot and mouth disease, meningitis, measles, etc.

[0179] In some embodiments, the pharmaceutical compositions of the present disclosure may be administered with one or more other infectious disease therapeutics known in the art.

[0180] The contents of all references, Genbank records, patents, and published patent applications cited in this disclosure are expressly incorporated herein by reference.

[0181] The present disclosure also provides the following specific implementation schemes, but the protection scope of the present disclosure is not limited thereto:

[0182] Embodiment 1. A pharmaceutical composition comprising: (a) an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) a buffer, (c) a surfactant, and (d) a stabilizer, wherein the anti-TIM-3 antibody or antigen-binding fragment thereof comprises: a HCDR1 of the amino acid sequence set forth in SEQ ID NO: 1, a HCDR2 of the amino acid sequence set forth in SEQ ID NO: 2, a HCDR3 of the amino acid sequence set forth in SEQ ID NO: 3, a LCDR1 of the amino acid sequence set forth in SEQ ID NO: 4, a LCDR2 of the amino acid sequence set forth in SEQ ID NO: 5, and a LCDR3 of the amino acid sequence set forth in SEQ ID NO: 6; or a HCDR1 of the amino acid sequence set forth in SEQ ID NO: 11, a HCDR2 of the amino acid sequence set forth in SEQ ID NO: 12, a HCDR3 of the amino acid sequence set forth in SEQ ID NO: 13, a LCDR1 of the amino acid sequence set forth in SEQ ID NO: 14, a LCDR2 of the amino acid sequence set forth in SEQ ID NO: 15, and a LCDR3 of the amino acid sequence set forth in SEQ ID NO: 16.

[0183] Embodiment 2. The pharmaceutical composition of embodiment 1, wherein the concentration of the anti-TIM-3 antibody or antigen-binding fragment thereof is 1 mg / mL to 120 mg / mL, 10 mg / mL to 100 mg / mL, 15 mg / mL to 90 mg / mL, 20 mg / mL to 80 mg / mL, 25 mg / mL to 70 mg / mL, 30 mg / mL to 60 mg / mL, 55 mg / mL to 65 mg / mL, or 59 mg / mL to 61 mg / mL.

[0184] Embodiment 3. The pharmaceutical composition of embodiment 1 or 2, wherein the concentration of the buffer is 1 mM to 100 mM, 2 mM to 80 mM, 5 mM to 60 mM, 8 mM to 40 mM, 10 mM to 30 mM, 10 mM to 20 mM, 20 mM to 30 mM, 15 mM to 25 mM, or 19 mM to 21 mM.

[0185] Embodiment 4. The pharmaceutical composition according to any one of Embodiments 1-3, wherein the buffer is a histidine buffer, a phosphate buffer, a succinate buffer, a citrate buffer and / or an acetate buffer; preferably, the buffer is a histidine buffer; more preferably, the buffer is a histidine-acetate buffer.

[0186] Embodiment 5. The pharmaceutical composition of any one of Embodiments 1-4, wherein the concentration of the histidine buffer is 1 mM to 100 mM, 2 mM to 80 mM, 5 mM to 60 mM, 8 mM to 40 mM, 10 mM to 30 mM, 10 mM to 20 mM, 20 mM to 30 mM, 15 mM to 25 mM, or 19 mM to 21 mM.

[0187] Embodiment 6. The pharmaceutical composition of any one of Embodiments 1-5, wherein the concentration of the surfactant is 0.01 mg / mL to 2 mg / mL, 0.05 mg / mL to 1.5 mg / mL, 0.1 mg / mL to 1 mg / mL, 0.2 mg / mL to 0.8 mg / mL, 0.4 mg / mL to 1.2 mg / mL, 0.4 mg / mL to 0.8 mg / mL, 0.3 mg / mL to 0.5 mg / mL, 0.6 mg / mL to 1 mg / mL, or 0.7 mg / mL to 0.9 mg / mL.

[0188] Embodiment 7. The pharmaceutical composition according to any one of Embodiments 1-6, wherein the surfactant is polysorbate; preferably, the surfactant is polysorbate 80 or polysorbate 20; more preferably, the surfactant is polysorbate 80.

[0189] Embodiment 8. The pharmaceutical composition of any one of Embodiments 1-7, wherein the concentration of polysorbate 80 is 0.01 mg / mL to 2 mg / mL, 0.05 mg / mL to 1.5 mg / mL, 0.1 mg / mL to 1 mg / mL, 0.2 mg / mL to 0.8 mg / mL, 0.4 mg / mL to 1.2 mg / mL, 0.4 mg / mL to 0.8 mg / mL, 0.3 mg / mL to 0.5 mg / mL, 0.6 mg / mL to 1 mg / mL, or 0.7 mg / mL to 0.9 mg / mL.

[0190] Embodiment 9. The pharmaceutical composition according to any one of Embodiments 1-8, wherein the stabilizer comprises mannitol, sorbitol, trehalose, sucrose, lysine, glycine, proline, arginine or a pharmaceutically acceptable salt thereof and / or sodium chloride.

[0191] Embodiment 10. The pharmaceutical composition of embodiment 9, wherein the stabilizer comprises sucrose.

[0192] Embodiment 11. The pharmaceutical composition of embodiment 9 or 10, wherein the concentration of sucrose is 30 mM to 960 mM, 60 mM to 720 mM, 120 mM to 480 mM, 240 mM to 480 mM, 210 mM to 270 mM, 230 mM to 250 mM, 450 mM to 510 mM, or 470 mM to 490 mM.

[0193] Embodiment 12. The pharmaceutical composition according to embodiment 10 or 11, wherein the stabilizer further comprises sodium chloride.

[0194] Embodiment 13. The pharmaceutical composition of embodiment 12, wherein the concentration of sodium chloride is 5 mM to 200 mM, 10 mM to 100 mM, 20 mM to 100 mM, or 50 mM to 100 mM.

[0195] Embodiment 14. The pharmaceutical composition of any one of Embodiments 1-13, wherein the pH of the pharmaceutical composition is 4 to 7, 5 to 7, 5.5 to 7, 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5.

[0196] Embodiment 15. A pharmaceutical composition according to any one of Embodiments 1-14, wherein the pharmaceutical composition comprises: (a) 1 mg / mL to 120 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 1 mM to 100 mM histidine buffer, (c) 0.01 mg / mL to 2 mg / mL of polysorbate 80, and (d) 30 mM to 960 mM sucrose; and the pH of the pharmaceutical composition is 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5.

[0197] Embodiment 16. A pharmaceutical composition according to any one of Embodiments 1-15, wherein the pharmaceutical composition comprises: (a) 10 mg / mL to 100 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 2 mM to 80 mM histidine buffer, (c) 0.05 mg / mL to 1.5 mg / mL polysorbate 80, and (d) 60 mM to 720 mM sucrose; and the pH of the pharmaceutical composition is 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5.

[0198] Embodiment 17. A pharmaceutical composition according to any one of Embodiments 1-16, wherein the pharmaceutical composition comprises: (a) 15 mg / mL to 90 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 5 mM to 60 mM histidine buffer, (c) 0.1 mg / mL to 1 mg / mL polysorbate 80, and (d) 120 mM to 480 mM sucrose; and the pH of the pharmaceutical composition is 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5.

[0199] Embodiment 18. A pharmaceutical composition according to any one of Embodiments 1-17, wherein the pharmaceutical composition comprises: (a) 20 mg / mL to 80 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 8 mM to 40 mM histidine buffer, (c) 0.2 mg / mL to 0.8 mg / mL polysorbate 80, and (d) 120 mM to 480 mM sucrose; and the pH of the pharmaceutical composition is 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5.

[0200] Embodiment 19. A pharmaceutical composition according to any one of Embodiments 1-18, wherein the pharmaceutical composition comprises: (a) 30 mg / mL to 60 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 10 mM to 30 mM histidine buffer, (c) 0.2 mg / mL to 0.8 mg / mL of polysorbate 80, and (d) 120 mM to 480 mM sucrose; and the pH of the pharmaceutical composition is 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5; preferably, the pH of the pharmaceutical composition is 6 to 7.

[0201] Embodiment 20. A pharmaceutical composition according to any one of Embodiments 1-19, wherein the pharmaceutical composition comprises: (a) 30 mg / mL to 60 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 10 mM to 30 mM histidine buffer, (c) 0.4 mg / mL to 0.8 mg / mL of polysorbate 80, and (d) 240 mM to 480 mM sucrose; and the pH of the pharmaceutical composition is 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5; preferably, the pH of the pharmaceutical composition is 6 to 7.

[0202] Embodiment 21. A pharmaceutical composition according to any one of Embodiments 1-20, wherein the pharmaceutical composition comprises: (a) 55 mg / mL to 65 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 15 mM to 25 mM histidine buffer, (c) 0.4 mg / mL to 0.8 mg / mL of polysorbate 80, and (d) 240 mM to 480 mM sucrose; and the pH of the pharmaceutical composition is 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5; preferably, the pH of the pharmaceutical composition is 6 to 7.

[0203] Embodiment 22. A pharmaceutical composition according to any one of Embodiments 1-21, wherein the pharmaceutical composition comprises: (a) 60 mg / mL of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 20 mM histidine buffer, (c) 0.4 mg / mL to 0.8 mg / mL of polysorbate 80, and (d) 240 mM to 480 mM sucrose; and the pH of the pharmaceutical composition is 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5; preferably, the pH of the pharmaceutical composition is 6 to 7.

[0204] Embodiment 23. A pharmaceutical composition according to any one of Embodiments 1-22, wherein the pharmaceutical composition comprises: (a) 55 mg / mL to 65 mg / mL of an anti-TIM-3 antibody or antigen-binding fragment thereof, (b) 15 mM to 25 mM histidine buffer, (c) 0.7 mg / mL to 0.9 mg / mL of polysorbate 80, and (d) 450 mM to 510 mM sucrose; and the pH of the pharmaceutical composition is 5 to 7, 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5; preferably, the pH of the pharmaceutical composition is 6 to 7.

[0205] Embodiment 24. A pharmaceutical composition according to any one of Embodiments 1-23, wherein the pharmaceutical composition comprises: (a) 60 mg / mL of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 20 mM histidine buffer, (c) 0.4 mg / mL of polysorbate 80, and (d) 240 mM sucrose; and the pH of the pharmaceutical composition is 5.5 to 6.5, preferably 6.

[0206] Embodiment 25. A pharmaceutical composition according to any one of Embodiments 1-24, wherein the pharmaceutical composition comprises: (a) 60 mg / mL of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 20 mM histidine buffer, (c) 0.8 mg / mL of polysorbate 80, and (d) 480 mM sucrose; and the pH of the pharmaceutical composition is 6 to 7, preferably 6.5.

[0207] Embodiment 26. The pharmaceutical composition of any one of Embodiments 1-25, wherein the anti-TIM-3 antibody or antigen-binding fragment thereof comprises: a heavy chain variable region having an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 7, and a light chain variable region having an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 8; or a heavy chain variable region having an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 17, and a light chain variable region having an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 18.

[0208] Embodiment 27. The pharmaceutical composition of any one of Embodiments 1-26, wherein the anti-TIM-3 antibody or antigen-binding fragment thereof comprises: a heavy chain having an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 9, and a light chain having an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 10; or a heavy chain having an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 19, and a light chain having an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 20.

[0209] Embodiment 28. The pharmaceutical composition according to any one of Embodiments 1-27, wherein the pharmaceutical composition is an injection; preferably, the pharmaceutical composition is an aqueous injection.

[0210] Embodiment 29. An article of manufacture comprising a container containing the pharmaceutical composition of any one of embodiments 1-28; optionally, the article of manufacture further comprises instructions for using the pharmaceutical composition.

[0211] Embodiment 30. Use of the pharmaceutical composition of any one of Embodiments 1-28 or the preparation of Embodiment 29 in the preparation of a medicament for treating cancer.

[0212] Embodiment 31. The use according to embodiment 30, wherein the cancer is recurrent, refractory, metastatic and / or advanced cancer.

[0213] Embodiment 32. The use according to embodiment 30 or 31, wherein the cancer is biliary tract cancer, carcinosarcoma, soft tissue sarcoma, esophageal cancer, gastroesophageal junction cancer, breast cancer, gastric cancer, gallbladder cancer, pancreatic cancer, head and neck cancer, colorectal cancer, kidney cancer, cervical cancer, ovarian cancer, endometrial cancer, uterine carcinosarcoma, uterine cancer, fallopian tube cancer, melanoma, pharyngeal cancer, laryngeal cancer, nasopharyngeal cancer, oral cancer, skin cancer, lung cancer, urethra cancer, prostate cancer, bladder cancer, anal cancer, testicular cancer, vaginal cancer, gastrointestinal stromal tumor, squamous cell carcinoma, peritoneal cancer, liver cancer, uterine cancer, salivary gland cancer, vulvar cancer, thyroid cancer, penile cancer, Merkel cell carcinoma, leukemia, malignant lymphoma, glioblastoma, bone cancer, plasmacytoma, myeloma, myelodysplastic syndrome, myelofibrosis, or Paget's disease. BRIEF DESCRIPTION OF THE DRAWINGS

[0214] FIG1 shows an ELISA assay for the binding of h50B5 to CHO-S-HuTIM-3 cells that highly express human TIM-3;

[0215] FIG2 shows the promoting effect of h50B5 on IFN-γ secretion in MLR;

[0216] FIG3 shows the blood concentration curve of h50B5 after a single intravenous injection of h50B5 in a subcutaneous transplanted tumor model of lung cancer HCC827 human immune reconstituted mice (PBMC-NCG).

[0217] Definition and Explanation

[0218] Unless otherwise indicated, the following terms used in this disclosure have the following meanings. A particular term should not be construed as undefined or unclear unless specifically defined, but rather should be understood according to its ordinary meaning in the art. When a trade name appears herein, it is intended to refer to the corresponding commercial product or its active ingredient.

[0219] The term "antibody" is used in the broadest sense to cover natural antibodies and artificial antibodies of various structures, including but not limited to monoclonal antibodies, polyclonal antibodies, monospecific antibodies and multispecific antibodies (e.g., bispecific antibodies, trispecific antibodies), and various antibody structures of single-chain antibodies, so long as they exhibit the desired antigen-binding activity.

[0220] The term "antigen-binding fragment" refers to one or more fragments of an antibody that retain the function of specifically binding to an antigen. Examples encompassed by the term "antigen-binding fragment" include: (i) Fab fragment: a monovalent fragment consisting of the VL, VH, CL, and CH1 domains; (ii) F(ab')2 fragment, a bivalent fragment comprising two Fab fragments connected by a disulfide bridge at the hinge region; (iii) Fd fragment consisting of the VH and CH1 domains; (iv) Fv fragment consisting of the VL and VH domains of a single antibody arm; (v) dAb fragment consisting of the VH domain (see Ward et al., Nature. 341: 544-546 (1989)); (vi) isolated complementarity-determining regions (CDRs); (vii) nanobodies; and single-chain Fv (scFv).

[0221] The term "variable domain" or "variable region" refers to the domain of an antibody that is involved in binding the antibody to the antigen. For example, a natural four-chain antibody (e.g., derived from humans, mice, etc.) has a heavy chain variable region (also referred to as a heavy chain variable domain, VH or VH domain) and a light chain variable region (also referred to as a light chain variable domain, VL or VL domain). In most cases, each variable domain of a natural antibody is essentially composed of four "framework regions (FR)" and three "complementarity determining regions (CDRs)." The four framework regions are respectively referred to as framework region 1 (or FR1), framework region 2 (or FR2), framework region 3 (or FR3), and framework region 4 (or FR4); the framework regions are separated by three complementary determining regions, which are respectively referred to in the art and hereinafter as complementary determining region 1 (or CDR1), complementary determining region 2 (or CDR2), and complementary determining region 3 (or CDR3). Therefore, the general structure of a variable domain can be represented as follows: FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4. The variable domain confers specificity to the antibody for the antigen by providing the antigen-binding site.

[0222] The term "complementarity determining region" (CDR), also known as "hypervariable region" (HVR). Natural four-chain antibodies usually contain six CDRs, three in the heavy chain variable region, namely heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3); and three in the light chain variable region, namely light chain CDR1 (LCDR1), light chain CDR2 (LCDR2) and light chain CDR3 (LCDR3).

[0223] The term "buffer" refers to a pharmaceutically acceptable buffer and refers to a substance that helps maintain the pH of a pharmaceutical composition within a desired pH range. Buffers suitable for use in the pharmaceutical compositions of the present disclosure include, but are not limited to, histidine buffers, phosphate buffers, succinate buffers, citrate buffers, and / or acetate buffers. Furthermore, mixtures of two or more buffers may also be used. Preferably, the buffer suitable for use in the present disclosure is a histidine buffer.

[0224] "Histidine buffer" is a buffer comprising histidine ions. Examples of histidine buffers include histidine-histidine hydrochloride buffer, histidine-hydrochloride buffer, histidine-acetate buffer, histidine-phosphate buffer, and the like. Histidine-hydrochloride buffer can be prepared using histidine (e.g., L-histidine) and further adjusted to pH with hydrochloric acid. Histidine-acetate buffer can be prepared using histidine (e.g., L-histidine) and further adjusted to pH with acetic acid. Histidine-phosphate buffer can be prepared using histidine (e.g., L-histidine) and further adjusted to pH with phosphoric acid. Histidine-histidine hydrochloride buffer can be prepared using histidine (e.g., L-histidine) and histidine hydrochloride or a hydrate thereof (e.g., histidine hydrochloride monohydrate) or can be prepared using histidine hydrochloride or a hydrate thereof (e.g., histidine hydrochloride monohydrate).

[0225] "Phosphate buffer" is a buffer containing phosphate ions. Examples of phosphate buffers include sodium phosphate buffer, potassium phosphate buffer, etc., and a preferred phosphate buffer is sodium phosphate buffer. Sodium phosphate buffer can be prepared using disodium hydrogen phosphate or a hydrate thereof (e.g., disodium hydrogen phosphate dodecahydrate) and sodium dihydrogen phosphate or a hydrate thereof (e.g., sodium dihydrogen phosphate dihydrate). Potassium phosphate buffer can be prepared using dipotassium hydrogen phosphate or a hydrate thereof and potassium dihydrogen phosphate or a hydrate thereof.

[0226] A "succinate buffer" is a buffer containing succinate ions. Examples of succinate buffers include sodium succinate buffer, potassium succinate buffer, and the like. Sodium succinate buffer can be prepared using succinic acid and sodium succinate or a hydrate thereof. Potassium succinate buffer can be prepared using succinic acid and potassium succinate or a hydrate thereof.

[0227] "Citrate buffer" is a buffer containing citrate ions. Examples of citrate buffers include sodium citrate buffer, potassium citrate buffer, etc., and a preferred citrate buffer is sodium citrate buffer. Sodium citrate buffer can be prepared using citric acid and sodium citrate or a hydrate thereof (e.g., trisodium citrate dihydrate). Potassium citrate buffer can be prepared using citric acid and potassium citrate or a hydrate thereof (e.g., tripotassium citrate monohydrate).

[0228] An "acetate buffer" is a buffer containing acetate ions. Examples of acetate buffers include potassium acetate buffer, ammonium acetate buffer, sodium acetate buffer, and the like. Potassium acetate buffer can be prepared using acetic acid and potassium acetate or a hydrate thereof. Ammonium acetate buffer can be prepared using acetic acid and ammonium acetate or a hydrate thereof. Sodium acetate buffer can be prepared using acetic acid and sodium acetate or a hydrate thereof (e.g., anhydrous sodium acetate or sodium acetate trihydrate).

[0229] "Surfactant" refers to a substance having a hydrophilic group and a hydrophobic group and used as one of the components of a pharmaceutical composition. The surfactant in the present disclosure is preferably polysorbate, such as polysorbate 80 (i.e., Tween 80) and polysorbate 20 (i.e., Tween 20).

[0230] The term "stabilizer" refers to a substance that helps maintain the structural integrity of the active ingredient (e.g., antibody) in a pharmaceutical composition, particularly during freezing, lyophilization, storage, and / or transportation (especially when exposed to stress). In this context, a stabilizer may also function as a viscosity reducer, an ionic strength modifier, an antioxidant, and / or an isotonicity agent.

[0231] The term "antioxidant" refers to a substance that can prevent, slow down or reduce the oxidation of an active ingredient (eg, an antibody) in a pharmaceutical composition.

[0232] The term "isotonic agent" refers to a substance that helps maintain the osmotic pressure of a pharmaceutical composition at or near physiological osmotic pressure.

[0233] The term "excipient" refers to any ingredient other than the active ingredient. The choice of excipient will depend, to a large extent, on factors such as the particular mode of administration, the effect of the excipient on solubility and stability, and the nature of the dosage form. As used herein, "pharmaceutically acceptable excipients" include, but are not limited to, excipients, diluents, fillers, binders, disintegrants, solubilizers, stabilizers, colorants, flavorings, surfactants, emulsifiers, buffers, or encapsulating materials.

[0234] The term "pharmaceutically acceptable" refers to those compounds, materials, compositions and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0235] In this disclosure, "pharmaceutical composition" and "formulation" are used interchangeably.

[0236] In some embodiments, the pharmaceutical composition is an injection. The term "injection" refers to a pharmaceutical composition that is administered to biological tissue (e.g., intravenously, intradermally, subcutaneously, intramuscularly, and intraperitoneally) with a needle. In the injection of the present disclosure, not only liquid injections are included, but also solid injections that are dissolved in a solvent when used. The solvent can be water, or a solvent other than water (e.g., sodium chloride injection, Ringer's solution, or glucose injection), or a mixed solvent of water and a solvent other than water.

[0237] In some embodiments, the pharmaceutical composition is an aqueous preparation; preferably, the aqueous preparation is an aqueous injection. The term "aqueous preparation" refers to a pharmaceutical composition that is in liquid form and whose solvent is water (when the aqueous preparation is an aqueous injection, it is preferably water for injection), and the aqueous preparation includes but is not limited to an unlyophilized aqueous preparation or an aqueous preparation reconstituted from a lyophilized powder.

[0238] In other embodiments, the pharmaceutical composition is a lyophilized preparation; preferably, the lyophilized preparation is a lyophilized injection. The term "lyophilized preparation" is a solid preparation that can be used by dissolving in water (preferably water for injection when the lyophilized preparation is a lyophilized injection) at the time of use, which is obtained by lyophilizing an aqueous solution containing a predetermined amount of the pharmaceutical component.

[0239] The term "primary drying" is one of the production processes of freeze-dried preparations and refers to a process for sublimating free water in the frozen body. The term "secondary drying" is one of the production processes of freeze-dried preparations and refers to a process for sublimating water bound to solutes (bound water).

[0240] A "stable" pharmaceutical composition refers to a pharmaceutical composition in which the active substance (e.g., an anti-TIM-3 antibody or antigen-binding fragment thereof) substantially maintains its physical stability and / or chemical stability and / or biological activity when stored therein. Preferably, the pharmaceutical composition substantially maintains its physical stability, chemical stability, and biological activity when stored therein.

[0241] “High molecular weight impurities” refers to a general term for impurities with a molecular weight greater than that of the target active ingredient (e.g., an anti-TIM-3 antibody or an antigen-binding fragment thereof).

[0242] The term "treating" means administering a pharmaceutical composition or preparation of the present disclosure to prevent, ameliorate, or eliminate a disease or one or more symptoms associated with a particular disease, and includes but is not limited to:

[0243] (i) preventing a disease or disease state from occurring in a mammal, particularly where such mammal is susceptible to the disease state but has not yet been diagnosed as having the disease state;

[0244] (ii) inhibiting the disease or disease state, i.e., curbing its development;

[0245] (iii) alleviate the disease or condition, even if the disease or condition regresses;

[0246] (iv) reducing any direct or indirect pathological consequences of the disease or disease state.

[0247] The term "therapeutically effective amount" means an amount of a pharmaceutical composition or product of the present disclosure that (i) treats or prevents a particular disease, condition, or disorder, (ii) alleviates, ameliorates, or eliminates one or more symptoms of a particular disease, condition, or disorder, or (iii) prevents or delays the onset of one or more symptoms of a particular disease, condition, or disorder. The amount of a pharmaceutical composition or product of the present disclosure that constitutes a "therapeutically effective amount" may vary depending on factors such as the ability of the pharmaceutical composition to elicit a desired response in an individual, the disease state and its severity, the mode of administration, and the age, sex, and weight of the mammal to be treated. A therapeutically effective amount may also be routinely determined by one skilled in the art based on their own knowledge and the present disclosure.

[0248] The term "EC 50 The half-maximal effect concentration (HEC), also known as the half-maximal effect concentration, is the antibody concentration that induces a response halfway between baseline and maximum after a specific exposure time.

[0249] The term "subject" includes any human or non-human animal. The term "non-human animal" includes all vertebrates, for example, mammals and non-mammals, such as non-human primates, sheep, dogs, cats, horses, cows, chickens, amphibians, reptiles, etc. Preferably, the subject of the present disclosure is a human. Unless otherwise indicated, the terms "patient" or "subject" can be used interchangeably. "Subjects in need thereof" include those who already have a disease or condition, those who are at risk of developing a disease or condition, and those who may develop a disease or condition and for which the purpose is to prevent, delay or attenuate the disease or condition.

[0250] The term "identity" is also known as consistency. The "percentage (%) identity" of an amino acid sequence refers to the percentage of amino acid residues in the sequence to be aligned that are identical to the amino acid residues in the specific amino acid sequence shown in this article, after comparing the sequence to be aligned and, if necessary, introducing gaps to achieve maximum sequence identity, and not considering any conservative substitutions as part of sequence identity. The alignment of amino acid sequences for identity can be performed in a variety of ways within the scope of the art, such as BLAST, BLAST-2, ALIGN or Megalign (DNASTAR) software. Those skilled in the art can determine the appropriate parameters for aligning sequences, including any algorithm needed to obtain maximum alignment over the full length of the comparison sequence.

[0251] The articles "a" and "an" are used herein to refer to one or more than one (i.e., at least one) of the grammatical object of the article. For example, "a pharmaceutical composition" refers to one pharmaceutical composition or more than one pharmaceutical composition.

[0252] The terms "comprise," "comprises," or "comprising" and their equivalents (for example, contain, contains, containing, include, includes, and including) should be understood as "including but not limited to," meaning that in addition to the listed elements, components, and steps, other unspecified elements, components, and steps may also be included.

[0253] "About" means within the acceptable error range for a particular value as determined by one of ordinary skill in the art, which depends in part on how the value is measured or determined, i.e., on the limitations of the measurement system. For example, "about" can mean within one or more standard deviations, as practiced in the art. Alternatively, "about" can mean a range of up to ±5%, for example, within ±2%, ±1%, or ±0.5% of the specific value range given. Whenever specific values ​​or parameters are given herein or in the claims, they are assumed to be modified by "about" unless otherwise indicated.

[0254] As described herein, any concentration range, percentage range, ratio range, or integer range should be understood to include the value of any integer within the range, and where appropriate, fractions thereof (such as tenths and hundredths of integers), unless otherwise indicated.

[0255] Herein, singular terms encompass plural referents and vice versa unless the context clearly dictates otherwise. DETAILED DESCRIPTION

[0256] For the sake of clarity, the present disclosure is further illustrated by examples, but the examples do not limit the scope of the present disclosure. Those skilled in the art will readily recognize a variety of non-critical parameters that can be changed or modified to produce substantially similar results.

[0257] The heavy chain amino acid sequence of the anti-TIM-3 antibody h50B5 in the Examples is shown in SEQ ID NO: 9, and the light chain amino acid sequence is shown in SEQ ID NO: 10. Examples 1 and 2 used the anti-TIM-3 antibody TSR022 as a benchmark, whose heavy chain amino acid sequence is shown in SEQ ID NO: 22, and light chain amino acid sequence is shown in SEQ ID NO: 23.

[0258] Example 1 Binding of Anti-TIM-3 Antibodies to Human TIM-3

[0259] Human TIM-3 (R&D) recombinant protein was coated in an ELISA 96-well plate (Greiner), and different concentrations of anti-TIM-3 antibody h50B5 (100,000 ng / mL, 30,000 ng / mL, 10,000 ng / mL, 3,000 ng / mL, 1,000 ng / mL, 300 ng / mL, 100 ng / mL, 30 ng / mL, 10 ng / mL, 30 ng / mL, 10 ng / mL, 3 ng / mL) were added and incubated at 37°C for 1 hour. After washing the 96-well plate with PBS (pH 7.4), Goat anti-human Ig kappachain-HRP conjugate (Millipore) was added and incubated at 37°C for 1 hour. HRP substrate o-phenylenediamine dihydrochloride (OPD) was added, and the reaction was terminated with H2SO4. The OD value was measured at a wavelength of 450 nm using a full-function microplate reader SynergyH4 (BioTek). The binding EC value was calculated based on the OD value. 50 .

[0260] The results showed that the anti-TIM-3 antibody h50B5 had a strong binding affinity with human TIM-3 protein and 50 was 291.0±14.1ng / mL, and the maximum binding E max (OD 450 ) is 0.52±0.03.

[0261] The human TIM-3 gene was transfected into CHO-S cells and cultured in Dynamis AGT culture medium (containing 4 mM glutamate and 3 μg / mL Puromycin) at 37° C. in an incubator containing 5% CO 2 in air to obtain CHO-S-HuTIM-3 cells. Different concentrations of h50B5 or TSR022 (300000ng / mL, 100000ng / mL, 30000ng / mL, 10000ng / mL, 3000ng / mL, 1000ng / mL, 300ng / mL, 100ng / mL, 30ng / mL, 10ng / mL) were incubated with CHO-S-HuTIM-3 cells at 4°C for 1 hour, and then Goatanti-humanIgkappachain-HRPconjugate was added and incubated at 37°C for 1 hour. Finally, HRP substrate OPD was added, and the reaction was terminated with H2SO4. The OD value was measured at a wavelength of 450nm using a full-function microplate reader SynergyH4. The binding EC was calculated based on the OD value. 50 .

[0262] As shown in Figure 1, h50B5 binds strongly to CHO-S-HuTIM-3 cells that highly express human TIM-3 and binds to EC 50 and the maximum combined E max (OD 450 ) were 1456.0±338.0 ng / mL and 0.21±0.03; the EC values ​​of TSR022 binding to CHO-S-HuTIM-3 cells were 50 and the maximum combined E max (OD 450 ) were 6815.5±166.2 ng / mL and 0.26±0.05, respectively. The binding activity of h50B5 was stronger than that of TSR022.

[0263] Example 2 Effect of anti-TIM-3 antibodies on IFN-γ secretion in mixed lymphocyte reaction (MLR)

[0264] CD14 was isolated from human PBMC (prepared in-house) using a CD14 magnetic bead isolation kit (Miltenyi Biotec). + After the mononuclear cells were cultured, they were induced with GM-CSF (Xiamen Tebao Bioengineering Co., Ltd.) for 7 days to differentiate into mature dendritic cells (DCs); CD4 + CD4 T cell magnetic bead separation kit (Miltenyi Biotec) was used to separate CD4 T cells from human PBMCs. + T cells, then CD4 + T cells were mixed with mature DCs and incubated for 5 days with varying concentrations of h50B5 or TSR022 (100 μg / mL, 10 μg / mL, and 1 μg / mL). A blank control (no antibody) was also established. The cells were then aspirated and the IFN-γ content was measured using an IFN-γ assay kit (Cisbio).

[0265] As shown in Figure 2, after DC cells were induced to mature, they were incubated with allogeneic CD4 + T cells interact with each other, and h50B5 can significantly promote the secretion of IFN-γ in MLR, with activity superior to TSR022.

[0266] Example 3 Effect of anti-TIM-3 antibodies on tumor-infiltrating lymphocytes in subcutaneous transplanted tumors of lung cancer HCC827 human immune reconstitution mice (PBMC-NCG)

[0267] NCG mice were subcutaneously inoculated with HCC827 cells (Cell Bank of the Chinese Academy of Sciences), 8×10 6 cells / mouse. When the tumor grows to 100-150 mm 3 , intravenous injection of human PBMC (Shanghai Ausells Biotechnology (Shanghai) Co., Ltd., catalog number PB004F), 6×106 cells / mouse. On the second day, h50B5 or hIgG4 (dosage of 10 mg / kg) was injected intravenously. Blood was collected from the mouse orbits at different time points (5 min, 4 h, 24 h, 48 h, 72 h, 96 h, 168 h) after administration, and serum was separated to detect drug concentration. Orbital blood was collected from 6 mice at each time point. 72 hours after administration, groups 1 and 3 were dissected; 168 hours after administration, groups 2 and 4 were dissected. Tumor tissues were taken from each group, tumor infiltrating lymphocytes (TILs) were separated, and human CD45 was analyzed by flow cytometry. + 、CD3 + 、CD4 + 、CD8 + Cell ratio. The experimental design is shown in Table 2-1. min represents minutes, and h represents hours.

[0268] Table 2-1. Experimental design

[0269] As shown in Table 2-2, 168 hours after tumor-bearing mice were intravenously injected with h50B5 (10 mg / kg), CD8 + / CD3 + The proportion of T cells was significantly higher in the hIgG4 group (46.9±6.9% vs 31.1±6.9%, P<0.01), and CD8 + / CD4 + The proportion of T cells was significantly higher than that in the hIgG4 group (112.7±32.1% vs 60.2±11.3%, P<0.01), indicating that h50B5 can promote the proliferation of CD8 T cells in HCC827 subcutaneous transplanted tumor tissues. + T cell infiltration, significantly increased CD3 + CD8 T cells + As shown in Figure 3, after a single intravenous injection of 50B5 (10 mg / kg) into tumor-bearing mice, the serum concentration of h50B5 gradually decreased over time. 168 hours after administration, the serum concentration of h50B5 still reached 29.4±2.3 μg / mL. The Cmax was 150.3 μg / mL, and the AUC 0-168h 8142.0 h·μg·mL -1 .

[0270] Table 2-2. Effects of a single intravenous injection of h50B5 or hIgG4 on the proportions of various T cell subsets in TILs of subcutaneous transplanted tumors in human immune reconstituted mice (PBMC-NCG) with lung cancer HCC827

[0271] Mean ± SD; **P < 0.01 vs hIgG4 group.

[0272] Example 4 Detection Method of Pharmaceutical Composition and Preparation of Buffer

[0273] The detection methods used in Examples 5-7 are as follows:

[0274] Size exclusion chromatography (SEC-HPLC): used to determine the purity of the antibody in the sample, using Thermo Vanquish F high performance liquid chromatography, TSKgel G3000 SW XL (5μm) as the chromatographic column, TSKgel guardcolumn G3000 SW XL A 7 μm precolumn was used, and elution was performed using a 50 mM phosphate-300 mM sodium chloride solution (pH 7.5) as the mobile phase. The detection wavelength was 280 nm. The peak area percentages of high molecular weight impurities and immunoglobulin monomers were calculated by the area normalization method.

[0275] Weak cation chromatography (CEX-HPLC): used to determine the charge variants of antibodies in samples, using a Thermo U3000 high-performance liquid chromatograph on a Sepax Antibodix WCX-NP5 (4.6 mm × 250 mm) column, with 10 mM sodium dihydrogen phosphate (pH 7.0) as mobile phase A and 10 mM sodium dihydrogen phosphate-300 mM sodium chloride (pH 5.5) as mobile phase B. Detection was at a wavelength of 280 nm. Peak area percentages of the main peak, acidic region, and basic region were calculated using the area normalization method.

[0276] Unfolding temperature (Tm): The unfolding temperature (Tm) of the samples was analyzed using MicroCal VP-Capillary DSC (Malvern). The samples were diluted to a concentration of 1 mg / mL with the following program: scan start temperature at 20°C, scan end temperature at 110°C, and heating rate at 60°C / h.

[0277] Aggregation temperature (Tagg): The aggregation temperature (Tagg) of the sample was analyzed using Dynaproplate Reader III (Wyatt). The sample was added to a 384-well sample plate, sealed with a sealing film, and then the 384-well sample plate was centrifuged to remove bubbles. The test was then performed using the following procedure: the temperature was raised from 35°C to 85°C, and the scanning time was 5 seconds.

[0278] Capillary electrophoresis with reduced sodium dodecyl sulfate (CE-SDS) was performed using a Beckman Coulter PA 800Plus Biopharmaceutical Analysis System with uncoated fused-silica capillaries (50 μm inner diameter, 31 cm total length, and 21 cm effective length) and a PDA detector at a wavelength of 220 nm. The corrected peak area percentages for the light chain, heavy chain, and aglycosylated heavy chain were calculated using the area normalization method.

[0279] Non-reduced CE-SDS analysis was performed using a Beckman Coulter PA 800Plus Biopharmaceutical Analyzer system, using uncoated fused silica capillaries with an inner diameter of 50 μm, a total length of 31 cm, and an effective length of 21 cm. Detection was performed using a PDA detector at a wavelength of 220 nm. The corrected peak area percentage of the main peak was calculated using the area normalization method.

[0280] Biological activity was assessed using an enzyme-linked immunosorbent assay (ELISA): 2 μg / mL TIM-3 protein (ACRO, TM3-H5229) was coated onto a 96-well plate at 100 μL / well and incubated overnight at 2-8°C. After washing the 96-well plate, 250 μL of blocking buffer (3% BSA in PBS) was added to each well and incubated at 25°C for 2 hours. After washing the 96-well plate, serial dilutions of the reference and test samples (initial concentration: 1000 ng / mL, 4-fold serial dilutions, 7 dilutions total) were added, along with a blank control, and incubated at 25°C for 2 hours. After washing the 96-well plate, 100 μL / well of HRP-conjugated goat anti-human antibody (PekinElmer, NEF802001EA, diluted according to the manufacturer's instructions) was added and incubated at 25°C for 1 hour. After washing the 96-well plate, add 100 μL of TMB to each well and incubate at 25°C in the dark for 5 minutes. Finally, add 1M H2SO4 to stop color development. After standing at room temperature for 5 minutes, detect using a microplate reader. Use 650 nm as the reference wavelength and measure the absorbance at 450 nm. Record the measurement results. 50 Value / test sample EC 50 value)×100%.

[0281] Dynamic light scattering (DLS) was used to detect particle size. A nanoparticle size analyzer (Malvern, ZEN3600) was used for detection. The sample was slowly added to a clean sample cell and allowed to stand for 15 minutes to remove bubbles. The test was then performed. The test type was size. Each sample was measured three times in a cycle, and the average value was taken as the result.

[0282] Preparation of buffer:

[0283] (1) The histidine-acetate buffer used in the examples was prepared with histidine and adjusted to the target pH with acetic acid. For example, a 20 mM histidine-acetate buffer (pH 6.0) was prepared with approximately 3.1 g / L histidine and the pH was adjusted to 6.0 with acetic acid.

[0284] (2) The sodium phosphate buffer used in the example is composed of disodium hydrogen phosphate and sodium dihydrogen phosphate, wherein the 20 mM sodium phosphate buffer (pH 6.0) is prepared from about 6.3 g / L disodium hydrogen phosphate dodecahydrate and about 0.4 g / L sodium dihydrogen phosphate dihydrate.

[0285] (3) The sodium citrate buffer used in the examples is composed of citric acid and sodium citrate, wherein 20 mM sodium citrate buffer (pH 6.0) is prepared from about 0.7 g / L citric acid and about 4.8 g / L sodium citrate dihydrate.

[0286] Example 5 Screening of pharmaceutical compositions

[0287] The anti-TIM-3 antibody h50B5 was replaced by ultrafiltration into the buffers listed in Table 3. After the replacement, the mixture was concentrated and then other excipients were added according to the pharmaceutical composition listed in Table 3. After mixing, the mixture was sterilized by filtration through a 0.22 μm filter membrane.

[0288] The initial screening and evaluation of the pharmaceutical composition was performed using Tm, Tagg, and SEC-HPLC as indicators. Tm characterizes the conformational stability of the antibody in the pharmaceutical composition, Tagg characterizes the colloidal stability, and SEC-HPLC detects antibody aggregates in the pharmaceutical composition.

[0289] Table 3. Pharmaceutical composition and its Tm, Tagg, SEC-HPLC test results

[0290] The results are shown in Table 3. The conformational and colloidal stability of the antibody in the pharmaceutical composition was better when the pH value was in the range of 6.0 to 7.0. Compared with the use of sodium phosphate buffer or sodium citrate buffer, the thermal stability of the formulation was better when the histidine-acetate buffer was used.

[0291] The anti-TIM-3 antibody h50B5 was replaced with the buffer listed in Table 4. After replacement, the mixture was concentrated and other excipients were added according to the pharmaceutical composition listed in Table 4. After mixing, the mixture was sterilized by filtration through a 0.22 μm filter. The Tm and Tagg indicators of the pharmaceutical composition were evaluated. The results are shown in Table 4. When the antibody concentration in the pharmaceutical composition was increased to 60 mg / mL, the antibody conformation and colloidal stability in the pharmaceutical composition remained good.

[0292] Table 4. Drug composition and its Tm, Tagg test results

[0293] Example 6 Screening of pharmaceutical compositions

[0294] The anti-TIM-3 antibody h50B5 was exchanged into the buffers listed in Table 5. After exchange, the mixture was concentrated, and then other excipients were added according to the pharmaceutical compositions listed in Table 5. After uniform mixing, the mixture was sterilized by filtration through a 0.22 μm filter. The pharmaceutical compositions were screened using Tm, Tagg, and SEC-HPLC as indicators. The experimental results are shown in Table 6. Antibodies in pharmaceutical compositions with pH values ​​between 6.0 and 7.0 and a histidine-acetate buffer concentration between 10 and 30 mM exhibited good conformational and colloidal stability, consistent with the results in Example 5. The addition of sucrose improved the conformational and colloidal stability of the antibody, while the addition of sodium chloride had the opposite effect.

[0295] Table 5. Pharmaceutical composition

[0296] Table 6. Tm, Tagg and SEC-HPLC test results of the pharmaceutical composition

[0297] Example 7 Stability Study of Pharmaceutical Composition

[0298] Pharmaceutical compositions were prepared according to Table 7 and stability studies were conducted. Candidate pharmaceutical compositions were evaluated based on physical, chemical, and biological parameters, including long-term stability at 2-8°C, accelerated stability at 25±2°C, and shaking stability at 25°C. The anti-TIM-3 antibody h50B5 was replaced by ultrafiltration into the buffer listed in Table 7. After replacement, the mixture was concentrated and the other excipients were added according to the pharmaceutical compositions listed in Table 7. After mixing, the mixture was sterilized by filtration through a 0.22μm filter. The stability study time points and test items are shown in Table 8.

[0299] Table 7. Pharmaceutical composition

[0300] Table 8. Stability study plan for pharmaceutical compositions

[0301] Among them, "A" represents Tm and Tagg detection, "X" represents appearance, DLS, SEC-HPLC, CEX-HPLC, CE-SDS and biological activity detection, "Z" represents osmotic pressure molar concentration detection, and " / " represents no detection.

[0302] The detection results of Tm and Tagg are shown in Table 9.

[0303] Table 9. Tm and Tagg test results

[0304] The results of the accelerated stability study are shown in Tables 10-1 and 10-2. The data in Tables 10-1 and 10-2 indicate that both F28 and F25 appear as colorless, slightly opalescent solutions. After one month of storage at 25±2°C, no significant changes in appearance, particle size, osmolarity, SEC-HPLC, CEX-HPLC, or CE-SDS were observed for F28 and F25. These results demonstrate that both F28 and F25 are relatively stable at 25°C.

[0305] Table 10-1. Results of accelerated stability investigation

[0306] Table 10-2 Results of investigation on accelerated stability

[0307] The results of the shaking stability study are shown in Tables 11-1 and 11-2. After shaking at 250 rpm for two weeks at 25°C, no significant changes were observed in the appearance, osmolarity, CEX-HPLC, SEC-HPLC, and CE-SDS performance of F28 and F25. These results demonstrate that both F28 and F25 are relatively stable at 25°C and 250 rpm.

[0308] Table 11-1. Results of investigation on shaking stability

[0309] Table 11-2. Results of investigation on shaking stability

[0310] The results of the long-term stability study are shown in Tables 12-1 and 12-2. The data in Tables 12-1 and 12-2 indicate that, after six months of storage at 2-8°C, the appearance, osmolality, and biological activity of F28 and F25 remained unchanged. The data in Table 12-2 also indicate that the purity indicators (CEX-HPLC, SEC-HPLC, and CE-SDS) for F28 and F25 showed similar trends. These results demonstrate that pharmaceutical compositions F28 and F25 can maintain stability at 2-8°C.

[0311] Table 12-1. Results of long-term stability investigation

[0312] “ / ” means not detected.

[0313] Table 12-2. Results of long-term stability investigation

[0314] Although the present disclosure has been described in conjunction with one or more embodiments, it should be understood that the present disclosure not only covers these embodiments, but is also intended to cover all alternatives, modifications and equivalents included in the spirit and scope of the appended claims. All documents cited herein are fully incorporated herein by reference.

Claims

1. A pharmaceutical composition comprising: (a) an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) a buffer, (c) a surfactant, and (d) a stabilizer, wherein: The anti-TIM-3 antibody or antigen-binding fragment thereof comprises: HCDR1 of the amino acid sequence shown in SEQ ID NO:1, HCDR2 of the amino acid sequence shown in SEQ ID NO:2, HCDR3 of the amino acid sequence shown in SEQ ID NO:3, LCDR1 of the amino acid sequence shown in SEQ ID NO:4, LCDR2 of the amino acid sequence shown in SEQ ID NO:5, and LCDR3 of the amino acid sequence shown in SEQ ID NO:6; or HCDR1 of the amino acid sequence shown in SEQ ID NO:11, HCDR2 of the amino acid sequence shown in SEQ ID NO:12, HCDR3 of the amino acid sequence shown in SEQ ID NO:13, LCDR1 of the amino acid sequence shown in SEQ ID NO:14, LCDR2 of the amino acid sequence shown in SEQ ID NO:15, and LCDR3 of the amino acid sequence shown in SEQ ID NO:

16.

2. The pharmaceutical composition according to any one of claims 1, wherein The concentration of the anti-TIM-3 antibody or antigen-binding fragment thereof is 1 mg / mL to 120 mg / mL, 10 mg / mL to 100 mg / mL, 15 mg / mL to 90 mg / mL, 20 mg / mL to 80 mg / mL, 25 mg / mL to 70 mg / mL, 30 mg / mL to 60 mg / mL, 55 mg / mL to 65 mg / mL, or 59 mg / mL to 61 mg / mL.

3. The pharmaceutical composition according to claim 1 or 2, wherein The concentration of the buffer is 1 mM to 100 mM, 2 mM to 80 mM, 5 mM to 60 mM, 8 mM to 40 mM, 10 mM to 30 mM, 10 mM to 20 mM, 20 mM to 30 mM, 15 mM to 25 mM, or 19 mM to 21 mM.

4. The pharmaceutical composition according to any one of claims 1 to 3, wherein The buffer is a histidine buffer, a phosphate buffer, a succinate buffer, a citrate buffer and / or an acetate buffer; preferably, the buffer is a histidine buffer; more preferably, the buffer is a histidine-acetate buffer.

5. The pharmaceutical composition according to any one of claims 1 to 4, wherein The concentration of the surfactant is 0.01 mg / mL to 2 mg / mL, 0.05 mg / mL to 1.5 mg / mL, 0.1 mg / mL to 1 mg / mL, 0.2 mg / mL to 0.8 mg / mL, 0.4 mg / mL to 1.2 mg / mL, 0.4 mg / mL to 0.8 mg / mL, 0.3 mg / mL to 0.5 mg / mL, 0.6 mg / mL to 1 mg / mL, or 0.7 mg / mL to 0.9 mg / mL.

6. The pharmaceutical composition according to any one of claims 1 to 5, wherein The surfactant is polysorbate; preferably, the surfactant is polysorbate 80 or polysorbate 20; more preferably, the surfactant is polysorbate 80.

7. The pharmaceutical composition according to any one of claims 1 to 6, wherein The stabilizer comprises mannitol, sorbitol, trehalose, sucrose, lysine, glycine, proline, arginine or pharmaceutically acceptable salts thereof and / or sodium chloride.

8. The pharmaceutical composition according to claim 7, wherein The stabilizer comprises sucrose.

9. The pharmaceutical composition according to claim 7 or 8, wherein The concentration of sucrose is 30 mM to 960 mM, 60 mM to 720 mM, 120 mM to 480 mM, 240 mM to 480 mM, 210 mM to 270 mM, 230 mM to 250 mM, 450 mM to 510 mM, or 470 mM to 490 mM.

10. The pharmaceutical composition according to any one of claims 1 to 9, wherein The pH of the pharmaceutical composition is 4 to 7, 5 to 7, 5.5 to 7, 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.

5.

11. The pharmaceutical composition according to any one of claims 1 to 10, wherein (1) The pharmaceutical composition comprises: (a) 1 mg / mL to 120 mg / mL of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 1 mM to 100 mM of a histidine buffer, (c) 0.01 mg / mL to 2 mg / mL of polysorbate 80, and (d) 30 mM to 960 mM of sucrose; and the pH of the pharmaceutical composition is 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5; (2) The pharmaceutical composition comprises: (a) 10 mg / mL to 100 mg / mL of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 2 mM to 80 mM of a histidine buffer, (c) 0.05 mg / mL to 1.5 mg / mL of polysorbate 80, and (d) 60 mM to 720 mM of sucrose; and the pH of the pharmaceutical composition is 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5; (3) The pharmaceutical composition comprises: (a) 15 mg / mL to 90 mg / mL of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 5 mM to 60 mM of a histidine buffer, (c) 0.1 mg / mL to 1 mg / mL of polysorbate 80, and (d) 120 mM to 480 mM of sucrose; and the pH of the pharmaceutical composition is 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5; (4) The pharmaceutical composition comprises: (a) 20 mg / mL to 80 mg / mL of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 8 mM to 40 mM of a histidine buffer, (c) 0.2 mg / mL to 0.8 mg / mL of polysorbate 80, and (d) 120 mM to 480 mM of sucrose; and the pH of the pharmaceutical composition is 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5; (5) The pharmaceutical composition comprises: (a) 30 mg / mL to 60 mg / mL of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 10 mM (c) 0.2 mg / mL to 0.8 mg / mL of polysorbate 80, and (d) 120 mM to 480 mM sucrose; and the pH of the pharmaceutical composition is 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5; preferably, the pH of the pharmaceutical composition is 6 to 7; (6) The pharmaceutical composition comprises: (a) 30 mg / mL to 60 mg / mL of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 10 mM to 30 mM of a histidine buffer, (c) 0.4 mg / mL to 0.8 mg / mL of polysorbate 80, and (d) 240 mM to 480 mM of sucrose; and the pH of the pharmaceutical composition is 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5; preferably, the pH of the pharmaceutical composition is 6 to 7; (7) The pharmaceutical composition comprises: (a) 55 mg / mL to 65 mg / mL of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 15 mM to 25 mM of a histidine buffer, (c) 0.4 mg / mL to 0.8 mg / mL of polysorbate 80, and (d) 240 mM to 480 mM of sucrose; and the pH of the pharmaceutical composition is 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5; preferably, the pH of the pharmaceutical composition is 6 to 7; (8) The pharmaceutical composition comprises: (a) 60 mg / mL of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 20 mM of a histidine buffer, (c) 0.4 mg / mL to 0.8 mg / mL of polysorbate 80, and (d) 240 mM to 480 mM of sucrose; and the pH of the pharmaceutical composition is 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5; preferably, the pH of the pharmaceutical composition is 6 to 7; (9) The pharmaceutical composition comprises: (a) 55 mg / mL to 65 mg / mL of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 15 mM to 25 mM of a histidine buffer, (c) 0.7 mg / mL to 0.9 mg / mL of polysorbate 80, and (d) 450 mM to 510 mM of sucrose; and the pH of the pharmaceutical composition is 5 to 7, 6 to 7, 6.5 to 7, 5.5 to 6.5, or 6 to 6.5; preferably, the pH of the pharmaceutical composition is 6 to 7; (10) The pharmaceutical composition comprises: (a) 60 mg / mL of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 20 mM of histidine buffer, (c) 0.4 mg / mL of polysorbate 80, and (d) 240 mM of sucrose; and the pH of the pharmaceutical composition is 5.5 to 6.5, preferably 6; or (11) The pharmaceutical composition comprises: (a) 60 mg / mL of an anti-TIM-3 antibody or an antigen-binding fragment thereof, (b) 20 mM of histidine buffer, (c) 0.8 mg / mL of polysorbate 80, and (d) 480 mM of sucrose; and the pH of the pharmaceutical composition is 6 to 7, preferably 6.

5.

12. The pharmaceutical composition according to any one of claims 1 to 11, wherein The anti-TIM-3 antibody or antigen-binding fragment thereof comprises: A heavy chain variable region having an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:7, and a light chain variable region having an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:8; or A heavy chain variable region having an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO:17, and a light chain variable region having an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO:

18.

13. The pharmaceutical composition according to any one of claims 1 to 12, wherein The anti-TIM-3 antibody or antigen-binding fragment thereof comprises: A heavy chain having an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:9, and a light chain having an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:10; or A heavy chain having an amino acid sequence at least 95% identical to the amino acid sequence shown in SEQ ID NO:19, and a light chain having an amino acid sequence at least 95% identical to the amino acid sequence shown in SEQ ID NO:

20.

14. The pharmaceutical composition according to any one of claims 1 to 13, wherein The pharmaceutical composition is an injection; preferably, the pharmaceutical composition is an aqueous injection.

15. Use of the preparation of the pharmaceutical composition according to any one of claims 1 to 14 in the preparation of a drug for treating cancer; Optionally, the cancer is relapsed, refractory, metastatic and / or advanced cancer; Optionally, the cancer is biliary tract cancer, carcinosarcoma, soft tissue sarcoma, esophageal cancer, gastroesophageal junction cancer, breast cancer, gastric cancer, gallbladder cancer, pancreatic cancer, head and neck cancer, colorectal cancer, kidney cancer, cervical cancer, ovarian cancer, endometrial cancer, uterine carcinosarcoma, uterine cancer, fallopian tube cancer, melanoma, pharyngeal cancer, laryngeal cancer, nasopharyngeal cancer, oral cancer, skin cancer, lung cancer, urethral cancer, prostate cancer, bladder cancer, anal cancer, testicular cancer, vaginal cancer, gastrointestinal stromal tumor, squamous cell carcinoma, peritoneal cancer, liver cancer, uterine cancer, salivary gland cancer, vulvar cancer, thyroid cancer, penile cancer, Merkel cell carcinoma, leukemia, malignant lymphoma, glioblastoma, bone cancer, plasmacytoma, myeloma, myelodysplastic syndrome, myelofibrosis, or Paget's disease.

Images