Stabilized formulation containing anti-CD20 x anti-CD3 bispecific antibody

Abstract

A vial for single-dose administration of a stable liquid pharmaceutical formulation comprising a human bispecific antibody that specifically binds to human CD20 and human CD3 is provided. [Solution] The single-dose vial contains a human bispecific antibody that specifically binds to human CD20 and human CD3, a buffer containing histidine at a concentration of 5 mM to 15 mM, polysorbate at a concentration of 0.05% to 0.15% w / v, and sucrose at a concentration of 8% to 12% w / v, and the formulation has a pH of 5.8±0.3.

Description

[Technical Field]

[0001] Sequence Listing Reference This application incorporates by reference the Sequence Listing, which was filed in computer readable format as file 10664WO01-Sequence.txt, created on November 20, 2020, and contains 25,368 bytes.

[0002] The present invention relates to the field of therapeutic antibody formulations, more particularly to the field of pharmaceutical formulations comprising a human bispecific antibody that specifically binds to human CD20 and CD3. [Background technology]

[0003] Therapeutic macromolecules (e.g., antibodies) must be formulated in a manner that not only renders the molecules suitable for administration to patients but also maintains their stability during storage and subsequent use. For example, therapeutic antibodies in solution are prone to degradation, aggregation, and / or undesirable chemical modification unless the solution is properly formulated. The stability of antibodies in liquid formulations depends not only on the types of excipients used in the formulation but also on the amounts and ratios of the excipients relative to each other. Furthermore, when preparing liquid antibody formulations, other considerations besides stability must be taken into account. Examples of such additional considerations include the concentration of antibody that can be accommodated by a given formulation, as well as the visual quality and appeal of the formulation. Therefore, when formulating therapeutic antibodies, great care must be taken to arrive at a formulation that maintains stability, contains an appropriate concentration of antibody, and retains other properties that allow the formulation to be conveniently administered to patients.

[0004] CD20 is a non-glycosylated phosphoprotein expressed on the plasma membrane of mature B cells. CD3 is a homodimeric or heterodimeric antigen expressed on T cells in conjunction with the T cell receptor complex (TCR) and is required for T cell activation.

[0005] Bispecific antibodies against human CD20 and human CD3 are an example of a therapeutically relevant macromolecule that requires appropriate formulation. Such antibodies are clinically useful, for example, in the treatment of cancer (e.g., B-cell cancers such as follicular lymphoma, diffuse large B-cell lymphoma, mantle cell lymphoma, marginal zone lymphoma, or other non-Hodgkin's lymphomas).

[0006] Although anti-CD20xanti-CD3 bispecific antibodies are known in the art (see, e.g., WO2014 / 047231), there remains a need for pharmaceutical formulations comprising anti-CD20xanti-CD3 bispecific antibodies that are sufficiently stable and suitable for administration to patients. Summary of the Invention

[0007] Stable liquid pharmaceutical formulations comprising an anti-CD20 x anti-CD3 bispecific antibody and one or more excipients are provided, as well as kits and unit dosage forms comprising such formulations and their uses.

[0008] In one aspect, the present invention provides a stable liquid pharmaceutical formulation, comprising: (a) a bispecific antibody comprising a first antigen-binding domain that specifically binds to human CD20 and a second antigen-binding domain that specifically binds to human CD3, wherein the first antigen-binding domain comprises three heavy chain complementarity-determining regions (CDRs) (A1-HCDR1, A1-HCDR2, and A1-HCDR3) comprised in a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 4 and three light chain CDRs (LCDR1, LCDR2, and LCDR3) comprised in a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 6; and the second antigen-binding domain comprises three heavy chain complementarity-determining regions (CDRs) (A1-HCDR1, A1-HCDR2, and A1-HCDR3) comprised in a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 5. and (d) a stabilizer comprising a sugar; (a) a bispecific antibody comprising three heavy chain CDRs (A2-HCDR1, A2-HCDR2, and A2-HCDR3) comprised in a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 6; and (b) a buffer comprising histidine; (c) an organic co-solvent comprising polysorbate; and (d) a stabilizer comprising a sugar; wherein the formulation has a pH of 5.8±0.3.

[0009] In one aspect, the present invention provides a stable liquid pharmaceutical formulation, comprising: (a) a bispecific antibody comprising a first antigen-binding domain that specifically binds to human CD20 and a second antigen-binding domain that specifically binds to human CD3, wherein the first antigen-binding domain comprises three heavy chain complementarity-determining regions (CDRs) (A1-HCDR1, A1-HCDR2, and A1-HCDR3) comprised in a heavy chain variable region (HCVR) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) comprised in a light chain variable region (LCVR); and the second antigen-binding domain comprises three heavy chain CDRs (A2-HCDR1, A2-HCDR2, and A2-HCDR3) comprised in the heavy chain variable region (HCVR) and three light chain CDRs (LCDR1, LCDR2, and LCDR3). and (d) a stabilizer comprising a sugar; and (b) a bispecific antibody comprising three light chain CDRs (LCDR1, LCDR2, and LCDR3) comprised in A1-HCDR1, A1-HCDR2, and A1-HCDR3, respectively, wherein A1-HCDR1, A1-HCDR2, and A1-HCDR3 comprise the amino acid sequences of SEQ ID NOs: 7, 8, and 9, A2-HCDR1, A2-HCDR2, and A2-HCDR3 comprise the amino acid sequences of SEQ ID NOs: 10, 11, and 12, respectively, and LCDR1, LCDR2, and LCDR3 comprise the amino acid sequences of SEQ ID NOs: 13, 14, and 15, respectively; and a stable liquid pharmaceutical formulation comprising: (a) a buffer comprising histidine; (c) an organic co-solvent comprising a polysorbate; and (d) a stabilizer comprising a sugar; wherein the formulation has a pH of 5.8±0.3.

[0010] In some embodiments, the antibody concentration is 1 mg / ml ± 0.1 mg / ml to 200 mg / ml ± 20 mg / ml. In some embodiments, the antibody concentration is 2 mg / ml ± 0.2 mg / ml. In some embodiments, the antibody concentration is 20 mg / ml ± 2 mg / ml. In some embodiments, the antibody concentration is 80 mg / ml ± 8 mg / ml. In some embodiments, the antibody concentration is 100 mg / ml ± 10 mg / ml. In some embodiments, the antibody concentration is 160 mg / ml ± 16 mg / ml. In some embodiments, the antibody concentration is about 2 mg / ml to about 100 mg / ml.

[0011] In some embodiments, the histidine buffer concentration is 5 mM ± 1 mM to 15 mM ± 3 mM. Optionally, the histidine buffer concentration is 10 mM ± 1 mM. In some embodiments, the histidine buffer comprises L-histidine and L-histidine monohydrochloride monohydrate. Optionally, the L-histidine concentration is 3.65 mM ± 0.5 mM and the L-histidine monohydrochloride monohydrate concentration is 6.35 mM ± 0.5 mM. Optionally, the histidine buffer comprises 0.57 ± 0.05 mg / ml L-histidine and 1.33 ± 0.13 mg / ml L-histidine monohydrochloride monohydrate.

[0012] In some embodiments, the polysorbate concentration is 0.01%±0.005% to 0.5%±0.25% w / v. In some embodiments, the polysorbate concentration is 0.1%±0.05% w / v. In some embodiments, the polysorbate concentration is 0.1%±0.01% w / v. In some embodiments, the surfactant is polysorbate 80.

[0013] In some embodiments, the sugar is sucrose. In some cases, the sucrose concentration is 5%±1% to 20%±4% w / v. In some cases, the sucrose concentration is 8%±0.5% to 12%±0.5% w / v. In some embodiments, the sucrose concentration is 10%±1% w / v.

[0014] In some embodiments, the pharmaceutical composition comprises: (a) 2 mg / ml ± 0.2 mg / ml antibody; (b) 5 mM ± 1 mM to 15 mM ± 3 mM histidine buffer; (c) 0.01% ± 0.005% to 0.5% ± 0.25% w / v polysorbate; and (d) 5% ± 1% to 20% ± 4% w / v sucrose at a pH of 5.8 ± 0.3.

[0015] In some embodiments, the pharmaceutical composition comprises (a) 2 mg / ml ± 0.2 mg / ml antibody, (b) 10 mM ± 1 mM histidine buffer, (c) 0.1% ± 0.01% w / v polysorbate, and (d) 10% ± 1% w / v sucrose at pH 5.8 ± 0.3.

[0016] In some embodiments, the pharmaceutical composition comprises (a) 2 mg / ml ± 0.2 mg / ml antibody, (b) 3.65 mM ± 0.2 mM L-histidine, (c) 6.35 mM ± 0.2 mM L-histidine monohydrochloride monohydrate, (d) 0.1% ± 0.01% w / v polysorbate, and (e) 10% ± 1% w / v sucrose at a pH of 5.8 ± 0.3.

[0017] In some embodiments, the pharmaceutical composition comprises (a) 2 mg / ml ± 0.2 mg / ml of antibody, (b) 0.57 mg / ml ± 0.05 mg / ml of L-histidine, (c) 1.33 mg / ml ± 0.13 mg / ml of L-histidine monohydrochloride monohydrate, (d) 0.1% ± 0.01% w / v of polysorbate, and (e) 10% ± 1% w / v of sucrose, at a pH of 5.8 ± 0.3.

[0018] In some embodiments, the pharmaceutical composition comprises: (a) 20 mg / ml ± 2 mg / ml antibody; (b) 5 mM ± 1 mM to 15 mM ± 3 mM histidine buffer; (c) 0.01% ± 0.005% to 0.5% ± 0.25% w / v polysorbate; and (d) 5% ± 1% to 20% ± 4% w / v sucrose at pH 5.8 ± 0.3.

[0019] In some embodiments, the pharmaceutical composition comprises (a) 20 mg / ml ± 2 mg / ml antibody, (b) 10 mM ± 1 mM histidine buffer, (c) 0.1% ± 0.01% w / v polysorbate, and (d) 10% ± 1% w / v sucrose at pH 5.8 ± 0.3.

[0020] In some embodiments, the pharmaceutical composition comprises (a) 20 mg / ml ± 2 mg / ml antibody, (b) 3.65 mM ± 0.2 mM L-histidine, (c) 6.35 mM ± 0.2 mM L-histidine monohydrochloride monohydrate, (d) 0.1% ± 0.01% w / v polysorbate, and (e) 10% ± 1% w / v sucrose at a pH of 5.8 ± 0.3.

[0021] In some embodiments, the pharmaceutical composition comprises (a) 20 mg / ml ± 2 mg / ml of antibody, (b) 0.57 mg / ml ± 0.05 mg / ml of L-histidine, (c) 1.33 mg / ml ± 0.13 mg / ml of L-histidine monohydrochloride monohydrate, (d) 0.1% ± 0.01% w / v of polysorbate, and (e) 10% ± 1% w / v of sucrose, at a pH of 5.8 ± 0.3.

[0022] In some embodiments, the pharmaceutical composition comprises: (a) 80 mg / ml ± 8 mg / ml antibody; (b) 5 mM ± 1 mM to 15 mM ± 3 mM histidine buffer; (c) 0.01% ± 0.005% to 0.5% ± 0.25% w / v polysorbate; and (d) 5% ± 1% to 20% ± 4% w / v sucrose at pH 5.8 ± 0.3.

[0023] In some embodiments, the pharmaceutical composition comprises (a) 80 mg / ml ± 8 mg / ml antibody, (b) 10 mM ± 1 mM histidine buffer, (c) 0.1% ± 0.01% w / v polysorbate, and (d) 10% ± 1% w / v sucrose at pH 5.8 ± 0.3. Includes.

[0024] In some embodiments, the pharmaceutical composition comprises (a) 80 mg / ml ± 8 mg / ml of antibody, (b) 3.65 mM ± 0.2 mM L-histidine, (c) 6.35 mM ± 0.2 mM L-histidine monohydrochloride monohydrate, (d) 0.1% ± 0.01% w / v polysorbate, and (e) 10% ± 1% w / v sucrose at a pH of 5.8 ± 0.3.

[0025] In some embodiments, the pharmaceutical composition comprises (a) 80 mg / ml ± 8 mg / ml of antibody, (b) 0.57 mg / ml ± 0.05 mg / ml of L-histidine, (c) 1.33 mg / ml ± 0.13 mg / ml of L-histidine monohydrochloride monohydrate, (d) 0.1% ± 0.01% w / v of polysorbate, and (e) 10% ± 1% w / v of sucrose, at a pH of 5.8 ± 0.3.

[0026] In some embodiments, the pharmaceutical composition comprises: (a) 100 mg / ml ± 10 mg / ml antibody; (b) 5 mM ± 1 mM to 15 mM ± 3 mM histidine buffer; (c) 0.01% ± 0.005% to 0.5% ± 0.25% w / v polysorbate; and (d) 5% ± 1% to 20% ± 4% w / v sucrose at pH 5.8 ± 0.3.

[0027] In some embodiments, the pharmaceutical composition comprises (a) 100 mg / ml ± 10 mg / ml antibody, (b) 10 mM ± 1 mM histidine buffer, (c) 0.1% ± 0.01% w / v polysorbate, and (d) 10% ± 1% w / v sucrose at pH 5.8 ± 0.3.

[0028] In some embodiments, the pharmaceutical composition comprises (a) 100 mg / ml ± 10 mg / ml of antibody, (b) 3.65 mM ± 0.2 mM L-histidine, (c) 6.35 mM ± 0.2 mM L-histidine monohydrochloride monohydrate, (d) 0.1% ± 0.01% w / v polysorbate, and (e) 10% ± 1% w / v sucrose at a pH of 5.8 ± 0.3.

[0029] In some embodiments, the pharmaceutical composition comprises (a) 100 mg / ml ± 10 mg / ml of antibody, (b) 0.57 mg / ml ± 0.05 mg / ml of L-histidine, (c) 1.33 mg / ml ± 0.13 mg / ml of L-histidine monohydrochloride monohydrate, (d) 0.1% ± 0.01% w / v of polysorbate, and (e) 10% ± 1% w / v of sucrose, at a pH of 5.8 ± 0.3.

[0030] In some embodiments, the pharmaceutical composition comprises: (a) 160 mg / ml ± 16 mg / ml antibody; (b) 5 mM ± 1 mM to 15 mM ± 3 mM histidine buffer; (c) 0.01% ± 0.005% to 0.5% ± 0.25% w / v polysorbate; and (d) 5% ± 1% to 20% ± 4% w / v sucrose at pH 5.8 ± 0.3.

[0031] In some embodiments, the pharmaceutical composition comprises (a) 160 mg / ml ± 16 mg / ml antibody, (b) 10 mM ± 1 mM histidine buffer, (c) 0.1% ± 0.01% w / v polysorbate, and (d) 10% ± 1% w / v sucrose at pH 5.8 ± 0.3.

[0032] In some embodiments, the pharmaceutical composition comprises (a) 160 mg / ml ± 16 mg / ml of antibody, (b) 3.65 mM ± 0.2 mM L-histidine, (c) 6.35 mM ± 0.2 mM L-histidine monohydrochloride monohydrate, (d) 0.1% ± 0.01% w / v polysorbate, and (e) 10% ± 1% w / v sucrose at a pH of 5.8 ± 0.3.

[0033] In some embodiments, the pharmaceutical composition comprises (a) 160 mg / ml ± 16 mg / ml of antibody, (b) 0.57 mg / ml ± 0.05 mg / ml of L-histidine, (c) 1.33 mg / ml ± 0.13 mg / ml of L-histidine monohydrochloride monohydrate, (d) 0.1% ± 0.01% w / v of polysorbate, and (e) 10% ± 1% w / v of sucrose, at a pH of 5.8 ± 0.3.

[0034] In any of these embodiments, the polysorbate may be polysorbate 80.

[0035] In various embodiments of the pharmaceutical compositions described above or discussed herein, at least 90% of the antibodies have a native conformation as determined by size-exclusion ultra-performance liquid chromatography (SE-UPLC) after one month of storage at 45° C. In some cases, at least 93% of the antibodies have a native conformation as determined by SE-UPLC after one month of storage at 45° C. In some cases, at least 95% of the antibodies have a native conformation as determined by SE-UPLC after one month of storage at 45° C.

[0036] In various embodiments of the pharmaceutical compositions described above or discussed herein, at least 95% of the antibody has a native conformation after 3 months of storage at 25° C. and 60% relative humidity as determined by SE-UPLC. In some cases, at least 97% of the antibody has a native conformation after 3 months of storage at 25° C. and 60% relative humidity as determined by SE-UPLC.

[0037] In various embodiments of the pharmaceutical compositions described above or discussed herein, at least 95% of the antibodies have a native conformation as determined by SE-UPLC after 3 months of storage at 5° C. In some cases, at least 98% of the antibodies have a native conformation as determined by SE-UPLC after 3 months of storage at 5° C.

[0038] In various embodiments of the pharmaceutical compositions described above or discussed herein, the formulation contains 6% or less high molecular weight (HMW) species, as determined by SE-UPLC, after 1 month of storage at 45° C. In some cases, the formulation contains 5.6% or less HMW species, as determined by SE-UPLC, after 1 month of storage at 45° C.

[0039] In various embodiments of the pharmaceutical compositions described above or discussed herein, the formulation contains 2% or less HMW species after 3 months of storage at 25° C. and 60% relative humidity as determined by SE-UPLC. In some cases, the formulation contains 1% or less HMW species after 3 months of storage at 25° C. and 60% relative humidity as determined by SE-UPLC.

[0040] In various embodiments of the pharmaceutical compositions described above or discussed herein, the formulation contains 1.5% or less HMW species as determined by SE-UPLC after 3 months of storage at 5° C. In some cases, the formulation contains 1% or less HMW species as determined by SE-UPLC after 9 months of storage at 5° C.

[0041] In various embodiments of any of the pharmaceutical compositions described above or discussed herein, the antibody comprises A1-HCDR1, A1-HCDR2, and A1-HCDR3 comprising the amino acid sequences of SEQ ID NOs: 7, 8, and 9, respectively; A2-HCDR1, A2-HCDR2, and A2-HCDR3 comprising the amino acid sequences of SEQ ID NOs: 10, 11, and 12, respectively; and LCDR1, LCDR2, and LCDR3 comprising the amino acid sequences of SEQ ID NOs: 13, 14, and 15, respectively. Includes R3.

[0042] In some embodiments, the first antigen-binding domain comprises an HCVR having at least 90% identity to the amino acid sequence of SEQ ID NO: 4 and an LCVR having at least 90% identity to the amino acid sequence of SEQ ID NO: 6, and the second antigen-binding domain comprises an HCVR having at least 90% identity to the amino acid sequence of SEQ ID NO: 5 and an LCVR having at least 90% identity to the amino acid sequence of SEQ ID NO: 6. In some embodiments, the first antigen-binding domain comprises an HCVR having at least 95% identity to the amino acid sequence of SEQ ID NO: 4 and an LCVR having at least 95% identity to the amino acid sequence of SEQ ID NO: 6, and the second antigen-binding domain comprises an HCVR having at least 95% identity to the amino acid sequence of SEQ ID NO: 5 and an LCVR having at least 95% identity to the amino acid sequence of SEQ ID NO: 6. In some embodiments, the first antigen-binding domain comprises an HCVR having at least 99% identity to the amino acid sequence of SEQ ID NO: 4 and an LCVR having at least 99% identity to the amino acid sequence of SEQ ID NO: 6, and the second antigen-binding domain comprises an HCVR having at least 99% identity to the amino acid sequence of SEQ ID NO: 5 and an LCVR having at least 99% identity to the amino acid sequence of SEQ ID NO: 6.

[0043] In various embodiments of any of the pharmaceutical compositions described above or discussed herein, the first antigen-binding domain comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 4 and an LCVR comprising the amino acid sequence of SEQ ID NO: 6, and the second antigen-binding domain comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 5 and an LCVR comprising the amino acid sequence of SEQ ID NO: 6.

[0044] In some embodiments, the antibody comprises a human IgG heavy chain constant region attached to the HCVR of each of the first and second antigen-binding domains, respectively. Optionally, the heavy chain constant region is of isotype IgG1. Optionally, the heavy chain constant region is of isotype IgG4. In some embodiments, the heavy chain constant region attached to the HCVR of the first antigen-binding domain or the heavy chain constant region attached to the HCVR of the second antigen-binding domain, but not both, comprises an amino acid modification that reduces Protein A binding compared to a heavy chain of the same isotype without the modification. Optionally, the modification comprises an H435R substitution (EU numbering) in a heavy chain of isotype IgG1 or IgG4. Optionally, the modification comprises an H435R substitution and a Y436F substitution (EU numbering) in a heavy chain of isotype IgG1 or IgG4.

[0045] In some embodiments, the antibody comprises a chimeric hinge. For example, the chimeric hinge, in one embodiment, comprises a first amino acid sequence or "upper hinge" sequence derived from a human IgG1 hinge region or a human IgG4 hinge region, and a second amino acid sequence or "lower hinge" sequence derived from a human IgG2 hinge region. In certain embodiments, the first or "upper hinge" sequence comprises amino acid residues at positions 216-227 according to EU numbering. In some embodiments, the second or "lower hinge" sequence comprises amino acid residues at positions 228-236 according to EU numbering. In some cases where the antibody heavy chain constant region is of the IgG4 isotype, the chimeric hinge comprises an upper hinge sequence derived from human IgG4 (positions 216-227 according to EU numbering) and a lower hinge sequence derived from human IgG2 (positions 228-236 according to EU numbering). In some cases where the antibody heavy chain constant region is of isotype IgG1, the chimeric hinge comprises an upper hinge sequence derived from human IgG1 (positions 216-227 according to EU numbering) and a lower hinge sequence derived from human IgG2 (positions 228-236 according to EU numbering). In some embodiments where the heavy chain constant region is of isotype IgG1 and the antibody comprises a chimeric hinge, the CH2 domain of the IgG1 heavy chain constant region that is not is of isotype IgG4. Unless otherwise specified, reference to an IgG1 or IgG4 heavy chain constant region refers to a heavy chain constant region (e.g., For example, a reference to an IgG1 heavy chain constant region includes an IgG1 heavy chain constant region with an IgG2 lower hinge sequence, and a reference to an IgG4 heavy chain constant region includes an IgG4 heavy chain constant region with an IgG2 lower hinge sequence.

[0046] In some embodiments, the bispecific antibody comprises a heavy chain constant region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 16, 17, 18, and 19. In some embodiments, the bispecific antibody comprises a heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 16 and a heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 17. In some embodiments, the bispecific antibody comprises a heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 18 and a heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 19.

[0047] In various embodiments of any of the pharmaceutical compositions described above or discussed herein, the antibody comprises a first heavy chain containing the HCVR of the first antigen-binding domain and a second heavy chain containing the HCVR of the second antigen-binding domain, wherein the first heavy chain comprises residues 1-452 of the amino acid sequence of SEQ ID NO: 1 and the second heavy chain comprises residues 1-448 of the amino acid sequence of SEQ ID NO: 2. In some embodiments, the antibody further comprises a common light chain containing the LCVRs of the first and second antigen-binding domains, wherein the common light chain comprises the amino acid sequence of SEQ ID NO: 3.

[0048] In various embodiments of any of the pharmaceutical compositions described above or discussed herein, the antibody comprises a first heavy chain containing the HCVR of the first antigen-binding domain and a second heavy chain containing the HCVR of the second antigen-binding domain, wherein the first heavy chain comprises the amino acid sequence of SEQ ID NO: 1 and the second heavy chain comprises the amino acid sequence of SEQ ID NO: 2. In some embodiments, the antibody further comprises a common light chain containing the LCVRs of the first and second antigen-binding domains, wherein the common light chain comprises the amino acid sequence of SEQ ID NO: 3.

[0049] In one aspect, the invention provides a pharmaceutical formulation comprising: (a) 2 mg / ml ± 0.2 mg / ml of a bispecific antibody comprising a first antigen-binding domain that specifically binds to human CD20 and a second antigen-binding domain that specifically binds to human CD3, wherein the first antigen-binding domain comprises a HCVR comprising the amino acid sequence of SEQ ID NO: 4 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6, and the second antigen-binding domain comprises a HCVR comprising the amino acid sequence of SEQ ID NO: 5 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6; (b) 10 mM ± 1 mM histidine buffer at pH 5.8 ± 0.2; (c) 0.1% ± 0.01% w / v polysorbate 80; and (d) 10% ± 1% w / v sucrose.

[0050] In some embodiments, the pharmaceutical formulation consists of (a) 2 mg / ml ± 0.2 mg / ml antibody, (b) 3.65 mM ± 0.2 mM L-histidine, (c) 6.35 mM ± 0.2 mM L-histidine monohydrochloride monohydrate, and (d) 0.1% ± 0.01% w / v polysorbate 80 in water at pH 5.8 ± 0.2. In some embodiments, the pharmaceutical formulation consists of (a) 2 mg / ml ± 0.2 mg / ml antibody, (b) 0.57 mg / ml ± 0.1 mg / ml L-histidine, (c) 1.33 mg / ml ± 0.1 mg / ml L-histidine monohydrochloride monohydrate, (d) 1 mg / ml ± 0.1 mg / ml polysorbate 80, and (e) 100 mg / ml ± 10 mg / ml sucrose in water at pH 5.8 ± 0.2.

[0051] In one aspect, the invention provides a bispecific antibody at 20 mg / ml ± 2 mg / ml, comprising: (a) a first antigen-binding domain that specifically binds to human CD20 and a second antigen-binding domain that specifically binds to human CD3, wherein the first antigen-binding domain comprises a HCVR comprising the amino acid sequence of SEQ ID NO: 4 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6; and the second antigen-binding domain comprises a HCVR comprising the amino acid sequence of SEQ ID NO: 5 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6. and (b) a 10 mM ± 1 mM histidine buffer at pH 5.8 ± 0.2; (c) 0.1% ± 0.01% w / v polysorbate 80; and (d) 10% ± 1% w / v sucrose.

[0052] In some embodiments, the pharmaceutical formulation consists of (a) 20 mg / ml ± 2 mg / ml antibody, (b) 3.65 mM ± 0.2 mM L-histidine, (c) 6.35 mM ± 0.2 mM L-histidine monohydrochloride monohydrate, and (d) 0.1% ± 0.01% w / v polysorbate 80 in water at pH 5.8 ± 0.2. In some embodiments, the pharmaceutical formulation consists of (a) 20 mg / ml ± 2 mg / ml antibody, (b) 0.57 mg / ml ± 0.1 mg / ml L-histidine, (c) 1.33 mg / ml ± 0.1 mg / ml L-histidine monohydrochloride monohydrate, (d) 1 mg / ml ± 0.1 mg / ml polysorbate 80, and (e) 100 mg / ml ± 10 mg / ml sucrose in water at pH 5.8 ± 0.2.

[0053] In one aspect, the invention provides a pharmaceutical formulation comprising: (a) 80 mg / ml ± 8 mg / ml of a bispecific antibody comprising a first antigen-binding domain that specifically binds to human CD20 and a second antigen-binding domain that specifically binds to human CD3, wherein the first antigen-binding domain comprises a HCVR comprising the amino acid sequence of SEQ ID NO: 4 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6, and the second antigen-binding domain comprises a HCVR comprising the amino acid sequence of SEQ ID NO: 5 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6; (b) 10 mM ± 1 mM histidine buffer at pH 5.8 ± 0.2; (c) 0.1% ± 0.01% w / v polysorbate 80; and (d) 10% ± 1% w / v sucrose.

[0054] In some embodiments, the pharmaceutical formulation consists of (a) 80 mg / ml ± 8 mg / ml antibody, (b) 3.65 mM ± 0.2 mM L-histidine, (c) 6.35 mM ± 0.2 mM L-histidine monohydrochloride monohydrate, and (d) 0.1% ± 0.01% w / v polysorbate 80 in water at pH 5.8 ± 0.2. In some embodiments, the pharmaceutical formulation consists of (a) 80 mg / ml ± 8 mg / ml of antibody, (b) 0.57 mg / ml ± 0.1 mg / ml of L-histidine, (c) 1.33 mg / ml ± 0.1 mg / ml of L-histidine monohydrochloride monohydrate, (d) 1 mg / ml ± 0.1 mg / ml of polysorbate 80, and (e) 100 mg / ml ± 10 mg / ml of sucrose in water at pH 5.8 ± 0.2.

[0055] In one aspect, the invention provides a pharmaceutical formulation comprising: (a) 100 mg / ml ± 10 mg / ml of a bispecific antibody comprising a first antigen-binding domain that specifically binds to human CD20 and a second antigen-binding domain that specifically binds to human CD3, wherein the first antigen-binding domain comprises a HCVR comprising the amino acid sequence of SEQ ID NO: 4 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6, and the second antigen-binding domain comprises a HCVR comprising the amino acid sequence of SEQ ID NO: 5 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6; (b) 10 mM ± 1 mM histidine buffer at pH 5.8 ± 0.2; (c) 0.1% ± 0.01% w / v polysorbate 80; and (d) 10% ± 1% w / v sucrose.

[0056] In some embodiments, the pharmaceutical formulation consists of (a) 100 mg / ml ± 10 mg / ml antibody, (b) 3.65 mM ± 0.2 mM L-histidine, (c) 6.35 mM ± 0.2 mM L-histidine monohydrochloride monohydrate, and (d) 0.1% ± 0.01% w / v polysorbate 80 in water at pH 5.8 ± 0.2. In some embodiments, the pharmaceutical formulation comprises (a) 100 mg / ml ± 10 mg / ml of antibody, (b) 0.57 mg / ml ± 0.1 mg / ml of L-histidine, (c) 1.33 mg / ml ± 0.1 mg / ml of L-histidine monohydrochloride monohydrate, (d) 1 mg / ml ± 0.1 mg / ml of polysorbate 80, and (e) 100 mg / ml ± 10 mg / ml of L-histidine monohydrochloride monohydrate in water at pH 5.8 ± 0.2. It consists of l of sucrose.

[0057] In some embodiments, the pharmaceutical formulation comprises (a) 1-200 mg / ml of antibody (as discussed above or herein), (b) a buffer comprising acetate or phosphate, (c) an organic co-solvent comprising polysorbate, and (d) a stabilizer comprising a sugar, wherein the formulation has a pH of 5.8±0.3. In some embodiments, the pharmaceutical formulation comprises (a) 1-200 mg / ml of antibody (e.g., 2 mg / ml, 20 mg / ml, 80 mg / ml, or 100 mg / ml), (b) 1-50 mM acetate or phosphate buffer, and (d) 0.1%±0.01% w / v polysorbate 80 in water at a pH of 5.8±0.2.

[0058] In one aspect, the present invention provides a pharmaceutical composition, wherein the composition comprises a pharmaceutical formulation as described above or discussed herein, and the composition is contained in a container. In some embodiments, the container is a vial. Optionally, the vial is a 2 ml, 5 ml, 10 ml, or 20 ml Type 1 clear glass vial. In some embodiments, the container is a syringe. Optionally, the syringe is a low-tungsten syringe. In some embodiments, the syringe is a pre-filled syringe. In some embodiments, the composition is contained in an auto-injector.

[0059] In one aspect, the invention provides a kit comprising: (i) a container containing a composition comprising a pharmaceutical formulation as described above or discussed herein, and instructions for use of the composition. In some embodiments, the container is a glass vial. In some embodiments, the container is a pre-filled syringe. In some embodiments, the container is an auto-injector. Optionally, the instructions describe subcutaneous administration of the composition. Optionally, the instructions describe intravenous administration of the composition.

[0060] In one embodiment, the invention provides a container, optionally a glass vial containing 2 mg of antibody in a stable formulation, the formulation comprising, in water at pH 5.8±0.2, (a) 2 mg / ml±0.2 mg / ml of antibody, b) 0.57 mg / ml±0.1 mg / ml of L-histidine, (c) 1.33 mg / ml±0.1 mg / ml of L-histidine monohydrochloride monohydrate, (d) 1 mg / ml±0.1 mg / ml of polysorbate 80, and (e) 100 mg / ml±10 mg / ml of sucrose. In one embodiment, the invention provides a container, optionally a glass vial containing 20 mg, 80 mg, or 160 mg of antibody in a stable formulation, the formulation comprising, in water at pH 5.8±0.2, (a) 20 mg / ml±2 mg / ml of antibody, b) 0.57 mg / ml±0.1 mg / ml of L-histidine, (c) 1.33 mg / ml±0.1 mg / ml of L-histidine monohydrochloride monohydrate, (d) 1 mg / ml±0.1 mg / ml of polysorbate 80, and (e) 100 mg / ml±10 mg / ml of sucrose. In one embodiment, the invention provides a container, optionally a glass vial containing 100 mg, 200 mg, or 400 mg of antibody in a stable formulation, the formulation comprising, in water at pH 5.8±0.2, (a) 100 mg / ml±10 mg / ml of antibody; b) 0.57 mg / ml±0.1 mg / ml of L-histidine; (c) 1.33 mg / ml±0.1 mg / ml of L-histidine monohydrochloride monohydrate; (d) 1 mg / ml±0.1 mg / ml of polysorbate 80; and (e) 100 mg / ml±10 mg / ml of sucrose.

[0061] In one aspect, the invention provides a unit dosage form comprising a pharmaceutical formulation as described above or discussed herein, wherein the antibody is present in an amount of 0.1 mg to 500 mg. Optionally, the antibody is present in an amount of 2 to 2.5 mg. Optionally, the antibody is present in an amount of 10 to 11 mg. Optionally, the antibody is present in an amount of 20 to 25 mg. Optionally, the antibody is present in an amount of 80 to 90 mg. Optionally, the antibody is present in an amount of 10 to 15 mg. In some embodiments, the unit dosage form is a glass vial. In some embodiments, the syringe is a pre-filled syringe. In some embodiments, the unit dosage form is an auto-injector. In some embodiments, the antibody is present in an amount of 0-125 mg. In some embodiments, the antibody is present in an amount of 160-180 mg. In some embodiments, the antibody is present in an amount of 190-210 mg. In some embodiments, the antibody is present in an amount of 320-360 mg. In some embodiments, the unit dosage form is a glass vial. In some embodiments, the syringe is a pre-filled syringe. In some embodiments, the unit dosage form is an auto-injector.

[0062] In any of the pharmaceutical compositions, kits, or unit dosage forms described above or herein, the container containing the pharmaceutical formulation may contain a headspace containing a gas, wherein the gas contains less than 5% by volume of oxygen. In some cases, the gas contains less than 1% by volume of oxygen. In some cases, the gas contains 0.1% by volume or less of oxygen.

[0063] In one aspect, the present invention provides a container comprising a pharmaceutical composition, wherein the composition comprises a pharmaceutical formulation as described above or herein. Optionally, the container is a syringe. Optionally, the container is a pre-filled syringe. Optionally, the container is an auto-injector. Optionally, the container is a glass vial.

[0064] In various embodiments, any of the features or components of the embodiments described above or discussed herein may be combined, and such combinations are encompassed within the scope of the present disclosure. Any specific value described above or discussed herein may be combined with another related value described above or discussed herein to recite a range, with those values ​​representing the upper and lower limits of the range, and all values ​​within such ranges are encompassed within the scope of the present disclosure. Each of the values ​​described above or discussed herein may be expressed with a 1%, 5%, 10%, or 20% variation. For example, a concentration of 10 mM may be expressed as 10 mM ± 0.1 mM (1% variation), 10 mM ± 0.5 mM (5% variation), 10 mM ± 1 mM (10% variation), or 10 mM ± 2 mM (20% variation).

[0065] Other embodiments will be apparent from a review of the detailed description that follows. DETAILED DESCRIPTION OF THE INVENTION

[0066] Before the present invention is described, it is to be understood that the present invention is not limited to the particular methods and experimental conditions described, as such methods and conditions may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope of the present invention will be limited only by the appended claims.

[0067] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this invention belongs.As used herein, the term "about" means that when used in relation to a specific listed numerical value, the value can vary by 1% or less from the listed value.For example, as used herein, the expression "about 100" includes 99 and 101 and all values ​​therebetween (for example, 99.1, 99.2, 99.3, 99.4, etc.).

[0068] Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, exemplary methods and materials are now described. All patents, applications, and non-patent publications mentioned herein are incorporated by reference in their entirety.

[0069] Pharmaceutical preparations As used herein, the phrase "pharmaceutical formulation" refers to a pharmaceutical formulation containing at least one active ingredient (e.g., a bispecific "Anti-CD20xanti-human CD3" refers to a combination of an anti-CD20 antibody and at least one inactive ingredient that, when combined with an active ingredient or one or more additional inactive ingredients, is suitable for therapeutic administration to a human or non-human animal. As used herein, the term "formulation" refers to a "pharmaceutical formulation" unless specifically indicated otherwise. The present invention provides pharmaceutical formulations comprising at least one therapeutic polypeptide. According to certain embodiments of the present invention, the therapeutic polypeptide is a bispecific antibody that specifically binds to human CD20 and human CD3, or an antigen-binding fragment thereof. More specifically, the present invention includes pharmaceutical formulations comprising (i) a human bispecific antibody that specifically binds to human CD20 and human CD3, (ii) a buffer comprising histidine, (iii) an organic cosolvent comprising polysorbate, and (iv) a stabilizer comprising a sugar. Additional components may be included in the formulations of the present invention, provided that such components do not significantly interfere with the stability of the formulation. Specific exemplary components and formulations included in the present invention are described in detail below.

[0070] In certain embodiments, the pharmaceutical formulation of the present invention may be a fluid formulation. As used herein, the term "fluid formulation" refers to a mixture of at least two components that exists in a predominantly fluid state at about 2°C to about 45°C. Fluid formulations include, among others, liquid formulations. Fluid formulations may be of low, medium, or high viscosity, depending on their specific components.

[0071] A bispecific antibody that specifically binds human CD20 and human CD3 The pharmaceutical formulations of the invention can comprise a human bispecific antibody or antigen-binding fragment thereof that specifically binds to human CD20 and human CD3.

[0072] The term "antibody," as used herein, is intended to refer to immunoglobulin molecules, as well as multimers thereof (e.g., IgM), generally comprising four polypeptide chains: two heavy (H) chains and two light (L) chains interconnected by disulfide bonds; however, immunoglobulin molecules consisting solely of heavy chains (i.e., lacking light chains) are also encompassed within the definition of the term "antibody." Each heavy chain comprises a heavy chain variable region (abbreviated herein as HCVR or VH) and a heavy chain constant region. The heavy chain constant region comprises three domains, CH1, CH2, and CH3. Each light chain comprises a light chain variable region (abbreviated herein as LCVR or VL) and a light chain constant region. The light chain constant region comprises one domain (CL1). The VH and VL regions can be further subdivided into hypervariable regions called complementarity-determining regions (CDRs), interspersed with more conserved regions called framework regions (FRs). Each VH and VL is composed of three CDRs and four FRs arranged from the amino terminus to the carboxy terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.

[0073] In certain embodiments of the invention, the anti-CD20x anti-CD3 bispecific antibodies of the invention are human antibodies. As used herein, the term "human antibody" is intended to include antibodies having variable and constant regions derived from human germline immunoglobulin sequences. Human antibodies of the invention may include, for example, amino acid residues in the CDRs, particularly CDR3, that are not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-directed mutagenesis in vitro or by somatic mutation in vivo). However, as used herein, the term "human antibody" is not intended to include antibodies in which CDR sequences derived from the germline of another mammalian species, such as a mouse, have been grafted onto human framework sequences. In various embodiments, the anti-CD20x anti-CD3 bispecific antibodies are human IgG antibodies. In various embodiments, the anti-CD20x anti-CD3 bispecific antibodies are human antibodies of the isotype IgG1, IgG2, IgG3, or IgG4, or mixed isotypes. In some embodiments, the anti-CD20x anti-CD3 bispecific antibodies are human IgG1 antibodies (i.e., the antibodies have a first antigen-binding domain and a second antigen-binding domain). In some embodiments, the anti-CD20x anti-CD3 bispecific antibody is a human IgG4 antibody (i.e., the antibody comprises a human IgG4 heavy chain constant region attached to the HCVR of each of the first antigen-binding domain and the second antigen-binding domain, respectively). In any of the embodiments described above or discussed herein, the anti-CD20x anti-CD3 bispecific antibody may comprise a human kappa light chain. In any of the embodiments described above or discussed herein, the anti-CD20x anti-CD3 bispecific antibody may comprise a human lambda light chain.

[0074] In any embodiment, the bispecific antibody may comprise a modification in one or both heavy chains to facilitate purification of the bispecific antibody (i.e., heterodimer) from homodimeric impurities. In some embodiments, the bispecific antibody comprises first and second heavy chains (i.e., the heavy chain of the anti-CD20 binding arm and the heavy chain of the anti-CD3 binding arm) that are identical (e.g., both isotype IgG1 or IgG4) except for a modification in the CH3 domain of one or the other heavy chain that reduces binding of the bispecific antibody to Protein A compared to an antibody lacking the modification. In some cases, the CH3 domain of the first heavy chain (e.g., of the anti-CD20 binding arm) binds to Protein A, and the CH3 domain of the second heavy chain (e.g., of the anti-CD3 binding arm) comprises a mutation that reduces or eliminates Protein A binding. In some cases, the mutation is an H435R modification (according to EU numbering; H95R according to IMGT exon numbering). In some cases, the mutations are H435R modifications (according to EU numbering; H95R according to IMGT exon numbering) and Y436F modifications (according to EU numbering; Y96F according to IMGT). Additional modifications that may be found within the second CH3 domain include D356E, L358M, N384S, K392N, V397M, and V422I according to EU (D16E, L18M, N44S, K52N, V57M, and V82I according to IMGT) for IgG1 CH3 domains, and Q355R, N384S, K392N, V397M, R409K, E419Q, and V422I according to EU (Q15R, N44S, K52N, V57M, R69K, E79Q, and V82I according to IMGT) for IgG4 CH3 domains.

[0075] In any embodiment, a bispecific antibody may comprise a chimeric hinge. The term "chimeric hinge" is intended to include a chimeric protein comprising a first amino acid sequence derived from the hinge region of one Ig molecule and a second amino acid sequence derived from the hinge region of a different class or subclass of Ig molecule. For example, a chimeric hinge, in one embodiment, comprises a first amino acid sequence or "upper hinge" sequence derived from a human IgG1 hinge region or a human IgG4 hinge region and a second amino acid sequence or "lower hinge" sequence derived from a human IgG2 hinge region. In certain embodiments, the first or "upper hinge" sequence comprises amino acid residues at positions 216-227 according to EU numbering. In some embodiments, the second or "lower hinge" sequence comprises amino acid residues at positions 228-236 according to EU numbering.

[0076] The antibodies of the present invention may, in some embodiments, be recombinant human antibodies. As used herein, the term "recombinant human antibody" is intended to include all human antibodies prepared, expressed, created, or isolated by recombinant means, such as antibodies expressed using a recombinant expression vector transfected into a host cell, antibodies isolated from a recombinant combinatorial human antibody library, antibodies isolated from an animal (e.g., a mouse) that is transgenic for human immunoglobulin genes (see, e.g., Taylor et al. (1992) Nucl. Acids Res. 20:6287-6295), or antibodies prepared, expressed, created, or isolated by any other means, including splicing of human immunoglobulin gene sequences into other DNA sequences. Such recombinant human antibodies contain variable and constant sequences derived from human germline immunoglobulin sequences. However, in certain embodiments, such recombinant human antibodies are subjected to in vitro mutagenesis (or in vivo somatic mutagenesis when animals transgenic for human Ig sequences are used), thus modifying the V and V regions of the recombinant antibody. H and V L The amino acid sequence of the region is human germline V H and VL These are sequences that are derived from and related to sequences, but which may not naturally occur within the human antibody germline repertoire in vivo.

[0077] The term "antigen-binding portion" of an antibody or "antibody fragment," as used herein, refers to one or more fragments of an antibody that retain the ability to specifically bind to anti-CD20 or anti-CD3.

[0078] An "isolated antibody," as used herein, is intended to refer to an antibody that is substantially free of other antibodies having different antigen specificities (e.g., an isolated bispecific antibody that specifically binds anti-CD20 or anti-CD3 is substantially free of antibodies that specifically bind to antigens other than anti-CD20 or anti-CD3).

[0079] "Specifically binds" or similar terms means that an antibody or antigen-binding fragment thereof forms a complex with an antigen that is relatively stable under physiological conditions. Specific binding is at least about 1 x 10 -6 The specific binding of an antibody to an anti-CD20 or anti-CD3 molecule may be characterized by a dissociation constant of M or greater. Methods for determining whether two molecules specifically bind are well known in the art and include, for example, equilibrium dialysis, surface plasmon resonance, and the like. However, an isolated antibody that specifically binds to anti-CD20 or anti-CD3 may have cross-reactivity to other antigens, such as CD20 or CD3 molecules from other species (orthologs). In the context of the present invention, multispecific (e.g., bispecific) antibodies that bind to human CD20 and human CD3, as well as one or more additional antigens, are considered to "specifically bind" to human CD20 and human CD3. Furthermore, an isolated antibody may be substantially free of other cellular material and / or chemicals.

[0080] Exemplary anti-CD20 x anti-CD3 bispecific antibodies that can be included in the pharmaceutical formulations of the present invention are described in WO2014 / 047231, the disclosure of which is incorporated by reference in its entirety.

[0081] According to certain embodiments of the invention, an anti-CD20×anti-CD3 bispecific antibody, or antigen-binding fragment thereof, comprises a first antigen-binding domain that specifically binds to human CD20 and a second antigen-binding domain that specifically binds to human CD3, wherein the first antigen-binding domain comprises heavy chain complementarity-determining regions (CDRs) A1-HCDR1, A1-HCDR2, and A1-HCDR3 comprising the amino acid sequences of SEQ ID NOs: 7, 8, and 9, respectively, and the second antigen-binding domain comprises heavy chain CDRs A2-HCDR1, A2-HCDR2, and A2-HCDR3 comprising the amino acid sequences of SEQ ID NOs: 10, 11, and 12, respectively. According to certain embodiments of the invention, an anti-CD20×anti-CD3 bispecific antibody, or antigen-binding fragment thereof, comprises a common (for both the first and second antibody-binding domains) light chain complementarity-determining region LCDR1-LCDR2-LCDR3 comprising the amino acid sequences of SEQ ID NOs: 13, 14, and 15, respectively.

[0082] In certain embodiments, an anti-CD20x anti-CD3 bispecific antibody, or antigen-binding fragment thereof, comprises a first antigen-binding domain that specifically binds human CD20 and a second antigen-binding domain that specifically binds human CD3, wherein the first antigen-binding domain comprises a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 4, and the second antigen-binding domain comprises a HCVR comprising the amino acid sequence of SEQ ID NO: 5. In certain embodiments, an anti-CD20x anti-CD3 bispecific antibody, or antigen-binding fragment thereof, comprises a common light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 6. In certain embodiments, an anti-CD20x anti-CD3 bispecific antibody, or antigen-binding fragment thereof, comprises a common light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 4 / 6. and a second antigen-binding domain comprising an HCVR / LCVR amino acid sequence pair comprising the amino acid sequence of SEQ ID NO:5 / 6. In some embodiments, the anti-CD20x anti-CD3 bispecific antibody comprises the HCVR / LCVR sequence pair described above and a human IgG1 heavy chain constant region. In some embodiments, the anti-CD20x anti-CD3 bispecific antibody comprises the HCVR / LCVR sequence pair described above and a human IgG4 heavy chain constant region. In some embodiments, the anti-CD20x anti-CD3 bispecific antibody comprises the HCVR / LCVR sequence pair described above and a human IgG heavy chain constant region. In some embodiments, the anti-CD20x anti-CD3 bispecific antibody comprises the HCVR / LCVR sequence pair described above and a human IgG1 or IgG4 heavy chain constant region. In some embodiments, an anti-CD20x anti-CD3 bispecific antibody comprises a first heavy chain comprising the amino acid sequence of SEQ ID NO: 1, a second heavy chain comprising the amino acid sequence of SEQ ID NO: 2, and a common light chain comprising the amino acid sequence of SEQ ID NO: 3. An exemplary anti-CD20x anti-CD3 bispecific antibody having a first antigen-binding domain that specifically binds to human CD20 and comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 4 and an LCVR comprising the amino acid sequence of SEQ ID NO: 6, and a second antigen-binding domain that specifically binds to human CD3 and comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 5 and an LCVR comprising the amino acid sequence of SEQ ID NO: 6, is referred to herein as REGN1979. In some embodiments, the bispecific antibody has a first heavy chain (comprising an HCVR that specifically binds to human CD20) comprising the amino acid sequence of SEQ ID NO: 1, a second heavy chain (comprising an HCVR that specifically binds to human CD3) the amino acid sequence of SEQ ID NO: 2, and a common light chain comprising the amino acid sequence of SEQ ID NO: 3. In some cases, the mature form of the antibody may not comprise the C-terminal lysine residues of SEQ ID NOs: 1 and 2. Thus, in some cases, the anti-CD20 binding arm of the antibody comprises a heavy chain comprising residues 1-452 of SEQ ID NO:1, and the anti-CD3 binding arm of the antibody comprises a heavy chain comprising residues 1-448 of SEQ ID NO:2.

[0083] The amount of antibody or antigen-binding fragment thereof contained in the pharmaceutical formulation of the present invention can vary depending on the desired specific properties of the formulation, as well as the particular situation and purpose for which the formulation is intended to be used. In certain embodiments, the pharmaceutical formulation can contain about 0.1 mg / mL to about 500 mg / mL of antibody, about 0.5 mg / mL to about 400 mg / mL of antibody, about 1 mg / mL to about 200 mg / mL of antibody, about 2 mg / mL to about 100 mg / mL of antibody, about 1 mg / mL to about 5 mg / mL of antibody, about 10 mg / mL to about 30 mg / mL of antibody, about 75 mg / mL to about 125 mg / mL of antibody, about 5 mg / mL to about 50 mg / mL of antibody, or about 2 mg / mL to about 160 mg / mL of antibody.For example, the formulations of the present invention may contain at least one of the following: about 0.5 mg / mL, about 1 mg / mL, about 2 mg / mL, about 3 mg / mL, about 4 mg / mL, about 5 mg / mL, about 6 mg / mL, about 7 mg / mL, about 8 mg / mL, about 9 mg / mL, about 10 mg / mL, about 11 mg / mL, about 12 mg / mL, about 13 mg / mL, about 14 mg / mL, about 15 mg / mL, about 16 mg / mL, about 17 mg / mL, and about 18 mg / mL, respectively, that specifically bind to human CD20 and human CD3. g / mL, approximately 18 mg / mL, approximately 19 mg / mL, approximately 20 mg / mL, approximately 21 mg / mL, approximately 22 mg / mL, approximately 23 mg / mL, approximately 24 mg / mL, approximately 25 mg / mL, approximately 26 mg / mL, approximately 27 mg / m L, about 28 mg / mL, about 29 mg / mL, about 30 mg / mL, about 35 mg / mL, about 40 mg / mL, about 45 mg / mL, about 50 mg / mL, about 55 mg / mL, about 60 mg / mL, about 65 mg / mL, about 70mg / mL, about 75mg / mL, about 80mg / mL, about 85mg / mL, about 90mg / mL, about 95mg / mL, about 96mg / mL, about 97mg / mL, about 98mg / mL, about 99mg / mL, about 100 mg / mL, about 101 mg / mL, about 102 mg / mL, about 103 mg / mL, about 104 mg / mL, about 105 mg / mL, about 110 mg / mL, about 115 mg / mL, about 120 mg / mL, about 125 mg / mL The liquid formulation may comprise about 130 mg / mL, about 135 mg / mL, about 140 mg / mL, about 145 mg / mL, about 150 mg / mL, about 155 mg / mL, about 160 mg / mL, about 165 mg / mL, about 170 mg / mL, about 175 mg / mL, about 180 mg / mL, about 185 mg / mL, about 190 mg / mL, about 195 mg / mL, or about 200 mg / mL of the antibody or antigen-binding fragment thereof. In certain embodiments, the pharmaceutical formulation is a liquid formulation that can contain 1±0.1 mg / mL to 200±20 mg / mL of antibody, 2±0.2 mg / mL to 100±10 mg / mL of antibody, 1±0.5 mg / mL to 30±5 mg / mL of antibody, 10±1 mg / mL to 30±3 mg / mL of antibody, 1±0.1 mg / mL to 3±0.3 mg / mL of antibody, 15±1.5 mg / mL to 25±2.5 mg / mL of antibody, 90±5 mg / mL to 110±5 mg / mL of antibody, or 150±7.5 mg / mL to 170±7.5 mg / mL of antibody. In some embodiments, the pharmaceutical formulation contains 2±0.2 mg / mL of antibody. In some embodiments, the pharmaceutical formulation contains 20±2 mg / mL of antibody. In some embodiments, the pharmaceutical formulation contains 80±8 mg / mL of antibody. In some embodiments, the pharmaceutical formulation comprises 100±10 mg / ml of antibody.

[0084] biological equivalent The present invention encompasses antibodies having amino acid sequences that differ from those of the exemplary molecules disclosed herein but that retain the ability to bind to human CD20 and human CD3. Such variant molecules may contain one or more additions, deletions, or substitutions of amino acids when compared to the parent sequence, but may exhibit biological activity that is essentially equivalent to that of the antibodies discussed herein.

[0085] The present invention includes antigen-binding molecules that are biologically equivalent to any of the exemplary antibodies defined herein.For example, when administered at the same molar dose under similar experimental conditions, either in a single dose or multiple doses, two antibodies are considered to be pharmaceutical equivalents or pharmaceutical substitutes if they do not show significant differences in absorption rate and extent.Some antibodies will be considered to be equivalents or pharmaceutical substitutes if they are equivalent in their absorption level but not equivalent in their absorption rate, but such differences in absorption rate are intentional and reflected in labeling, and can be considered to be biologically equivalent because, for example, they are not necessary for achieving effective body drug concentration during chronic use and are not considered medically significant for the specific pharmaceutical product being tested.

[0086] In one embodiment, two antibodies are bioequivalent if there are no clinically meaningful differences in their safety, purity, and potency.

[0087] In one embodiment, two antibodies are bioequivalent if a patient can be switched one or more times between the reference product and the biological product without a clinically significant change in immunogenicity or an expected increase in risk of adverse effects, including attenuation of efficacy, compared to maintenance therapy without such switching.

[0088] Bioequivalence can be demonstrated by in vivo and in vitro methods. Bioequivalence measurements include, for example, (a) in vivo studies in humans or other mammals in which the concentration of an antibody or its metabolites is measured as a function of time in blood, plasma, serum, or other biological fluids, (b) in vitro studies that correlate with and reasonably predict human in vivo bioavailability data, (c) in vivo studies in humans or other mammals in which the relevant acute pharmacological effects of the antibody (or its target) are measured as a function of time, and (d) well-controlled clinical studies that establish the safety, efficacy, or bioavailability or bioequivalence of the antigen-binding protein.

[0089] Formulation excipients and pH The pharmaceutical formulations of the present invention include one or more excipients. As used herein, the term "excipient" refers to any non-therapeutic agent added to a formulation to provide a desired consistency, viscosity, or stabilizing effect.

[0090] In certain embodiments, the pharmaceutical formulations of the present invention have a viscosity of less than 15 cP, which is advantageous for delivering the composition from a pre-filled syringe or auto-injector. In some cases, the pharmaceutical formulations have a viscosity of less than 14 cP, less than 13 cP, less than 12 cP, less than 11 cP, less than 10 cP, less than 9 cP, less than 8 cP, less than 7 cP, less than 6 cP, less than 5 cP, less than 4 cP, less than 3 cP, less than 2 cP, or less than 1.5 cP at 20°C when measured using a capillary viscometer (e.g., as discussed in Example 3). In some embodiments, the pharmaceutical formulations have a viscosity of less than 15 cP at antibody concentrations up to 150 mg / ml when measured using a capillary viscometer (e.g., as discussed in Example 3) at 20°C when measured using a capillary viscometer (e.g., as discussed in Example 3). In some embodiments, the pharmaceutical formulations have a viscosity of less than 5 cP at antibody concentrations up to 100 mg / ml when measured using a capillary viscometer (e.g., as discussed in Example 3). In some embodiments, the pharmaceutical formulation has a viscosity of less than 2 cP at antibody concentrations up to 20 mg / ml at 20° C., as measured using a capillary viscometer (e.g., as discussed in Example 3).

[0091] In certain embodiments, the pharmaceutical formulations of the present invention contain one or more carbohydrates, e.g., one or more sugars. The sugars can be reducing or non-reducing sugars. "Reducing sugars" include, for example, sugars with a ketone or aldehyde group, which contain a reactive hemiacetal group that allows the sugar to function as a reducing agent. Specific examples of reducing sugars include fructose, glucose, glyceraldehyde, lactose, arabinose, mannose, xylose, ribose, rhamnose, galactose, and maltose. Non-reducing sugars are acetals and can contain an anomeric carbon that does not substantially react with amino acids or polypeptides to initiate the Maillard reaction. Specific examples of non-reducing sugars include sucrose, trehalose, sorbose, sucralose, melezitose, and raffinose. Sugar acids include, for example, saccharinic acid, gluconate, and other polyhydroxy sugars and their salts. In some embodiments, the sugar is sucrose. In some cases, a sugar (eg, sucrose) functions as a thermal stabilizer for the anti-CD20 x anti-CD3 bispecific antibody.

[0092] The amount of sugar (e.g., sucrose) contained in the pharmaceutical formulations of the present invention will vary depending on the specific context and intended purpose of the formulation. In certain embodiments, the formulation may contain about 0.1% to about 20% sugar, about 0.5% to about 20% sugar, about 1% to about 20% sugar, about 2% to about 15% sugar, about 5% to about 15% sugar, about 7.5% to about 12.5% ​​sugar, or about 9% to about 11% sugar. For example, the pharmaceutical formulations of the present invention may contain about 0.5%, about 1.0%, about 1.5%, about 2.0%, about 2.5%, about 3.0%, about 3.5%, about 4.0%, about 4.5%, about 5.0%, about 5.5%, about 6.0%, about 6.5%, about 7.0%, about 7.5%, about 8.0%, about 8.5%, about 9.0%, about 9.5%, about 10.0%, about 10.5%, about 11.0%, about 11.5%, about 12.0%, about 12.5%, about 13.0%, about 13.5%, about 14.0%, about 14.5%, about 15%, or about 20% sugar (e.g., sucrose). In some embodiments, the formulation contains about 10% sugar (e.g., sucrose). Each of the above percentages corresponds to a weight / volume (w / v) percent. In some cases, the formulation contains 5%±1% to 20%±4% w / v sucrose. In some cases, the formulation contains 8%±0.5% to 12%±0.5% w / v sucrose. In some cases, the formulation contains 10%±1% w / v sucrose.

[0093] The pharmaceutical formulations of the present invention may also include one or more organic co-solvents (or interfacial stabilizers) of a type and amount that stabilizes the anti-CD20 x anti-CD3 bispecific antibody under conditions of rough handling or agitation, such as, for example, orbital shaking. In some embodiments, the organic co-solvent is a surfactant. As used herein, the term "surfactant" refers to a substance that reduces the surface tension of the fluid in which it is dissolved and / or reduces the interfacial tension between oil and water. Surfactants can be ionic or non-ionic. Exemplary non-ionic surfactants that can be included in the formulations of the present invention are, for example, alkyl acrylates, ... Examples of suitable nonionic surfactants include poly(ethylene oxide), alkyl polyglucosides (e.g., octyl glucoside and decyl maltoside), fatty alcohols such as cetyl alcohol and oleyl alcohol, cocamide MEA, cocamide DEA, and cocamide TEA. Specific nonionic surfactants that can be included in the formulations of the present invention include polysorbates such as polysorbate 20, polysorbate 28, polysorbate 40, polysorbate 60, polysorbate 65, polysorbate 80, polysorbate 81, and polysorbate 85; poloxamers such as poloxamer 188 (also known as Pluronic F68) and poloxamer 407; polyethylene-polypropylene glycol; or polyethylene glycol (PEG). Polysorbate 20 is also known as TWEEN 20, sorbitan monolaurate, and polyoxyethylene sorbitan monolaurate. In some embodiments, the surfactant is polysorbate 80.

[0094] The amount of surfactant contained within the pharmaceutical formulations of the present invention can vary depending on the specific desired properties of the formulation, as well as the particular situation and purpose for which the formulation is intended to be used. In certain embodiments, the formulation may contain 0.01% to about 1% surfactant, about 0.01% to about 0.5%, about 0.05% to about 0.15%, about 0.08% to about 0.12%, or about 0.09% to about 0.11% surfactant. For example, the formulations of the present invention may contain about 0.01%, about 0.02%, about 0.03%, about 0.04%, about 0.05%, about 0.06%, about 0.07%, about 0.08%, about 0.09%, about 0.10%, about 0.11%, about 0.12%, about 0.13%, about 0.14%, about 0.15%, about 0.16%, about 0.17%, about 0.18%, about 0.19%, about 0.20%, about 0.21%, about 0.22%, about 0.23%, about 0.24%, about 0.25%, about 0.26%, about 0.27%, about 0.28%, about 0.29%, or about 0.30% surfactant (e.g., polysorbate 80). In some embodiments, the formulation contains about 0.1% surfactant (e.g., polysorbate 80). Each of the above percentages corresponds to a weight / volume (w / v) percent. In some embodiments, the formulation contains 0.01%±0.005% to 0.5%±0.25% w / v polysorbate 80. In some embodiments, the formulation contains 0.1%±0.05% w / v polysorbate 80. In some embodiments, the formulation contains 0.1%±0.01% w / v polysorbate 80.

[0095] The pharmaceutical formulations of the present invention may also include a buffer or buffer system that helps maintain a stable pH and stabilize the anti-CD20 x anti-CD3 bispecific antibody. In some embodiments, the buffer or buffer system includes at least one buffer with a buffer range that completely or partially overlaps the pH range of 5.5 to 6.1. In certain embodiments, the buffer includes a histidine buffer. In certain embodiments, the buffer (e.g., histidine) is present at a concentration of about 1 mM to about 40 mM, about 1 mM to about 30 mM, about 1 mM to about 20 mM, about 3 mM to about 18 mM, about 5 mM to about 15 mM, or about 8 mM to about 12 mM. In some embodiments, the buffer (e.g., histidine) is present at a concentration of about 1 mM, about 2 mM, about 3 mM, about 4 mM, about 5 mM, about 6 mM, about 7 mM, about 8 mM, about 9 mM, about 10 mM, about 11 mM, about 12 mM, about 13 mM, about 14 mM, about 15 mM, about 16 mM, about 17 mM, about 18 mM, about 19 mM, or about 20 mM. In some embodiments, the formulation contains a histidine buffer at a concentration of 5 mM ± 1 mM to 15 mM ± 3 mM. In some embodiments, the histidine buffer comprises L-histidine and L-histidine monohydrochloride monohydrate. In some embodiments, the histidine buffer comprises L-histidine at a concentration of 3.65 mM ± 0.5 mM and L-histidine monohydrochloride monohydrate at a concentration of 6.35 mM ± 0.5 mM. In some embodiments, the histidine buffer comprises L-histidine at a concentration of 3.65 mM ± 0.2 mM and L-histidine monohydrochloride monohydrate at a concentration of 6.35 mM ± 0.2 mM. In some embodiments, the formulation contains an acetate buffer (e.g., at any of the concentrations described above or discussed herein). In some embodiments, the formulation contains a phosphate buffer. The buffer solution contains (eg, at any of the concentrations described above or discussed herein).

[0096] During the antibody purification process, it may be desirable or necessary to exchange one buffer for another to achieve appropriate excipient concentrations, antibody concentrations, pH, etc. Buffer exchange can be achieved, for example, by ultrafiltration / diafiltration (UF / DF) using semipermeable tangential flow filtration membranes. However, using such techniques can result in the Gibbs-Donnan effect (Bolton et al., 2011, Biotechnol. Prog. 27(1):140-152). The buildup of positive charge on the product side of the membrane during protein concentration is electrically counterbalanced by the preferential migration of positive ions to the opposite side of the membrane. A potential consequence of this phenomenon is that the final concentration of certain components (e.g., histidine) may be lower than the intended target concentration of these components due to the electrostatic repulsion of positively charged diafiltration buffer excipients toward the positively charged antibody protein during the UF / DF step. Thus, the present invention includes formulations in which the concentration of histidine varies from the amounts or ranges set forth herein due to, for example, the Gibbs-Donnan effect.

[0097] Volume exclusion describes the behavior of highly concentrated samples in which a significant portion of the total volume of a solution is occupied by solutes, particularly large molecules such as proteins, which have excluded solvent from this space. This then reduces the total amount of solvent available for dissolving other solutes, potentially resulting in unequal distribution across the ultrafiltration membrane. Thus, the present invention includes formulations in which the concentration of, for example, histidine may vary from the amounts or ranges described herein due to volume exclusion effects.

[0098] During the manufacture of the formulations of the invention, variations in the composition of the formulation may occur. These variations may include the concentration of the active ingredient, the concentration of the excipients, and / or the pH of the formulation. The invention includes formulations comprising anti-CD20xanti-CD3 bispecific antibodies that are stable and retain potency with up to at least a 10% variation in excipient concentration. For example, included herein are anti-CD20xanti-CD3 bispecific antibody formulations in which the stability and potency of the formulation are not affected by ±10% or ±20% variations in the concentrations of the antibody, sucrose, histidine buffer, and / or polysorbate.

[0099] Stability of pharmaceutical formulations The pharmaceutical formulations of the present invention exhibit a high level of stability. As used herein with respect to pharmaceutical formulations, the term "stable" means that the antibody in the pharmaceutical formulation retains an acceptable degree of structure and / or function and / or biological activity after storage for a defined amount of time. A formulation can be stable even if the antibody contained therein does not maintain 100% of its structure and / or function and / or biological activity after storage for a defined amount of time. Under certain circumstances, maintenance of about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% of the structure and / or function and / or biological activity of the antibody after storage for a defined amount of time can be considered "stable".

[0100] Stability can be measured, among other things, by determining the percentage of native antibody remaining in the formulation after storage at a given temperature for a defined amount of time. The percentage of native antibody can be determined, among other things, by size exclusion chromatography (e.g., size exclusion high performance liquid chromatography [SE-HPLC]). "Acceptable degree of stability," as the phrase is used herein, means that at least 90% of the native form of the antibody can be detected in the formulation after storage at a given temperature for a defined amount of time. In certain embodiments, at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% of the native form of the antibody can be detected in the formulation after storage at a given temperature for a defined amount of time. Stability can be measured. The defined amount of time after the determination can be at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 12 months, at least 18 months, at least 24 months, at least 30 months, at least 36 months, or more. The temperature at which the pharmaceutical formulation can be stored when assessing stability can be any temperature between about -80°C and about 45°C, e.g., storage at about -80°C, about -30°C, about -20°C, about 0°C, about 4°C to 8°C, about 5°C, about 25°C, about 35°C, about 37°C, or about 45°C. For example, a pharmaceutical formulation can be considered stable if greater than about 90%, 95%, 96%, or 97% of the native antibody is detected by SE-HPLC after 3 months of storage at 5°C. A pharmaceutical formulation may also be considered stable if greater than about 90%, 95%, 96%, or 97% of the native antibody is detected by SE-HPLC after 6 months of storage at 5° C. A pharmaceutical formulation may also be considered stable if greater than about 90%, 95%, 96%, 96.5%, 97%, 97.5%, 98%, 98.5%, 99%, or 99.5% of the native antibody is detected by SE-HPLC after 9 months of storage at 5° C. A pharmaceutical formulation may also be considered stable if greater than about 90%, 95%, 96%, 96.5%, 97%, 97.5%, 98%, 98.5%, 99%, or 99.5% of the native antibody is detected by SE-HPLC after 12 months of storage at 5° C. A pharmaceutical formulation may also be considered stable if greater than about 90%, 95%, 96%, 96.5%, 97%, 97.5%, 98%, 98.5%, 99%, or 99.5% of the native antibody is detected by SE-HPLC after 24 months of storage at 5° C. A pharmaceutical formulation may also be considered stable if greater than about 90%, 95%, 96%, 96.5%, 97%, 97.5%, 98%, 98.5%, 99%, or 99.5% of the native antibody is detected by SE-HPLC after 36 months of storage at 5° C.A pharmaceutical formulation may also be considered stable if greater than about 90%, 95%, 96%, 96.5%, 97%, 97.5%, 98%, 98.5%, 99%, or 99.5% of the native antibody is detected by SE-HPLC after three months of storage at 25° C. A pharmaceutical formulation may also be considered stable if greater than about 90%, 95%, 96%, 96.5%, 97%, 97.5%, 98%, 98.5%, 99%, or 99.5% of the native antibody is detected by SE-HPLC after six months of storage at 25° C. A pharmaceutical formulation may also be considered stable if greater than about 90%, 95%, 96%, 96.5%, 97%, 97.5%, 98%, 98.5%, 99%, or 99.5% of the native antibody is detected by SE-HPLC after nine months of storage at 25° C. A pharmaceutical formulation may also be considered stable if greater than about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97% or 98% of the native antibody is detected by SE-HPLC after 1 month of storage at 45°C.

[0101] Other methods, such as differential scanning calorimetry (DSC) to determine thermal stability, controlled stirring to determine mechanical stability, and absorbance at about 350 nm or about 405 nm to determine solution turbidity, can be used to assess the stability of the formulations of the invention. For example, the OD of a formulation after storage at about 5°C to about 25°C for 6 months or more can be measured. 405 The change in OD of the formulation at t=0 405 to less than about 0.05 (e.g., 0.04, 0.03, 0.02, 0.01, or less), the formulations of the present invention can be considered stable.

[0102] Measuring the binding affinity of an antibody to a target can also be used to assess stability. For example, a formulation of the invention is considered stable if, after storage for a defined amount of time (e.g., 14 days to 9 months) at, e.g., -80°C, -30°C, -20°C, 5°C, 25°C, 37°C, 45°C, etc., the anti-CD20 x anti-CD3 bispecific antibody contained within the formulation binds to human CD20 and human CD3 with an affinity that is at least 80%, 85%, 90%, 95%, or greater than the binding affinity of the antibody prior to storage. Binding affinity can be determined by any method, e.g., ELISA or plasmon resonance. Biology Antibody activity can be determined by a CD20 or CD3 activity assay, such as by contacting cells expressing CD20 or CD3 with a formulation containing an anti-CD20 x anti-CD3 bispecific antibody. Binding of the antibody to such cells can be measured directly, such as via FACS analysis.

[0103] Stability can be measured, inter alia, by determining the percentage of antibody that forms aggregates (high molecular weight (HMW) species within the formulation) after storage for a defined amount of time at a defined temperature, with stability being inversely proportional to the percent aggregates formed. The percentage of aggregated antibody can be determined, inter alia, by size exclusion chromatography (e.g., size exclusion high performance liquid chromatography [SE-HPLC] or size exclusion ultra performance liquid chromatography [SE-UPLC]). "Acceptable degree of stability," as that phrase is used herein, means that up to 6% of the antibody is in a detectable aggregated form in the formulation after storage for a defined amount of time at a given temperature. In certain embodiments, acceptable degree of stability means that up to about 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of the antibody can be detected in aggregates in the formulation after storage for a defined amount of time at a given temperature. The defined amount of time after which stability is measured can be at least 2 weeks, at least 28 days, at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 12 months, at least 18 months, at least 24 months, at least 30 months, at least 36 months, or more. The temperature at which the pharmaceutical formulation can be stored when assessing stability can be any temperature between about -80°C and about 45°C, for example, about -80°C, about -30°C, about -20°C, about 0°C, about 4°C to 8°C, about 5°C, about 25°C, about 35°C, about 37°C, or about 45°C. For example, a pharmaceutical formulation may be considered stable if less than about 2%, 1.75%, 1.5%, 1.25%, 1%, 0.75%, 0.5%, 0.25%, or 0.1% of the antibody is detected in aggregate form after 9 months of storage at 5° C. A pharmaceutical formulation may also be considered stable if less than about 2%, 1.75%, 1.5%, 1.25%, 1%, 0.75%, 0.5%, 0.25%, or 0.1% of the antibody is detected in aggregate form after 3 months of storage at 25° C. and 60% relative humidity.A pharmaceutical formulation may also be considered stable if less than about 6%, 5.9%, 5.8%, 5.7%, 5.6%, 5.5%, 5%, 4.5%, 4%, 3.5%, 3%, 2.5%, 2%, 1.5%, 1%, 0.5%, or 0.1% of the antibody is detected in aggregated form after 1 month of storage at 45° C. A pharmaceutical formulation may also be considered stable if less than about 2%, 1.9%, 1.8%, 1.7%, 1.6%, 1.5%, 1%, 0.5%, or 0.1% of the antibody is detected in aggregated form after 3 months of storage at −20° C., −30° C., or −80° C.

[0104] Stability can be measured, inter alia, by determining the percentage of antibody that migrates in the acidic fraction ("acidic form") during ion exchange rather than the main fraction ("mainly charged form") of the antibody, with stability being inversely proportional to the fraction of antibody in acidic form. Without wishing to be bound by theory, deamidation of an antibody can make the antibody more negatively charged and therefore more acidic relative to non-deamidated antibody (e.g., Robinson, N., Protein Deamidation, PNAS, April 16, 2002, 99(8):5283-5288). The percentage of "acidified" antibody can be determined by ion exchange chromatography (e.g., cation exchange high-performance liquid chromatography [CEX-HPLC] or cation exchange ultra-performance liquid chromatography [CEX-UPLC]). "Acceptable degree of stability," as that phrase is used herein, means that up to 52% of the antibody is in the more acidic form detected in the formulation after storage at a defined temperature for a defined amount of time. In certain embodiments, an acceptable degree of stability is achieved by up to about 52%, 50%, 45%, 40%, 35%, 30%, 29%, 28%, 27%, 26%, 25%, 20%, 15%, 10%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 1.5% of the antibody. This means that at least 0.1%, or 0.1%, of the hydroxybenzoates can be detected in the acidic form in the formulation after storage at a given temperature for a defined amount of time. The defined amount of time after which stability is measured can be at least 2 weeks, at least 28 days, at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 12 months, at least 18 months, at least 24 months, at least 30 months, at least 36 months, or more. The temperature at which the pharmaceutical formulation can be stored when assessing stability can be any temperature between about -80°C and about 45°C, such as about -80°C, about -30°C, about -20°C, about 0°C, about 4°C to 8°C, about 5°C, about 25°C, or about 45°C. For example, a pharmaceutical formulation can be considered stable if less than about 29%, 28%, 27%, 26%, 25%, 24%, 23%, 22%, 21%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of the antibody is in the more acidic form after 3 months of storage at -80°C, -30°C, or -20°C. A pharmaceutical formulation may also be considered stable if less than about 28%, 27%, 26%, 25%, 24%, 23%, 22%, 21%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of the antibody is in the more acidic form after 9 months of storage at 5°C. A pharmaceutical formulation may also be considered stable if less than about 30%, 29%, 28%, 27%, 26%, 25%, 24%, 23%, 22%, 21%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of the antibody is in the more acidic form after 28 days of storage at 25°C.A pharmaceutical formulation may also be considered stable if less than about 37%, 36%, 35%, 34%, 33%, 32%, 31%, 30%, 29%, 28%, 27%, 26%, 25%, 24%, 23%, 22%, 21%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of the antibody is in the more acidic form after 28 days of storage at 37°C. A pharmaceutical formulation is also considered stable if less than about 52%, 51%, 50%, 49%, 48%, 47%, 46%, 45%, 44%, 43%, 42%, 41%, 40%, 39%, 38%, 37%, 36%, 35%, 34%, 33%, 32%, 31%, 30%, 29%, 28%, 27%, 26%, 25%, 24%, 23%, 22%, 21%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of the antibody can be detected in the more acidic form after 28 days of storage at 45°C.

[0105] Reference to stability of a pharmaceutical formulation "after" a particular period of time is intended to mean that measurements of the stability parameter (e.g., % native form, % HMW species, or % acidic form) were made at or around the end of the particular period of time, and is not intended to mean that the pharmaceutical formulation will necessarily maintain the same degree of stability for the parameter measured thereafter. For example, a reference to a particular stability after 9 months means that the stability measurements were made at the start of the study or after about 9 months. Additional methods for assessing the stability of an antibody in a formulation are demonstrated in the Examples provided below.

[0106] As demonstrated in the Examples below, the present invention is based, in part, on the discovery that the combination of the claimed excipients with a bispecific anti-CD20 x anti-CD3 antibody produces a formulation that is stable.

[0107] Exemplary Formulations According to one aspect of the present invention, the pharmaceutical formulation comprises: (i) a human anti-CD20×anti-CD3 bispecific antibody that specifically binds human CD20 and human CD3; and (ii) a buffer containing histidine. (iii) an organic co-solvent comprising a polysorbate; and (iv) a stabilizer comprising a sugar.

[0108] In some cases, the stable liquid pharmaceutical formulation comprises: (i) a human bispecific antibody that specifically binds to human CD20 and human CD3, and comprises a first antigen-binding domain that specifically binds to human CD20, the first antigen-binding domain comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 4 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6, and a second antigen-binding domain that specifically binds to human CD3, the second antigen-binding domain comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 5 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6 at a concentration of about 2 mg / ml to about 200 mg / ml; (ii) histidine at a concentration of about 5 mM to about 15 mM; (iii) polysorbate 80 at a concentration of about 0.05% w / v to about 0.15% w / v; and (iv) sucrose at a concentration of about 5% w / v to about 15% w / v, wherein the formulation has a pH of 5.8±0.3.

[0109] In some cases, the stable liquid pharmaceutical formulation comprises: (i) a human bispecific antibody that specifically binds to human CD20 and human CD3, and comprises a first antigen-binding domain that specifically binds to human CD20, the first antigen-binding domain comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 4 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6, and a second antigen-binding domain that specifically binds to human CD3, the second antigen-binding domain comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 5 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6 at a concentration of about 2 mg / ml to about 200 mg / ml, wherein the antibody (ii) histidine at a concentration of about 5 mM to about 15 mM; (iii) polysorbate 80 at a concentration of about 0.05% w / v to about 0.15% w / v; and (iv) sucrose at a concentration of about 5% w / v to about 15% w / v, wherein the formulation has a pH of 5.8±0.3.

[0110] In some cases, the stable liquid pharmaceutical formulation comprises: (i) a human bispecific antibody that specifically binds to human CD20 and human CD3, and comprises a first antigen-binding domain that specifically binds to human CD20, the first antigen-binding domain comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 4 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6, and a second antigen-binding domain that specifically binds to human CD3, the second antigen-binding domain comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 5 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6 at a concentration of about 2 mg / ml to about 200 mg / ml; (ii) histidine at a concentration of about 5 mM to about 15 mM; (iii) polysorbate 80 at a concentration of about 0.05% w / v to about 0.15% w / v; and (iv) sucrose at a concentration of about 5% w / v to about 15% w / v, wherein the formulation has a pH of 5.8±0.3.

[0111] In some cases, the stable liquid pharmaceutical formulation comprises: (i) a human bispecific antibody that specifically binds to human CD20 and human CD3, and comprises a first heavy chain comprising the amino acid sequence of SEQ ID NO: 1, a second heavy chain comprising the amino acid sequence of SEQ ID NO: 2, and a common light chain comprising the amino acid sequence of SEQ ID NO: 3 at a concentration of about 2 mg / ml to about 200 mg / ml; (ii) histidine at a concentration of about 5 mM to about 15 mM; (iii) polysorbate 80 at a concentration of about 0.05% w / v to about 0.15% w / v; and (iv) sucrose at a concentration of about 5% w / v to about 15% w / v, wherein the formulation has a pH of 5.8±0.3.

[0112] In some cases, the stable liquid pharmaceutical formulation comprises: (i) a first antigen-binding domain that specifically binds to human CD20 and human CD3, and that specifically binds to human CD20, comprising an HCVR comprising the amino acid sequence of SEQ ID NO:4 and an LCVR comprising the amino acid sequence of SEQ ID NO:6; a human bispecific antibody comprising a second antigen-binding domain that specifically binds to human CD3, the second antigen-binding domain comprising an HCVR comprising the amino acid sequence of SEQ ID NO: 5 and an LCVR comprising the amino acid sequence of SEQ ID NO: 6 at a concentration of about 2 mg / ml to about 200 mg / ml; (ii) histidine at a concentration of about 8 mM to about 12 mM; (iii) polysorbate 80 at a concentration of about 0.075% w / v to about 0.125% w / v; and (iv) sucrose at a concentration of about 8% w / v to about 12% w / v; wherein the formulation has a pH of 5.8±0.3.

[0113] In some cases, the stable liquid pharmaceutical formulation comprises: (i) a human bispecific antibody that specifically binds to human CD20 and human CD3, and comprises a first antigen-binding domain that specifically binds to human CD20, the first antigen-binding domain comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 4 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6, and a second antigen-binding domain that specifically binds to human CD3, the second antigen-binding domain comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 5 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6 at a concentration of about 2 mg / ml to about 200 mg / ml, wherein the antibody is (ii) histidine at a concentration of about 8 mM to about 12 mM; (iii) polysorbate 80 at a concentration of about 0.075% w / v to about 0.125% w / v; and (iv) sucrose at a concentration of about 8% w / v to about 12% w / v; wherein the formulation has a pH of 5.8±0.3.

[0114] In some cases, the stable liquid pharmaceutical formulation comprises: (i) a human bispecific antibody that specifically binds to human CD20 and human CD3, and comprises a first antigen-binding domain that specifically binds to human CD20, the first antigen-binding domain comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 4 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6, and a second antigen-binding domain that specifically binds to human CD3, the second antigen-binding domain comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 5 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6 at a concentration of about 2 mg / ml to about 200 mg / ml, wherein the antibody (ii) histidine at a concentration of about 8 mM to about 12 mM; (iii) polysorbate 80 at a concentration of about 0.075% w / v to about 0.125% w / v; and (iv) sucrose at a concentration of about 8% w / v to about 12% w / v, wherein the formulation has a pH of 5.8±0.3.

[0115] In some cases, the stable liquid pharmaceutical formulation comprises: (i) a human bispecific antibody that specifically binds to human CD20 and human CD3, and comprises a first heavy chain comprising the amino acid sequence of SEQ ID NO: 1, a second heavy chain comprising the amino acid sequence of SEQ ID NO: 2, and a common light chain comprising the amino acid sequence of SEQ ID NO: 3 at a concentration of about 2 mg / ml to about 200 mg / ml; (ii) histidine at a concentration of about 8 mM to about 12 mM; (iii) polysorbate 80 at a concentration of about 0.075% w / v to about 0.125% w / v; and (iv) sucrose at a concentration of about 8% w / v to about 12% w / v, wherein the formulation has a pH of 5.8±0.3.

[0116] In some cases, the stable liquid pharmaceutical formulation comprises: (i) a human bispecific antibody that specifically binds to human CD20 and human CD3, and comprises a first antigen-binding domain that specifically binds to human CD20, the first antigen-binding domain comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 4 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6, and a second antigen-binding domain that specifically binds to human CD3, the second antigen-binding domain comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 5 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6 at a concentration of about 2 mg / ml to about 200 mg / ml; (ii) histidine at a concentration of 10 mM; (iii) polysorbate 80 at a concentration of 0.1% w / v; and (iv) sucrose at a concentration of 10% w / v, wherein the formulation has a pH of 5.8±0.3.

[0117] In some cases, the stable liquid pharmaceutical formulation comprises: (i) a human bispecific antibody that specifically binds to human CD20 and human CD3, and comprises a first antigen-binding domain that specifically binds to human CD20, the first antigen-binding domain comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 4 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6; and a second antigen-binding domain that specifically binds to human CD3, the second antigen-binding domain comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 5 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6 at a concentration of about 2 mg / ml to about 200 mg / ml, wherein the antibody has a heavy chain constant region of the IgG1 isotype (optionally, one of the two heavy chains has reduced Protein A binding compared to an unmodified heavy chain of the same isotype, and optionally one or both of the two heavy chains have a chimeric hinge); (ii) histidine at a concentration of 10 mM; (iii) polysorbate 80 at a concentration of 0.1% w / v; and (iv) sucrose at a concentration of 10% w / v; wherein the formulation has a pH of 5.8±0.3.

[0118] In some cases, the stable liquid pharmaceutical formulation comprises: (i) a human bispecific antibody that specifically binds to human CD20 and human CD3, and that specifically binds to human CD20, the first antigen-binding domain comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 4 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6; and a second antigen-binding domain that specifically binds to human CD3, the second antigen-binding domain comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 5 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6 at a concentration of about 2 mg / ml to about 200 mg / ml, wherein the antibody has a heavy chain constant region of the IgG4 isotype (optionally, one of the two heavy chains has reduced Protein A binding compared to an unmodified heavy chain of the same isotype, and optionally one or both of the two heavy chains have a chimeric hinge); (ii) histidine at a concentration of 10 mM; (iii) polysorbate 80 at a concentration of 0.1% w / v; and (iv) sucrose at a concentration of 10% w / v; wherein the formulation has a pH of 5.8±0.3.

[0119] In some cases, the stable liquid pharmaceutical formulation comprises: (i) a human bispecific antibody that specifically binds human CD20 and human CD3, and comprises a first heavy chain comprising the amino acid sequence of SEQ ID NO: 1, a second heavy chain comprising the amino acid sequence of SEQ ID NO: 2, and a common light chain comprising the amino acid sequence of SEQ ID NO: 3 at a concentration of about 2 mg / ml to about 200 mg / ml; (ii) histidine at a concentration of 10 mM; (iii) polysorbate 80 at a concentration of 0.1% w / v; and (iv) sucrose at a concentration of 10% w / v, wherein the formulation has a pH of 5.8±0.3.

[0120] In some cases, the stable liquid pharmaceutical formulation comprises: (i) a human bispecific antibody that specifically binds to human CD20 and human CD3, and that comprises a first antigen-binding domain that specifically binds to human CD20, the first antigen-binding domain comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 4 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6; and a second antigen-binding domain that specifically binds to human CD3, the HCVR comprising the amino acid sequence of SEQ ID NO: 5 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6 at a concentration of about 2 mg / ml ± 0.2 mg / ml to about 200 mg / ml ± 20 mg / ml; (ii) histidine at a concentration of 10 mM ± 1 mM; (iii) polysorbate 80 at a concentration of 0.1% ± 0.01 w / v; and (iv) sucrose at a concentration of 10% ± 1% w / v; wherein the formulation has a pH of 5.8 ± 0.3.

[0121] In some cases, the stable liquid pharmaceutical formulation comprises: (i) a human bispecific antibody that specifically binds to human CD20 and human CD3, and comprises a first antigen-binding domain that specifically binds to human CD20, the first antigen-binding domain comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 4 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6, and a second antigen-binding domain that specifically binds to human CD3, the HCVR comprising the amino acid sequence of SEQ ID NO: 5 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6 at a concentration of 2 mg / ml ± 0.2 mg / ml; (ii) histidine at a concentration of 10 mM ± 1 mM; (iii) polysorbate 80 at a concentration of 0.1% ± 0.01 w / v; and (iv) sucrose at a concentration of 10% ± 1% w / v, wherein the formulation has a pH of 5.8 ± 0.3.

[0122] In some cases, the stable liquid pharmaceutical formulation comprises: (i) a human bispecific antibody that specifically binds to human CD20 and human CD3, and comprises a first antigen-binding domain that specifically binds to human CD20, the first antigen-binding domain comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 4 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6, and a second antigen-binding domain that specifically binds to human CD3, the HCVR comprising the amino acid sequence of SEQ ID NO: 5 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6 at a concentration of 20 mg / ml ± 2 mg / ml; (ii) histidine at a concentration of 10 mM ± 1 mM; (iii) polysorbate 80 at a concentration of 0.1% ± 0.01 w / v; and (iv) sucrose at a concentration of 10% ± 1% w / v, wherein the formulation has a pH of 5.8 ± 0.3.

[0123] In some cases, the stable liquid pharmaceutical formulation comprises: (i) a human bispecific antibody that specifically binds to human CD20 and human CD3, and comprises a first antigen-binding domain that specifically binds to human CD20, the first antigen-binding domain comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 4 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6, and a second antigen-binding domain that specifically binds to human CD3, the HCVR comprising the amino acid sequence of SEQ ID NO: 5 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6 at a concentration of 100 mg / ml ± 10 mg / ml; (ii) histidine at a concentration of 10 mM ± 1 mM; (iii) polysorbate 80 at a concentration of 0.1% ± 0.01 w / v; and (iv) sucrose at a concentration of 10% ± 1% w / v, wherein the formulation has a pH of 5.8 ± 0.3.

[0124] In some cases, the stable liquid pharmaceutical formulation comprises: (i) a human bilayer comprising a first antigen-binding domain that specifically binds to human CD20 and human CD3, and that specifically binds to human CD20, the first antigen-binding domain comprising an HCVR comprising the amino acid sequence of SEQ ID NO: 4 and an LCVR comprising the amino acid sequence of SEQ ID NO: 6; and a second antigen-binding domain that specifically binds to human CD3, the second antigen-binding domain comprising an HCVR comprising the amino acid sequence of SEQ ID NO: 5 and an LCVR comprising the amino acid sequence of SEQ ID NO: 6 at a concentration of 2 mg / ml ± 0.2 mg / ml to 200 mg / ml ± 20 mg / ml. (ii) histidine at a concentration of 10 mM ± 1 mM; (iii) polysorbate 80 at a concentration of 0.1% ± 0.01 w / v; and (iv) sucrose at a concentration of 10% ± 1% w / v; wherein the formulation has a pH of 5.8 ± 0.3.

[0125] In some cases, the stable liquid pharmaceutical formulation is a human bispecific antibody that specifically binds to (i) human CD20 and human CD3, and comprises a first antigen-binding domain that specifically binds to human CD20, the first antigen-binding domain comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 4 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6, and a second antigen-binding domain that specifically binds to human CD3, the second antigen-binding domain comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 5 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6 at a concentration of 2 mg / ml ± 0.2 mg / ml. (ii) histidine at a concentration of 10 mM ± 1 mM; (iii) polysorbate 80 at a concentration of 0.1% ± 0.01 w / v; and (iv) sucrose at a concentration of 10% ± 1% w / v; wherein the formulation has a pH of 5.8 ± 0.3.

[0126] In some cases, the stable liquid pharmaceutical formulation comprises: (i) a first antigen-binding domain that specifically binds to human CD20 and human CD3, and that specifically binds to human CD20, comprising an HCVR comprising the amino acid sequence of SEQ ID NO:4 and an LCVR comprising the amino acid sequence of SEQ ID NO:6; and a second antigen-binding domain that specifically binds to human CD3, the second antigen-binding domain comprising an HCVR comprising the amino acid sequence of number 5 and an LCVR comprising the amino acid sequence of SEQ ID NO: 6 at a concentration of 20 mg / ml ± 2 mg / ml, wherein the antibody has a heavy chain constant region of the isotype IgG4 (optionally, one of the two heavy chains has reduced Protein A binding compared to an unmodified heavy chain of the same isotype, and optionally one or both of the two heavy chains have a chimeric hinge); (ii) histidine at a concentration of 10 mM ± 1 mM; (iii) polysorbate 80 at a concentration of 0.1% ± 0.01 w / v; and (iv) sucrose at a concentration of 10% ± 1% w / v, wherein the formulation has a pH of 5.8 ± 0.3.

[0127] In some cases, the stable liquid pharmaceutical formulation is a human bispecific antibody (i) that specifically binds to human CD20 and human CD3, and comprises a first antigen-binding domain that specifically binds to human CD20, the first antigen-binding domain comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 4 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6, and a second antigen-binding domain that specifically binds to human CD3, the second antigen-binding domain comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 5 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6 at a concentration of 100 mg / ml ± 10 mg / ml. (ii) histidine at a concentration of 10 mM ± 1 mM; (iii) polysorbate 80 at a concentration of 0.1% ± 0.01 w / v; and (iv) sucrose at a concentration of 10% ± 1% w / v; wherein the formulation has a pH of 5.8 ± 0.3.

[0128] In some cases, the stable liquid pharmaceutical formulation comprises: (i) a human bispecific antibody that specifically binds to human CD20 and human CD3, comprising a first antigen-binding domain that specifically binds to human CD20, the first antigen-binding domain comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 4 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6, and a second antigen-binding domain that specifically binds to human CD3, the second antigen-binding domain comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 5 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6 at a concentration of 2 mg / ml ± 0.2 mg / ml to 200 mg / ml ± 20 mg / ml, wherein the antibody has a heavy chain constant region of isotype IgG4, one of the two heavy chains having modifications in the CH3 domain (e.g., H435R and Y436F according to EU numbering), which is the same as unmodified IgG4 a human bispecific antibody that has reduced binding to Protein A compared to the CH3 domain; (ii) histidine at a concentration of 10 mM ± 1 mM; (iii) polysorbate 80 at a concentration of 0.1% ± 0.01 w / v; and (iv) sucrose at a concentration of 10% ± 1% w / v, wherein the formulation has a pH of 5.8 ± 0.3.

[0129] In some cases, the stable liquid pharmaceutical formulation comprises: (i) a human bispecific antibody that specifically binds to human CD20 and human CD3, comprising a first antigen-binding domain that specifically binds to human CD20, the first antigen-binding domain comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 4 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6, and a second antigen-binding domain that specifically binds to human CD3, the second antigen-binding domain comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 5 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6 at a concentration of 2 mg / ml ± 0.2 mg / ml, wherein the antibody has a heavy chain constant region of isotype IgG4, one of the two heavy chains having modifications in the CH3 domain (e.g., H435R and Y436F according to EU numbering), which is the same as unmodified IgG4 a human bispecific antibody that has reduced binding to Protein A compared to the CH3 domain; (ii) histidine at a concentration of 10 mM ± 1 mM; (iii) polysorbate 80 at a concentration of 0.1% ± 0.01 w / v; and (iv) sucrose at a concentration of 10% ± 1% w / v, wherein the formulation has a pH of 5.8 ± 0.3.

[0130] In some cases, the stable liquid pharmaceutical formulation comprises: (i) an antibody specific for human CD20 and human CD3; and (iv) sucrose at a concentration of 10% ± 1% w / v, wherein the formulation has a pH of 5.8 ± 0.3.

[0131] In some cases, the stable liquid pharmaceutical formulation comprises: (i) a human bispecific antibody that specifically binds to human CD20 and human CD3, comprising a first antigen-binding domain that specifically binds to human CD20, the first antigen-binding domain comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 4 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6, and a second antigen-binding domain that specifically binds to human CD3, the second antigen-binding domain comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 5 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6 at a concentration of 100 mg / ml ± 10 mg / ml, wherein the antibody has a heavy chain constant region of isotype IgG4, one of the two heavy chains having modifications in the CH3 domain (e.g., H435R and Y436F according to EU numbering), which is the same as unmodified IgG4 a human bispecific antibody that has reduced binding to Protein A compared to the CH3 domain; (ii) histidine at a concentration of 10 mM ± 1 mM; (iii) polysorbate 80 at a concentration of 0.1% ± 0.01 w / v; and (iv) sucrose at a concentration of 10% ± 1% w / v, wherein the formulation has a pH of 5.8 ± 0.3.

[0132] In some cases, the stable liquid pharmaceutical formulation comprises: (i) a human bispecific antibody that specifically binds human CD20 and human CD3, and comprises a first heavy chain comprising the amino acid sequence of SEQ ID NO: 1, a second heavy chain comprising the amino acid sequence of SEQ ID NO: 2, and a common light chain comprising the amino acid sequence of SEQ ID NO: 3 at a concentration of 2 mg / ml ± 0.2 mg / ml to about 100 mg / ml ± 10 mg / ml; (ii) histidine at a concentration of 10 mM ± 1 mM; (iii) polysorbate 80 at a concentration of 0.1% ± 0.01 w / v; and (iv) sucrose at a concentration of 10% ± 1% w / v, wherein the formulation has a pH of 5.8 ± 0.3.

[0133] In some cases, the stable liquid pharmaceutical formulation comprises: (i) a human bispecific antibody that specifically binds human CD20 and human CD3, and comprises a first heavy chain comprising the amino acid sequence of SEQ ID NO: 1, a second heavy chain comprising the amino acid sequence of SEQ ID NO: 2, and a common light chain comprising the amino acid sequence of SEQ ID NO: 3 at a concentration of 2 mg / ml ± 0.2 mg / ml; (ii) histidine at a concentration of 10 mM ± 1 mM; (iii) polysorbate 80 at a concentration of 0.1% ± 0.01 w / v; and (iv) sucrose at a concentration of 10% ± 1% w / v, wherein the formulation has a pH of 5.8 ± 0.3.

[0134] In some cases, the stable liquid pharmaceutical formulation comprises: (i) a human bispecific antibody that specifically binds human CD20 and human CD3, and comprises a first heavy chain comprising the amino acid sequence of SEQ ID NO: 1, a second heavy chain comprising the amino acid sequence of SEQ ID NO: 2, and a common light chain comprising the amino acid sequence of SEQ ID NO: 3 at a concentration of 20 mg / ml ± 2 mg / ml; (ii) histidine at a concentration of 10 mM ± 1 mM; (iii) polysorbate 80 at a concentration of 0.1% ± 0.01 w / v; and (iv) sucrose at a concentration of 10% ± 1% w / v, wherein the formulation has a pH of 5.8 ± 0.3.

[0135] In some cases, the stable liquid pharmaceutical formulation comprises: (i) an antibody specific for human CD20 and human CD3; a human bispecific antibody specifically binding to a first heavy chain comprising the amino acid sequence of SEQ ID NO: 1, a second heavy chain comprising the amino acid sequence of SEQ ID NO: 2, and a common light chain comprising the amino acid sequence of SEQ ID NO: 3 at a concentration of 100 mg / ml ± 10 mg / ml; (ii) histidine at a concentration of 10 mM ± 1 mM; (iii) polysorbate 80 at a concentration of 0.1% ± 0.01 w / v; and (iv) sucrose at a concentration of 10% ± 1% w / v, wherein the formulation has a pH of 5.8 ± 0.3.

[0136] In some cases, the stable liquid pharmaceutical formulation comprises in water: (i) a human bispecific antibody that specifically binds to human CD20 and human CD3, and that specifically binds to human CD20, the first antigen-binding domain comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 4 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6; and a second antigen-binding domain that specifically binds to human CD3, the HCVR comprising the amino acid sequence of SEQ ID NO: 5 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6 at a concentration of 2 mg / ml ± 0.2 mg / ml; (ii) L-histidine at a concentration of 0.57 mg / mL ± 0.1 mg / mL; (iii) L-histidine monohydrochloride monohydrate at a concentration of 1.33 mg / mL ± 0.1 mg / mL; (iv) polysorbate 80 at a concentration of 1 mg / mL ± 0.1 mg / mL; and (v) sucrose at a concentration of 100 mg / mL ± 10 mg / mL; wherein the formulation has a pH of 5.8 ± 0.3.

[0137] In some cases, the stable liquid pharmaceutical formulation comprises, in water, (i) a human bispecific antibody comprising: a first antigen-binding domain that specifically binds to human CD20 and human CD3, and that specifically binds to human CD20, comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 4 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6; and a second antigen-binding domain that specifically binds to human CD3, comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 5 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6 at a concentration of 2 mg / ml ± 0.2 mg / ml, wherein the antibody has a heavy chain constant region of isotype IgG1 (optional). (ii) L-histidine at a concentration of 0.57 mg / mL ± 0.1 mg / mL; (iii) L-histidine monohydrochloride monohydrate at a concentration of 1.33 mg / mL ± 0.1 mg / mL; (iv) polysorbate 80 at a concentration of 1 mg / mL ± 0.1 mg / mL; and (v) sucrose at a concentration of 100 mg / mL ± 10 mg / mL, wherein the formulation has a pH of 5.8 ± 0.3.

[0138] In some cases, the stable liquid pharmaceutical formulation comprises, in water, (i) a human bispecific antibody comprising: a first antigen-binding domain that specifically binds to human CD20 and human CD3, and that specifically binds to human CD20, comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 4 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6; and a second antigen-binding domain that specifically binds to human CD3, comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 5 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6 at a concentration of 2 mg / ml ± 0.2 mg / ml, wherein the antibody has a heavy chain constant region of isotype IgG4 (optionally (ii) L-histidine at a concentration of 0.57 mg / mL ± 0.1 mg / mL; (iii) L-histidine monohydrochloride monohydrate at a concentration of 1.33 mg / mL ± 0.1 mg / mL; (iv) polysorbate 80 at a concentration of 1 mg / mL ± 0.1 mg / mL; and (v) sucrose at a concentration of 100 mg / mL ± 10 mg / mL, wherein the formulation has a pH of 5.8 ± 0.3.

[0139] In some cases, the stable liquid pharmaceutical formulation comprises, in water, (i) a human bispecific antibody that specifically binds human CD20 and human CD3, and that comprises a first heavy chain comprising the amino acid sequence of SEQ ID NO: 1, a second heavy chain comprising the amino acid sequence of SEQ ID NO: 2, and a common light chain comprising the amino acid sequence of SEQ ID NO: 3 at a concentration of 2 mg / ml ± 0.2 mg / ml; and (ii) a human bispecific antibody that specifically binds human CD20 and human CD3, and that comprises a first heavy chain comprising the amino acid sequence of SEQ ID NO: 1, a second heavy chain comprising the amino acid sequence of SEQ ID NO: 2, and a common light chain comprising the amino acid sequence of SEQ ID NO: 3 at a concentration of 2 mg / ml ± 0.2 mg / ml. (iii) L-histidine at a concentration of 1.33 mg / mL ± 0.1 mg / mL, (iv) polysorbate 80 at a concentration of 1 mg / mL ± 0.1 mg / mL, and (v) sucrose at a concentration of 100 mg / mL ± 10 mg / mL, wherein the formulation has a pH of 5.8 ± 0.3.

[0140] In some cases, the stable liquid pharmaceutical formulation comprises in water: (i) a human bispecific antibody that specifically binds to human CD20 and human CD3, and that specifically binds to human CD20, the first antigen-binding domain comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 4 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6; and a second antigen-binding domain that specifically binds to human CD3, the HCVR comprising the amino acid sequence of SEQ ID NO: 5 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6 at a concentration of 20 mg / ml ± 2 mg / ml; (ii) L-histidine at a concentration of 0.57 mg / mL ± 0.1 mg / mL; (iii) L-histidine monohydrochloride monohydrate at a concentration of 1.33 mg / mL ± 0.1 mg / mL; (iv) polysorbate 80 at a concentration of 1 mg / mL ± 0.1 mg / mL; and (v) sucrose at a concentration of 100 mg / mL ± 10 mg / mL; wherein the formulation has a pH of 5.8 ± 0.3.

[0141] In some cases, the stable liquid pharmaceutical formulation comprises, in water, (i) a human bispecific antibody comprising: a first antigen-binding domain that specifically binds to human CD20 and human CD3, and that specifically binds to human CD20, comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 4 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6; and a second antigen-binding domain that specifically binds to human CD3, comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 5 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6 at a concentration of 20 mg / ml ± 2 mg / ml, wherein the antibody has a heavy chain constant region of isotype IgG1 (optionally (ii) L-histidine at a concentration of 0.57 mg / mL ± 0.1 mg / mL; (iii) L-histidine monohydrochloride monohydrate at a concentration of 1.33 mg / mL ± 0.1 mg / mL; (iv) polysorbate 80 at a concentration of 1 mg / mL ± 0.1 mg / mL; and (v) sucrose at a concentration of 100 mg / mL ± 10 mg / mL, wherein the formulation has a pH of 5.8 ± 0.3.

[0142] In some cases, the stable liquid pharmaceutical formulation comprises, in water, (i) a human bispecific antibody comprising: a first antigen-binding domain that specifically binds to human CD20 and human CD3, and that specifically binds to human CD20, comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 4 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6; and a second antigen-binding domain that specifically binds to human CD3, comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 5 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6 at a concentration of 20 mg / ml ± 2 mg / ml, wherein the antibody has a heavy chain constant region of isotype IgG4 (optionally (ii) L-histidine at a concentration of 0.57 mg / mL ± 0.1 mg / mL; (iii) L-histidine monohydrochloride monohydrate at a concentration of 1.33 mg / mL ± 0.1 mg / mL; (iv) polysorbate 80 at a concentration of 1 mg / mL ± 0.1 mg / mL; and (v) sucrose at a concentration of 100 mg / mL ± 10 mg / mL, wherein the formulation has a pH of 5.8 ± 0.3.

[0143] In some cases, the stable liquid pharmaceutical formulation comprises a liquid pharmaceutical composition comprising: (i) a human bispecific antibody that specifically binds to human CD20 and human CD3, and that comprises a first heavy chain comprising the amino acid sequence of SEQ ID NO: 1, a second heavy chain comprising the amino acid sequence of SEQ ID NO: 2, and a common light chain comprising the amino acid sequence of SEQ ID NO: 3 at a concentration of 20 mg / ml ± 2 mg / ml; (ii) L-histidine at a concentration of 0.57 mg / ml ± 0.1 mg / ml; (iii) L-histidine monohydrochloride monohydrate at a concentration of 1.33 mg / ml ± 0.1 mg / ml; and (iv) 1 mg / ml ± 0.1 mg / ml of L-histidine monohydrochloride monohydrate. g / mL, and (v) sucrose at a concentration of 100 mg / mL ± 10 mg / mL, and the formulation has a pH of 5.8 ± 0.3.

[0144] In some cases, the stable liquid pharmaceutical formulation comprises in water: (i) a human bispecific antibody that specifically binds to human CD20 and human CD3, and that specifically binds to human CD20, the first antigen-binding domain comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 4 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6; and a second antigen-binding domain that specifically binds to human CD3, the HCVR comprising the amino acid sequence of SEQ ID NO: 5 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6 at a concentration of 100 mg / ml ± 10 mg / ml; (ii) L-histidine at a concentration of 0.57 mg / mL ± 0.1 mg / mL; (iii) L-histidine monohydrochloride monohydrate at a concentration of 1.33 mg / mL ± 0.1 mg / mL; (iv) polysorbate 80 at a concentration of 1 mg / mL ± 0.1 mg / mL; and (v) sucrose at a concentration of 100 mg / mL ± 10 mg / mL; wherein the formulation has a pH of 5.8 ± 0.3.

[0145] In some cases, the stable liquid pharmaceutical formulation comprises, in water, (i) a human bispecific antibody comprising: a first antigen-binding domain that specifically binds to human CD20 and human CD3, and that specifically binds to human CD20, comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 4 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6; and a second antigen-binding domain that specifically binds to human CD3, comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 5 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6 at a concentration of 100 mg / ml ± 10 mg / ml, wherein the antibody has a heavy chain constant region of isotype IgG1 (optional). (ii) L-histidine at a concentration of 0.57 mg / mL ± 0.1 mg / mL; (iii) L-histidine monohydrochloride monohydrate at a concentration of 1.33 mg / mL ± 0.1 mg / mL; (iv) polysorbate 80 at a concentration of 1 mg / mL ± 0.1 mg / mL; and (v) sucrose at a concentration of 100 mg / mL ± 10 mg / mL, wherein the formulation has a pH of 5.8 ± 0.3.

[0146] In some cases, the stable liquid pharmaceutical formulation comprises, in water, (i) a human bispecific antibody comprising: a first antigen-binding domain that specifically binds to human CD20 and human CD3, and that specifically binds to human CD20, comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 4 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6; and a second antigen-binding domain that specifically binds to human CD3, comprising a HCVR comprising the amino acid sequence of SEQ ID NO: 5 and a LCVR comprising the amino acid sequence of SEQ ID NO: 6 at a concentration of 100 mg / ml ± 10 mg / ml, wherein the antibody has a heavy chain constant region of isotype IgG4 (optionally (ii) L-histidine at a concentration of 0.57 mg / mL ± 0.1 mg / mL; (iii) L-histidine monohydrochloride monohydrate at a concentration of 1.33 mg / mL ± 0.1 mg / mL; (iv) polysorbate 80 at a concentration of 1 mg / mL ± 0.1 mg / mL; and (v) sucrose at a concentration of 100 mg / mL ± 10 mg / mL, wherein the formulation has a pH of 5.8 ± 0.3.

[0147] In some cases, the stable liquid pharmaceutical formulation comprises in water: (i) a human bispecific antibody that specifically binds human CD20 and human CD3, and comprises a first heavy chain comprising the amino acid sequence of SEQ ID NO: 1, a second heavy chain comprising the amino acid sequence of SEQ ID NO: 2, and a common light chain comprising the amino acid sequence of SEQ ID NO: 3 at a concentration of 100 mg / ml ± 10 mg / ml; (ii) L-histidine at a concentration of 0.57 mg / mL ± 0.1 mg / mL; (iii) L-histidine monohydrochloride monohydrate at a concentration of 1.33 mg / mL ± 0.1 mg / mL; (iv) polysorbate 80 at a concentration of 1 mg / mL ± 0.1 mg / mL; and (v) sucrose at a concentration of 100 mg / mL ± 10 mg / mL; wherein the formulation has a pH of 5.8 ± 0.3.

[0148] For any of these exemplary formulations, "stable" may be defined as follows: (a) at least 90% of the antibody possesses a native conformation after 1 month of storage at 45°C as determined by size exclusion-ultra performance liquid chromatography (SE-UPLC); (b) at least 93% of the antibody possesses a native conformation after 1 month of storage at 45°C as determined by SE-UPLC; (c) at least 95% of the antibody exhibits a native conformation after 1 month of storage at 45°C as determined by SE-UPLC; (d) at least 95% of the antibody possesses a native conformation after 3 months of storage at 25°C and 60% relative humidity as determined by SE-UPLC; (e) at least 97% of the antibody possesses a native conformation after 3 months of storage at 25°C and 60% relative humidity as determined by SE-UPLC; (f) at least 95% of the antibody possesses a native conformation after 5 months of storage at 25°C and 60% relative humidity as determined by SE-UPLC. (g) at least 98% of the antibody has a native conformation after 3 months of storage at 5°C as determined by SE-UPLC; (h) the formulation contains 6% or less high molecular weight (HMW) species after 1 month of storage at 45°C as determined by SE-UPLC; (i) the formulation contains 5.6% or less HMW species after 1 month of storage at 45°C as determined by SE-UPLC; (j) the formulation contains 5.6% or less HMW species after 1 month of storage at 45°C as determined by SE-UPLC. (k) the formulation contains 2% or less HMW species after 3 months of storage at 25°C and 60% relative humidity as determined by SE-UPLC; (l) the formulation contains 1.5% or less HMW species after 3 months of storage at 5°C as determined by SE-UPLC; and / or (m) the formulation contains 1% or less HMW species after 9 months of storage at 5°C as determined by SE-UPLC.

[0149] In any of these exemplary formulations, the bispecific antibody may contain a modification in one or both heavy chains that facilitates purification of the bispecific antibody (i.e., the heterodimer) from homodimeric impurities. In some embodiments, the bispecific antibody contains first and second heavy chains (i.e., the heavy chain of the anti-CD20 binding arm and the heavy chain of the anti-CD3 binding arm) that are identical (e.g., both isotype IgG1 or IgG4) except for a modification in the CH3 domain of one or the other heavy chain that reduces binding of the bispecific antibody to Protein A compared to an antibody lacking the modification. In some cases, the CH3 domain of the first heavy chain (e.g., of the anti-CD20 binding arm) binds to Protein A, and the CH3 domain of the second heavy chain (e.g., of the anti-CD3 binding arm) contains a mutation that reduces or eliminates Protein A binding. In some cases, the mutation is an H435R modification (according to EU numbering; H95R according to IMGT exon numbering). In some cases, the mutations are H435R modifications (according to EU numbering; H95R according to IMGT exon numbering) and Y436F modifications (according to EU numbering; Y96F according to IMGT). Additional modifications that may be found within the second CH3 domain include D356E, L358M, N384S, K392N, V397M, and V422I according to EU (D16E, L18M, N44S, K52N, V57M, and V82I according to IMGT) for IgG1 CH3 domains, and Q355R, N384S, K392N, V397M, R409K, E419Q, and V422I according to EU (Q15R, N44S, K52N, V57M, R69K, E79Q, and V82I according to IMGT) for IgG4 CH3 domains.

[0150] Additional non-limiting examples of pharmaceutical formulations encompassed by the present invention are set forth elsewhere herein, including in the Examples provided below.

[0151] Container and method of administration The pharmaceutical formulations of the present invention may be contained in any container suitable for storing drugs and other therapeutic compositions. For example, the pharmaceutical formulation may be contained in a sealed and sterilized plastic or glass container having a defined volume, such as a vial, an ampoule, a syringe, a cartridge, a bottle, or an IV bag. For example, different types of vials, including transparent and opaque (e.g., amber) glass or plastic vials, may be used to contain the formulations of the present invention. Similarly, any type of syringe may be used to contain and / or administer the pharmaceutical formulations of the present invention. In some embodiments, the pharmaceutical formulation is contained in a pre-filled syringe. In some embodiments, the pharmaceutical formulation is contained in a pre-filled fixed-needle syringe.

[0152] The pharmaceutical formulations of the present invention can be contained in "normal tungsten" syringes or "low tungsten" syringes. As will be understood by those skilled in the art, the process of making glass syringes generally involves the use of a hot tungsten rod, which functions to puncture the glass, thereby creating a hole through which liquid can be drawn and expelled from the syringe. This process results in the deposition of trace amounts of tungsten on the interior surface of the syringe. Subsequent cleaning and other processing steps can be used to reduce the amount of tungsten in the syringe. As used herein, the term "normal tungsten" means that the syringe contains more than 500 parts per billion (ppb) of tungsten. The term "low tungsten" means that the syringe contains less than 500 ppb of tungsten. For example, a low tungsten syringe according to the present invention can contain less than about 490, 480, 470, 460, 450, 440, 430, 420, 410, 390, 350, 300, 250, 200, 150, 100, 90, 80, 70, 60, 50, 40, 30, 20, 10, or even less ppb of tungsten.

[0153] The rubber plunger used in a syringe and the rubber stopper used to close the opening of a vial can be coated to prevent contamination of the pharmaceutical contents of the syringe or vial and / or maintain their stability.Therefore, according to certain embodiments, the pharmaceutical formulation of the present invention can be contained in a syringe with a coated plunger or in a vial sealed with a coated rubber stopper.For example, the plunger or stopper can be coated with a fluorocarbon film.Examples of coated stoppers and / or plungers suitable for use with vials and syringes containing the pharmaceutical formulations of the present disclosure are mentioned in, for example, U.S. Patent Nos. 4,997,423, 5,908,686, 6,286,699, 6,645,635, and 7,226,554, the contents of which are incorporated herein by reference in their entirety. Certain exemplary coated rubber stoppers and plungers that may be used in the context of the present invention are commercially available under the trade name "FluroTec®" available from West Pharmaceutical Services, Inc. (Lionville, PA). FluroTec® is an example of a fluorocarbon coating used to minimize or prevent pharmaceutical agents from adhering to rubber surfaces. According to certain embodiments of the present invention, pharmaceutical formulations may be contained within low-tungsten syringes that include fluorocarbon-coated plungers.

[0154] The pharmaceutical formulations may be administered to a subject by injection (e.g., subcutaneous, intravenous, intramuscular, intraperitoneal, etc.), or by oral routes such as transdermal, mucosal, nasal, pulmonary, and / or oral administration. Numerous reusable pen and / or autoinjector delivery devices may be used to subcutaneously deliver the pharmaceutical formulations of the present invention. Examples include the AUTOPEN™ (Owen Mumford, Inc., Woodstock, UK), the DISETRONIC™ pen (Disetronic Medical Systems, Bergdorf, Switzerland), the HUMALOG MIX 75 / 25™ pen, the HUMALOG™ pen, the HUMALIN 70 / 30™ pen (Eli Lilly and Company). OPTIPEN™, OPTIPEN PRO™, OPTIPEN STARLET™, and OPTICLIK™ (sanofi-aventis, Frankfurt, Germany), to name just a few. Examples of disposable pen and / or autoinjector delivery devices having application in the subcutaneous delivery of pharmaceutical compositions of the present invention include, but are not limited to, the SOLOSTAR™ pen (sanofi-aventis), FLEXPEN™ (Novo Nordisk), and KWIKPEN™ (Eli Lilly), the SURECLICK™ autoinjector (Amgen, Thousand Oaks, CA), PENLET™ (Haselmeier, Stuttgart, Germany), EPIPEN (Dey, LP), and the HUMIRA™ pen (Abbott Labs, Abbott Park, IL), to name just a few. In some cases, the pharmaceutical formulation is contained in a syringe specifically adapted for use with the autoinjector.

[0155] The use of a microinfuser to deliver the pharmaceutical formulations of the present invention is also contemplated herein. As used herein, the term "microinfuser" refers to a subcutaneous delivery device designed to slowly administer a large amount (e.g., up to about 2.5 mL, about 3.0 mL, or more) of a therapeutic formulation over an extended period of time (e.g., about 10, 15, 20, 25, 30 minutes, or more). See, for example, US Pat. No. 6,629,949, US Pat. No. 6,659,982, and Meehan et al., J. Controlled Release 46:107-116 (1996). Microinfusers are particularly useful for delivering large amounts of therapeutic proteins contained in highly concentrated (e.g., about 100, 125, 150, 175, 200 mg / mL, or more) and / or viscous solutions.

[0156] In certain embodiments, the pharmaceutical formulation is administered via IV infusion, such that the formulation is diluted in an IV bag containing a physiologically acceptable solution. In one embodiment, the pharmaceutical composition is a sterile preparation compounded in an intravenous infusion bag, such that a single dose of the pharmaceutical is diluted in 100 mL, 250 mL (or other similar volume suitable for intravenous delivery) of a physiological buffer solution (e.g., 0.9% saline).

[0157] The pharmaceutical formulations of the present invention may also be contained in unit dosage forms. As used herein, the term "unit dosage form" refers to physically discrete units suitable as unitary dosages for a patient to be treated, each unit containing a predetermined amount of active compound calculated to produce a desired therapeutic effect in association with the required pharmaceutical carrier, diluent, or excipient. In various embodiments, the unit dosage form is contained within a container as discussed herein. The actual dosage level of the active ingredient (e.g., an anti-CD20 x anti-CD3 bispecific antibody) in the formulations of the present invention may be varied to obtain an amount of active ingredient that is effective to achieve a desired therapeutic response for a particular patient, composition, and mode of administration without adverse effects on the patient. The selected dosage level will depend on various pharmacokinetic factors, including the activity of the particular composition of the present invention employed, the route of administration, the time of administration, the rate of excretion of the particular compound employed, the duration of treatment, other drugs, compounds, and / or materials used in combination with the particular composition employed, the age, sex, weight, condition, general health, and previous medical history of the patient being treated, and similar factors well known in the medical field. As used herein, the term "diluent" refers to a solution suitable for modifying or achieving the exemplary or suitable concentrations described herein.

[0158] In various embodiments, the unit dosage form contains an amount of active ingredient (e.g., an anti-CD20 x anti-CD3 bispecific antibody) intended for a single use. In various embodiments, the amount of active ingredient in the unit dosage form is about 0.1 mg to about 5000 mg, about 100 mg to about 1000 mg, and about 100 mg to about 500 mg, about 100 mg to about 400 mg, about 100 mg to about 200 mg, about 250 mg to about 350 mg, about 125 mg to about 175 mg, about 275 mg to about 325 mg, about 1 mg to about 250 mg, about 1 mg to about 100 mg, about 1 mg to about 50 mg, about 1 mg to about 25 mg, about 1 mg to about 10 mg, about 1 mg to about 5 mg, or a range or interval therein. Ranges intermediate to the above-listed amounts, e.g., about 2 mg to about 100 mg or 2 mg to about 20 mg, are also contemplated as part of the present invention. For example, ranges of values ​​using any combination of the above-recited values ​​(or values ​​included within the above-recited ranges) as upper and / or lower limits are intended to be included. In certain embodiments, the formulations are often supplied as liquid unit dosage forms. In some embodiments, the unit dosage forms contain 2-2.5 mg, or 10-11 mg, 20-25 mg, 80-90 mg, 100-125 mg, 160-180 mg, 200-225 mg, or 320-360 mg. In some embodiments, the unit dosage forms provided herein are suitable for subcutaneous administration to a patient (e.g., the unit dosage forms contain the antibody at a concentration of about 100 mg / ml or about 160 mg / ml).

[0159] In one embodiment, the invention provides a unit dosage form comprising about 2 mg of antibody in a stable formulation, the formulation comprising, in water, the antibody at a concentration of 2 mg / ml ± 0.2 mg / ml; (ii) L-histidine at a concentration of 0.57 mg / mL ± 0.1 mg / mL; (iii) L-histidine monohydrochloride monohydrate at a concentration of 1.33 mg / mL ± 0.1 mg / mL; (iv) polysorbate 80 at a concentration of 1 mg / mL ± 0.1 mg / mL; and (v) sucrose at a concentration of 100 mg / mL ± 10 mg / mL; and the formulation has a pH of 5.8 ± 0.3. In one embodiment, the invention provides a unit dosage form comprising about 20 mg, about 80 mg, about 160 mg, or about 320 mg of an antibody in a stable formulation, the formulation comprising, in water, the antibody at a concentration of 20 mg / ml ± 2 mg / ml; (ii) L-histidine at a concentration of 0.57 mg / mL ± 0.1 mg / mL; (iii) L-histidine monohydrochloride monohydrate at a concentration of 1.33 mg / mL ± 0.1 mg / mL; (iv) polysorbate 80 at a concentration of 1 mg / mL ± 0.1 mg / mL; and (v) sucrose at a concentration of 100 mg / mL ± 10 mg / mL; and the formulation has a pH of 5.8 ± 0.3. In one embodiment, the invention provides a unit dosage form comprising about 100 mg, about 200 mg, about 300 mg, or about 400 mg of an antibody in a stable formulation, the formulation comprising, in water, the antibody at a concentration of 100 mg / ml ± 10 mg / ml; (ii) L-histidine at a concentration of 0.57 mg / mL ± 0.1 mg / mL; (iii) L-histidine monohydrochloride monohydrate at a concentration of 1.33 mg / mL ± 0.1 mg / mL; (iv) polysorbate 80 at a concentration of 1 mg / mL ± 0.1 mg / mL; and (v) sucrose at a concentration of 100 mg / mL ± 10 mg / mL; and the formulation has a pH of 5.8 ± 0.3.

[0160] The present invention also includes methods of preparing unit dosage forms. In exemplary embodiments, the method for preparing a pharmaceutical unit dosage form includes combining the formulations of any of the foregoing embodiments in a suitable container (e.g., those containers discussed herein).

[0161] In various embodiments, the pharmaceutical formulation is contained in a container (e.g., a vial or pre-filled syringe) that can contain a headspace gas containing less than 5% by volume of an oxidizing gas (e.g., oxygen). The concentration of oxidizing gas (e.g., oxygen) in the headspace of the container can be less than 4.5%, less than 4%, less than 3.5%, less than 3%, less than 2.5%, less than 2%, or less than 1.5%, in various embodiments. In one embodiment, the concentration of oxidizing gas (e.g., oxygen) in the headspace is less than about 1%. In one embodiment, the concentration of oxidizing gas (e.g., oxygen) in the headspace is less than about 1%. The concentration of oxidizing gas (e.g., oxygen) is about 0.5% or less. In one embodiment, the concentration of oxidizing gas (e.g., oxygen) in the headspace is about 0.1% or less. In various embodiments, the concentration of oxidizing gas (e.g., oxygen) in the headspace of the pharmaceutical container is less than 0.9%, less than 0.8%, less than 0.7%, less than 0.6%, less than 0.5%, less than 0.4%, less than 0.3%, less than 0.2%, or less than 0.1%. In some cases, the concentration of oxygen in the headspace gas is about 0.01% to about 1.5%. In some cases, the concentration of oxygen in the headspace gas is about 0.75% to about 1.25%. In some cases, the concentration of oxygen in the headspace gas is about 0.05% to about 0.15%. In various embodiments, the oxidizing gas (e.g., oxygen) in the headspace is replaced or substantially replaced with an inert gas such as nitrogen, argon, helium, xenon, neon, krypton, or radon. In one embodiment, the non-oxidizing gas is nitrogen. In one embodiment, the non-oxidizing gas is argon.

[0162] Therapeutic Uses of Pharmaceutical Preparations The pharmaceutical formulations of the present invention are useful, inter alia, for the treatment, prevention, and / or amelioration of any disease or disorder associated with cells expressing human CD20. Exemplary, non-limiting diseases and disorders that can be treated by administration of the pharmaceutical formulations of the present invention include B-cell cancers such as non-Hodgkin's lymphoma, e.g., follicular lymphoma, diffuse large B-cell lymphoma, mantle cell lymphoma, and marginal zone lymphoma.

[0163] The therapeutic method of the present invention includes administering to a subject any formulation comprising an anti-CD20xCD3 bispecific antibody as disclosed herein. The subject to which the pharmaceutical formulation is administered may be, for example, any human or non-human animal in need of such treatment. For example, the subject may be an individual diagnosed with or considered at risk of suffering from any of the aforementioned diseases or disorders. The present invention further includes the use of any of the pharmaceutical formulations disclosed herein in the manufacture of a medicament for the treatment of any disease or disorder associated with cell-expressed human CD20, including any of the above-mentioned exemplary diseases, disorders, and conditions.

[0164] In some embodiments, the present invention provides kits that include a pharmaceutical formulation (e.g., a container having a formulation or unit dosage form) as discussed herein and packaging or labeling (e.g., a package insert) with instructions for using the pharmaceutical formulation for the treatment of a disease or disorder as discussed above. In some cases, the instructions provide for the use of the unit dosage form for the treatment of a disease or disorder as discussed herein.

[0165] A summary of the sequences referenced herein and their corresponding SEQ ID NOs are provided in Table 1 below. [Table 1] [Example]

[0166] The following examples are presented to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the methods and compositions of the present invention, and are not intended to limit the scope of what the inventors regard as the invention. Efforts have been made to ensure accuracy with respect to numbers used (e.g., amounts, temperatures, etc.), but some experimental error and deviation should be accounted for. Unless otherwise indicated, parts are parts by weight, molecular weight is average molecular weight, temperature is in degrees Celsius, and pressure is at or near atmospheric.

[0167] Example 1: Development of a stable liquid anti-CD20xanti-CD3 bispecific antibody formulation Formulation development activities included evaluation of buffers, pH, organic cosolvents, surfactants, and thermal stabilizers to identify excipients that enhance protein stability. The results from these studies, and those discussed in Example 2, were used to develop a stable liquid formulation suitable for clinical use.

[0168] The effect of buffer and pH on the thermal stability of REGN1979 was investigated in liquid formulations by incubating 15 mg / mL REGN1979 at 45°C for 28 days in a series of buffer systems across a varying pH range. The following pHs and buffer systems were tested: acetate (pH 4.5-5.5), histidine (pH 5.5-6.5), and phosphate (pH 6.0-7.0). Based on the results from SE-UPLC analysis, maximum protein stability was observed when REGN1979 was formulated in histidine buffer at pH 5.0-6.0 (Table 2). Based on the results from CEX-UPLC analysis, the greatest protein stability was observed when REGN1979 was formulated in histidine buffer at pH 5.0-6.0 or acetate buffer at pH 4.0-7.0. Maximum protein stability was observed when the protein was formulated at pH 0.5 to 5.0. These analyses also revealed that fragmentation (i.e., formation of low molecular weight species), formation of HMW species, and charge variants were the major degradation pathways. Histidine buffer was selected as the formulation buffer for the drug product (DP) formulation because it minimized the formation of high molecular weight species, which are more concerning degradation pathways. A pH of 5.8 was selected for the DP formulation because the formation of high molecular weight species and charge variants was minimized at this pH. Based on these results and those discussed in Example 2, a 10 mM histidine buffer at pH 5.8 was selected for the REGN1979 DP formulation.

[0169] To enhance the thermal stability of proteins, thermostabilizers such as sucrose are often added to antibody formulations. REGN1979 at 25 mg / mL in a liquid formulation showed improved stability when formulated with 10% sucrose and incubated under accelerated conditions, as shown in Table 3. After 28 days of incubation at 45°C, HMW species increased by 0.6% in the formulation containing 10% sucrose, compared to a 1.4% increase in the formulation without sucrose. Based on these results and those described in Example 2, sucrose was selected as the thermostabilizer for the REGN1979 DP formulation.

[0170] Organic cosolvents, such as surfactants, are often added to antibody formulations to protect proteins from agitation-induced aggregation. The effects of surfactants on the agitation stress and thermal stability of 25 mg / mL REGN1979 were investigated. The following surfactants were evaluated: in this example, 0.1% polysorbate 20 and 0.1% polysorbate 80. The results of the agitation stress and thermal stability tests are summarized in Tables 4 and 5, respectively. REGN1979 was unstable when agitated by vortexing for 120 minutes in the absence of surfactant. In the absence of surfactant, the formulation showed a 1.5% increase in HMW species as measured by SE-UPLC (Table 4). Both surfactants tested protected REGN1979 from agitation-induced instability to a similar extent (Table 4). Furthermore, both surfactants tested reduced the thermal stability of REGN1979 to a similar extent, which was manifested as an increase in HMW species, LMW species, and basic charged species (Table 5). Based on these results, and those discussed in Example 2, polysorbate 80 was selected as the surfactant for the REGN1979 DP formulation.

[0171] REGN1979 demonstrated the greatest stability when formulated in the presence of histidine, polysorbate 80, and sucrose at pH 5.8. The primary degradation pathways identified during the development of REGN1979 liquid formulations were the formation of high and low molecular weight species and charge variants. Based on the results of these experiments and those discussed in Example 2, aqueous buffer formulations containing 10 mM histidine, 0.1% (w / v) polysorbate 80, 10% (w / v) sucrose, and 2-160 mg / mL (e.g., 2 mg / ml, 20 mg / ml, or 100 mg / ml) REGN1979 at pH 5.8 were determined to be the most stable. [Table 2] [Table 3] [Table 4] [Table 5]

[0172] Example 2: Storage and stress stability of formulations An investigational stability study was conducted to evaluate the storage, accelerated, and stress stability of the REGN1979 drug product (DP) formulation. The DP used in the investigational stability study was generated by filling 1.2 mL or 5.5 mL of drug substance into 2 mL or 10 mL Type 1 glass vials, respectively, followed by a nitrogen overlay process. The DP was incubated under several elevated temperature conditions. These accelerated conditions simulated conditions that the DP may experience during manufacturing and handling, and the REGN1979 drug product formulation with nitrogen overlay was It was selected to elucidate the degradation pathway of DP.

[0173] Storage Stability: Nine months of research stability data are currently available for the 2 mL and 10 mL vials of DP. In both DP presentations, REGN1979 DP was physically and chemically stable when stored at 5°C for nine months (Tables 6 and 7). No discernible changes in physical or chemical stability were detected in any of the monitored attributes.

[0174] Accelerated Stability: Results from analysis of REGN1979 2 mL and 10 mL vial DPs after incubation under accelerated conditions are shown in Tables 8 and 9, respectively. For both DP presentations, no appreciable degradation was observed when the protein was incubated at 25°C for one month, indicating that both REGN1979 DPs can be exposed to short periods at room temperature. After 28 days of incubation at 45°C, significant formation of HMW, LMW, and charge variants was detected. Results from this accelerated condition indicated that the formation of HMW, LMW, and charge variants was the primary degradation pathway for the 2 mL and 10 mL vials of DPs.

[0175] Stress Stability: Stress stability results for the 2 mL and 10 mL vial DPs of REGN1979 are shown in Tables 10 and 11, respectively. Both DP presentations were physically and chemically stable when agitated (vortexed at ambient temperature) for 120 minutes or subjected to eight freeze / thaw cycles (freezing at -30°C and thawing at room temperature). No discernible changes in physical or chemical stability were detected in any of the monitored attributes. [Table 6] [Table 7] [Table 8] [Table 9] [Table 10] [Table 11]

[0176] Results from storage, accelerated, and stress stability studies indicate that REGN1979 is stable during manufacturing (formulation, fill / finish, and labeling operations) and can withstand brief exposure to room temperature without physical or chemical stability.

[0177] Additional stability experiments were performed on formulations containing 2 mg / ml, 20 mg / ml, 100 mg / ml, and 160 mg / ml of bispecific antibody (REGN1979) with 10 mM histidine, 10% w / v sucrose, 0.1% w / v polysorbate 80, and pH 5.8. The results are shown in Tables 12-27 below. NR = not reported; ND = too degraded to analyze. [Table 12] [Table 13] [Table 14] [Table 15] [Table 16] [Table 17] [Table 18] [Table 19] [Table 20] [Table 21] [Table 22] [Table 23] [Table 24] [Table 25] [Table 26] [Table 27]

[0178] Additional stability studies were conducted using various other excipients with a bispecific antibody (REGN1979) concentration of 100 mg / ml. The various formulations (F1-F13) are shown in Table 28 below. The results of these stability experiments are shown in Tables 29-41. In each of these experiments, the volume The container / closure is made of 20mm FluroTec® coated West V2-F451W 4432 / 50 GRY 5mL type with B2-TR stopper 1 borosilicate glass vial. [Table 28] [Table 29] [Table 30] [Table 31] [Table 32] [Table 33] [Table 34] [Table 35] [Table 36] [Table 37] [Table 38] [Table 39] [Table 40] [Table 41]

[0179] Example 3: Viscosity of Formulations Viscosity measurements were performed at 20°C using a Rheosense m-VROC capillary viscometer (Rheosense, San Ramon, CA). Samples of various REGN1979 concentrations ranging from 79.9 to 184.9 mg / ml were prepared in formulation buffer containing 10 mM histidine and 5% sucrose (pH 5.8). All samples were filtered using 2 μm centrifugal spin filters prior to measurement. The results of the measurements are shown below in Table 42. [Table 42]

[0180] A pharmaceutical formulation containing specific concentrations of REGN1979, L-histidine (0.57 mg / ml), L-histidine monohydrochloride monohydrate (1.33 mg / ml), sucrose (100 mg / ml), polysorbate 80 (1 mg / ml), and water for injection (USP) was determined to have the viscosity shown in Table 43 below. [Table 43]

[0181] The low viscosity (eg, less than 15 cP) observed in formulations containing as much as 154.8 mg / ml of antibody is advantageous for use in pre-filled syringes or auto-injectors.

[0182] The present invention is not to be limited in scope by the specific embodiments described herein. Indeed, various modifications of the invention in addition to those described herein will become apparent to those skilled in the art from the foregoing description. Such modifications are intended to be included within the scope of the appended claims.

Claims

1. A single-dose vial containing a pharmaceutical formulation comprising a bispecific antibody, wherein the vial contains 2 mg to 2.5 mg of the bispecific antibody, and the pharmaceutical formulation comprises, (a) A bispecific antibody in a concentration of 2 mg / ml ± 0.2 mg / ml comprising a first antigen-binding domain that specifically binds to human CD20 and a second antigen-binding domain that specifically binds to human CD3, wherein the first antigen-binding domain comprises a heavy chain variable region (HCVR) and a light chain variable region (LCVR), the second antigen-binding domain comprises the HCVR and LCVR, the bispecific antibody comprises a first heavy chain comprising the HCVR of the first antigen-binding domain, a second heavy chain comprising the HCVR of the second antigen-binding domain, and two copies of a common light chain, wherein the first heavy chain comprises residues 1 to 452 of the amino acid sequence of SEQ ID NO: 1, the second heavy chain comprises residues 1 to 448 of the amino acid sequence of SEQ ID NO: 2, and the common light chain comprises the amino acid sequence of SEQ ID NO: 3, (b) 10 mM ± 1 mM histidine buffer at pH 5.8 ± 0.2, (c) 0.1% ± 0.01% w / v polysorbate 80, (d) containing 10% ± 1% w / v sucrose, The formulation, as determined using a capillary viscometer, has a viscosity of less than 2 centipoise (cP) at 20°C, and as determined by SE-UPLC, at least 95% of the antibody has native conformation after storage at 45°C for one month, in the single-dose vial.

2. The aforementioned pharmaceutical preparation (a) 2 mg / ml antibody, (b) 0.57 mg / ml of L-histidine and 1.3 mg / ml of L-histidine monohydrochloride monohydrate, (c) 1 mg / ml of polysorbate 80, and (d) 100 mg / ml sucrose A single-dose vial according to claim 1, comprising:

3. The vial contains 2 mg of bispecific antibody, as described in claim 1. 。

4. The single-dose vial according to claim 2, wherein the vial contains 2 mg of bispecific antibody.

5. A single-dose vial containing a pharmaceutical formulation comprising a bispecific antibody, wherein the vial contains 80 mg to 90 mg of the bispecific antibody, and the pharmaceutical formulation comprises, (a) A bispecific antibody in a concentration of 20 mg / ml ± 2 mg / ml comprising a first antigen-binding domain that specifically binds to human CD20 and a second antigen-binding domain that specifically binds to human CD3, wherein the first antigen-binding domain comprises a heavy chain variable region (HCVR) and a light chain variable region (LCVR), the second antigen-binding domain comprises the HCVR and LCVR, the bispecific antibody comprises a first heavy chain comprising the HCVR of the first antigen-binding domain, a second heavy chain comprising the HCVR of the second antigen-binding domain, and two copies of a common light chain, wherein the first heavy chain comprises residues 1 to 452 of the amino acid sequence of SEQ ID NO: 1, the second heavy chain comprises residues 1 to 448 of the amino acid sequence of SEQ ID NO: 2, and the common light chain comprises the amino acid sequence of SEQ ID NO: 3, (b) 10 mM ± 1 mM histidine buffer at pH 5.8 ± 0.2, (c) 0.1% ± 0.01% w / v polysorbate 80, (d) containing 10% ± 1% w / v sucrose, The formulation, as determined using a capillary viscometer, has a viscosity of less than 2 cP at 20°C, and as determined by SE-UPLC, at least 95% of the antibody has native conformation after storage at 45°C for one month, in the single-dose vial.

6. The aforementioned pharmaceutical preparation (a) 20 mg / ml antibody, (b) 0.57 mg / ml L-histidine and 1.3 mg / ml L-histidine monohydrochloride monohydrate, (c) 1 mg / ml of polysorbate 80, and (d) 100 mg / ml sucrose A single-dose vial according to claim 5, comprising:

7. The single-dose vial according to claim 5, wherein the vial contains 80 mg of bispecific antibody.

8. The single-dose vial according to claim 6, wherein the vial contains 80 mg of bispecific antibody.

9. A single-dose vial containing a pharmaceutical formulation comprising a bispecific antibody, wherein the vial contains 320 mg to 360 mg of the bispecific antibody, and the pharmaceutical formulation comprises, (a) A bispecific antibody in a concentration of 20 mg / ml ± 2 mg / ml comprising a first antigen-binding domain that specifically binds to human CD20 and a second antigen-binding domain that specifically binds to human CD3, wherein the first antigen-binding domain comprises a heavy chain variable region (HCVR) and a light chain variable region (LCVR), the second antigen-binding domain comprises the HCVR and LCVR, the bispecific antibody comprises a first heavy chain comprising the HCVR of the first antigen-binding domain, a second heavy chain comprising the HCVR of the second antigen-binding domain, and two copies of a common light chain, wherein the first heavy chain comprises residues 1 to 452 of the amino acid sequence of SEQ ID NO: 1, the second heavy chain comprises residues 1 to 448 of the amino acid sequence of SEQ ID NO: 2, and the common light chain comprises the amino acid sequence of SEQ ID NO: 3, (b) 10 mM ± 1 mM histidine buffer at pH 5.8 ± 0.2, (c) 0.1% ± 0.01% w / v polysorbate 80, (d) containing 10% ± 1% w / v sucrose, The formulation, as determined using a capillary viscometer, has a viscosity of less than 2 cP at 20°C, and as determined by SE-UPLC, at least 95% of the antibody has native conformation after storage at 45°C for one month, in the single-dose vial.

10. The aforementioned pharmaceutical preparation (a) 20 mg / ml antibody, (b) 0.57 mg / ml L-histidine and 1.3 mg / ml L-histidine monohydrochloride monohydrate, (c) 1 mg / ml of polysorbate 80, and (d) 100 mg / ml sucrose A single-dose vial according to claim 9, comprising:

11. The single-dose vial according to claim 9, wherein the vial contains 320 mg of bispecific antibody.

12. The single-dose vial according to claim 10, wherein the vial contains 320 mg of bispecific antibody.

13. A single-dose vial containing a pharmaceutical formulation comprising a bispecific antibody, wherein the vial contains 1 mg to 360 mg of the bispecific antibody, and the pharmaceutical formulation comprises, (a) A bispecific antibody having a concentration of 2 ± 0.2 mg / ml or 20 ± 2 mg / ml, wherein the bispecific antibody comprises a first antigen-binding domain that specifically binds to human CD20 and a second antigen-binding domain that specifically binds to human CD3, wherein the first antigen-binding domain comprises a heavy chain variable region (HCVR) and a light chain variable region (LCVR), and the second antigen-binding domain comprises the HCVR and LCVR, and the bispecific antibody comprises a first heavy chain containing the HCVR of the first antigen-binding domain, a second heavy chain containing the HCVR of the second antigen-binding domain, and two copies of a common light chain, wherein the first heavy chain comprises residues 1 to 452 of the amino acid sequence of SEQ ID NO: 1, the second heavy chain comprises residues 1 to 448 of the amino acid sequence of SEQ ID NO: 2, and the common light chain comprises the amino acid sequence of SEQ ID NO:

3. (b) A buffer solution containing histidine at a concentration of 5 mM to 15 mM, (c) Polysorbate with a concentration of 0.05% to 0.15% w / v, (d) Sucrose at a concentration of 8% to 12% w / v, The formulation contains, and the formulation has a pH of 5.8 ± 0.

3. A single-dose vial having native conformation after storage at 45°C for one month, provided that at least 95% of the antibody is determined by SE-UPLC.

14. The single-dose vial according to claim 13, wherein the histidine buffer comprises L-histidine and L-histidine monohydrochloride monohydrate, and the polysorbate is polysorbate 80.

15. A single-dose vial according to any one of claims 1 to 14, wherein the first heavy chain comprises the amino acid sequence of SEQ ID NO: 1, the second heavy chain comprises the amino acid sequence of SEQ ID NO: 2, and the common light chain comprises the amino acid sequence of SEQ ID NO: 3.