Stable formulations containing bispecific BCMA / CD3 antibodies

Abstract

Provided herein are stable aqueous pharmaceutical compositions comprising formulations of bispecific BCMA / CD3 antibodies or antigen-binding fragments thereof, and methods for preparing the same. Also provided herein are methods of treating cancer in a subject in need thereof by administering to the subject a stable aqueous pharmaceutical composition disclosed herein. Further provided herein are kits and articles of manufacture comprising the stable aqueous pharmaceutical compositions disclosed herein.

Description

[Technical field]

[0001] (CROSS REFERENCE TO RELATED APPLICATIONS) This application claims the benefit of U.S. Provisional Application No. 63 / 277,885, filed November 10, 2021, which is incorporated by reference in its entirety.

[0002] (Reference to electronically submitted sequence listing) This application contains a Sequence Listing that has been submitted electronically in XML format, which is incorporated by reference in its entirety. The XML copy, created on November 8, 2022, has the file name "258199.061002_PRD4191WOPCT1_SL.xml" and is 22.6 kilobytes in size.

[0003] FIELD OF THEINVENTION Compositions and methods for formulating stable pharmaceutical compositions comprising bispecific BCMA / CD3 antibodies are disclosed.

[0004] BACKGROUND OF THEINVENTION B-cell maturation antigen (BCMA), also known as CD269 and tumor necrosis factor (TNF) receptor superfamily member 17, is a receptor that plays a key role in the maturation of B lymphocytes (B cells) and their subsequent differentiation into plasma cells. BCMA binds to two ligands, namely A proliferation-inducing ligand (APRIL, CD256) and BAFF. APRIL and BAFF are type II transmembrane proteins that are readily cleaved by furin and secreted as soluble trimers by many cells (B cells [autocrine], monocytes, dendritic cells, T cells, osteoclasts, etc.) and can bind to the BCMA receptor. Unlike other surface markers, BCMA is exclusively expressed on B lineage cells and selectively induced during plasma cell differentiation.

[0005] The human BCMA receptor is a 184 amino acid protein with neither a secretory signal sequence nor a specific protease cleavage site in the N-terminal 54 amino acid extracellular domain. However, the N-terminal fragment is observed as a soluble protein in serum as a result of gamma secretase activity, which cleaves the BCMA protein at the transmembrane domain. Inhibition of gamma secretase processing leads to a marked increase in BCMA surface protein in human primary B cells. High levels of soluble BCMA (sBCMA) were measured in serum samples from patients with multiple myeloma (data not shown) and correlated with plasma cell counts.

[0006] BCMA mRNA and protein were universally detected by the applicants in all malignant plasma cells from MM cell lines and multiple myeloma patients (data not shown). Similarly, in multiple myeloma cell lines and patient samples, BCMA is more stably expressed compared to the major plasma cell marker (CD138), which is also expressed in normal fibroblasts and epithelial cells. BCMA expression was selective for the B cell lineage and was not detected in any major tissues, except in infiltrating plasma cells, as determined by immunohistochemistry (IHC). Taken together, the selective expression of BCMA on the B cell lineage makes it an attractive target for T cell-mediated therapy to treat plasma cell disorders such as multiple myeloma.

[0007] T cell redirection killing is a desired mode of action in many therapeutic areas. Typically, T cell redirection molecules are engineered to have at least two antigen binding sites, one site that binds to a surface antigen on a target cell and the other site that binds to a T cell surface antigen. Among the T cell surface antigens, the human CD3 epsilon subunit of the TCR protein complex is the one that is most targeted to redirect T cell killing. Various bispecific antibody formats have been shown to mediate T cell redirection in both preclinical and clinical studies.

[0008] The role of both BCMA and CD3 in cancer is well established, making these targets attractive for combination therapy. The use of anti-BCMA antibodies for the treatment of lymphoma and multiple myeloma is mentioned in WO2002066516 and WO2010104949. Antibodies against BCMA are described, for example, in Gras MP.et al.Int Immunol.1997;7:1093-1106, WO200124811 and WO200124812. Bispecific antibodies against BCMA and CD3 are described, for example, in WO2017 / 031104.

[0009] Although anti-BCMA / CD3 antibodies have shown promising results, there remains a need in the art for pharmaceutical compositions comprising such antibodies that are stable for extended periods at refrigerated temperatures (2-8° C.) and at ambient temperatures.

[0010] (Summary of the invention) Disclosed herein are stable aqueous pharmaceutical compositions comprising particular formulations of bispecific B-cell maturation antigen (BCMA) / cluster of differentiation 3 (CD3) antibodies.

[0011] In one aspect, provided herein is a stable aqueous pharmaceutical composition comprising: (a) a bispecific B cell maturation antigen (BCMA) / cluster of differentiation 3 (CD3) antibody or antigen-binding fragment thereof at a concentration of about 7.5 mg / mL to about 12.5 mg / mL, wherein the bispecific BCMA / CD3 antibody or antigen-binding fragment thereof is (1) a first heavy chain (HC1) comprising an HC1 variable region 1 (VH1); wherein VH1 comprises heavy chain complementarity determining region 1 (HCDR1), HCDR2 and HCDR3 having the amino acid sequences of SEQ ID NOs: 1, 2 and 3, respectively; (2) a first light chain (LC1) comprising an LC1 variable region (VL1); wherein VL1 comprises a light chain complementarity determining region 1 (LCDR1), LCDR2 and LCDR3 having the amino acid sequences of SEQ ID NOs: 4, 5 and 6, respectively; (3) a second heavy chain (HC2) comprising an HC2 variable region 2 (VH2); and wherein VH2 comprises heavy chain complementarity determining region 1 (HCDR1), HCDR2 and HCDR3 having the amino acid sequences of SEQ ID NOs: 11, 12 and 13, respectively; (4) a second light chain (LC2) comprising an LC2 variable region 2 (VL2); and wherein VL2 comprises a light chain complementarity determining region 1 (LCDR1), LCDR2 and LCDR3 having the amino acid sequences of SEQ ID NOs: 14, 15 and 16, respectively; Including, (b) about 10 mM to about 20 mM acetate and / or a pharma- ceutically acceptable acetate salt; (c) about 6% (w / v) to about 10% (w / v) sucrose; (d) about 16 μg / mL to about 24 μg / mL of ethylenediaminetetraacetic acid (EDTA); (e) about 0.01% to about 0.07% polysorbate 20; and (f) a pH of about 4.7 to about 5.7; A stable aqueous pharmaceutical composition is provided comprising:

[0012] In one aspect, provided herein is a stable aqueous pharmaceutical composition comprising: (a) a bispecific B cell maturation antigen (BCMA) / cluster of differentiation 3 (CD3) antibody or antigen-binding fragment thereof at a concentration of about 76.5 mg / mL to about 103.5 mg / mL, wherein the bispecific BCMA / CD3 antibody or antigen-binding fragment thereof is (1) a first heavy chain (HC1) comprising an HC1 variable region 1 (VH1); wherein VH1 comprises heavy chain complementarity determining region 1 (HCDR1), HCDR2 and HCDR3 having the amino acid sequences of SEQ ID NOs: 1, 2 and 3, respectively; (2) a first light chain (LC1) comprising an LC1 variable region (VL1); wherein VL1 comprises a light chain complementarity determining region 1 (LCDR1), LCDR2 and LCDR3 having the amino acid sequences of SEQ ID NOs: 4, 5 and 6, respectively; (3) a second heavy chain (HC2) comprising an HC2 variable region 2 (VH2); and wherein VH2 comprises heavy chain complementarity determining region 1 (HCDR1), HCDR2 and HCDR3 having the amino acid sequences of SEQ ID NOs: 11, 12 and 13, respectively; (4) a second light chain (LC2) comprising an LC2 variable region 2 (VL2); and wherein VL2 comprises a light chain complementarity determining region 1 (LCDR1), LCDR2 and LCDR3 having the amino acid sequences of SEQ ID NOs: 14, 15 and 16, respectively; Including, (b) about 10 mM to about 20 mM acetate and / or a pharma- ceutically acceptable acetate salt; (c) about 6% (w / v) to about 10% (w / v) sucrose; (d) about 16 μg / mL to about 24 μg / mL of ethylenediaminetetraacetic acid (EDTA); (e) about 0.01% to about 0.07% polysorbate 20; and (f) a pH of about 4.7 to about 5.7; A stable aqueous pharmaceutical composition is provided comprising:

[0013] Also provided herein is a method of treating cancer in a subject in need thereof, the method comprising administering to the subject a stable aqueous pharmaceutical composition disclosed herein.

[0014] Also provided are methods for preparing stable aqueous pharmaceutical compositions of bispecific B cell maturation antigen (BCMA) / cluster of differentiation 3 (CD3) antibodies or antigen-binding fragments thereof. The bispecific BCMA / CD3 antibodies or antigen-binding fragments thereof can be prepared, for example, by (1) a first heavy chain (HC1) comprising an HC1 variable region 1 (VH1); wherein VH1 comprises heavy chain complementarity determining region 1 (HCDR1), HCDR2 and HCDR3 having the amino acid sequences of SEQ ID NOs: 1, 2 and 3, respectively; (2) a first light chain (LC1) comprising an LC1 variable region (VL1); wherein VL1 comprises a light chain complementarity determining region 1 (LCDR1), LCDR2 and LCDR3 having the amino acid sequences of SEQ ID NOs: 4, 5 and 6, respectively; (3) a second heavy chain (HC2) comprising an HC2 variable region 2 (VH2); and wherein VH2 comprises heavy chain complementarity determining region 1 (HCDR1), HCDR2 and HCDR3 having the amino acid sequences of SEQ ID NOs: 11, 12 and 13, respectively; (4) a second light chain (LC2) comprising an LC2 variable region 2 (VL2); and wherein VL2 comprises a light chain complementarity determining region 1 (LCDR1), LCDR2 and LCDR3 having the amino acid sequences of SEQ ID NOs: 14, 15 and 16, respectively; may include.

[0015] The method can include, for example, combining a composition comprising about 10 mg / mL or about 90 mg / mL of a bispecific BCMA / CD3 antibody, about 15 mM acetate and / or a pharma- ceutically acceptable acetate salt, about 8% (w / v) sucrose, about 20 mg / mL EDTA, and about 0.04% polysorbate (PS) 20, wherein the stable aqueous pharmaceutical composition has a pH of about 5.2.

[0016] Also provided herein is a kit comprising the stable aqueous pharmaceutical composition disclosed herein and instructions for its use.

[0017] Further provided herein is an article of manufacture comprising a container holding a stable aqueous pharmaceutical composition disclosed herein. [Brief description of the drawings]

[0018] The above summary, as well as the following detailed description of preferred embodiments of the present application, will be better understood when read in conjunction with the appended drawings. It should be understood, however, that the present application is not limited to the precise embodiments shown in the drawings. [Figure 1] 1 shows a graph showing purity by SE-HPLC for teclistamab acetate and histidine buffer formulations.

[0019] DETAILED DESCRIPTION OF THE PRESENT EMBODIMENT The compositions and methods of the present disclosure may be more readily understood by reference to the following detailed description taken in conjunction with the accompanying drawings, which form a part of this disclosure: It is to be understood that the disclosed compositions and methods are not limited to the specific compositions and methods described and / or illustrated herein, and that the terminology used herein is for the purpose of describing particular embodiments, by way of example only, and is not intended to limit the compositions and methods claimed.

[0020] Unless otherwise stated, any description of possible mechanisms or modes of operation or reasons for improvement is intended to be illustrative only, and the compositions and methods of the present disclosure are not limited by the merits or demerits of any such proposed mechanisms or modes of operation or reasons for improvement.

[0021] When a range of numerical values ​​is recited or established herein, the range includes its endpoints, and all individual integers and rational numbers within the range, and also includes each of the narrower ranges formed by all the various possible combinations of these endpoints and internal integers and rational numbers, forming a subgroup of the larger group of values ​​within the recited range, as if each of the narrower ranges were explicitly recited. When a range of numerical values ​​is recited herein as being greater than the recited value, the range is nevertheless finite, and its upper limit is defined by a value that is operable within the context of the invention described herein. When a range of numerical values ​​is recited herein as being less than the recited value, the range is nevertheless defined by a lower limit by a non-zero value. It is not intended that the scope of the invention be limited to the specific values ​​recited in defining the range. All ranges are inclusive and combinable.

[0022] When values ​​are expressed as approximations, by use of the antecedent "about," it is to be understood that the particular value forms another embodiment. Reference to a particular numerical value is intended to include at least the particular value unless the context clearly indicates otherwise.

[0023] It is also understood that certain features of the disclosed compositions and methods that are, for clarity, described herein in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the disclosed compositions and methods that are, for brevity, described in the context of a single embodiment, may also be provided separately or in any subcombination.

[0024] As used herein, the singular forms "a," "an," and "the" are intended to include plurals.

[0025] Various terms relating to the embodiments of the present specification are used throughout the specification and claims. Unless otherwise indicated, such terms are to be given their ordinary meaning in the art. Other specifically defined terms are to be interpreted in a manner consistent with the definitions provided herein.

[0026] As used herein, "about" is used when used in connection with a numerical range, cutoff, or specific value to indicate that the recited value may vary by up to 10% from the recited value. Since many of the numerical values ​​used herein are experimentally determined, those skilled in the art will understand that such determinations may, and often will, vary between different experiments. The values ​​used herein should not be considered unduly limited by this inherent variation. Thus, the term "about" is used to encompass a variation of ±10% or less, a variation of ±5% or less, a variation of ±1% or less, a variation of ±0.5% or less, or a variation of ±0.1% or less from the stated value.

[0027] Similarly, the term "comprising" is intended to include examples encompassed by the terms "consisting essentially of" and "consisting of." Similarly, the term "consisting essentially of" is intended to include examples encompassed by the term "consisting of." Unless the context clearly requires otherwise, throughout the specification and claims, the words "comprise," "comprising," and the like, are to be construed in an inclusive sense, i.e., "including, but not limited to," as opposed to an exclusive or exclusive sense.

[0028] The term "antibody" and similar terms are intended in a broad sense to include immunoglobulin molecules or fragments thereof, including monoclonal antibodies (such as murine, human, human-adapted, humanized, and chimeric monoclonal antibodies), antibody fragments, bispecific or multispecific antibodies, dimeric, tetrameric, or multimeric antibodies, and single-chain antibodies.

[0029] Immunoglobulins can be assigned to five major classes, namely IgA, IgD, IgE, IgG, and IgM, depending on the amino acid sequence of the heavy chain constant domain. IgA and IgG are further classified into isotypes IgA1, IgA2, IgG1, IgG2, IgG3, IgG4. Antibody light chains of any vertebrate species can be assigned to one of two clearly distinct types, kappa (κ) and lambda (λ), based on the amino acid sequence of their constant domain.

[0030] "Antibody fragment" refers to a portion of an immunoglobulin molecule that retains the antigen-binding properties of the parent full-length antibody. Exemplary antibody fragments are heavy chain complementarity determining regions (HCDRs) 1, 2, and 3, light chain complementarity determining regions (LCDRs) 1, 2, and 3, heavy chain variable region (VH), or light chain variable region (VL). Antibody fragments include Fab fragments, which are monovalent fragments consisting of the VL, VH, constant light chain (CL), and constant heavy chain 1 (CH1) domains; F(ab')2 fragments, which are bivalent fragments containing two Fab fragments linked by a disulfide bridge at the hinge region; Fd fragments consisting of the VH and CH1 domains; Fv fragments consisting of the VL and VH domains of a single arm of an antibody; and domain antibody (dAb) fragments consisting of the VH domain (Ward et al., Nature, 341:544-546, 1989). The VH and VL domains can be engineered and linked together via synthetic linkers to form a variety of single chain antibody designs, where the VH / VL domains pair intramolecularly or, when the VH and VL domains are expressed by separate single chain antibody constructs, pair intermolecularly to form a monovalent antigen binding site, such as a single chain Fv (scFv) or diabody. For example, as described in WO 1998 / 44001, WO 1988 / 01649, WO 1994 / 13804, and WO 1992 / 01047. These antibody fragments are obtained using techniques known to those skilled in the art, and the fragments are screened for utility in the same manner as full length antibodies.

[0031] Antibody variable regions consist of a "framework" region interrupted by three "antigen-binding sites", which are defined using various terms: (i) the complementarity determining regions (CDRs), three in VH (HCDR1, HCDR2, HCDR3) and three in VL (LCDR1, LCDR2, LCDR3), are based on sequence diversity (Wu and Kabat J Exp Med 132:211-50, 1970; Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md., 1991), and (ii) the "hypervariable regions" ("HVRs" or "HVs"), three in VH (H1, H2, H3) and three in VL (L1, L2, L3), are based on sequence diversity (Wu and Kabat J Exp Med 132:211-50, 1970; Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md., 1991), which ... and (ii) the "hypervariable regions" ("HVRs" or "HVs"), three in VH (H1, H2, H3) and three in VL (L1, L2, 196:901-17,1987). Other terms include "IMGT-CDR" (Lefranc et al., Dev Comparat Immunol 27:55-77,2003) and "Specificity Determining Residue Usage" (SDRU) (Almagro Mol Recognit 17:132-43,2004). The International ImMunoGeneTics (IMGT) database (www_imgt_org) provides standard numbering and definitions for antigen binding sites. The correspondence between CDR, HV and IMGT outlines is provided in Lefranc et al., Dev Comparat Immunol 27:55-77,2003.

[0032] "Monoclonal antibody" refers to a preparation of antibody molecules of a single molecular composition. A monoclonal antibody composition exhibits a single binding specificity and affinity for a particular epitope, or, in the case of bispecific monoclonal antibodies, dual binding specificity for two distinct epitopes. Thus, a monoclonal antibody refers to an antibody population in which the amino acid composition of each heavy and each light chain is uniform, except for possible known modifications such as removal of the C-terminal lysine from the antibody heavy chain. Monoclonal antibodies may have heterogeneous glycosylation within the antibody population. Monoclonal antibodies may be monospecific or multispecific, or monovalent, bivalent, or multivalent. Bispecific antibodies are included in the term monoclonal antibody.

[0033] "Bispecific" refers to an antibody that specifically binds to two different antigens or two different epitopes on the same antigen. Bispecific antibodies can be cross-reactive to other related antigens, e.g., the same antigen (homologs) from other species, such as humans or monkeys, e.g., cynomolgus monkeys (Macaca cynomolgus, cyno) or chimpanzees (Pan troglodytes), or can bind to epitopes shared between two or more different antigens.

[0034] "Human antibody" refers to an antibody that is optimized to have a minimal immune response when administered to a human subject. The variable regions of a human antibody are derived from human immunoglobulin sequences. If the human antibody contains a constant region or a portion of a constant region, the constant region is also derived from a human immunoglobulin sequence. A human antibody contains heavy and light chain variable regions that are "derived" from sequences of human origin when the variable regions of the human antibody are derived from a system that uses human germline immunoglobulin or rearranged immunoglobulin genes. Exemplary such systems are phage-displayed human immunoglobulin gene libraries and transgenic non-human animals, such as mice or rats, that carry human immunoglobulin loci. "Human antibodies" typically contain amino acid differences when compared to immunoglobulins expressed in humans, due to differences in the system used to obtain the human antibodies and human immunoglobulin loci, the intentional introduction of somatic mutations or substitutions into the framework or CDRs, or both. Typically, a "human antibody" is at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical in amino acid sequence to the amino acid sequence encoded by a human germline immunoglobulin or rearranged immunoglobulin gene. Optionally, a "human antibody" can comprise a consensus framework sequence obtained from human framework sequence analysis as described, for example, in Knappik et al., (2000) J Mol Biol 296:57-86, or a synthetic HCDR3 incorporated into a library of human immunoglobulin genes displayed on phage as described, for example, in Shi et al., (2010) J Mol Biol 397:385-96 and WO 2009 / 085462. Antibodies in which at least one CDR is derived from a non-human species are not included in the definition of "human antibody."

[0035] "Humanized antibody" refers to an antibody in which at least one CDR is derived from a non-human species and at least one framework is derived from a human immunoglobulin sequence. Humanized antibodies can contain substitutions in the framework, such that the framework may not be an exact copy of an expressed human immunoglobulin or human immunoglobulin germline gene sequence.

[0036] "Isolated" refers to a protein that has been subjected to at least one purification or isolation step, as well as to a homogenous population of molecules (e.g., a protein such as a synthetic polynucleotide or an antibody) that have been substantially separated and / or purified from other components associated with the system in which the molecule is produced, such as a recombinant cell. An "isolated antibody" refers to an antibody that is substantially free of other cellular material and / or chemicals, and includes antibodies that have been isolated to greater degrees of purity, e.g., 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% purity.

[0037] "BCMA / CD3 bispecific antibody" refers to a bispecific antibody that specifically binds to BCMA and CD3. BCMA / CD3 bispecific antibodies are described in U.S. Patent No. 10,072,088, which is incorporated herein by reference in its entirety.

[0038] "BCMA" refers to human B cell maturation antigen, which is also known as CD269 or TNFRSF17 (UniProt Q02223). The extracellular domain of BCMA encompasses residues 1-54 of Q02223. Human BCMA comprises the amino acid sequence of SEQ ID NO:21. MLQMAGQCSQNEYFDSLLHACIPCQLRCSSNTPPLTCQRYCNASVTNSVKGTNAILWTCLGLSLIISLAVFVLMFLLRKINSEPLKDEFKNTGSGLLGMANIDLEKSRTGDEIILPRGLEYTVEECTCEDCIKSKPKVDSDHCFPLPAMEEGATILVTTKTNDYCKSLPAALSATEIEKSISAR (SEQ ID NO: 21)

[0039] "CD3" refers to a human antigen expressed on T cells as part of the multimolecular T cell receptor (TCR) complex and consisting of a homodimer or heterodimer formed from the association of two or four receptor chains: CD3 epsilon, CD3 delta, CD3 zeta, and CD3 gamma. Human CD3 epsilon comprises the amino acid sequence of SEQ ID NO: 22. SEQ ID NO: 23 shows the extracellular domain of CD3 epsilon. MQSGTHWRVLGLCLLSVGVWGQDGNEEMGGITQTPYKVSISGTTVILTCPQYPGSEILWQHNDKNIGGDEDDKNIGSDEDHLSLKEFSELEQSGYYVCYPRGSKPEDANFYLYLRARVCENCMEMDVMSVATIVIVDICITGGLLLLVYYWSKNRKAKAKPVTRGAGAGGRQRGQNKERPPPVPNPDYEPIRKGQRDLYSGLNQRRI (SEQ ID NO: 22) DGNEEMGGITQTPYKVSISGTTVILTCPQYPGSEILWQHNDKNIGGDEDDKNIGSDEDHLSLKEFSELEQSGYYVCYPRGSKPEDANFYLYLRARVCENCMEMD (SEQ ID NO: 23)

[0040] "Epitope" refers to a portion of an antigen to which an antibody specifically binds. Epitopes usually consist of chemically active (such as polar, non-polar, or hydrophobic) surface groups of moieties such as amino acids or polysaccharide side chains and may have specific three-dimensional structural characteristics as well as specific charge characteristics. Epitopes may be composed of contiguous and / or discontinuous amino acids that form a conformational spatial unit. In discontinuous epitopes, amino acids in different parts of the linear sequence of the antigen are brought into close proximity in three-dimensional space due to folding of the protein molecule.

[0041] "Variant" refers to a polypeptide or polynucleotide that differs from a reference polypeptide or polynucleotide by one or more modifications, such as, for example, a substitution, insertion, or deletion.

[0042] By "in combination with" it is meant that two or more therapeutic agents can be administered to a subject together in a mixture, simultaneously as single agents, or sequentially in any order as single agents.

[0043] "Treat", "treatment" and similar terms refer to both therapeutic and prophylactic or preventative treatments, including reducing the severity and / or frequency of a symptom, eliminating a symptom and / or the underlying cause of a symptom, reducing the frequency or likelihood of a symptom and / or the underlying cause of a symptom, ameliorating or repairing damage caused directly or indirectly by a malignancy. Treatment also includes prolonging survival as compared to the expected survival of a subject not receiving treatment. Subjects to be treated include those suffering from a condition or disease, as well as those susceptible to a condition or disease, or those in which a condition or disease is to be prevented.

[0044] "Therapeutically effective amount" refers to an amount of the disclosed composition effective to achieve the desired treatment at the required dosage and for the required period of time. The therapeutically effective amount may vary depending on factors such as the subject's condition, age, sex, and weight, as well as the ability of the combined therapy to induce the desired response in the subject. Exemplary indicators of a therapeutically effective amount include, for example, an improvement in the patient's health, a reduction in tumor burden, the cessation or slowing of tumor growth, and / or the absence of metastasis of cancer cells to other locations in the body.

[0045] A "pharmaceutical composition" refers to a composition comprising an active ingredient and a pharma- ceutically acceptable carrier.

[0046] A "pharmaceutically acceptable carrier" or "excipient" refers to an ingredient in a pharmaceutical composition, other than an active ingredient, that is not toxic to a subject.

[0047] The term "cancer," as used herein, is defined as a disease characterized by the rapid and uncontrolled growth of abnormal cells. Cancer cells can spread locally or via the bloodstream and lymphatic system to other parts of the body. In certain embodiments, the cancer is a hematological malignancy or a solid tumor. In some embodiments, the hematological malignancy is selected from the group consisting of multiple myeloma, smoldering multiple myeloma, monoclonal gammopathy of undetermined significance (MGUS), acute lymphoblastic leukemia (ALL), diffuse large B-cell lymphoma (DLBCL), Burkitt's lymphoma (BL), follicular lymphoma (FL), mantle-cell lymphoma (MCL), Waldenstrom's hypergammaglobulinemia, plasma cell leukemia, light chain amyloidosis (AL), precursor B-cell lymphoblastic leukemia, precursor B-cell lymphoblastic leukemia, acute myeloid leukemia (AML), myelodysplastic syndrome (MDS), myelodysplastic leukemia, myelodysplastic syndrome (MS ... and malignant lymphomas such as myeloid leukemia (CML), erythroblastic lymphoid tumors (ALT), leukemia, myelogenous leukemia (MDS), chronic lymphocytic leukemia (CLL), B-cell malignancies, chronic myeloid leukemia (CML), hairy cell leukemia (HCL), blastic plasmacytoid dendritic cell neoplasms, Hodgkin's lymphoma, non-Hodgkin's lymphoma, marginal zone B-cell lymphoma (MZL), mucosa-associated lymphatic tissue (MALT), plasma cell leukemia, anaplastic large-cell lymphoma (ALCL), leukemia, or lymphoma.

[0048] "Tumor cell" or "cancer cell" refers to a cancerous, precancerous, or transformed cell that has spontaneous or induced phenotypic changes, either in vivo, ex vivo, or in tissue culture. These changes do not necessarily involve the incorporation of new genetic material. Transformation may occur by infection with a transforming virus and incorporation of new genomic nucleic acid, incorporation of exogenous nucleic acid, or may occur spontaneously or after exposure to a carcinogen, thereby mutating endogenous genes. Transformation / cancer is exemplified by morphological changes, cellular immortalization, aberrant growth control, formation of foci, proliferation, malignant lesions, modulation of tumor-specific marker levels, invasiveness, tumor growth in suitable animal hosts such as nude mice, and the like, in vitro, in vivo, and ex vivo.

[0049] "T cell redirecting therapeutic" refers to a molecule that contains two or more binding regions, where one of the binding regions specifically binds to a cell surface antigen on a target cell or tissue, and where a second binding region of the molecule specifically binds to a T cell antigen. Examples of cell surface antigens include tumor-associated antigens such as BCMA. Examples of T cell antigens include, for example, CD3. This dual / multiple target binding ability recruits T cells to the target cell or tissue, resulting in eradication of the target cell or tissue.

[0050] A "subject" includes any human or non-human animal. A "non-human animal" includes any vertebrate, e.g., mammals and non-mammals, such as non-human primates, sheep, dogs, cats, horses, cows, chickens, amphibians, reptiles, etc. The terms "subject" and "patient" may be used interchangeably herein.

[0051] explanation Throughout this specification, the numbering of amino acid residues in antibody constant regions is according to the EU index as set forth in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD. (1991), unless otherwise expressly stated herein. The numbering of antibody constant chains can be found, for example, in the IMGT Web resource, IMGT Scientific charts, on the ImMunoGeneTics website.

[0052] The conventional one-letter and three-letter amino acid codes are used herein as shown in Table 1.

[0053] [Table 1]

[0054] BCMA×CD3 Bispecific Antibodies and Uses Thereof The present invention is based, at least in part, on the discovery that the therapeutic agent teclistamab, a bispecific BCMA / CD3 antibody, can be used to treat multiple myeloma in subjects that are relapsed or refractory to treatment with previous anti-cancer therapeutic agents.

[0055] B-cell maturation antigen (BCMA) is a cell membrane-bound tumor necrosis factor receptor family member involved in the differentiation of B cells into plasma cells. BCMA expression is restricted to the B-cell lineage, where it is expressed primarily within the intrafollicular regions of germinal centers and on differentiated plasma cells and plasmablasts. BCMA is virtually absent on naive and memory B cells (Tai and Anderson, Immunotherapy 7:1187-99, 2015).

[0056] Thus, disclosed herein is a stable aqueous pharmaceutical composition comprising a bispecific BCMA / CD3 antibody.

[0057] In one aspect, provided herein is a stable aqueous pharmaceutical composition comprising: (a) a bispecific B cell maturation antigen (BCMA) / cluster of differentiation 3 (CD3) antibody or antigen-binding fragment thereof at a concentration of about 7.5 mg / mL to about 12.5 mg / mL, wherein the bispecific BCMA / CD3 antibody or antigen-binding fragment thereof is (1) a first heavy chain (HC1) comprising an HC1 variable region 1 (VH1); wherein VH1 comprises heavy chain complementarity determining region 1 (HCDR1), HCDR2 and HCDR3 having the amino acid sequences of SEQ ID NOs: 1, 2 and 3, respectively; (2) a first light chain (LC1) comprising an LC1 variable region (VL1); wherein VL1 comprises a light chain complementarity determining region 1 (LCDR1), LCDR2 and LCDR3 having the amino acid sequences of SEQ ID NOs: 4, 5 and 6, respectively; (3) a second heavy chain (HC2) comprising an HC2 variable region 2 (VH2); and wherein VH2 comprises heavy chain complementarity determining region 1 (HCDR1), HCDR2 and HCDR3 having the amino acid sequences of SEQ ID NOs: 11, 12 and 13, respectively; (4) a second light chain (LC2) comprising an LC2 variable region 2 (VL2); and wherein VL2 comprises a light chain complementarity determining region 1 (LCDR1), LCDR2 and LCDR3 having the amino acid sequences of SEQ ID NOs: 14, 15 and 16, respectively; Including, (b) about 10 mM to about 20 mM acetate and / or a pharma- ceutically acceptable acetate salt; (c) about 6% (w / v) to about 10% (w / v) sucrose; (d) about 16 μg / mL to about 24 μg / mL of ethylenediaminetetraacetic acid (EDTA); (e) about 0.01% to about 0.07% polysorbate 20; and (f) a pH of about 4.7 to about 5.7; A stable aqueous pharmaceutical composition is provided comprising:

[0058] In another general aspect, provided herein is a stable aqueous pharmaceutical composition comprising: (a) a bispecific B cell maturation antigen (BCMA) / cluster of differentiation 3 (CD3) antibody or antigen-binding fragment thereof at a concentration of about 76.5 mg / mL to about 103.5 mg / mL, wherein the bispecific BCMA / CD3 antibody or antigen-binding fragment thereof is (1) a first heavy chain (HC1) comprising an HC1 variable region 1 (VH1); wherein VH1 comprises heavy chain complementarity determining region 1 (HCDR1), HCDR2 and HCDR3 having the amino acid sequences of SEQ ID NOs: 1, 2 and 3, respectively; (2) a first light chain (LC1) comprising an LC1 variable region (VL1); wherein VL1 comprises a light chain complementarity determining region 1 (LCDR1), LCDR2 and LCDR3 having the amino acid sequences of SEQ ID NOs: 4, 5 and 6, respectively; (3) a second heavy chain (HC2) comprising an HC2 variable region 2 (VH2); and wherein VH2 comprises heavy chain complementarity determining region 1 (HCDR1), HCDR2 and HCDR3 having the amino acid sequences of SEQ ID NOs: 11, 12 and 13, respectively; (4) a second light chain (LC2) comprising an LC2 variable region 2 (VL2); and wherein VL2 comprises a light chain complementarity determining region 1 (LCDR1), LCDR2 and LCDR3 having the amino acid sequences of SEQ ID NOs: 14, 15 and 16, respectively; Including, (b) about 10 mM to about 20 mM acetate and / or a pharma- ceutically acceptable acetate salt; (c) about 6% (w / v) to about 10% (w / v) sucrose; (d) about 16 μg / mL to about 24 μg / mL of ethylenediaminetetraacetic acid (EDTA); (e) about 0.01% to about 0.07% polysorbate 20; and (f) a pH of about 4.7 to about 5.7; A stable aqueous pharmaceutical composition is provided comprising:

[0059] In certain embodiments, the bispecific BCMA / CD3 antibody comprises a VH1 having the amino acid sequence of SEQ ID NO: 7 and a VL1 having the amino acid sequence of SEQ ID NO: 8. In certain embodiments, the bispecific BCMA / CD3 antibody comprises an HC1 having the amino acid sequence of SEQ ID NO: 9 and an LC1 having the amino acid sequence of SEQ ID NO: 10. In certain embodiments, the bispecific BCMA / CD3 antibody comprises a VH2 having the amino acid sequence of SEQ ID NO: 17 and a VL2 having the amino acid sequence of SEQ ID NO: 18. In certain embodiments, the bispecific BCMA / CD3 antibody comprises an HC2 having the amino acid sequence of SEQ ID NO: 19 and an LC2 having the amino acid sequence of SEQ ID NO: 20. The bispecific BCMA / CD3 antibody may be, for example, teclistamab.

[0060] In certain embodiments, the bispecific BCMA / CD3 antibody has a concentration of about 7 mg / mL to about 13 mg / mL, about 7.5 mg / mL to about 12.5 mg / mL, about 8 mg / mL to about 12 mg / mL, or about 9 mg / mL to about 11 mg / mL. The bispecific BCMA / CD3 antibody can have a concentration of, for example, about 7 mg / mL, about 7.5 mg / mL, about 8 mg / mL, about 9 mg / mL, about 10 mg / mL, about 11 mg / mL, about 12 mg / mL, about 12.5 mg / mL, or about 13 mg / mL, or any value therebetween. In a preferred embodiment, the bispecific BCMA / CD3 antibody has a concentration of about 10 mg / mL.

[0061] In certain embodiments, the bispecific BCMA / CD3 antibody has a concentration of about 76.5 mg / mL to about 103.5 mg / mL, about 81 mg / mL to about 99 mg / mL, about 85 mg / mL to about 95 mg / mL, or about 87 mg / mL to about 93 mg / mL. Bispecific BCMA / CD3 antibodies can have a concentration of, for example, about 76.5 mg / mL, about 77 mg / mL, about 78 mg / mL, about 79 mg / mL, about 80 mg / mL, about 81 mg / mL, about 82 mg / mL, about 83 mg / mL, about 84 mg / mL, about 85 mg / mL, about 86 mg / mL, about 87 mg / mL, about 88 mg / mL, about 89 mg / mL, about 90 mg / mL, about 91 mg / mL, about 92 mg / mL, about 93 mg / mL, about 94 mg / mL, about 95 mg / mL, about 96 mg / mL, about 97 mg / mL, about 98 mg / mL, about 99 mg / mL, about 100 mg / mL, about 101 mg / mL, about 102 mg / mL, about 103 mg / mL, or about 103.5 mg / mL, or any value in between. In a preferred embodiment, the bispecific BCMA / CD3 antibody has a concentration of about 90 mg / mL.

[0062] In certain embodiments, the composition comprises about 10 mM to about 20 mM, about 12 mM to about 18 mM, or about 14 mM to about 16 mM acetate and / or a pharma- ceutically acceptable acetate salt. The composition may comprise, for example, 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 acetate and / or a pharma- ceutically acceptable acetate salt, or any value therebetween. In a preferred embodiment, the composition comprises about 15 mM acetate or a pharma- ceutically acceptable acetate salt.

[0063] In certain embodiments, the composition comprises about 6% (w / v) to about 10% (w / v) or about 7% (w / v) to about 9% (w / v) sucrose. The composition may comprise, for example, about 6% (w / v), about 7% (w / v), about 8% (w / v), about 9% (w / v), or about 10% (w / v) sucrose, or any value therebetween.

[0064] In certain embodiments, the composition comprises about 16 mg / mL to about 24 mg / mL or about 18 mg / mL to about 22 mg / mL of EDTA. The composition may comprise, for example, about 16 mg / mL, about 17 mg / mL, about 18 mg / mL, about 19 mg / mL, about 20 mg / mL, about 21 mg / mL, about 22 mg / mL, about 23 mg / mL, or about 24 mg / mL of EDTA, or any value therebetween. In a preferred embodiment, the composition comprises about 20 mg / mL of EDTA.

[0065] In certain embodiments, the composition comprises about 0.01% to about 0.07%, about 0.02% to about 0.06%, or about 0.03% to about 0.05% polysorbate 20 (PS 20). The composition may comprise, for example, about 0.01%, about 0.02%, about 0.03%, about 0.04%, about 0.05%, about 0.06%, or about 0.07%, or any value therebetween, of PS 20. In a preferred embodiment, the composition comprises about 0.04% PS 20.

[0066] In certain embodiments, the pH of the composition is about 4.7 to about 5.7, about 4.8 to about 5.6, or about 4.9 to about 5.5. The pH of the composition can be, for example, about 4.7, about 4.8, about 4.9, about 5.0, about 5.1, about 5.2, about 5.3, about 5.4, about 5.5, about 5.6, or about 5.7, or any value therebetween. In a preferred embodiment, the pH of the composition is about 5.2.

[0067] The stability of the aqueous pharmaceutical compositions of the present disclosure, also referred to as drug products (DP), is determined based on the particular amounts or proportions of the pathlength BCMAxCD3 antibodies and other components of the DP provided herein, including but not limited to acetate and / or pharma- ceutically acceptable acetate salts, sucrose, PS20, and EDTA, as well as an evaluation of a variety of factors, including but not limited to solution color, pH, turbidity, percentage of purity, number of particles subvisible to the naked eye, percentage of new peaks, percentage of major components, percentage of high molecular weight species (HMWS), percentage of low molecular weight species (LMWS), percentage of total acidic peaks, percentage of total basic peaks, protein concentration, percentage of T cell activation, and / or percentage of PS20.

[0068] A stable DP disclosed herein should not be construed as requiring all factors listed herein, but rather as requiring at least one, at least two, or at least three or more of those factors. In some embodiments, a stable disclosed DP exhibits the following results for at least one, at least two, at least three, or more of the factors detailed below in this specification. In a preferred embodiment, a stable DP exhibits the following results for most of the factors detailed below in this specification. In a most preferred embodiment, a stable DP exhibits the following results for all factors detailed below in this specification.

[0069] Solution color The color of the DP solution can be monitored and evaluated to verify that the appearance of the solution is consistent with previous batches upon release and over shelf life. The color of the DP solution can reflect stability. In one embodiment, the stability of the DP is defined as having a solution color ranging from colorless to about BY2 or less, about BY4 or less, about B2 or less, about B4 or less, about Y2 or less, or about Y4 or less, as described in European Pharmacopoeia 2.2.2, Degree of Coloration of Liquids European Pharmacopoeia (Ph.Eur.) 10th Edition Monograph No. 20202, July 2019.

[0070] In one embodiment, stability is defined as having a solution color that is colorless to about BY2 or less, about B2 or less, to about Y2 or less after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, stability is defined as having a solution color that is colorless to about BY4 or less, about B4 or less, to about Y4 or less after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a most preferred embodiment, stability is defined as having a solution color that is colorless to about BY5 or less, about B5 or less, to about Y5 or less after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more.

[0071] pH Measuring the pH of the DP solution can ensure that the pH is consistent with previous DP batches upon release and over the shelf life. In one embodiment, the stability of a DP is defined as when its pH is about 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, or 7.0. In one embodiment, the pH of the DP is about 5.2 after about 12 months or more of storage at a temperature of about 5° C., after about 12 months or more of storage at a temperature of about 25° C., and / or after about 2 years or more of storage at a temperature of about 5° C. In one embodiment, the pH is in the range of about 4.7 to about 5.7 after about 12 months or more of storage at a temperature of about 5° C., after about 12 months or more of storage at a temperature of about 25° C., and / or after about 2 years or more of storage at a temperature of about 5° C. In a preferred embodiment, the stability of the DP is defined as a pH in the range of about 4.8 to about 5.6 after storage for about 12 months or more at a temperature of about 5° C., after storage for about 12 months or more at a temperature of about 25° C., and / or after storage for about 2 years or more at a temperature of about 5° C. In a most preferred embodiment, the stability of the DP is defined as a pH in the range of about 4.9 to about 5.5 after storage for about 12 months or more at a temperature of about 5° C., after storage for about 12 months or more at a temperature of about 25° C., and / or after storage for about 2 years or more at a temperature of about 5° C.

[0072] Turbidity Turbidity allows for the measurement of the presence of particles in the DP solution to ensure consistency with previous DP batches and allows for compendial guidance applicable at the time of release and over the shelf life. In one embodiment, the stability of a DP is defined as having a turbidity value of about 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 nephelometric turbidity units (NTU) after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In one embodiment, the stability of a DP is defined as having a turbidity value of about 18 NTU or less after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, the stability of a DP is defined as its turbidity value being about 13 NTU or less after storage for about 12 months or more at a temperature of about 5° C., after storage for about 12 months or more at a temperature of about 25° C., and / or after storage for about 2 years or more at a temperature of about 5° C. In a most preferred embodiment, the stability of a DP is defined as its turbidity value being about 8 NTU or less after storage for about 12 months or more at a temperature of about 5° C., after storage for about 12 months or more at a temperature of about 25° C., and / or after storage for about 2 years or more at a temperature of about 5° C.

[0073] particle analysis The stability of the DP is set to a certain threshold of particle contamination based on the average number of particles that are not visible to the naked eye. In one embodiment, the average number of particles present in the DP unit tested should not exceed 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 2000, 3000, 4000, 5000, or 6000 per container for a particle size of 10 μm or more. In one embodiment, the average number of particles present in the DP unit tested should not exceed 6000 per container for a particle size of 10 μm or more. In one embodiment, the average number of particles present in the DP unit tested should not exceed 100, 200, 300, 400, 500, or 600 per container for a particle size of 25 μm or more. In one embodiment, the average number of particles present in the DP unit tested should not exceed 600 per container for a particle size of 25 μm or more.

[0074] cSDS conditions Capillary SDS-PAGE (cSDS) is a method that separates denatured proteins based on their molecular weight, similar to gel-based SDS-PAGE. This process allows for quantifying DP purity and monitoring its stability upon release and over shelf life.

[0075] In one embodiment, DP stability is defined based on the results of various SDS variables (e.g., percent purity or presence of new peaks) when cSDS is performed under reducing or non-reducing conditions after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more.

[0076] In one embodiment, DP stability is defined as having a percent purity of about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or about 100% or equal to 100%, or any range therebetween.

[0077] In one embodiment, DP stability is defined as the cSDS result showing no new peaks greater than 0.5%, 0.8%, 0.9%, 1.0%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, or greater than 2% compared to an untreated reference material.

[0078] In one embodiment, DP stability is defined as having a purity percentage of about 90% or more and no new peaks of more than 1.5% compared to the reference material. In a preferred embodiment, DP stability is defined as having a purity percentage of about 95% or more and no new peaks of more than 1.2% compared to the reference material. In a most preferred embodiment, DP stability is defined as having a purity percentage of about 97% or more and no new peaks of more than 1.0% compared to the reference material.

[0079] Size-exclusion HPLC (SE-HPLC) results consistent with stability The SE-HPLC procedure makes it possible to assess the purity of the DP and to monitor its stability under non-denaturing conditions upon release and over its shelf life.

[0080] In one embodiment, DP stability is defined based on the results of various SE-HPLC variables such as major components (MC), high molecular weight species (HMWS), or low molecular weight species (LMWS) after storage of the DP at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more.

[0081] In one embodiment, DP stability is defined as having an MC equal to about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or about 100%, or any range therebetween. In one embodiment, DP stability is defined as having an MC of about 90% or more. In a preferred embodiment, DP stability is defined as having an MC of about 95% or more. In a most preferred embodiment, DP stability is defined as having an MC of about 97% or more.

[0082] In one embodiment, DP stability is defined as having an HMWS of about 0.1%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, or any range therebetween. In one embodiment, DP stability is defined as having an HMWS of about 10% or less. In a preferred embodiment, DP stability is defined as having an HMWS of about 5% or less. In a most preferred embodiment, DP stability is defined as having an HMWS of about 3% or less.

[0083] In one embodiment, DP stability is defined as having an LMWS of about 0.1%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, or 10%. In one embodiment, DP stability is defined as having an LMWS of about 5% or less. In a preferred embodiment, DP stability is defined as having an LMWS of about 2% or less. In a most preferred embodiment, DP stability is defined as having an LMWS of about 1% or less.

[0084] Capillary isoelectric focusing (cIEF) cIEF separates proteins based on their overall charge or isoelectric point (pI), similar to isoelectric gel electrophoresis (IEF). This procedure allows for monitoring the distribution of charge-based isoforms of a drug upon release and over shelf life. In one embodiment, DP stability is defined based on the results of various cIEF variables, such as Main Peak (MP), sum of acidic peaks, or sum of basic peaks, after storage of the DP at a temperature of about 25° C. for about 12 months or more and / or after storage at a temperature of about 5° C. for about 2 years or more.

[0085] In one embodiment, DP stability is defined as having a cIEF with MP of about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, or any range therebetween, after storage of the DP at a temperature of about 25° C. for about 12 months or more and / or after storage at a temperature of about 5° C. for about 2 years or more. In one embodiment, DP stability is defined as having a cIEF of MP≧60% after storage of the DP at a temperature of about 25° C. for about 12 months or more and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, DP stability is defined as having a cIEF of MP≧65% after storage of the DP at a temperature of about 25° C. for about 12 months or more and / or after storage at a temperature of about 5° C. for about 2 years or more. In a most preferred embodiment, DP stability is defined as having a cIEF of MP≧70% after storage of the DP at a temperature of about 25° C. for about 12 months or more and / or after storage at a temperature of about 5° C. for about 2 years or more.

[0086] In one embodiment, DP stability is defined as having a cIEF with a sum of acidic peaks totaling about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, or any range therebetween after storage of the DP at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In one embodiment, DP stability is defined as having a cIEF with acidic peaks totaling ≦40% after storage of the DP at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, DP stability is defined as having a cIEF with acidic peaks totaling about ≦30% after storage of the DP at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a most preferred embodiment, DP stability is defined as having a cIEF with acidic peaks totaling about ≦25% after storage of the DP at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more.

[0087] In one embodiment, DP stability is defined as having a cIEF with basic peaks totaling about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or 20%, or any range therebetween, after storage of the DP at a temperature of about 25° C. for about 12 months or more and / or after storage at a temperature of about 5° C. for about 2 years or more. In one embodiment, DP stability is defined as having a cIEF with basic peaks totaling about 15% or less after storage of the DP at a temperature of about 25° C. for about 12 months or more and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, DP stability is defined as having a cIEF with basic peaks totaling about 10% or less after storage of the DP at a temperature of about 25° C. for about 12 months or more and / or after storage at a temperature of about 5° C. for about 2 years or more. In a most preferred embodiment, DP stability is defined as having a cIEF with a total of basic peaks totaling less than about 8% after storage of the DP at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more.

[0088] Protein concentration The protein concentration of the DP allows verification that it matches previous DP batches at the time of shipment and over the shelf life. Protein concentration can be quantified by measuring the UV light absorbance of the pharmaceutical solution at 280 nm (A280).

[0089] 10mg / mL DP formulation In one embodiment, DP stability is defined as having a protein concentration of about 5 mg / mL, 6 mg / mL, 7 mg / mL, 8 mg / mL, 9 mg / mL, 10 mg / mL, 11 mg / mL, 12 mg / mL, 13 mg / mL, 14 mg / mL, 15 mg / mL, or any value therebetween, after storage of the DP at a temperature of about 25° C. for about 12 months or more and / or after storage at a temperature of about 5° C. for about 2 years or more. In one embodiment, DP stability is defined as having a protein concentration of about 7 mg / mL to about 13 mg / mL after storage of the DP at a temperature of about 25° C. for about 12 months or more and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, DP stability is defined as having a protein concentration of about 8 mg / mL to about 12 mg / mL after storage of the DP at a temperature of about 25° C. for about 12 months or more and / or after storage at a temperature of about 5° C. for about 2 years or more. In a most preferred embodiment, DP stability is defined as having a protein concentration of about 9 mg / mL to 11 mg / mL after storage of the DP at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more.

[0090] 90mg / mL DP formulation In one embodiment, DP stability is defined as having a protein concentration of 75 mg / mL, 76 mg / mL, 77 mg / mL, 78 mg / mL, 79 mg / mL, 80 mg / mL, 81 mg / mL, 82 mg / mL, 83 mg / mL, 84 mg / mL, 85 mg / mL, 86 mg / mL, 87 mg / mL, 88 mg / mL, 89 mg / mL, 90 mg / mL, 91 mg / mL, 92 mg / mL, 93 mg / mL, 94 mg / mL, 95 mg / mL, 96 mg / mL, 97 mg / mL, 98 mg / mL, 99 mg / mL, 100 mg / mL, 101 mg / mL, 102 mg / mL, 103 mg / mL, 104 mg / mL or 105 mg / mL, or any value therebetween, after storage of the DP at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In one embodiment, DP stability is defined as having a protein concentration of about 81 mg / mL to about 99 mg / mL after storage of the DP at a temperature of about 25° C. for about 12 months or more and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, DP stability is defined as having a protein concentration of about 85 mg / mL to about 95 mg / mL after storage of the DP at a temperature of about 25° C. for about 12 months or more and / or after storage at a temperature of about 5° C. for about 2 years or more. In a most preferred embodiment, DP stability is defined as having a protein concentration of about 87 mg / mL to about 93 mg / mL after storage of the DP at a temperature of about 25° C. for about 12 months or more and / or after storage at a temperature of about 5° C. for about 2 years or more.

[0091] Peptide mapping Post-translational modifications (PTMs), such as oxidation, deamidation, and isomerization, are enzymatic modifications that can be detected in the structure of an antibody. In some embodiments, PD stability is evaluated based on the level of PTMs in the antibody. The test article is enzymatically digested to obtain peptide segments. These peptides are then evaluated, for example, by mass spectrometry (MS), tandem mass spectrometry (MS-MS), or Ultra High-Performance Liquid Chromatography Mass Spectroscopy (UPLC-MS). Each analyzed peptide sequence is identified relative to its known location in the overall antibody structure. Post-translational modifications are measured by comparing the measured mass of the identified peptide sequence with its expected mass.

[0092] Drug efficacy An in vitro T cell activation assay allows one to assess the level of DP stability. This activation can be assessed, but is not limited to, by using the nuclear factor of activated T cells response element (NFAT-RE)-mediated luminescence assay.

[0093] T cell activation by BCMA×CD3 In one embodiment, DP stability is defined as having about 40%, 50%, 60%, 70%, 80%, 90%, 100%, 110%, 120%, 130%, 140%, 150%, or 160%, or any range therebetween, of BMCA×CD3-mediated T cell activation activity, relative to a reference, after storage of the DP at a temperature of about 5° C. for about 12 months or more, after storage of the DP at a temperature of about 25° C. for about 12 months or more, and / or after storage of the DP at a temperature of about 5° C. for about 2 years or more. In one embodiment, DP stability is defined as having about 50% to about 150% of BMCA×CD3-mediated T cell activation activity, relative to a reference, after storage of the DP at a temperature of about 25° C. for about 12 months or more, and / or after storage of the DP at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, DP stability is defined as having a BMCA×CD3-mediated T cell activation activity in the range of about 60% to about 140% compared to a reference after storage of the DP for about 12 months or more at a temperature of about 25° C. and / or after storage for about 2 years or more at a temperature of about 5° C. In a most preferred embodiment, DP stability is defined as having a BMCA×CD3-mediated T cell activation activity in the range of about 80% to about 120% compared to a reference after storage of the DP for about 12 months or more at a temperature of about 25° C. and / or after storage for about 2 years or more at a temperature of about 5° C.

[0094] Polysorbate 20 (PS20) In one embodiment, DP stability is defined by a PS20 concentration of about 0.005%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.10%, or any range therebetween, weight to volume percentage, after storage of the DP at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In one embodiment, DP stability is defined by a PS20 concentration of about 0.02% to about 0.1% after storage of the DP at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In one embodiment, DP stability is defined by a PS20 concentration of about 0.01% to about 0.07%. In a preferred embodiment, DP stability is defined by a PS20 concentration of about 0.02% to about 0.06% after storage of the DP for about 12 months or more at a temperature of about 25° C. and / or after storage for about 2 years or more at a temperature of about 5° C. In a most preferred embodiment, DP stability is defined by a PS20 concentration of about 0.03% to about 0.05% after storage of the DP for about 12 months or more at a temperature of about 25° C. and / or after storage for about 2 years or more at a temperature of about 5° C.

[0095] Total Volume of Stable Aqueous Pharmaceutical Composition 10 mg / mL formulation In one embodiment, the total volume of the stable aqueous pharmaceutical composition (or DP) is in the range of about 5 mL to about 10 mL. In one embodiment, the total volume of the stable aqueous pharmaceutical composition (or DP) is in the range of about 0.5 mL to about 20 mL, about 1 mL to about 15 mL, about 5 mL to about 10 mL, or about 6 mL to about 8 mL. In one embodiment, the total volume of the stable aqueous pharmaceutical composition is about 0.5 mL, 0.6 mL, 0.7 mL, 0.8 mL, 0.9 mL, 1 mL, 2 mL, 3 mL, 4 mL, 5 mL, 6 mL, 8 mL, 9 mL, 10 mL, 11 mL, 12 mL, 13 mL, 14 mL, 15 mL, 16 mL, 18 mL, 19 mL, 20 mL, 25 mL, or 30 mL, or any range therebetween. In one embodiment, the total volume of the stable aqueous pharmaceutical composition is 2 mL.

[0096] 90 mg / mL formulation In one embodiment, the total volume of the stable aqueous pharmaceutical composition (or DP) is in the range of about 5 mL to about 10 mL. In one embodiment, the total volume of the stable aqueous pharmaceutical composition (or DP) is in the range of about 0.5 mL to about 20 mL, about 1 mL to about 15 mL, about 5 mL to about 10 mL, or about 6 mL to about 8 mL. In one embodiment, the total volume of the stable aqueous pharmaceutical composition is about 0.5 mL, 0.6 mL, 0.7 mL, 0.8 mL, 0.9 mL, 1 mL, 2 mL, 3 mL, 4 mL, 5 mL, 6 mL, 8 mL, 9 mL, 10 mL, 11 mL, 12 mL, 13 mL, 14 mL, 15 mL, 16 mL, 18 mL, 19 mL, 20 mL, 25 mL, or 30 mL, or any range therebetween. In one embodiment, the total volume of the stable aqueous pharmaceutical composition is 3.5 mL.

[0097] method Provided herein is a method for treating cancer in a subject in need of such treatment.The method comprises administering to the subject a stable aqueous pharmaceutical composition as disclosed herein.In certain embodiments, the administration is intravenous.

[0098] cancer In some embodiments, the cancer is a hematological malignancy or a solid tumor.

[0099] In some embodiments, the hematological malignancy is selected from the group consisting of multiple myeloma, smoldering multiple myeloma, monoclonal gammopathy of undetermined significance (MGUS), acute lymphoblastic leukemia (ALL), diffuse large B-cell lymphoma (DLBCL), Burkitt's lymphoma (BL), follicular lymphoma (FL), mantle-cell lymphoma (MCL), Waldenstrom's hypergammaglobulinemia, plasma cell leukemia, light chain amyloidosis (AL), precursor B-cell lymphoblastic leukemia, precursor B-cell lymphoblastic leukemia, acute myeloid leukemia (AML), myelodysplastic syndrome (MDS), myelodysplastic leukemia, myelodysplastic syndrome (MS ... and malignant lymphomas such as myeloid leukemia (CML), erythroblastic lymphoid tumors (ALT), leukemia, myelogenous leukemia (MDS), chronic lymphocytic leukemia (CLL), B-cell malignancies, chronic myeloid leukemia (CML), hairy cell leukemia (HCL), blastic plasmacytoid dendritic cell neoplasms, Hodgkin's lymphoma, non-Hodgkin's lymphoma, marginal zone B-cell lymphoma (MZL), mucosa-associated lymphatic tissue (MALT), plasma cell leukemia, anaplastic large-cell lymphoma (ALCL), leukemia, or lymphoma.

[0100] In preferred embodiments, the hematological malignancy is multiple myeloma. In some embodiments, the subject has newly diagnosed multiple myeloma. In some embodiments, the subject is relapsed or refractory to treatment with a previous anti-cancer therapy, such as a therapeutic agent used to treat multiple myeloma or other hematological malignancies.

[0101] In some embodiments, the subject is refractory or relapsed to treatment with one or more therapies or therapies, such as THALOMID® (thalidomide), REVLIMID® (lenalidomide), POMALYST® (pomalidomide), VELCADE® (bortezomib), NINLARO (ixazomib), KYPROLIS® (carfilzomib), FARADYK® (pambinostat), AREDIA® (pamidronate), ZOMETA® (zoledronic acid), DARZALEX® (daratumumab), elotozumab or melphalan, Xpovio® (selinexor), Venclexta® (venetoclax), GSK916, CAR-T therapy, other BCMA-directed therapies, etc.

[0102] A variety of qualitative and / or quantitative methods can be used to determine disease recurrence or refractoryness. Possible associated symptoms are, for example, a decline or plateau in the patient's health status, recurrence or worsening of various symptoms associated with solid tumors, and / or metastasis of cancerous cells from one location to other organs, tissues, or cells within the body.

[0103] In some embodiments, the multiple myeloma is relapsed or refractory to treatment with selinexor, venetoclax, anti-CD38 antibodies, lenalinomide, bortezomib, pomalidomide, carfilzomib, elotozumab, ixazomib, melphalan, or thalidomide, or any combination thereof.

[0104] In some embodiments, the multiple myeloma is high-risk multiple myeloma. Subjects with high-risk multiple myeloma are known to relapse early and have poor prognosis and outcome. Subjects that can be classified as having high-risk multiple myeloma have one or more of the following cytogenetic abnormalities: t(4;14)(p16;q32), t(14;16)(q32;q23), del17p, 1qAmp, t(4;14)(p16;q32) and t(14;16)(q32;q23), t(4;14)(p16;q32) and del17p, t(14;16)(q32;q23) and del17p, or t(4;14)(p16;q32), t(14;16)(q32;q23) and del17p. In some embodiments, subjects with high-risk multiple myeloma may have one or more chromosomal abnormalities including the following: t(4;14)(p16;q32), t(14;16)(q32;q23), del17p, 1qAmp, t(4;14)(p16;q32) and t(14;16)(q32;q23), t(4;14)(p16;q32) and del17p, t(14;16)(q32;q23) and del17p; or t(4;14)(p16;q32), t(14;16)(q32;q23) and del17p, or any combination thereof.

[0105] Cytogenetic abnormalities can be detected, for example, by fluorescent in situ hybridization (FISH). In chromosomal translocations, oncogenes are translocated to the IgH region on chromosome 14q32, leading to dysregulation of these genes. t(4;14)(p16;q32) involves translocation of fibroblast growth factor receptor 3 (FGFR3) and multiple myeloma SET domain-containing protein (MMSET) (also known as WHSC1 / NSD2), and t(14;16)(q32;q23) involves translocation of MAF transcription factor C-MAF. 17p deletion (del17p) involves loss of the p53 locus.

[0106] Chromosomal rearrangements can be identified using well-known methods, such as fluorescent in situ hybridization, karyotyping, pulsed-field gel electrophoresis, or sequencing.

[0107] 1. A method for preparing a stable aqueous pharmaceutical composition of a bispecific B-cell maturation antigen (BCMA) / cluster of differentiation 3 (CD3) antibody or antigen-binding fragment thereof, wherein the bispecific BCMA / CD3 antibody or antigen-binding fragment thereof comprises a first heavy chain (HC1) comprising an HC1 variable region 1 (VH1), wherein VH1 comprises a heavy chain complementarity determining region 1 (HCDR1), HCDR2, and HCDR3 having the amino acid sequences of SEQ ID NOs: 1, 2, and 3, respectively, and a first light chain (LC1) comprising an LC1 variable region 2 (VL1) comprising a heavy chain complementarity determining region 1 (HCDR1), HCDR2, and HCDR3 having the amino acid sequences of SEQ ID NOs: 4, 5, and 6, respectively. and a second heavy chain (HC2) comprising an HC2 variable region 2 (VH2) comprising light chain complementarity determining region 1 (LCDR1), LCDR2 and LCDR3 having the amino acid sequences of SEQ ID NOs: 11, 12 and 13, respectively, wherein VH2 comprises heavy chain complementarity determining region 1 (HCDR1), HCDR2 and HCDR3 having the amino acid sequences of SEQ ID NOs: 11, 12 and 13, respectively, and a second light chain (LC2) comprising an LC2 variable region 2 (VL2), wherein VL2 comprises light chain complementarity determining region 1 (LCDR1), LCDR2 and LCDR3 having the amino acid sequences of SEQ ID NOs: 14, 15 and 16, respectively; Also provided herein is a method comprising combining about 10 mg / mL or about 90 mg / mL of a bispecific BCMA / CD3 antibody, about 15 mM acetate and / or a pharma- ceutically acceptable acetate salt, about 8% (w / v) sucrose, about 20 mg / mL EDTA, and about 0.04% polysorbate (PS) 20, wherein the stable aqueous pharmaceutical composition has a pH of about 5.2.

[0108] In certain embodiments, the bispecific BCMA / CD3 antibody comprises a VH1 having the amino acid sequence of SEQ ID NO: 7 and a VL1 having the amino acid sequence of SEQ ID NO: 8. In certain embodiments, the bispecific BCMA / CD3 antibody comprises an HC1 having the amino acid sequence of SEQ ID NO: 9 and an LC1 having the amino acid sequence of SEQ ID NO: 10. In certain embodiments, the bispecific BCMA / CD3 antibody comprises a VH2 having the amino acid sequence of SEQ ID NO: 17 and a VL2 having the amino acid sequence of SEQ ID NO: 18. In certain embodiments, the bispecific BCMA / CD3 antibody comprises an HC2 having the amino acid sequence of SEQ ID NO: 19 and an LC2 having the amino acid sequence of SEQ ID NO: 20. The bispecific BCMA / CD3 antibody may be, for example, teclistamab.

[0109] The stable aqueous pharmaceutical compositions disclosed herein can be packaged in a kit, container, pack, dispenser, or vial.

[0110] Also provided herein is a kit comprising the stable aqueous medicament of the present disclosure and instructions for its use.

[0111] Also provided herein is an article of manufacture comprising a container holding a stable aqueous pharmaceutical composition of the present disclosure, hi some embodiments, the container is a vial having a stopper pierceable by a syringe.

[0112] Embodiment Exemplary embodiments of the disclosed technology are provided herein, which are for illustrative purposes only and are not intended to limit the scope of the disclosure or the claims appended hereto.

[0113] Embodiment 1 is a stable aqueous pharmaceutical composition comprising: a) a bispecific B cell maturation antigen (BCMA) / cluster of differentiation 3 (CD3) antibody or antigen-binding fragment thereof at a concentration of about 7.5 mg / mL to about 12.5 mg / mL, wherein the bispecific BCMA / CD3 antibody or antigen-binding fragment thereof is (1) a first heavy chain (HC1) comprising an HC1 variable region 1 (VH1); wherein VH1 comprises heavy chain complementarity determining region 1 (HCDR1), HCDR2 and HCDR3 having the amino acid sequences of SEQ ID NOs: 1, 2 and 3, respectively; (2) a first light chain (LC1) comprising an LC1 variable region (VL1); wherein VL1 comprises a light chain complementarity determining region 1 (LCDR1), LCDR2 and LCDR3 having the amino acid sequences of SEQ ID NOs: 4, 5 and 6, respectively; (3) a second heavy chain (HC2) comprising an HC2 variable region 2 (VH2); and wherein VH2 comprises heavy chain complementarity determining region 1 (HCDR1), HCDR2 and HCDR3 having the amino acid sequences of SEQ ID NOs: 11, 12 and 13, respectively; (4) a second light chain (LC2) comprising an LC2 variable region 2 (VL2); and wherein VL2 comprises a light chain complementarity determining region 1 (LCDR1), LCDR2 and LCDR3 having the amino acid sequences of SEQ ID NOs: 14, 15 and 16, respectively; Including, b) about 10 mM to about 20 mM acetate and / or a pharma- ceutically acceptable acetate salt; c) about 6% (w / v) to about 10% (w / v) sucrose; d) about 16 μg / mL to about 24 μg / mL of ethylenediaminetetraacetic acid (EDTA); e) about 0.01% to about 0.07% polysorbate 20; f) a pH of about 4.7 to about 5.7; A stable aqueous pharmaceutical composition comprising:

[0114] Embodiment 1a is a stable aqueous pharmaceutical composition comprising: a) a bispecific B cell maturation antigen (BCMA) / cluster of differentiation 3 (CD3) antibody or antigen-binding fragment thereof at a concentration of about 76.5 mg / mL to about 103.5 mg / mL, wherein the bispecific BCMA / CD3 antibody or antigen-binding fragment thereof is (1) a first heavy chain (HC1) comprising an HC1 variable region 1 (VH1); wherein VH1 comprises heavy chain complementarity determining region 1 (HCDR1), HCDR2 and HCDR3 having the amino acid sequences of SEQ ID NOs: 1, 2 and 3, respectively; (2) a first light chain (LC1) comprising an LC1 variable region (VL1); wherein VL1 comprises a light chain complementarity determining region 1 (LCDR1), LCDR2 and LCDR3 having the amino acid sequences of SEQ ID NOs: 4, 5 and 6, respectively; (3) a second heavy chain (HC2) comprising an HC2 variable region 2 (VH2); and wherein VH2 comprises heavy chain complementarity determining region 1 (HCDR1), HCDR2 and HCDR3 having the amino acid sequences of SEQ ID NOs: 11, 12 and 13, respectively; (4) a second light chain (LC2) comprising an LC2 variable region 2 (VL2); and wherein VL2 comprises a light chain complementarity determining region 1 (LCDR1), LCDR2 and LCDR3 having the amino acid sequences of SEQ ID NOs: 14, 15 and 16, respectively; Including, b) about 10 mM to about 20 mM acetate and / or a pharma- ceutically acceptable acetate salt; c) about 6% (w / v) to about 10% (w / v) sucrose; d) about 16 μg / mL to about 24 μg / mL of ethylenediaminetetraacetic acid (EDTA); e) about 0.01% to about 0.07% polysorbate 20; f) a pH of about 4.7 to about 5.7; A stable aqueous pharmaceutical composition comprising:

[0115] Embodiment 2 is a stable aqueous pharmaceutical composition according to embodiment 1 or embodiment 1a, wherein the bispecific BCMA / CD3 antibody comprises a VH1 having the amino acid sequence of SEQ ID NO:7 and a VL1 having the amino acid sequence of SEQ ID NO:8.

[0116] Embodiment 3 is a stable aqueous pharmaceutical composition according to Embodiment 1 or Embodiment 1a, wherein the bispecific BCMA / CD3 antibody comprises a HC having the amino acid sequence of SEQ ID NO: 9 and an LC1 having the amino acid sequence of SEQ ID NO: 10.

[0117] Embodiment 4 is a stable aqueous pharmaceutical composition according to embodiment 1 or embodiment 1a, wherein the bispecific BCMA / CD3 antibody comprises a VH2 having the amino acid sequence of SEQ ID NO: 17 and a VL2 having the amino acid sequence of SEQ ID NO: 18.

[0118] Embodiment 5 is a stable aqueous pharmaceutical composition according to Embodiment 1 or Embodiment 1a, wherein the bispecific BCMA / CD3 antibody comprises an HC2 having the amino acid sequence of SEQ ID NO: 19 and an LC2 having the amino acid sequence of SEQ ID NO: 20.

[0119] Embodiment 6 is the stable aqueous pharmaceutical composition of Embodiment 1 or 1a, wherein the bispecific BCMA / CD3 antibody is teclistamab.

[0120] Embodiment 7 is the stable aqueous pharmaceutical composition of Embodiment 1 or 1a, wherein the bispecific BCMA / CD3 antibody has a concentration of about 8 mg / mL to about 12 mg / mL.

[0121] Embodiment 8 is the stable aqueous pharmaceutical composition of embodiment 7, wherein the bispecific BCMA / CD3 antibody has a concentration of about 9 mg / mL to about 11 mg / mL.

[0122] Embodiment 9 is a stable aqueous pharmaceutical composition according to embodiment 8, wherein the bispecific BCMA / CD3 antibody has a concentration of about 10 mg / mL.

[0123] Embodiment 10 is the stable aqueous pharmaceutical composition of Embodiment 1 or 1a, wherein the bispecific BCMA / CD3 antibody has a concentration of about 85 mg / mL to about 95 mg / mL.

[0124] Embodiment 11 is the stable aqueous pharmaceutical composition of Embodiment 10, wherein the bispecific BCMA / CD3 antibody has a concentration of about 87 mg / mL to about 93 mg / mL.

[0125] Embodiment 12 is the stable aqueous pharmaceutical composition of embodiment 11, wherein the bispecific BCMA / CD3 antibody has a concentration of about 90 mg / mL.

[0126] Embodiment 13 is a stable aqueous pharmaceutical composition according to embodiment 1 or 1a, comprising about 12 mM to about 18 mM acetate and / or a pharma- ceutically acceptable acetate salt.

[0127] Embodiment 14 is a stable aqueous pharmaceutical composition according to embodiment 1 or 1a, comprising about 14 mM to about 16 mM acetate and / or a pharma- ceutically acceptable acetate salt.

[0128] Embodiment 15 is a stable aqueous pharmaceutical composition according to embodiment 1 or 1a, comprising about 15 mM acetate and / or a pharma- ceutically acceptable acetate salt.

[0129] Embodiment 16 is a stable aqueous pharmaceutical composition according to embodiment 1 or 1a, comprising about 7% (w / v) to about 9% (w / v) sucrose.

[0130] Embodiment 17 is a stable aqueous pharmaceutical composition according to embodiment 16, wherein the composition comprises about 8% (w / v) sucrose.

[0131] Embodiment 18 is a stable aqueous pharmaceutical composition according to embodiment 1 or 1a, comprising about 18 μg / mL to about 22 μg / mL of EDTA.

[0132] Embodiment 19 is a stable aqueous pharmaceutical composition according to embodiment 1 or 1a, comprising about 20 μg / mL EDTA.

[0133] Embodiment 20 is a stable aqueous pharmaceutical composition according to embodiment 1 or 1a, comprising about 0.02% to about 0.06% PS-20.

[0134] Embodiment 21 is the stable aqueous pharmaceutical composition according to embodiment 20, comprising about 0.03 to about 0.05% PS-20.

[0135] Embodiment 22 is a stable aqueous pharmaceutical composition according to embodiment 21, comprising about 0.04% PS-20.

[0136] Embodiment 23 is a stable aqueous pharmaceutical composition according to embodiment 1 or 1a, having a pH of about 4.8 to about 5.6.

[0137] Embodiment 24 is the stable aqueous pharmaceutical composition of embodiment 23, having a pH of about 4.9 to about 5.5.

[0138] Embodiment 25 is a stable aqueous pharmaceutical composition according to embodiment 24, wherein the pH is about 5.2.

[0139] Embodiment 26 is a stable aqueous pharmaceutical composition according to embodiment 1 or 1a comprising 10 mg / mL of the bispecific BCMA / CD3 antibody, 15 mM acetate and / or a pharma- ceutically acceptable acetate salt, 8% (w / v) sucrose, 20 μg / mL EDTA, 0.04% PS 20, and a pH of 5.2.

[0140] Embodiment 27 is a stable aqueous pharmaceutical composition according to embodiment 1 or 1a, which is stable at a temperature of about 2-8° C. for at least 2 years.

[0141] Embodiment 28 is a stable aqueous pharmaceutical composition according to embodiment 1 or 1a, wherein the stability is defined based on solution color, pH, turbidity, percentage of purity, percentage of new peaks, percentage of major components, percentage of high molecular weight species (HWMS), percentage of low molecular weight species (LMWS), percentage of sum of acidic peaks, percentage of sum of basic peaks, protein concentration, percentage of T cell activation, percentage of PS 20 (w / v), or any combination thereof.

[0142] Embodiment 29 is a method of treating cancer in a subject in need thereof, comprising administering to the subject the stable aqueous pharmaceutical composition of embodiment 1 or 1a.

[0143] Embodiment 30 is the method of embodiment 29, wherein the administering is subcutaneously.

[0144]

[0033] Embodiment 31 is a method for preparing a stable aqueous pharmaceutical composition of a bispecific B-cell maturation antigen (BCMA) / cluster of differentiation 3 (CD3) antibody or antigen-binding fragment thereof, comprising: (1) a first heavy chain (HC1) comprising an HC1 variable region 1 (VH1); wherein VH1 comprises heavy chain complementarity determining region 1 (HCDR1), HCDR2 and HCDR3 having the amino acid sequences of SEQ ID NOs: 1, 2 and 3, respectively; (2) a first light chain (LC1) comprising an LC1 variable region (VL1); wherein VL1 comprises a light chain complementarity determining region 1 (LCDR1), LCDR2 and LCDR3 having the amino acid sequences of SEQ ID NOs: 4, 5 and 6, respectively; (3) a second heavy chain (HC2) comprising an HC2 variable region 2 (VH2); and wherein VH2 comprises heavy chain complementarity determining region 1 (HCDR1), HCDR2 and HCDR3 having the amino acid sequences of SEQ ID NOs: 11, 12 and 13, respectively; (4) a second light chain (LC2) comprising an LC2 variable region 2 (VL2); and wherein VL2 comprises a light chain complementarity determining region 1 (LCDR1), LCDR2 and LCDR3 having the amino acid sequences of SEQ ID NOs: 14, 15 and 16, respectively; Including, The method includes combining a composition comprising about 10 mg / mL of a bispecific BCMA / CD3 antibody, about 15 mM acetate and / or a pharma- ceutically acceptable acetate salt, about 8% (w / v) sucrose, about 20 mg / mL EDTA, and about 0.04% polysorbate (PS) 20, wherein the stable aqueous pharmaceutical composition has a pH of about 5.2.

[0145]

[0033] Embodiment 31a is a method for preparing a stable aqueous pharmaceutical composition of a bispecific B-cell maturation antigen (BCMA) / cluster of differentiation 3 (CD3) antibody or antigen-binding fragment thereof, the bispecific BCMA / CD3 antibody or antigen-binding fragment thereof comprising: (1) a first heavy chain (HC1) comprising an HC1 variable region 1 (VH1); wherein VH1 comprises heavy chain complementarity determining region 1 (HCDR1), HCDR2 and HCDR3 having the amino acid sequences of SEQ ID NOs: 1, 2 and 3, respectively; (2) a first light chain (LC1) comprising an LC1 variable region (VL1); wherein VL1 comprises a light chain complementarity determining region 1 (LCDR1), LCDR2 and LCDR3 having the amino acid sequences of SEQ ID NOs: 4, 5 and 6, respectively; (3) a second heavy chain (HC2) comprising an HC2 variable region 2 (VH2); and wherein VH2 comprises heavy chain complementarity determining region 1 (HCDR1), HCDR2 and HCDR3 having the amino acid sequences of SEQ ID NOs: 11, 12 and 13, respectively; (4) a second light chain (LC2) comprising an LC2 variable region 2 (VL2); and wherein VL2 comprises a light chain complementarity determining region 1 (LCDR1), LCDR2 and LCDR3 having the amino acid sequences of SEQ ID NOs: 14, 15 and 16, respectively; Including, The method comprises combining about 90 mg / mL of a bispecific BCMA / CD3 antibody, about 15 mM acetate and / or a pharma- ceutically acceptable acetate salt, about 8% (w / v) sucrose, about 20 mg / mL EDTA, and about 0.04% polysorbate (PS) 20, wherein the stable aqueous pharmaceutical composition has a pH of about 5.2.

[0146] Embodiment 32 is the method of embodiment 31 or 31a, wherein the bispecific BCMA / CD3 antibody comprises a VH1 having the amino acid sequence of SEQ ID NO:7 and a VL1 having the amino acid sequence of SEQ ID NO:8.

[0147] Embodiment 33 is the method of embodiment 31 or 31a, wherein the bispecific BCMA / CD3 antibody comprises a HC having the amino acid sequence of SEQ ID NO: 9 and an LC1 having the amino acid sequence of SEQ ID NO: 10.

[0148] Embodiment 34 is the method of embodiment 31 or 31a, wherein the bispecific BCMA / CD3 antibody comprises a VH2 having the amino acid sequence of SEQ ID NO: 17 and a VL2 having the amino acid sequence of SEQ ID NO: 18.

[0149] Embodiment 35 is the method of embodiment 31 or 31a, wherein the bispecific BCMA / CD3 antibody comprises an HC2 having the amino acid sequence of SEQ ID NO: 19 and an LC2 having the amino acid sequence of SEQ ID NO: 20.

[0150] Embodiment 36 is the method of embodiment 31 or 31a, wherein the bispecific BCMA / CD3 antibody is teclistamab.

[0151] Embodiment 37 is a kit comprising the stable aqueous pharmaceutical composition of embodiment 1 and instructions for use.

[0152] Embodiment 38 is an article of manufacture comprising a container holding a stable aqueous pharmaceutical composition of embodiment 1 or 1a.

[0153] Embodiment 39 is the article of manufacture of embodiment 38, wherein the container is a vial having a stopper pierceable by a syringe.

[0154] Embodiment 40 is a stable aqueous pharmaceutical composition according to embodiment 1 or 1a for use in the treatment of cancer.

[0155] Embodiment 41 is a stable aqueous pharmaceutical composition according to embodiment 1 or 1a for use in the preparation of a medicament for the treatment of cancer.

[0156] Embodiment 42 is a use of the stable aqueous pharmaceutical composition of embodiment 1 or 1a for treating a subject in need of treatment, comprising administering the stable aqueous pharmaceutical composition to a subject in need of treatment.

[0157] Embodiment 43 is the use according to embodiment 42, wherein the administration is subcutaneous. EXAMPLES

[0158] The following examples are provided to further illustrate some of the embodiments disclosed herein and are intended to be illustrative and not limiting of the embodiments of the present disclosure.

[0159] Description of analytical tests used herein Analytical Tests - General Characteristics Evaluation Solution color Solution color is monitored for drug products to assess appearance and ensure consistency with previous batches upon release and over shelf life. Solution color can also be an indicator of product stability. To determine solution color, test samples are visually compared to a set of defined reference solutions.

[0160] A defined amount of the liquid contents is transferred into a pre-scored ampoule of the same dimensions as the reference solution. The contents of the ampoule are then visually compared to the European Pharmacopoeia color reference solutions. The degree of color is judged in diffuse daylight observed against a white background.

[0161] Solution material color and method Materials and methods are described in European Pharmacopoeia 2.2.2, Degree of Coloration of Liquids European Pharmacopoeia (Ph.Eur.) 10th Edition Monograph No. 20202, July 2019. Briefly, test articles are compared to a set of B (brown), BY (brownish yellow), and Y (yellow) color reference solutions.

[0162] The solution color results are consistent with stability. In one embodiment, stability is defined as having a solution color that is colorless to about BY2 or less, about B2 or less, to about Y2 or less after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, stability is defined as having a solution color that is colorless to about BY4 or less, about B4 or less, to about Y4 or less after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a most preferred embodiment, stability is defined as having a solution color that is colorless to about BY5 or less, about B5 or less, to about Y5 or less after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more.

[0163] pH pH Materials and Methods - The pH of the test articles is measured using a daily calibrated electronic pH meter with a standardized pH electrode. All calibration solutions, reference buffers, and test articles are equilibrated to 25°C prior to testing and maintained at 25°C during testing.

[0164] The pH results are consistent with stability. In one embodiment, stability is defined as having a pH range of 4.7 to about 5.7 after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, stability is defined as having a pH range of 4.8 to about 5.6 after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a most preferred embodiment, stability is defined as having a pH range of 4.9 to about 5.5 after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more.

[0165] Turbidity Turbidity Materials and Methods - Materials and methods are based on European Pharmacopoeia 2.2.1, Clarity and Degree of Opalescence of Liquids.

[0166] Turbidity results are consistent with stability. Test results are reported in nephelometric turbidity units (NTU). In one embodiment, stability is defined as having a turbidity value of about 18 NTU or less after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, stability is defined as having a turbidity value of about 13 NTU or less after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a most preferred embodiment, stability is defined as having a turbidity value of about 8 NTU or less after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more.

[0167] Analytical Testing - Particulate Matter Particulate matter (not visible to the naked eye) Materials and Methods - All materials and methods conform to the United States Pharmacopoeia <788> Compendial liquid particle counters equipped with compendial volumetric sampler devices are used. Test articles are equilibrated to room temperature for at least 60 minutes but not more than 10 hours prior to testing. Test article vials are measured using a method consistent with the United States Pharmacopeia (USP) Particulate Matter (EPM) standard. <788> Pool in a manner consistent with Particulate Matter. <788> Remove four portions of appropriate volume of each of the pooled test articles as instructed by Particulate Matter and count the number of particles 10 μm and 25 μm or larger per portion. Disregard the result obtained for the first portion and use the results of the remaining three to calculate the average particle count for the preparation tested.

[0168] Particle Analysis (not visible to the naked eye) Compendial Results - Test results are in accordance with the United States Pharmacopoeia <788> Particulate Matter, in accordance with European Pharmacopoeia 2.9.19, and Japanese Pharmacopoeia XVII / 6.07 Particulate Contamination: Particles invisible to the naked eye. Therefore, the average number of particles present in the tested unit should not exceed 6000 particles per container for particles of 10 μm or greater, and 600 particles per container for particles of 25 μm or greater.

[0169] Analytical Testing - Purity Capillary electrophoresis sodium dodecyl sulfate (cSDS)-reduced form Reduced cSDS Materials and Methods - The analysis uses a commercially available capillary electrophoresis system with a bare fused silica capillary, 50 μm ID x 30.2 cm length, in a temperature-controlled cartridge. The capillary is equipped with a detection window transparent to UV light. The capillary is electrokinetically rinsed before each injection. A sieving matrix consisting of an entangled polymer solution is loaded into the capillary prior to each sample analysis. The method utilizes SDS-MW gel running buffer and protein molecular weight authentication standards ranging from approximately 10 to 148 kDa. The UV absorption spectrophotometer detector of the instrument is set to a wavelength of 220 nm and the capillary temperature is set to 25°C. For reduced sample processing conditions, the test article (in duplicate) is mixed with SDS and 2-mercaptoethanol and then heated for a specified time and temperature to completely denature and reduce the protein. The reduced sample is electrokinetically injected by applying a voltage of 5 kV to the capillary for approximately 20 seconds, and then analyzed by applying a larger electric field for approximately 35 minutes. Detection is achieved by absorbance in the far UV region of the spectrum at 220 nm. Percent of total signal data is collected for the light chain, heavy chain, and aglycosylated heavy chain (AG HC).

[0170] The reduced cSDS results are consistent with stability. In one embodiment, stability is defined as having a percent purity of ≥ 90.0% and no new peaks of > 1.5% compared to a validated stock of teclistamab reference material after storage at a temperature of about 5°C for about 12 months or more, after storage at a temperature of about 25°C for about 12 months or more, and / or after storage at a temperature of about 5°C for about 2 years or more. In a preferred embodiment, stability is defined as having a percent purity of ≥ 95.0% and no new peaks of > 1.2% compared to the reference material after storage at a temperature of about 5°C for about 12 months or more, after storage at a temperature of about 25°C for about 12 months or more, and / or after storage at a temperature of about 5°C for about 2 years or more. In a most preferred embodiment, stability is defined as having a percent purity of ≥ 97.0% and no new peaks of > 1.0% compared to the reference material after storage at a temperature of about 5°C for about 12 months or more, after storage at a temperature of about 25°C for about 12 months or more, and / or after storage at a temperature of about 5°C for about 2 years or more.

[0171] Capillary electrophoresis sodium dodecyl sulfate (cSDS) - non-reducing Non-reduced cSDS Materials and Methods - The analysis uses a commercially available capillary electrophoresis system with a bare fused silica capillary, 50 μm ID x 30.2 cm length, in a temperature-controlled cartridge. The capillary is equipped with a detection window transparent to UV light. The capillary is electrokinetically rinsed before each injection. A sieving matrix consisting of an entangled polymer solution is loaded into the capillary prior to each sample analysis. The method utilizes SDS-MW gel running buffer, protein molecular weight authentication standards spanning the approximate range of 10-148 kDa, and a validated teclistamab reference material sample. The instrument's UV absorption spectrophotometer detector is set to a wavelength of 220 nm and the capillary temperature is set to 25 °C. For non-reduced sample processing conditions, the test article (in duplicate) is mixed with SDS and an alkylating reagent (N-ethylmaleimide, to prevent shuffling or reformation of disulfide bonds). It is then heated for a specified time and temperature to completely denature the protein and minimize the formation of fragments and artifact bands. The non-reduced sample is electrokinetically injected by applying a voltage of 5 kV to the capillary for about 20 seconds, and then analyzed by applying a larger electric field for about 35 minutes. Detection is achieved by absorbance in the far-UV region of the spectrum at 220 nm. Percentage of total signal data is collected. Data is also analyzed for the presence of new peaks compared to the teclistamab reference material. Percentage purity is defined as percent heavy chain + percent light chain.

[0172] The results for non-reduced cSDS are consistent with stability. In one embodiment, stability is defined as having a percent purity of about 90.0% or greater and no new peaks greater than 1.5% compared to the reference material after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, stability is defined as having a percent purity of about 95.0% or greater and no new peaks greater than 1.2% compared to the reference material after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a most preferred embodiment, stability is defined as having a percent purity of about 97.0% or greater and no new peaks greater than 1.0% compared to the reference material after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more.

[0173] Size Exclusion High Performance Liquid Chromatography (SE-HPLC) SE-HPLC Materials and Methods - Reference materials and test articles are diluted to the target protein concentration. A volume of 20 μL of analyte is injected into a 7.8 mm x 30 cm size exclusion column with a silica base of 5 μm particle size, with a fractionation range of 10-500 kDa. Aqueous phosphate buffer is used as the mobile phase at a flow rate of 0.7 mL / min, and the absorbance of the eluate is continuously monitored at 280 nm. Monomers (major components or main peaks), aggregates (high molecular weight species, or HMWS), and fragments (low molecular weight species, or LMWS) are separated on the column and elute at different retention times. The amounts of these species are measured by monitoring the peak absorbance at 280 nm.

[0174] SE-HPLC results are consistent with stability Major Components - In one embodiment, stability is defined as having about 90.0% or greater major components after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, stability is defined as having about 95.0% or greater major components after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a most preferred embodiment, stability is defined as having about 97.0% major components after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more.

[0175] High Molecular Weight Species (HMWS) - In one embodiment, stability is defined as having about 10.0% or less HMWS after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, stability is defined as having about 5.0% or less HMWS after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a most preferred embodiment, stability is defined as having about 3.0% or less HMWS after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more.

[0176] Low Molecular Weight Species (LMWS) - In one embodiment, stability is defined as having an LMWS of about 5.0% or less after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, stability is defined as having an LMWS of about 2.0% or less after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a most preferred embodiment, stability is defined as having an LMWS of about 1.0% or less after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more.

[0177] Capillary isoelectric focusing (cIEF) cIEF Materials and Methods - The analytical procedure is performed on a commercial imaging cIEF analyzer equipped with an autosampler. The analysis uses 100 μm inner wall coated silica capillaries with an outer wall polyimide coating. In addition, an analyte solution of dilute phosphoric acid and methylcellulose, a catholyte solution of sodium hydroxide and methylcellulose, and ampholytes of predetermined type and amount are used. The specimens are treated with carboxypeptidase B (CPB) to remove the C-terminal lysine and eliminate ambiguity introduced by the presence of multiple C-terminal variants for each charged species. The instrument's autosampler is set at 4° C. for both pre-run and run. The pre-run voltage and time are 1500 V and 1 min, respectively. The run voltage and time are 3000 V and 7 min, respectively.

[0178] cIEF results are consistent with stability Main Peak - In one embodiment, stability is defined as having ≦60% of the main peak after about 12 months or more of storage at a temperature of about 5° C., after about 12 months or more of storage at a temperature of about 25° C., and / or after about 2 years or more of storage at a temperature of about 5° C. In a preferred embodiment, stability is defined as having ≦65% of the main peak after about 12 months or more of storage at a temperature of about 5° C., after about 12 months or more of storage at a temperature of about 25° C., and / or after about 2 years or more of storage at a temperature of about 5° C. In a most preferred embodiment, stability is defined as having ≦70% of the main peak after about 12 months or more of storage at a temperature of about 5° C., after about 12 months or more of storage at a temperature of about 25° C., and / or after about 2 years or more of storage at a temperature of about 5° C.

[0179] Sum of Acidic Peaks - In one embodiment, stability is defined as having a sum of ≦40% of the sum of acidic peaks after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, stability is defined as having a sum of ≦30% of the sum of acidic peaks after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a most preferred embodiment, stability is defined as having a sum of ≦25% of the sum of acidic peaks after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more.

[0180] Sum of basic peaks - In one embodiment, stability is defined as having a sum of ≦15% of the sum of basic peaks after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, stability is defined as having a sum of ≦10.0% of the sum of basic peaks after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a most preferred embodiment, stability is defined as having a sum of ≦8% of the sum of basic peaks after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more.

[0181] Analytical Test - Quantity Protein Concentration by A280 The protein concentration of the preparation is determined by quantification of absorbance at 280 nm (A280).

[0182] Protein Concentration by A280 Materials and Methods Protein concentration is measured using a qualified and calibrated double beam UV-Vis spectrophotometer. Test articles are diluted 1:125 using 0.9% (w / v) NaCl. Samples are measured using quartz semi-microcuvette (1.4 mL) with 1 cm path length and black or matte surface. The spectrophotometer is set at 280 nm wavelength, 1 nm slit width, and 1 second response. 0.9% (w / v) NaCl is used as a blank control. Protein concentration (mg / mL) is calculated by multiplying the absorbance of the test article by the dilution factor and the extinction constant of the antibody and the path length of the instrument (e.g., but not limited to, the extinction constant of 1.58 (mg / mL) for teclistamab). -1 cm -1 It is calculated by dividing the measurement time by the product of the measurement time and the path length of the instrument (1 cm).

[0183] Results at 10mg / mL formulation protein concentration are consistent with stability In one embodiment, stability is defined as having a protein concentration of 7.5-12.5 mg / mL after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, stability is defined as having a protein concentration of 8-12 mg / mL after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a most preferred embodiment, stability is defined as having a protein concentration of 9 mg / mL-11 mg / mL after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more.

[0184] Results at 90mg / mL formulation protein concentration are consistent with stability In one embodiment, stability is defined as having a protein concentration of 76.5-103.5 mg / mL after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, stability is defined as having a protein concentration of 85-95 mg / mL after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a most preferred embodiment, stability is defined as having a protein concentration of 87 mg / mL-93 mg / mL after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more.

[0185] Analytical Testing - Potency Efficacy (BCMA×CD3 T cell activation) In vitro T cell activation by BCMAxCD3 is demonstrated using a nuclear factor of activated T cells-response element (NFAT-RE)-mediated luminescence assay.

[0186] This reporter assay uses luminescence induced by activation of the NFAT (nuclear factor of activated T cells) pathway in engineered effector cells expressing CD3 as a readout for target cell / effector cell co-engagement and is a surrogate measure of target cell killing. Daudi B lymphoblast cells expressing BCMA on the cell surface are used as target cells. Both engagement of the anti-BCMA Fab region with BCMA-expressing target cells and engagement of the anti-CD3 Fab region with engineered CD3+ T cells are required for T cell activation and subsequent NFAT-RE-mediated luminescence.

[0187] Materials and methods for BCMA×CD3 T cell activation. Qualified teclistamab is used as the reference material and control. 1.0×10 per mL of culture medium (4% heat-inactivated fetal bovine serum in RPMI) 6 Solution of viable Jurkat TCR / CD3 effector cells and 4 x 10 per mL of culture medium 5 A solution of Daudi B-lymphoblast target cells at 1000x the viable cells is prepared. Equal amounts of effector and target cells are aliquoted into individual wells on a 96-well cell culture plate. The plate is then incubated for 16-24 hours at 37°C (±2°C) with 5% (±2%) CO2. After incubation, Bio-Glo™ Luciferase Substrate is added to each well and gently agitated for at least 5 minutes on a plate shaker. Within 30 minutes of adding the substrate, luminescence (relative light unit (RLU) values) is measured using a microtiter luminescence plate reader. Data are reported as percent bioactivity relative to the reference material.

[0188] The BCMAxCD3 activity results are consistent with stability. In one embodiment, stability is defined as 50% to 150% of the biological activity of the reference material after storage at a temperature of about 5°C for about 12 months or more, after storage at a temperature of about 25°C for about 12 months or more, and / or after storage at a temperature of about 5°C for about 2 years or more. In a preferred embodiment, stability is defined as 60% to 140% of the biological activity of the reference material after storage at a temperature of about 5°C for about 12 months or more, after storage at a temperature of about 25°C for about 12 months or more, and / or after storage at a temperature of about 5°C for about 2 years or more. In a most preferred embodiment, stability is defined as a biological activity in the range of about 80% to 120% of the biological activity of the reference material after storage at a temperature of about 5°C for about 12 months or more, after storage at a temperature of about 25°C for about 12 months or more, and / or after storage at a temperature of about 5°C for about 2 years or more.

[0189] Analytical Tests - Surfactants Quantitative determination of polysorbate 20 Polysorbate 20 is quantitatively determined by mixed mode ion exchange / hydrophobic HPLC.

[0190] PS20 Materials and Methods. The analysis is performed on a gradient HPLC equipped with a 2.1 x 20 mm online column containing 30 μm water-wettable mixed-mode polymeric spherical sorbent particles, an ELSD, and a temperature-controlled column section at 30°C. The flow rate is set at 1 mL / min and the ELSD evaporator temperature is set at 50°C. Mobile phase A is 2% v / v formic acid in water and mobile phase B is 2% v / v formic acid in isopropyl alcohol. Neat polysorbate 20 is used to make calibration and check standards. Test article samples are injected undiluted.

[0191] The polysorbate 20 results are consistent with stability. In one embodiment, stability is defined as a PS20 concentration of 0.01-0.07% after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, stability is defined as a PS20 concentration of 0.02-0.06% after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a most preferred embodiment, stability is defined as a PS20 concentration of 0.03-0.05% after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more.

[0192] Analytical Testing - Routine Characterization Peptide Map The purpose of this test is to measure the level of post-translational modifications such as oxidation, deamidation, and isomerization that may be present in the antibody structure. The test article is enzymatically digested to obtain peptide segments. These peptides are then evaluated by ultra-performance liquid chromatography mass spectrometry (UPLC-MS). Each peptide sequence analyzed is identified relative to its known location within the overall antibody structure. Post-translational modifications are measured by comparing the measured mass of the identified peptide sequence with its expected mass.

[0193] Peptide Mapping Materials and Methods. Samples are denatured with 6M guanidine, 50mM Tris pH 8.0, 5mM EDTA and filtered using a 30kDa centrifugal filter device (flow-through discarded). Denatured samples are reduced with 1M dithiothreitol (DTT) followed by alkylation with 1M sodium iodoacetate and further treatment with DTT to quench the reaction. The reaction mixture is exchanged into digestion buffer (50mM Tris pH 7.0 with 1mM CaCl2) through a Sephadex G-25 column with separate columns used for blanks, reference material, and test articles. An aliquot of 1mg / mL trypsin stock solution is added to the sample in digestion buffer to give a trypsin concentration of 20μL / mL. The solution is incubated at 37°C for 2 hours ± 30 minutes. The trypsinized solution is cooled to room temperature and the enzyme is inactivated with trifluoroacetic acid. Processed samples are evaluated by ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) equipped with a Waters Acquity BEH (Ethylene Bridged Hybrid) C18, 2.1 x 100 mm, 1.7 μm, 130 Å column and attached autosampler. Mobile phase A is 0.1% formic acid in water and mobile phase B is 0.1% FA in acetonitrile (mobile phase B). The autosampler is set to 2-8 °C, the column is set to 40 °C, and the flow rate is set to 500 μL / min. Eluted peptides are subjected to electrospray ionization and detected using a calibrated online mass spectrometry.

[0194] Example 1: Initial formulation screening studies Initial formulation screening studies were designed to evaluate the impact of pH, buffer type, surfactants, and stabilizers on the chemical, physical, and biological stability of teclistamab solutions. Development work identified a larger design space for the formulation, including acetate buffers in the pH range of 4.5-5.4 and histidine buffers in the pH range of 5.4-6.4. The formulations were targeted at pH 5.2 in acetate buffers and pH 6.0 in histidine buffers to have sufficient buffering capacity in subsequent confirmatory stability studies. During the initial feasibility evaluation of the lead BCMA×CD3 antibody, it was noted that exposure to excessive levels of iron correlated with increased aggregation as measured by SEC. Therefore, EDTA was added to the formulation.

[0195] Based on the formulation screening study, storage stability was evaluated for the two formulations listed below. 1. 10 mg / mL teclistamab in 10 mM acetate, 8% (w / v) sucrose, 20 mg / mL EDTA, 0.04% (w / v) PS 20, pH 5.2 2. 10 mg / mL teclistamab in 10 mM histidine, 8% (w / v) sucrose, 20 mg / mL EDTA, 0.04% (w / v) PS20, pH 6.0

[0196] These two formulations were subjected to storage stability studies at 2-8°C, 25°C, and 37°C. The results of the study showed that there was no significant change in stability for either formulation by cSDS (reduced and non-reduced) and cIEF (data not shown). Size exclusion chromatography (SE-HPLC) results showed that there was no loss of major component content after 6 months of storage at 2-8°C in either formulation. However, differences were observed at higher storage temperatures, especially at 37°C for more than 3 months and at 25°C for more than 6 months. The histidine buffer formulation showed a greater loss of major components at 25°C and 37°C compared to the acetate buffer formulation (Figure 1). These results indicated that teclistamab was more stable in the acetate buffer formulation at pH 5.2.

[0197] To investigate the stability of formulations with higher teclistamab concentrations, two formulations containing 30 mg / mL and 90 mg / mL teclistamab, respectively, were placed in a stability chamber at 2-8°C and 25°C for up to 2 years. The target composition was 10 mM acetate, 8% (w / v) sucrose, 20 μg / mL disodium EDTA, 0.04% (w / v) PS 20, pH 5.2 for both the 30 mg / mL and 90 mg / mL formulations. Data were compared to a 10 mg / mL formulation containing the same target composition. Formulations were analyzed for major components, aggregates, and fragment content by SE-HPLC, purity by cSDS (reduced and non-reduced), charge heterogeneity by cIEF, and subvisible particulate matter by light obscuration (LO).

[0198] No substantial changes were observed in the cIEF and cSDS (reduced and non-reduced) data and subvisible particle counts (data not shown).

[0199] The SE-HPLC data (Table 2) show a slight but present correlation between increased protein concentration and decreased major components, however the magnitude of the decrease occurred over a narrow range and all protein concentrations showed major component values ​​consistent with the most preferred embodiment.

[0200] [Table 2] a The 10 mg / mL formulation samples were generated at the 6 month time point. NA=Not Applicable

[0201] At the end of the initial formulation screening studies, the acetate concentration was changed to 10-15 mM.

[0202] Example 2: Shaking a range of polysorbate concentrations Two shaking stress studies were conducted to identify polysorbate 20 (PS 20) levels in acetate formulations. The first pilot study evaluated mechanical stress on 1 mg / mL teclistamab in 10 mM acetate, 8% (w / v) sucrose, 20 μg / mL EDTA (pH 5.2) supplemented with either 0, 0.01, 0.02, 0.04, or 0.06% (w / v) PS20. The test formulations were placed on an orbital platform shaker and shaken at 250 rpm for 120 hours at ambient temperature. SEC analysis of the shaken samples showed that samples without PS20 exhibited less than 30% monomer and more than 70% aggregates. This demonstrates that the research test conditions used produced sufficient mechanical stress to induce catastrophic loss of DP stability. The addition of PS20 sharply increases monomer and reduces aggregate formation, with 0.02% (w / v) PS20 demonstrating monomer and aggregate levels consistent with the preferred embodiment for DP stability.

[0203] The same shaking study design was used in a second study, augmented with an additional time point of 72 hours, a control sample of 0.04% (w / v) PS20 held at ambient conditions for 120 hours without shaking, and a broader orthogonal battery of analytical assays. The second study evaluated 10 mg / mL and 90 mg / mL teclistamab in 15 mM acetate, 8% (w / v) sucrose, 20 μg / mL EDTA (pH 5.2) supplemented with either 0.02, 0.04, or 0.06% (w / v) PS20. The 10 mg / mL formulation was aliquoted into 6 mL type 1 glass vials in a fill volume of 3.5 mL, and the 90 mg / mL formulation was aliquoted into 2 mL type 1 glass vials in a fill volume of 2 mL.

[0204] The results are shown in Tables 3 and 4. IE-HPLC, cSDS reduced and non-reduced, and post-shaking samples showed essentially no change in values ​​relative to the T=0 values ​​for both the 10 mg / mL and 90 mg / mL formulations. Similarly, protein concentration, solution color, pH, and turbidity also remained unchanged. The Sub-Vis results, in the context of the magnitude of their official acceptance criteria and the overall methodology, can be considered essentially the same relative to T=0 within a given time point. The post-shaking SEC results showed a slight trend towards increased aggregation in the lowest (0.02% w / v) PS20 concentration sample. However, these values ​​are consistent with the preferred embodiment of DP stability.

[0205] [Table 3]

[0206] [Table 4]

[0207] Taken together, these studies demonstrate that 0.02–0.08% (w / v) PS20 protects teclistamab from mechanical stress-induced instability.

[0208] Example 3: Freeze-thaw stress test A study was performed to demonstrate the robustness of the formulation against physicochemical stress induced by repeated freeze-thaw cycles.

[0209] In this study, 10 mg / mL and 90 mg / mL formulations (15 mM, 8% (w / v) sucrose, 20 μg / mL EDTA, 0.04% (w / v) PS20, pH 5.2) were aliquoted into 5 mL polycarbonate containers and placed upright in a -70°C freezer. After freezing was complete, the vials were removed from the freezer and placed at ambient temperature for thawing. Once fully thawed, the containers were gently swirled to ensure homogeneity of the solution, and then additional freeze-thaw (F / T) cycles were repeated until a total of 5 cycles were completed. The control sample did not undergo any freeze-thaw cycles. Both the control and F / T samples were analyzed.

[0210] As shown in Table 5, the quality attributes of the samples that underwent five F / T cycles were nearly identical to the control samples stored at 2-8 °C and not subjected to freeze-thaw conditions. Furthermore, no significant changes in protein concentration, solution color, pH, or turbidity were observed.

[0211] [Table 5]

[0212] The results of this study demonstrate the robustness of the 10 mg / mL and 90 mg / mL formulations to freeze-thaw induced stress.

[0213] Example 4: Metal Spiking Studies A study was conducted to evaluate the effect of metal ions potentially present or introduced into pharmaceutical products during the manufacturing process.

[0214] Test formulations consisted of 10 mg / mL or 90 mg / mL teclistamab, 15 mM acetate, 8% sucrose, 20 μg / mL EDTA, and 0.04% PS 20 (pH 5.2) with or without added iron (Fe3+), chromium (Cr3+), copper (Cu2+), nickel (Ni2+), and molybdenum (Mo5+). Metal selection was based on the composition of metal alloy components potentially present in the manufacturing process. To demonstrate robustness, exaggerated stress conditions were created by utilizing metal concentrations 2-4 times higher than the highest values ​​found in a commercially available GMP drug manufacturing process.

[0215] The 10 mg / mL test formulation was aliquoted into 6R vials with a fill volume of 3.5 mL and the 90 mg / mL test formulation was aliquoted into 2 mL vials with a fill volume of 2.0 mL. All vials were stoppered, capped and crimp sealed. The vials were kept stable at recommended (5° C.) and accelerated (25° C.) conditions for up to 6 months. Samples were taken and assayed at the designated time points.

[0216] As shown in Table 6, the quality attributes of the metal ion stressed 10 mg / mL and 90 mg / mL formulations after 6 months of storage at 2-8° C. were nearly identical to their respective control formulations. Furthermore, there was little or no change in the quality attribute values ​​at T=6 months compared to the T=0 values ​​for all test formulations stored at 2-8° C. No significant changes in protein concentration, solution color, pH, or turbidity were observed for any of the formulations tested over the storage time. Finally, the quality attribute values ​​of all metal loaded formulations stored at 2-8° C. for 6 months were consistent with the most favorable embodiment of stability.

[0217] As shown in Table 7, the quality attributes of the metal ion stressed 10 mg / mL and 90 mg / mL formulations after 6 months of storage at 25° C. were nearly identical to the respective control formulations. For all test formulations, slight to minimal changes in several quality attribute values ​​were seen at T=6 months compared to the T=0 values. The extent of change was consistent with expected degradation of these monoclonal antibody quality attributes upon storage at 25° C. for 6 months. No significant changes in protein concentration, solution color, pH, or turbidity were observed for any of the formulations tested over the storage time. Finally, the quality attribute values ​​of all metal spiked formulations stored at 25° C. for 6 months were consistent with the preferred embodiment of stability.

[0218] Taken together, the data demonstrate the robustness of the formulations at 10 mg / mL and 90 mg / mL against potential oxidative stress experienced during normal and / or exaggerated pharmaceutical manufacturing processes and storage conditions.

[0219] [Table 6]

[0220] [Table 7]

[0221] Example 5: Formulation robustness development Study design A stable monoclonal antibody formulation is the result of the net effect of complex interactions between the components of the formulation. To evaluate these interactions, a Design of Experiments (DOE) methodology was used to create a multifactorial stability study in which a statistically determined number of experimental test formulations are generated that simultaneously vary parameters of the formulation components. Statistical analysis of the study results provides a quantitative understanding of how the formulation parameters interact to affect stability attributes, thus demonstrating the robustness of the formulation. Experimental test formulations of BCMA x CD3 drug product were held at recommended conditions (5°C) and accelerated conditions (25°C) for up to 12 months and up to 6 months, respectively. The formulation components evaluated were protein concentration, acetate concentration, sucrose concentration, polysorbate 20 concentration, EDTA concentration, and pH. The concentration ranges of the factors tested are listed in Table 8 (target formulation values ​​are included in the table for context). The study encompassed both 10 mg / mL and 90 mg / mL DP formulations. Typical protein concentration ranges for evaluation are + / - 12% of the target value. Thus, the protein concentration values ​​selected for the study represent 88% for the 10 mg / mL formulation and 111% for the 90 mg / mL formulation.

[0222] [Table 8]

[0223] Based on this criteria, a fractional factorial design was created using JMP® statistical software (Table 9).

[0224] [Table 9]

[0225] The 16 formulations were filled into 6R vials with a fill volume of 2 mL per vial (largest vial size and smallest fill volume among the 10 mg / mL and 90 mg / mL DP offerings) to reflect worst case headspace. Table 10 shows the full range of formulation parameters tested. Acetate concentrations measured after UF / DF during sample preparation were higher than the nominal levels before UF / DF due to Gibbs-Donnan effect limits.

[0226] [Table 10]

[0227] The vials were stoppered, capped and crimp sealed. The vials were placed on stability at recommended conditions (5° C.) and accelerated conditions (25° C.). At the designated time points, samples were withdrawn and assayed.

[0228] Test results The test results for each attribute of the 16 formulations at the start of the study (time 0), after 6 months at accelerated temperature (25° C.), and after 6 and 12 months at recommended storage conditions (5° C.) are shown in Table 11. Data are reported as ranges, means, and standard deviations for the eight formulations for each attribute.

[0229] Analytical results for all formulations held at 5°C for 6 and 12 months demonstrated little change in assay test values, indicating stability. The ability of all formulations with a multivariate range of excipient concentrations to produce a narrow range of assay test result values ​​demonstrates the robustness of the formulations within the boundaries and storage conditions tested. Furthermore, the full range of values ​​observed for all but one assay in this study was consistent with those held at 2-8°C.

[0230] With regard to turbidity, the calculated mean turbidity value (6.4 NTU) for all formulations held at 5°C for 12 months was consistent with the definition of a stable aqueous pharmaceutical composition provided herein. However, the range of observed assay values ​​and the calculated standard deviation were relatively larger than those observed for the other assays. Further analysis of the data showed that three of the 16 test formulations consistently reported turbidity values ​​greater than 15 NTU but less than or equal to 20 NTU at all time points and temperatures. These formulations were three of the four test formulations in the study formulated at a high pH value (5.8) and a high protein concentration (100 mg / mL). When the turbidity values ​​for the four high pH value (5.8) and high protein concentration (100 mg / mL) test formulations are omitted, the calculated mean, standard deviation, and range of turbidity assay values ​​for the remaining 12 formulations held at 5°C for 12 months were 3.5 NTU, 1.6 NTU, and 1.4 to 6.3 NTU, respectively, consistent with the most preferred embodiment of stability.

[0231] Analytical results for all formulations held at accelerated storage conditions (25° C.) for six months showed degradation effects consistent with the stability profile of BCMA×CD3 antibodies exposed to long-term accelerated storage conditions, as demonstrated by the increased magnitude of the range of result values. However, the average values ​​calculated for seven of the nine assay results were consistent with the most favorable embodiment of stability when held at accelerated storage conditions (25° C.) for six months.

[0232] [Table 11]

[0233] Statistical analysis of study results A general linear model was used to statistically analyze the 12-month 2-8°C data to determine which factors had a statistically significant effect on a given response. The analysis was performed using the actual measurements for each factor in each test formulation after UF / DF treatment. Table 12 lists the p-values ​​calculated for each factor for a given response, and the corresponding adjusted R-squared values ​​generated by the model. p-values ​​less than 0.05 are considered statistically significant.

[0234] [Table 12]

[0235] API concentration API concentration showed a statistically significant negative correlation with %monomer while also showing a corresponding statistically significant positive correlation with %aggregate. These trends indicate that increasing API concentration correlates with decreased stability as measured by SEC. However, within the concentration range evaluated in this study (8-100 mg / mL), the range of %monomer and %aggregate assay results observed were consistent with the definition of a stable aqueous pharmaceutical composition provided herein.

[0236] API concentration also showed a statistically significant negative correlation with % Acidic Peak, while showing a statistically significant positive correlation with % Basic Peak. However, as shown in Table 12, the range of result values ​​for % Acidic Peak and % Basic Peak appears to be very narrow for these two attributes. Therefore, although calculated to be statistically significant, API concentrations between 8 and 100 mg / mL are not expected to have a practical impact on % Acidic Peak and % Basic Peak.

[0237] pH of the formulation Formulation pH showed a statistically significant negative correlation with %monomer while also showing a corresponding statistically significant positive correlation with %aggregate. These trends indicate that increased pH correlates with decreased stability as measured by SEC. However, within the pH range evaluated in this study (4.6-5.8), the range of %monomer and %aggregate assay results observed was consistent with the most favorable embodiment of stability.

[0238] The pH of the formulations showed a statistically significant positive correlation with turbidity, indicating that increasing pH correlates with decreasing stability as measured by turbidity. However, the r-squared (adjusted) value of the linear model was relatively low (54.28%). This may be related to the observations mentioned above where turbidity values ​​above 15 NTU were consistently observed in the three test formulations formulated at high pH values ​​(5.8) and high protein concentrations (100 mg / mL). In turn, the three turbidity result values ​​may have distorted the linear model used for this analysis. However, within the pH range evaluated in this study (4.6-5.8), the range of turbidity results for the low protein concentration test formulations observed was consistent with the most favorable embodiment of stability.

[0239] pH also showed a statistically significant negative correlation with %Main Peak and %Purity by cSDS reduction, while it also showed a statistically significant positive correlation with %Basic Peak. However, similar to API concentration, the range of result values ​​for %Acid Peak, %Main Peak, and %Purity by cSDS reduction appears to be quite narrow for these three attributes, as shown in Table 12. Therefore, although calculated to be statistically significant, a pH of 4.6-5.8 is not expected to have a practical effect on %Acid, %Basic Peak, and %Purity by cSDS reduction.

[0240] Acetate Concentration The acetate formulations showed a statistically significant negative correlation with %monomer while also showing a corresponding statistically significant positive correlation with %aggregates. These trends indicate that increasing acetate concentrations correlate with decreased stability as measured by SEC. However, within the acetate concentration range evaluated in this study (10-20 mM), the range of observed %monomer and %aggregates assay results was consistent with the most favorable embodiment of stability.

[0241] Example 6: Bulk production of formulated drug Process Description Processing Solution

[0242] [Table 13]

[0243] Ultrafiltration / Diafiltration (UF / DF) Ultrafiltration / diafiltration (UF / DF) was performed to reformulate the teclistamab planova filtrate intermediate manufacturing solution into a pre-formulated bulk (pFB) solution consisting of 90 mg / mL teclistamab, 10 mM acetate, 8% sucrose, pH 4.9.

[0244] Preparation of Teclistamab Formulated Bulk (FB) Polysorbate 20 (4.0% w / v) and EDTA (2 mg / mL) stock solutions were added to the pFB at a 1:100 dilution to obtain a final concentration of 0.04% (w / v) polysorbate 20 and 20 μg / mL EDTA, resulting in a formulated bulk (FB) consisting of 90 mg / mL teclistamab in 15 mM acetate, 8% (w / v) sucrose, 0.04% polysorbate 20, 20 μg / mL EDTA, pH 5.2. The FB solution was then mixed homogeneously. Final filtration of the formulated bulk was achieved using a sterile 0.45 / 0.22 μm filter followed immediately by an in-line 0.22 μm filter.

[0245] Final Bulk Fill After the final filtration, the FB was packed into polycarbonate biotainers, with the packing volume ranging from 20% to 90% of the listed volume of the biotainer.

[0246] Final bulk storage and transportation Storage and transportation conditions for the formulated bulk prior to drug manufacturing were 5°C ± 3°C, protected from light, if the FB was stored for approximately 1 week or less, or -40°C ± 10°C, protected from light, if the FB was stored for more than 1 week.

[0247] Example 7: 90 mg / mL pharmaceutical formulation: composition and components of primary packaging Provided herein is Table 14, a summary of the composition of the 90 mg / mL teclistamab pharmaceutical formulation.

[0248] [Table 14]

[0249] The primary packaging of the 90 mg / mL teclistamab drug product (DP) consists of a glass vial, a polymeric vial stopper, and an aluminum seal. Table 15 provides the specific components of the primary packaging material of the 90 mg / mL DP presentation.

[0250] [Table 15]

[0251] Example 8: Preparation of 10 mg / mL DP solution Dilution of Teclistamab Formulated Bulk (FB) Dilutions were performed to reformulate the bulk intermediate manufacturing solution containing teclistamab into a drug product solution consisting of 10 mg / mL teclistamab, 15 mM acetate, 8% (w / v) sucrose, 20 μg / mL, EDTA, pH 5.2.

[0252] Dilution Buffer Provided herein is a tabular summary of the composition of the dilution buffer in Table 16. Final filtration of the dilution buffer was achieved using a sterile 0.22 μm filter.

[0253] [Table 16]

[0254] Preparation of teclistamab 10 mg / mL drug solution Dilution buffer was added to 90 mg / mL FB to obtain a 10 mg / mL drug product solution consisting of 10 mg / mL teclistamab in 15 mM acetate, 8% (w / v) sucrose, 0.04% polysorbate 20, 20 μg / mL EDTA, pH 5.2.

[0255] Example 9: 10 mg / mL pharmaceutical formulation: composition and components of primary packaging Provided herein is Table 17, a summary of the composition of the 10 mg / mL teclistamab pharmaceutical formulation.

[0256] [Table 17]

[0257] The primary packaging of the 10 mg / mL teclistamab drug product (DP) consists of a glass vial, a polymeric vial stopper, and an aluminum seal. Table 18 lists the specific components of the primary packaging materials for the 30 mg DP presentation.

[0258] [Table 18]

[0259] Example 10: Description of 90 mg / mL Stability Study This study was conducted to monitor the drug product attributes of stable teclistamab 90 mg / mL under various environmental conditions and time lengths. Investigational test articles were prepared by aliquoting the formulated bulk into 2 mL vials with a fill volume of 2.0 mL. The vials were stoppered, capped, and crimp sealed.

[0260] All tests were performed with the vials inverted.

[0261] [Table 19]

[0262] Stability test results Listed below are stability results for the teclistamab DP held under recommended, accelerated, and stressed conditions. At all time points for the DP held under recommended storage conditions, all test parameter result values ​​observed per assay study exceeded criteria consistent with the most preferred embodiment of stability when held after storage at a temperature of about 5° C. for about 12 months or more and / or after storage at a temperature of about 5° C. for about 2 years or more. Similarly, peptide map results showed little or no consequential change over time in the measured rates of post-translational modifications.

[0263] Results for the teclistamab DP held under accelerated conditions (25±2° C. / 60% RH for 12 months) and stressed conditions (40±2° C. / 75% RH for 6 months) demonstrated the expected degradation rates for drug products exposed to long-term accelerated and stressed storage conditions. Of particular note, the majority of the test parameter result values ​​observed per assay study for the DP held at accelerated conditions (25° C.) for 12 months were consistent with or exceeded the most preferred embodiment of stability, with all but one of the remaining results showing results consistent with the preferred or described embodiment.

[0264] 5℃ data

[0265] [Table 20]

[0266] [Table 21]

[0267] [Table 22]

[0268] [Table 23] * Scheduled exams for when they are cancelled.

[0269] 25℃ data

[0270] [Table 24]

[0271] [Table 25]

[0272] [Table 26]

[0273] [Table 27] * Scheduled exams for when they are cancelled.

[0274] 40℃ data

[0275] [Table 28]

[0276] [Table 29]

[0277] [Table 30]

[0278] Example 11: Description of 10 mg / mL Stability Study This study was conducted to monitor the drug product attributes of stable teclistamab 10 mg / mL under various environmental conditions and time lengths. Investigational test articles were prepared by aliquoting the formulated bulk into 6R vials at 3.5 mL fill volumes. The vials were stoppered, capped, and crimp sealed.

[0279] All tests were performed with the vials inverted.

[0280] [Table 31]

[0281] Stability test results Listed below are stability results for the teclistamab DP held under recommended, accelerated, and stressed conditions. At all time points for the DP held under recommended storage conditions, all test parameter result values ​​observed per assay study exceeded criteria consistent with the most preferred embodiment of stability when held after storage at a temperature of about 5° C. for about 12 months or more and / or after storage at a temperature of about 5° C. for about 2 years or more. Similarly, peptide map results showed little or no consequential change over time in the measured rates of post-translational modifications.

[0282] Results for the teclistamab DP held at accelerated (25±2°C / 60% RH) and stressed (40±2°C / 75% RH) conditions demonstrated the expected degradation rates for drug products exposed to long-term accelerated and stressed storage conditions. Of particular note, the majority of the test parameter result values ​​observed per assay study for the DP held at accelerated (25°C) conditions for 12 months were consistent with or exceeded the most preferred embodiment for stability, with all but one of the remaining results showing results consistent with the preferred or described embodiment.

[0283] 5℃ data

[0284] [Table 32]

[0285] [Table 33]

[0286] [Table 34]

[0287] [Table 35]

[0288] 25℃ data

[0289] [Table 36]

[0290] [Table 37]

[0291] [Table 38]

[0292] [Table 39] * Scheduled exams for which time has been cancelled

[0293] 40℃ data

[0294] [Table 40]

[0295] [Table 41]

[0296] [Table 42]

[0297] [Table 43]

Claims

1. A stable aqueous pharmaceutical composition comprising: a) a bispecific B-cell maturation antigen (BCMA) / cluster of differentiation 3 (CD3) antibody at a concentration of (i) about 7.5 mg / mL to about 12.5 mg / mL, or (ii) about 76.5 mg / mL to about 103.5 mg / mL; wherein the bispecific BCMA / CD3 antibody is (1) a first heavy chain (HC1) comprising an HC1 variable region 1 (VH1); wherein the VH1 comprises heavy chain complementarity determining region 1 (HCDR1), HCDR2 and HCDR3 comprising the amino acid sequences of SEQ ID NOs: 1, 2 and 3, respectively; (2) a first light chain (LC1) comprising an LC1 variable region (VL1); and wherein the VL1 comprises a light chain complementarity determining region 1 (LCDR1), a LCDR2 and a LCDR3 comprising the amino acid sequences of SEQ ID NOs: 4, 5 and 6, respectively; (3) a second heavy chain (HC2) comprising an HC2 variable region 2 (VH2); and wherein the VH2 comprises heavy chain complementarity determining region 1 (HCDR1), HCDR2 and HCDR3 comprising the amino acid sequences of SEQ ID NOs: 11, 12 and 13, respectively; (4) a second light chain (LC2) comprising an LC2 variable region 2 (VL2); and wherein the VL2 comprises a light chain complementarity determining region 1 (LCDR1), LCDR2 and LCDR3 comprising the amino acid sequences of SEQ ID NOs: 14, 15 and 16, respectively; Including, b) about 10 mM to about 20 mM acetate and / or a pharmaceutically acceptable acetate salt; c) about 6% (w / v) to about 10% (w / v) sucrose; and d) about 16 μg / mL to about 24 μg / mL ethylenediaminetetraacetic acid (EDTA); e) about 0.01% to about 0.07% polysorbate 20 (PS-20); f) a pH of about 4.7 to about 5.7; A stable aqueous pharmaceutical composition comprising:

2. 2. The stable aqueous pharmaceutical composition of claim 1, wherein the bispecific BCMA / CD3 antibody comprises an HC1 comprising the amino acid sequence of SEQ ID NO: 9 and an LC1 comprising the amino acid sequence of SEQ ID NO:

10.

3. 2. The stable aqueous pharmaceutical composition of claim 1, wherein the bispecific BCMA / CD3 antibody comprises an HC2 comprising the amino acid sequence of SEQ ID NO: 19 and an LC2 comprising the amino acid sequence of SEQ ID NO:

20.

4. 2. The stable aqueous pharmaceutical composition of claim 1, wherein the bispecific BCMA / CD3 antibody is teclistamab.

5. 10. The stable aqueous pharmaceutical composition of claim 1, wherein the bispecific BCMA / CD3 antibody has a concentration of about 8 mg / mL to about 12 mg / mL.

6. 6. The stable aqueous pharmaceutical composition of claim 5, wherein the bispecific BCMA / CD3 antibody has a concentration of about 9 mg / mL to about 11 mg / mL.

7. 7. The stable aqueous pharmaceutical composition of claim 6, wherein the bispecific BCMA / CD3 antibody has a concentration of about 10 mg / mL.

8. 10. The stable aqueous pharmaceutical composition of claim 1, wherein the bispecific BCMA / CD3 antibody has a concentration of about 85 mg / mL to about 95 mg / mL.

9. 9. The stable aqueous pharmaceutical composition of claim 8, wherein the bispecific BCMA / CD3 antibody has a concentration of about 87 mg / mL to about 93 mg / mL.

10. 10. The stable aqueous pharmaceutical composition of claim 9, wherein the bispecific BCMA / CD3 antibody has a concentration of about 90 mg / mL.

11. The stable aqueous pharmaceutical composition of claim 1, wherein the composition comprises about 12 mM to about 18 mM acetate and / or a pharmaceutically acceptable acetate salt.

12. The stable aqueous pharmaceutical composition of claim 11, wherein the composition comprises about 14 mM to about 16 mM acetate and / or a pharmaceutically acceptable acetate salt.

13. The stable aqueous pharmaceutical composition of claim 12, wherein the composition comprises about 15 mM acetate and / or a pharmaceutically acceptable acetate salt.

14. The stable aqueous pharmaceutical composition of claim 1, wherein the composition comprises about 7% (w / v) to about 9% (w / v) sucrose.

15. The stable aqueous pharmaceutical composition of claim 14, wherein the composition comprises about 8% (w / v) sucrose.

16. The stable aqueous pharmaceutical composition of claim 1, wherein the composition contains about 18 μg / mL to about 22 μg / mL of EDTA.

17. The stable aqueous pharmaceutical composition of claim 16, wherein the composition contains about 20 μg / mL of EDTA.

18. The stable aqueous pharmaceutical composition of claim 1, wherein the composition comprises about 0.02% to about 0.06% PS-20.

19. The stable aqueous pharmaceutical composition of claim 18, wherein the composition comprises about 0.03 to about 0.05% PS-20.

20. The stable aqueous pharmaceutical composition of claim 19, wherein the composition comprises about 0.04% PS-20.

21. 2. The stable aqueous pharmaceutical composition of claim 1, wherein the pH is from about 4.8 to about 5.

6.

22. 22. The stable aqueous pharmaceutical composition of claim 21, wherein the pH is from about 4.9 to about 5.

5.

23. 23. The stable aqueous pharmaceutical composition of claim 22, wherein the pH is about 5.

2.

24. The stable aqueous pharmaceutical composition of claim 1, wherein the composition comprises 10 mg / mL of the bispecific BCMA / CD3 antibody, 15 mM acetate and / or a pharmaceutically acceptable acetate salt, 8% (w / v) sucrose, 20 μg / mL EDTA, 0.04% PS20, and pH 5.

2.

25. The stable aqueous pharmaceutical composition of claim 1, wherein the composition is stable at a temperature of about 2-8°C for at least two years.

26. 2. The stable aqueous pharmaceutical composition of claim 1, wherein stability is defined based on solution color, pH, turbidity, percentage of purity, percentage of new peaks, percentage of major components, percentage of high molecular weight species (HMWS), percentage of low molecular weight species (LMWS), percentage of sum of acidic peaks, percentage of sum of basic peaks, protein concentration, percentage of T cell activation, percentage of PS20 (w / v), or any combination thereof.

27. 10. The stable aqueous pharmaceutical composition of claim 1, for use in a method of treating cancer in a subject in need thereof, the method comprising administering the stable aqueous pharmaceutical composition to the subject.

28. 28. The stable aqueous pharmaceutical composition of claim 27, wherein the administering is subcutaneous administration.

29. 1. A method for preparing a stable aqueous pharmaceutical composition of a bispecific B-cell maturation antigen (BCMA) / cluster of differentiation 3 (CD3) antibody, comprising: the bispecific BCMA / CD3 antibody (1) a first heavy chain (HC1) comprising an HC1 variable region 1 (VH1); wherein the VH1 comprises heavy chain complementarity determining region 1 (HCDR1), HCDR2 and HCDR3 comprising the amino acid sequences of SEQ ID NOs: 1, 2 and 3, respectively; (2) a first light chain (LC1) comprising an LC1 variable region (VL1); and wherein the VL1 comprises a light chain complementarity determining region 1 (LCDR1), a LCDR2 and a LCDR3 comprising the amino acid sequences of SEQ ID NOs: 4, 5 and 6, respectively; (3) a second heavy chain (HC2) comprising an HC2 variable region 2 (VH2); and wherein the VH2 comprises heavy chain complementarity determining region 1 (HCDR1), HCDR2 and HCDR3 comprising the amino acid sequences of SEQ ID NOs: 11, 12 and 13, respectively; (4) a second light chain (LC2) comprising an LC2 variable region 2 (VL2); and wherein the VL2 comprises a light chain complementarity determining region 1 (LCDR1), LCDR2 and LCDR3 comprising the amino acid sequences of SEQ ID NOs: 14, 15 and 16, respectively; Including, 10. The method of claim 1, wherein the stable aqueous pharmaceutical composition has a pH of about 5.

2.

30. 30. The method of claim 29, wherein the bispecific BCMA / CD3 antibody comprises an HC1 comprising the amino acid sequence of SEQ ID NO: 9 and an LC1 comprising the amino acid sequence of SEQ ID NO:

10.

31. 30. The method of claim 29, wherein the bispecific BCMA / CD3 antibody comprises an HC2 comprising the amino acid sequence of SEQ ID NO: 19 and an LC2 comprising the amino acid sequence of SEQ ID NO:

20.

32. 30. The method of claim 29, wherein the bispecific BCMA / CD3 antibody is teclistamab.

33. A kit comprising the stable aqueous pharmaceutical composition of claim 1 and instructions for use.

34. 10. An article of manufacture comprising a container holding the stable aqueous pharmaceutical composition of claim 1.

35. 35. The article of manufacture of claim 34, wherein the container is a vial having a stopper pierceable by a syringe.

36. 10. The stable aqueous pharmaceutical composition of claim 1 for use in the treatment of cancer.

37. 10. The stable aqueous pharmaceutical composition of claim 1 for use in preparing a medicament for the treatment of cancer.

38. 10. Use of the stable aqueous pharmaceutical composition of claim 1 in the preparation of a medicament for treating a subject in need thereof, wherein said treating comprises administering said medicament to said subject in need thereof.

39. 39. The use of claim 38, wherein the administering is subcutaneous administration.

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