Combination of FcgammaRIIB and tumor antibodies for treatment of FcgammaRIIB negative cancers

Abstract

The present invention describes: the use of a first antibody molecule that specifically binds to Fc [gamma] RIIB via its Fab region but lacks or has reduced binding to the Fc [gamma] receptor via its Fc region for use in combination with a second antibody molecule in the treatment of Fc [gamma] RIIB negative cancer in a patient, a second antibody molecule that specifically binds to a receptor present on a tumor cell, the second antibody molecule having an Fc region that binds to at least one activated Fc [gamma] receptor; as well as pharmaceutical compositions and kits comprising these to antibody molecules; and methods of using these two antibodies to treat cancer.

Description

Technical Field

[0001] The present invention relates to the combined use of 1) an antibody molecule that specifically binds to FcγRIIB via its Fab region and lacks an Fc region or has reduced binding to Fcγ receptors via its Fc region in the treatment of FcγRIIB-negative cancers, and 2) an antibody molecule that specifically binds to a receptor present on tumor cells, the second antibody molecule having an Fc region that binds to at least one activating Fcγ receptor. Background Art

[0002] It has long been recognized that the inhibitory Fcγ receptor (FcγR) IIB expressed by many cells of the immune system negatively regulates innate and adaptive immunity through the engagement of immune complexes (IC). Similarly, the knowledge that FcγRIIB negatively regulates monoclonal antibody-mediated immunotherapy has been known for more than a decade. Therefore, when treated with therapeutic monoclonal antibodies (mAbs), FcγRIIB-deficient mice are able to clear tumors more effectively than wild-type (WT) mice, indicating that FcγRIIB expression on effector cells (i.e., macrophages and monocytes) causes inhibition of its in vivo phagocytic and cytotoxic potential. In addition, FcγRIIB regulates the antigen presentation potential of dendritic cells (DCs), and FcγRIIB negative DCs have improved ability to activate naive T cells (van Montfoor et al., Journal of Immunology (J Immunol.) July 1, 2012; 189 (1): 92-101). Recently, antagonist antibodies that block FcγRIIB signaling and internalization in B cells have been developed. Such antibodies show effective depletion of B cells expressing FcγRIIB, and effectively promote rituximab-mediated depletion of normal and malignant B cells, demonstrating utility in hematological cancers (WO 2012 / 022985). FcγRIIB blocking antibodies with wild-type IgG1 Fc proficient in FcγR binding function and Fc (IgG1 N297Q) engineered for impaired FcγR binding show similar ability to enhance rituximab-mediated B cell depletion, indicating that the rituximab-promoting effect is independent of anti-FcγRIIB Fc. However, it has not been examined or demonstrated whether such antibodies will also have utility in enhancing the therapeutic activity of tumor-directed targeting antibodies (e.g., anti-HER2 or anti-EGFR) in the treatment of FcγRIIB-negative cancers (such as most solid cancers).

[0003] Recently, we demonstrated that Fc-FcγR proficient and impaired anti-FcγRIIB antibodies differentially enhanced the therapeutic activity of immunomodulatory antibodies against the immunosuppressive checkpoints CTLA-4 and PD-1 expressed on T cells. Specifically, the strongest anti-tumor enhancement was observed with Fc-FcγR impaired anti-FcγRIIB antibodies in the context of anti-CTLA-4 (WO 2019 / 138005). Conversely, anti-FcγRIIB enhanced therapeutic anti-tumor activity in the context of combination immunotherapy with Fc-proficient but not Fc-impaired anti-PD-1 antibodies (WO 2021 / 009358). Although studies in knockout animals have shown that anti-FcγRIIB antibodies have potential therapeutic enhancement with tumor-directed targeting antibodies in both FcγRIIB+ (e.g., B cell lymphoma when combined with anti-CD20 antibodies) and FcγRIIB- cancers (e.g., solid cancers in combination with anti-HER2 antibodies) (Clynes et al., Nat Med. 2000 Apr;6(4):443-6), it remains unclear whether anti-FcγRIIB antibodies (e.g., anti-HER2 in the treatment of FcγRIIB cancers) will enhance the therapeutic activity of tumor-directed targeting antibodies and, if so, what type of FcγRIIB antibodies will enhance the therapeutic activity of tumor-directed targeting antibodies. Summary of the invention

[0004] Herein, we demonstrate that only anti-FcγRIIB antibodies lacking an Fc region or whose Fc region exhibits reduced or impaired binding to FcγRs (e.g., F(ab)'2 antibodies or deglycosylated antibodies) are able to enhance the therapeutic activity of tumor-directed targeting antibodies (e.g., anti-HER2 and anti-EGFR for the treatment of FcγRIIB-negative cancers, including solid cancers). This is in contrast to our previous patent applications describing Fc:FcγR-proficient and Fc:FcγR-impaired anti-FcγRIIB antibodies in promoting activity and overcoming resistance to B-cell-directed targeting antibodies (e.g., anti-CD20 for the treatment of NHL (WO 2012 / 022985)), as well as the widespread use of the differential Fc:FcγR dependency of anti-FcγRIIB described in patent applications WO 2021 / 009358 and WO 2019 / 138005 for enhancing the therapeutic activity of immunomodulatory (as opposed to tumor cell-directed targeting) anti-PD-1 and anti-CTLA-4 antibodies. Furthermore, our data demonstrate that combination treatment with an anti-FcγRIIB antibody that lacks an Fc region or whose Fc region exhibits reduced or impaired binding to FcγRs (e.g., a F(ab)'2 antibody or a deglycosylated antibody) enables anti-HER2 treatment of cancers with low HER2 expression that are not amenable to treatment with currently used clinically approved anti-HER2 regimens.

[0005] Disclosed herein is a first antibody molecule that specifically binds to FcγRIIB via (or through) its Fab region and lacks an Fc region or has reduced binding to an Fcγ receptor via (or through) its Fc region, for use in combination with a second antibody molecule in the treatment of an FcγRIIB-negative cancer in a patient,

[0006] The second antibody molecule specifically binds to a receptor present on tumor cells, and the second antibody molecule has an Fc region that binds to at least one activated Fcγ receptor.

[0007] The present invention also discloses a pharmaceutical composition comprising:

[0008] (i) a first antibody molecule that specifically binds to FcγRIIB via its Fab region and lacks an Fc region or has reduced binding to an Fcγ receptor via its Fc region, and

[0009] (ii) a second antibody molecule that specifically binds to a receptor present on tumor cells, the second antibody molecule having an Fc region that binds to at least one activated Fcγ receptor;

[0010] The pharmaceutical composition is for use in the treatment of FcγRIIB-negative cancer in a patient.

[0011] Further disclosed herein is a kit for use in the treatment of FcγRIIB-negative cancers, comprising:

[0012] (i) a first antibody molecule that specifically binds to FcγRIIB via its Fab region and lacks an Fc region or has reduced binding to an Fcγ receptor via its Fc region, and

[0013] (ii) a second antibody molecule that specifically binds to a receptor present on tumor cells, wherein the second antibody molecule has an Fc region that binds to at least one activated Fcγ receptor.

[0014] The present invention further discloses the following:

[0015] (i) a first antibody molecule that specifically binds to FcγRIIB via its Fab region and lacks an Fc region or has reduced binding to an Fcγ receptor via its Fc region, and

[0016] (ii) a second antibody molecule that specifically binds to a receptor present on tumor cells, the second antibody molecule having an Fc region that binds to at least one activated Fcγ receptor;

[0017] Use in the manufacture of a medicament for use in the treatment of an FcγRIIB-negative cancer in a patient.

[0018] Also disclosed herein is a method for treating an FcγRIIB-negative cancer in a patient comprising administering:

[0019] (i) a first antibody molecule that specifically binds to FcγRIIB via its Fab region and lacks an Fc region or has reduced binding to an Fcγ receptor via its Fc region, and

[0020] (ii) a second antibody molecule that specifically binds to a receptor present on tumor cells, and has an Fc region that can activate at least one activated Fcγ receptor. DETAILED DESCRIPTION

[0021] Therefore, the present invention relates to the combined use of:

[0022] (i) a first antibody molecule that specifically binds to FcγRIIB via its Fab region and lacks an Fc region or has reduced binding to an Fcγ receptor via its Fc region, and

[0023] (ii) a second antibody molecule that specifically binds to a receptor present on tumor cells, and has an Fc region that can activate at least one activated Fcγ receptor.

[0024] Therefore, the second antibody molecule is a tumor direct targeting antibody, or it is also referred to as a direct tumor targeting antibody. The therapeutic activity of the antibody depends on the engagement of FcγR. The binding of the second antibody molecule to the receptor on the tumor cell and the subsequent binding of the FcγR on the immune effector cell triggers the redirected FcγR-dependent immune effector cell-mediated killing (e.g., by macrophage-dependent ADCC or ADCP) of the antibody-coated targeted tumor cell. Tumor direct targeting antibodies may or may not provide tumor cell killing by additional mechanisms (e.g., by blocking tumor growth factor signaling, as is believed to be the case for some anti-HER2 antibodies). In any case, the present invention is applicable to any tumor direct targeting antibody whose mechanism covers FcγR-dependent tumor cell killing. Therefore, the present invention is about maximizing therapeutic activity by optimizing FcγR-dependent tumor cell killing.

[0025] The combination is intended for use in treating FcγRIIB-negative cancers in patients with the goal of improving the therapeutic efficacy of the second antibody molecule through enhanced binding of its Fc portion to activating FcγRs and reduced binding / activation to inhibitory FcγRs.

[0026] Fc receptors are membrane proteins present on the cell surface of immune effector cells (such as macrophages). The name comes from their binding specificity to the Fc region of antibodies, which is the usual way for antibodies to bind to receptors. However, in the case where antibodies specifically bind to one or more Fc receptors, some antibodies may also bind to Fc receptors via the CDR sequences of the antibodies.

[0027] A subgroup of Fc receptors are the Fcγ receptors (Fc-gamma receptors, FcgammaR, FcgR), which are specific for IgG antibodies. There are two types of Fcγ receptors: activating Fcγ receptors (also denoted activated Fcγ receptors) and inhibitory Fcγ receptors. Activating and inhibitory receptors transmit signals through immunoreceptor tyrosine-based activation motifs (ITAMs) or immunoreceptor tyrosine-based inhibition motifs (ITIMs), respectively. In humans, FcγRIIB (FcγRIIb, FcgRIIB, CD32b) is an inhibitory Fcγ receptor, while FcγRI (CD64), FcγRIIA (CD32a), FcγRIIC (CD32c), FcγRIIIA (CD16a), and FcγRIV are activating Fcγ receptors. FcγgRIIIB is a GPI-linked receptor expressed on neutrophils that lacks an ITAM motif, but is also considered activating through its ability to cross-link lipid rafts and engage with other receptors. In mice, the activating receptors are FcγRI, FcγRIII, and FcγRIV.

[0028] It is well known that antibodies regulate immune cell activity by interacting with Fcγ receptors. Specifically, how antibody immune complexes regulate immune cell activation depends on the relative engagement of their activating and inhibitory Fcγ receptors. Different antibody isotypes bind to activating and inhibitory Fcγ receptors with different affinities, resulting in different A:I ratios (activation: inhibition ratios) (Nimmerjahn et al.; Science. December 2, 2005; 310(5753):1510-2).

[0029] By binding to inhibitory Fcγ receptors, antibodies can inhibit, block and / or downregulate the function of effector cells.

[0030] By binding to activating Fcγ receptors, antibodies can activate effector cell functions and thereby trigger mechanisms such as antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP), cytokine release and / or antibody-dependent endocytosis and neutrophil extracellular trap generation (NETosis) in the case of neutrophils (i.e., activation and release of NETs (neutrophil extracellular traps)). Antibodies that bind to activating Fcγ receptors can also lead to an increase in certain activation markers, such as CD40, MHCII, CD38, CD80 and / or CD86.

[0031] The antibody molecule according to the present invention that specifically binds to FcγRIIB (i.e., the first antibody) binds or interacts with the Fcγ receptor via the Fab region of the antibody (i.e., via the antigen binding region on the antibody that binds to the antigen, the antigen binding region consisting of a constant domain and a variable domain of each of the heavy chain and the light chain). In particular, it binds to FcγRIIB present on immune effector cells, and in particular binds to FcγRIIB present on the surface of immune effector cells. If the antibody will have a usual or common Fc region, the antibody may also have been bound to the activated Fcγ receptor by the normal interaction between the Fc region and the Fc receptor. However, according to the present invention, the antibody molecule that specifically binds to FcγRIIB completely lacks the Fc region or has reduced binding to the Fcγ receptor, which means that the antibody molecule that specifically binds to FcγRIIB binds poorly to the Fcγ receptor or cannot bind or interact with it at all. This seems to have at least two therapeutically important consequences:

[0032] 1) The lack of Fc-mediated binding to activating FcγRs makes a greater number of activating Fcγ receptors available for Fc binding of (other) therapeutic anti-cancer antibodies. This is important because clustering of increased numbers of activating FcγRs (vs. inhibitory FcγRs; Nimmerjahn et al.; Science 2005 Dec 2;310(5753):1510-2) is known to increase effector cell-mediated target cell depletion, a potential mechanism for the activity of both checkpoint inhibitors, immune agonists, and other immunomodulatory antibodies (such as anti-IL-2R).

[0033] 2) Lack of Fc-mediated binding to inhibitory FcγRs or reduced Fc-mediated binding to inhibitory FcγRs is shown to reduce inhibitory signaling in FcγR-expressing immune effector cells. Therefore, lack of Fc-mediated binding to FcγRs of FcγRIIB-targeting antibodies or reduced Fc-mediated binding to FcγRs of FcγRIIB-targeting antibodies may improve therapeutic efficacy through at least two mechanisms involving both improved activating FcγRs and reduced inhibitory Fcγ signaling in immune effector cells in response to a second immunomodulatory anti-cancer antibody.

[0034] "Reduced binding" or "binding with reduced affinity" in this context means that the antibody molecule has reduced Fc-mediated binding to the Fcγ receptor, or in other words, the Fc region of the antibody molecule that specifically binds to FcγRIIB binds to the activated Fcγ receptor with a lower affinity than the Fc region of normal human IgG1. The reduction in binding can be assessed using techniques such as surface plasmon resonance. In this article, "normal IgG1" refers to a conventionally produced IgG1 with an unmutated Fc region, the production of which does not change its glycosylation. As a reference for this "normal IgG1", rituximab produced in CHO cells without any modification can be used (Tipton et al., Blood 2015 125: 1901-1909; for example, rituximab is described in EP 0 605 442).

[0035] By "reduced binding" is meant that the binding of the Fc region of an antibody molecule that specifically binds FcγRIIB to an activating Fcγ receptor is reduced by at least 10-fold less than the binding of the Fc region of a normal human IgG1 to the same receptor for all Fc receptors. In some embodiments, it is reduced by at least 20-fold less. In some embodiments, it is reduced by at least 30-fold less. In some embodiments, it is reduced by at least 40-fold less. In some embodiments, it is reduced by at least 50-fold less. In some embodiments, it is reduced by at least 60-fold less. In some embodiments, it is reduced by at least 70-fold less.

[0036] In some embodiments of the invention, the antibody molecule that specifically binds to FcγRIIB is not bound to its Fc region at all, and in some such cases, the antibody has no Fc region; it may then be a Fab, Fab'2, scFv or PEGylated versions thereof.

[0037] In some embodiments, the antibody molecule that specifically binds to FcγRIIB can be an alpaca antibody, and in particular an alpaca hcIgG. Like all mammals, camelids produce conventional antibodies (IgG1) consisting of two heavy chains and two light chains bound together in a Y-shape with disulfide bonds. However, they also produce two unique subclasses of immunoglobulin G: IgG2 and IgG3, also known as heavy chain IgG (hcIgG). These antibodies consist of only two heavy chains that lack the CH1 region but still carry an antigen binding domain at their N-terminus, known as V HH. Conventional Ig requires the association of variable regions from both heavy and light chains to allow for a high degree of diversity in antigen-antibody interactions. Although isolated heavy and light chains still exhibit this ability, they exhibit very low affinity when compared to paired heavy and light chains. The unique feature of hcIgG is the ability of their monomeric antigen-binding regions to bind antigen with specificity, affinity, and especially diversity comparable to conventional antibodies without the need for pairing with another region.

[0038] In some embodiments, decreased binding means that the antibody has a 20-fold lower affinity for binding to FcγRI.

[0039] In order to obtain reduced binding of IgG1 antibodies (such as IgG1 antibodies) to Fc receptors, the Fc region of IgG antibodies can be modified by deglycosylation. For example, such deglycosylation of IgG1 antibodies can be achieved, for example, by amino acid substitution of asparagine (N297X) in position 297 in the antibody chain. The substitution can be carried out with glutamine (N297Q), or with alanine (N297A), or with glycine (N297G), or with asparagine (N297D) or by serine (N297S).

[0040] The Fc region can be modified by additional substitutions, for example, as described by Jacobsen FW et al., JBC 2017, 292, 1865-1875 (see, e.g., Table 1). Such additional substitutions include L242C, V259C, A287C, R292C, V302C, L306C, V323C, I332C and / or K334C. Such modifications also include the following combinations of substitutions in IgG1:

[0041] L242C, N297G, K334C,

[0042] A287C, N297G, L306C,

[0043] R292C, N297G, V302C,

[0044] N297G, V323C, I332C, and

[0045] V259C, N297G, L306C.

[0046] Alternatively, carbohydrates in the Fc region may be enzymatically cleaved and / or cells used to produce the antibody may be grown in a medium that impairs carbohydrate addition and / or cells engineered to lack the ability to add sugars may be used for antibody production, or by production of the antibody in host cells (e.g., prokaryotes, including E. coli) that do not glycosylate the antibody or do not functionally glycosylate the antibody, as described above.

[0047] Reduced affinity for Fcγ receptors can be further achieved by engineering amino acids in the Fc region of the antibody (such modifications have been previously described by, e.g., Xencor, Macrogenics, and Genentech) or by production of the antibody in host cells (e.g., prokaryotes, including E. coli) that do not glycosylate the antibody or do not functionally glycosylate the antibody.

[0048] In addition to having reduced binding to Fcγ receptors via the Fc region, it is also preferred in some embodiments that an antibody molecule that specifically binds to FcγRIIB does not cause phosphorylation of FcγRIIB upon target binding. ITIM phosphorylation of FcγRIIB is an inhibitory event that blocks activity in immune cells.

[0049] The immune effector cells expressing Fcγ receptors herein mainly refer to innate effector cells, and specifically include macrophages, neutrophils, monocytes, natural killer (NK) cells, basophils, eosinophils, mast cells and platelets. Cytotoxic T cells and memory T cells do not usually express FcγR, but may express in certain circumstances. In certain embodiments, the immune effector cells are innate immune effector cells. In certain embodiments, the immune effector cells are macrophages.

[0050] In contrast to antibody molecules that specifically bind to FcγRIIB, antibody molecules that specifically bind to or interact with receptors present on tumor cells (i.e., second antibody molecules or tumor-directed targeting antibodies) have an Fc region that binds to or interacts with activated Fcγ receptors to a degree that is not reduced or at least not significantly reduced. Binding of the second antibody to tumor cells causes activation of Fc receptor-dependent anti-tumor activity (such as depletion), antibody-dependent cellular cytotoxicity (ADCC) and / or antibody-dependent cellular phagocytosis (ADCP). When referring to depletion herein, we mean that depletion, deletion or elimination of tumor cells by physical removal of cells causes depletion of the tumor cells.

[0051] To determine whether an antibody molecule is a tumor depleting antibody molecule within the meaning of the present invention, an in vitro ADCC or ADCP assay may be used. To determine whether an antibody molecule is a tumor cell depleting antibody molecule, the same assay will be performed in the presence and absence of the depleting antibody, which will show whether the depleting antibody to be tested is actually depleting.

[0052] ADCC assay can be performed by labeling target cells with calcein AM (acetyl methyl ester) and then adding dilute concentrations of antibodies. The target cells are then co-cultured with human peripheral blood mononuclear cells (PBMC) at a 50:1 effector: target (E: T) ratio at 37°C for 4 hours. The plate is centrifuged at 400 x g for 5 minutes to precipitate the cells, and the supernatant is transferred to a white 96-well plate. Calcein release is measured using Varioskan (Thermo Scientific) using an excitation wavelength of 485 nm and an emission wavelength of 530 nm. The maximum release percentage is calculated as follows: % maximum release = (treated sample / triton) * 100.

[0053] ADCP assays can be performed by labeling target cells with 5mM carboxyfluorescein succinimidyl ester (CFSE) at room temperature for 10 minutes, followed by washing in a culture medium containing fetal bovine serum. The CFSE-labeled target is then opsonized with a diluted concentration of antibody and then co-cultured with bone marrow-derived macrophages (BMDM) in a 96-well plate at 37°C for 1 hour at a 1:5E:T ratio. BMDMs are then labeled at room temperature for 15 minutes with anti-F4 / 80 allophycocyanin and washed twice with PBS. The plate is kept on ice, the wells are scraped to collect BMDMs, and phagocytosis is assessed by flow cytometry using a FACSCalibur (BD) to determine the percentage of F4 / 80+CFSE+ cells in the F4 / 80+ cell population.

[0054] The method as described by Cleary et al., J Immunol, April 12, 2017, 1601473, may also be used.

[0055] The tumor cell to which the second antibody molecule binds is an FcγRIIB negative cancer tumor, which means that it is a tumor that does not present any FcγRIIB receptors. This can be tested using anti-FcγRIIB specific antibodies in a variety of methods, including immunohistochemistry and flow cytometry, such as Tutt et al., Journal of Immunology (J Immunol) 2015, 195 (11) 5503-5516.

[0056] In addition to specifically binding to the target on the tumor cell, the second antibody molecule also binds to the activated Fcγ receptors present on the immune effector cells via its Fc region. In order to be able to bind to the activated Fcγ receptors, in some embodiments, the Fc region of the second antibody should be glycosylated at least at position 297. The carbohydrate residues in this position help to bind to the Fcγ receptors. In certain embodiments, it is preferred that these residues are biantennary carbohydrates containing GlnNAc, mannose with terminal galactose residues, and sialic acid. It should contain the CH2 portion of the Fc molecule.

[0057] Antibodies are well known to those skilled in the art of immunology and molecular biology. Typically, an antibody comprises two heavy (H) chains and two light (L) chains. In this article, we sometimes refer to such complete antibody molecules as full-size or full-length antibodies. The heavy chain of an antibody comprises one variable domain (VH) and three constant domains (CH1, CH2, and CH3), and the light chain of an antibody molecule comprises one variable domain (VL) and one constant domain (CL). The variable domains (sometimes collectively referred to as F V The constant domains are the heavy chain constant domains that bind to the antibody's target or antigen. Each variable domain contains three loops, called complementarity determining regions (CDRs), which are responsible for target binding. The constant domains are not directly involved in the binding of the antibody to the antigen, but exhibit various effector functions. Antibodies or immunoglobulins can be divided into different classes according to the amino acid sequence of their heavy chain constant domains. There are five major classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, and in humans, several of these classes are further divided into subclasses (isotypes), e.g., IgG1, IgG2, IgG3, and IgG4; IgA1 and IgA2.

[0058] Another part of the antibody is the Fc region (also called the fragment crystallizable domain), which comprises two constant domains of each of the antibody heavy chains. As mentioned above, the Fc region is responsible for the interaction between the antibody and the Fc receptor.

[0059] As used herein, the term antibody molecule encompasses full length or full size antibodies as well as functional fragments of full length antibodies and derivatives of such antibody molecules.

[0060] The functional fragment of a full-size antibody has the same antigen-binding properties as the corresponding full-size antibody and includes the same variable domains (i.e., VH and VL sequences) and / or the same CDR sequences as the corresponding full-size antibody. The functional fragment has the same antigen-binding properties as the corresponding full-size antibody, meaning that it binds to the same epitope on the full-size antibody target. Such functional fragments may correspond to the Fv portion of a full-size antibody. Alternatively, such fragments may be Fab (also expressed as F(ab), which is a monovalent antigen-binding fragment without an Fc portion) or F(ab')2 (also expressed as Fab'2 or Fab2, which is a divalent antigen-binding fragment containing two antigen-binding Fab portions linked together by a disulfide bond) or F(ab') (i.e., a monovalent variant of F(ab')2). Such fragments may also be single-chain variable fragments (scFv).

[0061] Functional fragments do not always contain all six CDRs of the corresponding full-size antibody. It should be understood that molecules containing three or fewer CDR regions (in some cases, even just a single CDR or a portion thereof) can retain the antigen-binding activity of the antibody from which the CDR is derived. For example, Gao et al., 1994, Journal of Biological Chemistry (J. Biol. Chem.), 269: 32389-93 describe that the entire VL chain (including all three CDRs) has a high affinity for its substrate.

[0062] Molecules containing two CDR regions are described, for example, by Vaughan and Sollazzo 2001, Combinatorial Chemistry & High Throughput Screening, 4: 417-430. On page 418 (right column - 3 Our design strategy) a minibody comprising only the H1 and H2 CDR hypervariable regions interspersed within the framework region is described. The minibody is described as being able to bind to the target. Pessi et al., 1993, Nature, 362: 367-9 and Bianchi et al., 1994, J. Mol. Biol., 236: 649-59 are cited by Vaughan and Sollazzo and describe the H1 and H2 minibodies and their properties in more detail. In Qiu et al., 2007, Nature Biotechnology, 25: 921-9 it was demonstrated that a molecule consisting of two linked CDRs is able to bind to an antigen. Quiocho 1993, Nature, 362:293-4 provides an overview of "minibody" technology. Ladner 2007, Nature Biotechnology, 25:875-7 reviews that molecules containing two CDRs can retain antigen binding activity.

[0063] For example, in Laune et al., 1997, Journal of Biological Chemistry (JBC), 272: 30937-44, antibody molecules containing a single CDR region are described, in which a series of hexapeptides derived from the CDR are shown to exhibit antigen binding activity, and it is noted that synthetic peptides of a complete single CDR exhibit strong binding activity. In Monnet et al., 1999, Journal of Biological Chemistry (JBC), 274: 3789-96, a series of 12-mer peptides and associated framework regions are shown to have antigen binding activity, and it is commented that individual CDR3-like peptides can bind to antigens. In Heap et al., 2005, Journal of General Virology (J. Gen. Virol.), 86: 1791-1800, it is reported that "miniantibodies" (molecules containing a single CDR) can bind to antigens, and it is shown that cyclic peptides from anti-HIV antibodies have antigen binding activity and function. It was shown in Nicaise et al., 2004, Protein Science, 13: 1882-91 that a single CDR can confer antigen binding activity and affinity to its lysozyme antigen.

[0064] Thus, antibody molecules with five, four, three or fewer CDRs are able to retain the antigen-binding properties of the full-length antibody from which they are derived.

[0065] The antibody molecule may also be a derivative of a full-length antibody or a fragment of such an antibody. When a derivative is used, it should have the same antigen binding characteristics as the corresponding full-length antibody in the sense that it binds to the same epitope on the target as the full-length antibody.

[0066] Therefore, as used herein, when referring to the term "antibody molecule", we include all types of antibody molecules and their functional fragments and derivatives thereof, including: monoclonal antibodies, polyclonal antibodies, synthetic antibodies, recombinantly produced antibodies, multispecific antibodies, bispecific antibodies, human antibodies, antibodies of human origin, humanized antibodies, chimeric antibodies, single-chain antibodies, single-chain Fv (scFv), Fab fragments, F(ab')2 fragments, F(ab') fragments, disulfide-linked Fv (sdFv), antibody heavy chains, antibody light chains, homodimers of antibody heavy chains, homodimers of antibody light chains, heterodimers of antibody heavy chains, heterodimers of antibody light chains, and antigen-binding functional fragments of such homodimers and heterodimers.

[0067] Furthermore, as used herein, the term "antibody molecule" includes all classes of antibody molecules and functional fragments, including: IgG, IgG1, IgG2, IgG3, IgG4, IgA, IgM, IgD and IgE, unless otherwise specified.

[0068] In some embodiments, the antibody is human IgG1. The skilled artisan will appreciate that mouse IgG2a and human IgG1 engage activating Fcγ receptors and share the ability to activate deletions of target cells through activation of immune cells bearing activating Fcγ receptors, such as ADCP and ADCC. Therefore, in embodiments where mouse IgG2a is the preferred isotype for deletions in mice, human IgG1 is the preferred isotype for deletions in humans in such embodiments.

[0069] As mentioned above, different types and forms of antibody molecules are encompassed by the present invention and are known to those skilled in the art of immunology.As is known, antibodies used for therapeutic purposes are often modified with additional components that modify the properties of the antibody molecule.

[0070] Thus, an antibody molecule comprising or used according to the invention (eg, a monoclonal antibody molecule, and / or a polyclonal antibody molecule, and / or a bispecific antibody molecule) comprises a detectable moiety and / or a cytotoxic moiety.

[0071] When referring to a "detectable moiety" we include one or more from the group consisting of: an enzyme; a radioactive atom; a fluorescent moiety; a chemiluminescent moiety; a bioluminescent moiety. The detectable moiety allows visualization of the antibody molecule in vitro, and / or in vivo, and / or ex vivo.

[0072] By "cytotoxic moiety" we include a radioactive moiety and / or an enzyme, wherein the enzyme is a caspase and / or a toxin, wherein the toxin is a bacterial toxin or venom; wherein the cytotoxic moiety is capable of inducing cell lysis.

[0073] Further included are antibody molecules that may be in isolated and / or purified form, and / or may be pegylated. Pegylation is a process by which polyethylene glycol polymers are added to a molecule such as an antibody molecule or derivative to modify its behavior, for example by increasing its hydrodynamic size to extend its half-life, thereby preventing renal clearance.

[0074] As described above, the CDRs of an antibody bind to an antibody target. The amino acid assignments for each CDR described herein are according to the definitions described in Kabat EA et al. 1991, Sequences of Proteins of Immunological Interest, Fifth Edition, NIH Publication No. 91-3242, pp. xv to xvii.

[0075] As will be known to those skilled in the art, there are other methods for assigning amino acids to each CDR, for example, the International ImMunoGeneTics information system (IMGT(R)) (http: / / www.imgt.org / and Lefranc and Lefranc, The Immunoglobulin Facts Book, Academic Press, 2001).

[0076] In a further embodiment, the antibody molecule of the present invention or used according to the present invention is an antibody molecule capable of competing with the specific antibody provided herein, for example, an antibody molecule for binding to a specific target comprising any of the amino acid sequences listed in, for example, SEQ ID NOs: 1 to 194.

[0077] By "capable of competing against" we mean that the competing antibody is capable of at least partially inhibiting or otherwise interfering with the binding of an antibody molecule as defined herein to a specific target.

[0078] For example, such competing antibody molecules may be able to inhibit the binding of an antibody molecule described herein by at least about 10%; e.g., at least about 20%, or at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, about 100%, and / or inhibit the ability of an antibody described herein to prevent or reduce binding to a specific target by at least about 10%; e.g., at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or about 100%.

[0079] Competitive binding can be determined by methods well known to those skilled in the art, such as enzyme-linked immunosorbent assay (ELISA).

[0080] ELISA assays can be used to assess epitope modification or blocking antibodies. Other methods suitable for identifying competing antibodies are disclosed in Antibodies: A Laboratory Manual, Harlow and Lane, which is incorporated herein by reference (e.g., see pages 567 to 569, 574 to 576, 583, and 590 to 612, 1988, CSHL, NY, ISBN 0-87969-314-2).

[0081] The targets of the antibodies according to the invention or of the antibodies used according to the invention are expressed on the surface of cells, i.e. they are cell surface antigens which will comprise epitopes for the antibodies (in this context otherwise referred to as cell surface epitopes). Cell surface antigens and epitopes are terms which will be readily understood by those skilled in the art of immunology or cell biology.

[0082] When referring to "cell surface antigens", we include that the cell surface antigen is exposed on the extracellular side of the cell membrane, but may be only transiently exposed on the extracellular side of the cell membrane. When referring to "transiently exposed", we include that the cell surface antigen may be internalized into the cell or released from the extracellular side of the cell membrane into the extracellular space. The cell surface antigen may be released from the extracellular side of the cell membrane by cleavage, which may be mediated by a protease.

[0083] We also include that the cell surface antigen may be attached to the cell membrane, but may only be transiently associated with the cell membrane. When referring to "transiently associated", we include that the cell surface antigen may be released from the extracellular side of the cell membrane into the extracellular space. The cell surface antigen may be released from the extracellular side of the cell membrane by cleavage, which may be mediated by a protease.

[0084] We further include that the cell surface antigen may be a peptide, or a polypeptide, or a carbohydrate, or an oligosaccharide chain, or a lipid; and / or an epitope present on a protein, or a glycoprotein, or a lipoprotein.

[0085] Methods for assessing protein binding are known to those skilled in the art in the fields of biochemistry and immunology. It will be appreciated by those skilled in the art that those methods can be used to assess the binding of an antibody to a target and / or the binding of an antibody's Fc region to an Fc receptor; and the relative strength, or specificity, or inhibition, or prevention or reduction in those interactions. Examples of methods that can be used to assess protein binding are, for example, immunoassays, BIAcore, Western blots, radioimmunoassays (RIA) and enzyme-linked immunosorbent assays (ELISA) (see Fundamental Immunology Second Edition, Raven Press, New York, pp. 332 to 336 (1989) for discussion of antibody specificity).

[0086] The meaning of an antibody that specifically binds or interacts with a defined target molecule or antigen is well known and means that the antibody preferentially and selectively binds to its target rather than to non-target molecules. In this context, the term "binds to" can be used interchangeably with "interacts with". Thus, when referring to a "specifically binding antibody molecule" or "target-specific antibody molecule", we include that the antibody molecule specifically binds to the target but not to the non-target, or binds to the non-target more weakly (such as with a lower affinity) than the target.

[0087] We also include the meaning that an antibody specifically binds to a target at least 2-fold, or at least 5-fold, or at least 10-fold, or at least 20-fold, or at least 50-fold, or at least 100-fold, or at least 200-fold, or at least 500-fold, or at least about 1000-fold more strongly than it specifically binds to a non-target.

[0088] In addition, we include the meaning that if the antibody -1 K d , or at least about 10 -2 K d , or at least about 10 -3 K d , or at least about 10 -4 K d , or at least about 10 -5 K d , or at least about 10 -6 K d , or at least about 10 -7 K d , or at least about 10 -8 K d , or at least about 10 -9 K d , or at least about 10 -10 K d , or at least about 10 -11 K d , or at least about 10 -12 K d , or at least about 10 -13 K d , or at least about 10 -14 K d or at least about 10 -15 K d K d When the antibody binds to a target, the antibody specifically binds to the target.

[0089] In some embodiments, the antibody molecule that specifically binds FcγRIIB is a human antibody.

[0090] In some embodiments, the antibody molecule that specifically binds FcγRIIB is an antibody of human origin, ie, an original human antibody that has been modified as described herein.

[0091] In some embodiments, the antibody molecule that specifically binds to FcγRIIB is a humanized antibody, ie, an original non-human antibody that has been modified to increase its similarity to a human antibody. The humanized antibody may be, for example, a murine antibody or an alpaca antibody.

[0092] In some embodiments, the antibody molecule that specifically binds to FcγRIIB comprises the following constant regions (CH and CL): IgG1-CH [SEQ ID NO: 1]

[0093] ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS

[0094] SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELL

[0095] GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPRE

[0096] EQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0097] IgG1-CL [SEQ ID NO: 2]

[0098] QPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSK

[0099] QSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS

[0100] These constant regions (SEQ ID NO: 1 and SEQ ID NO: 2) are of human origin. The Fc region is further modified for reduced binding to Fcγ receptors via its Fc region. As described herein, in some embodiments, preferably, SEQ ID NO: 1 has been deglycosylated by N297Q substitution, and IgG1-CH then has the following CH sequence [SEQ ID NO: 195], wherein the 297 Q residue is marked in bold:

[0101]

[0102] In some embodiments and / or examples, mouse antibody molecules are used. These can also be used for alternative antibodies. These can then contain the following constant regions (CH and CL):

[0103] CH [SEQ ID NO: 196]

[0104]

[0105] CL [SEQ ID NO: 197]

[0106] QPKSSPSVTLFPPSSEELETNKATLVCTITDFYPGVVTVDWKVDGTPVTQGMETTQPSK

[0107] QSNNKYMASSYLTLTARAWERHSSYSCQVTHEGHTVEKSLSRADCS

[0108] Therefore, these constant regions (SEQ ID NO: 196 and SEQ ID NO: 197) are of murine origin. SEQ ID NO: 196 comprises a N297A mutation (the 297 A residue is marked in bold in the above sequence). This N297A mutation in the mouse sequence corresponds to the N297Q mutation in the human sequence.

[0109] In some embodiments, an antibody molecule that specifically binds to FcγRIIB comprises one or more sequences of the following clones:

[0110] Antibody clone: ​​1A01

[0111] 1A01-VH [SEQ ID NO:3]

[0112] EVQLLESGGGLVQPGGSLRLSCAASGFTFSDYYMNWIRQTPGKGLEWVSLIGWDGGS

[0113] TYYADSVKGRFTISRDNSENTLYLQMNSLRAEDTAVYYCARAYSGYELDYWGQGTLV

[0114] TVSS

[0115] 1A01-VL [SEQ ID NO:27]

[0116] QSVLTQPPSASGTPGQRVTISCSGSSSNIGNNAVNWYQQLPGTAPKLLIYDNNNRPSGV

[0117] PDRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLNASIFGGGTKLTVLG CDR Area

[0118] CDRH1: DYYMN [ SEQ ID NO:51]

[0119] CDRH2: LIGWDGGSTYYADSVKG [ SEQ ID NO:52]

[0120] CDRH3: AYSGYELDY [ SEQ ID NO:53]

[0121] CDRL1: SGSSSNIGNNAVN [ SEQ ID NO:54]

[0122] CDRL2: DNNNRPS [ SEQ ID NO:55]

[0123] CDRL3: AAWDDSLNASI [ SEQ ID NO:56]

[0124] Antibody clone: 1B07

[0125] 1B07-VH [SEQ ID NO:4]

[0126] EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAFTRYDGSN

[0127] KYYADSVRGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARENIDAFDVWGQGTLVT

[0128] VSS

[0129] 1B07-VL [SEQ ID NO:28]

[0130] QSVLTQPPSASGTPGQRVTISCSGSSSNIGNNAVNWYQQLPGTAPKLLIYDNQQRPSGV

[0131] PDRFSGSKSGTSASLAISGLRSEDEADYYCEAWDDRLFGPVFGGGTKLTVLG CDR Area

[0132] CDRH1: SYGMH [ SEQ ID NO:57]

[0133] CDRH2: FTRYDGSNKYYADSVRG [ SEQ ID NO:58]

[0134] CDRH3: ENIDAFDV [ SEQ ID NO:59]

[0135] CDRL1: SGSSSNIGNNAVN [ SEQ ID NO:60]

[0136] CDRL2: DNQQRPS [ SEQ ID NO:61]

[0137] CDRL3: WDDRLFGPV [ SEQ ID NO:62]

[0138] Antibody clone: 1C04

[0139] 1C04-VH [SEQ ID NO:5]

[0140] EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSSISDSGAGR

[0141] YYADSVEGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARTHDSGELLDAFDIWGQGT

[0142] LVTVSS

[0143] 1C04-VL [SEQ ID NO:29]

[0144] QSVLTQPPSASGTPGQRVTISCSGSSSNIGSNHVLWYQQLPGTAPKLLIYGNSNRPSGVP

[0145] DRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLNGWVFGGGTKLTVLG CDR Area

[0146] CDRH1: SYAMS [ SEQ ID NO:63]

[0147] CDRH2: SISDSGAGRYYADSVEG [ SEQ ID NO:64]

[0148] CDRH3: THDSGELLDAFDI [ SEQ ID NO:65]

[0149] CDRL1: SGSSSNIGSNHVL [ SEQ ID NO:66]

[0150] CDRL2: GNSNRPS [ SEQ ID NO:67]

[0151] CDRL3: AAWDDSLNGWV [ SEQ ID NO:68]

[0152] Antibody clone: 1E05

[0153] 1E05-VH [SEQ ID NO:6]

[0154] EVQLLESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQVPGKGLEWVAVISYDGSN

[0155] KNYVDSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARNFDNSGYAIPDAFDIWG

[0156] QGTLVTVSS

[0157] 1E05-VL [SEQ ID NO:30]

[0158] QSVLTQPPSASGTPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYDNNSRPSG

[0159] VPDRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLGGPVFGGGTKLTVLG CDR Area

[0160] CDRH1: TYAMN [ SEQ ID NO:69]

[0161] CDRH2: VISYDGSNKNYVDSVKG [ SEQ ID NO:70]

[0162] CDRH3: NFDNSGYAIPDAFDI [ SEQ ID NO:71]

[0163] CDRL1: TGSSSNIGAGYDVH [ SEQ ID NO:72]

[0164] CDRL2: DNNSRPS [ SEQ ID NO:73]

[0165] CDRL3: AAWDDSLGGPV [ SEQ ID NO:74]

[0166] Antibody Clone: 2A09

[0167] 2A09-VH [SEQ ID NO:7]

[0168] EVQLLESGGGLVQPGGSLRLSCAASGFTFSNAWMSWVRQAPGKGLEWVAYISRDADIT

[0169] HYPASVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCTTGFDYAGDDAFDIWGQGTL

[0170] VTVSS

[0171] 2A09-VL [SEQ ID NO:31]

[0172] QSVLTQPPSASGTPGQRVTISCSGSSSNIGSNAVNWYQQLPGTAPKLLIYGNSDRPSGVP

[0173] DRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLNGRWVFGGGTKLTVLG CDR Area

[0174] CDRH1: NAWMS [ SEQ ID NO:75]

[0175] CDRH2: YISRDADITHYPASVKG [ SEQ ID NO:76]

[0176] CDRH3: GFDYAGDDAFDI [ SEQ ID NO:77]

[0177] CDRL1: SGSSSNIGSNAVN [ SEQ ID NO:78]

[0178] CDRL2: GNSDRPS [ SEQ ID NO:79]

[0179] CDRL3: AAWDDSLNGRWV [ SEQ ID NO:80]

[0180] Antibody clone: 2B08

[0181] 2B08-VH [SEQ ID NO:8]

[0182] EVQLLESGGGLVQPGGSLRLSCAASGFTFSDYYMSWVRQAPGKGLEWVALIGHDGNN

[0183] KYYLDSLEGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARATDSGYDLLYWGQGTLV

[0184] TVSS

[0185] 2B08-VL [SEQ ID NO:32]

[0186] QSVLTQPPSASGTPGQRVTISCSGSSSNIGNNAVNWYQQLPGTAPKLLIYYDDLLPSGVP

[0187] DRFSGSKSGTSASLAISGLRSEDEADYYCTTWDDSLSGVVFGGGTKLTVLG CDR Area

[0188] CDRH1: DYYMS [ SEQ ID NO:81]

[0189] CDRH2: LIGHDGNNKYYLDSLEG [ SEQ ID NO:82]

[0190] CDRH3: ATDSGYDLLY [ SEQ ID NO:83]

[0191] CDRL1: SGSSSNIGNNAVN [ SEQ ID NO:84]

[0192] CDRL2: YDDLLPS [ SEQ ID NO:85]

[0193] CDRL3: TTWDDSLSGVV [ SEQ ID NO:86]

[0194] Antibody clone: 2E8-VH

[0195] 2E8-VH [SEQ ID NO:9]

[0196] EVQLLESGGGLVQPGGSLRLSCAASGFTFSDYYMSWIRQAPGKGLEWVSAIGFSDDNT

[0197] YYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAGGDGSGWSFWGQGTLVTV

[0198] SS

[0199] 2E8-VL [SEQ ID NO:33]

[0200] QSVLTQPPSASGTPGQRVTISCSGSSSNIGNNAVNWYQQLPGTAPKLLIYDNNKRPSGV

[0201] PDRFSGSKSGTSASLAISGLRSEDEADYYCATWDDSLRGWVFGGGTKLTVLG CDR Area

[0202] CDRH1: DYYMS [ SEQ ID NO:87]

[0203] CDRH2: AIGFSDDNTYYADSVKG [ SEQ ID NO:88]

[0204] CDRH3: GDGSGWSF [ SEQ ID NO:89]

[0205] CDRL1: SGSSSNIGNNAVN [ SEQ ID NO:90]

[0206] CDRL2: DNNKRPS [ SEQ ID NO:91]

[0207] CDRL3: ATWDDSLRGWV [ SEQ ID NO:92]

[0208] Antibody Clone: 5C04

[0209] 5C04-VH [SEQ ID NO:10]

[0210] EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYGMHWVRQAPGKGLEWVAVISYDGSN

[0211] KYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAREWRDAFDIWGQGTLVT

[0212] VSS

[0213] 5C04-VL[ SEQ ID NO:34]

[0214] QSVLTQPPSASGTPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYSDNQRPSG

[0215] VPDRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLSGSWVFGGGTKLTVLG CDR Area

[0216] CDRH1: NYGMH [ SEQ ID NO:93]

[0217] CDRH2: VISYDGSNKYYADSVKG [ SEQ ID NO:94]

[0218] CDRH3: WRDAFDI [ SEQ ID NO:95]

[0219] CDRL1: TGSSSNIGAGYDVH [ SEQ ID NO:96]

[0220] CDRL2: SDNQRPS [ SEQ ID NO:97]

[0221] CDRL3: AAWDDSLSGSWV [ SEQ ID NO:98]

[0222] Antibody clone: 5C05

[0223] 5C05-VH [SEQ ID NO:11]

[0224] EVQLLESGGGLVQPGGSLRLSCAASGFTFSTYGMHWVRQAPGKGLEWVAVISYDGSN

[0225] KYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARENFDAFDVWGQGTLVT

[0226] VSS

[0227] 5C05-VL[ SEQ ID NO:35]

[0228] QSVLTQPPSASGTPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYSNSQRPSG

[0229] VPDRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLNGQVVFGGGTKLTVLG CDR Area

[0230] CDRH1: TYGMH [ SEQ ID NO:99]

[0231] CDRH2: VISYDGSNKYYADSVKG [ SEQ ID NO:100]

[0232] CDRH3: ENFDAFDV [ [SEQ ID NO:101]

[0233] CDRL1: TGSSSNIGAGYDVH [ [SEQ ID NO:102]

[0234] CDRL2: SNSQRPS [ [SEQ ID NO:103]

[0235] CDRL3: AAWDDSLNGQVV [ [SEQ ID NO:104]

[0236] Antibody Clone: 5D07

[0237] 5D07-VH [SEQ ID NO:12]

[0238] EVQLLESGGGLVQPGGSLRLSCAASGFTFSTYGMHWVRQAPGKGLEWVAVIAYDGSK

[0239] KDYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAREYRDAFDIWGQGTLVT

[0240] VSS

[0241] 5D07-VL[ [SEQ ID NO:36]

[0242] QSVLTQPPSASGTPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSG

[0243] VPDRFSGSKSGTTASLAISGLRSEDEADYYCAAWDDSVSGWMFGGGTKLTVLG CDR Area

[0244] CDRH1: TYGMH [ [SEQ ID NO:105]

[0245] CDRH2: VIAYDGSKKDYADSVKG [ [SEQ ID NO:106]

[0246] CDRH3: EYRDAFDI [ [SEQ ID NO:107]

[0247] CDRL1: TGSSSNIGAGYDVH [ SEQ ID NO:108]

[0248] CDRL2: GNSNRPS [ SEQ ID NO:109]

[0249] CDRL3: AAWDDSVSGWM [ SEQ ID NO:110]

[0250] Antibody clone: 5E12

[0251] 5E12-VH [SEQ ID NO:13]

[0252] EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVISYDGINK

[0253] DYADSMKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARERKDAFDIWGQGTLVTV

[0254] SS

[0255] 5E12-VL[ SEQ ID NO:37]

[0256] QSVLTQPPSASGTPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYSNNQRPSG

[0257] VPDRFSGSKSGTSASLAISGLRSEDEADYYCATWDDSLNGLVFGGGTKLTVLG CDR Area

[0258] CDRH1: SYGMH [ SEQ ID NO:111]

[0259] CDRH2: VISYDGINKDYADSMKG [ SEQ ID NO:112]

[0260] CDRH3: ERKDAFDI [ SEQ ID NO:113]

[0261] CDRL1: TGSSSNIGAGYDVH [ SEQ ID NO:114]

[0262] CDRL2: SNNQRPS [ SEQ ID NO:115]

[0263] CDRL3: ATWDDSLNGLV [ SEQ ID NO:116]

[0264] Antibody clone: 5G08

[0265] 5G08-VH [SEQ ID NO:14]

[0266] EVQLLESGGGLVQPGGSLRLSCAASGFTFNNYGMHWVRQAPGKGLEWVAVISYDGSN

[0267] RYYADSVKGRFTMSRDNSKNTLYLQMNSLRAEDTAVYYCARDRWNGMDVWGQGTL

[0268] VTVSS

[0269] 5G08-VL[ SEQ ID NO:38]

[0270] QSVLTQPPSASGTPGQRVTISCSGSSSNIGAGYDVHWYQQLPGTAPKLLIYANNQRPSG

[0271] VPDRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLNGPWVFGGGTKLTVLG CDR Area

[0272] CDRH1: NYGMH [ SEQ ID NO:117]

[0273] CDRH2: VISYDGSNRYYADSVKG [ SEQ ID NO:118]

[0274] CDRH3: DRWNGMDV [ SEQ ID NO:119]

[0275] CDRL1: SGSSSNIGAGYDVH [ SEQ ID NO:120]

[0276] CDRL2: ANNQRPS [ SEQ ID NO:121]

[0277] CDRL3: AAWDDSLNGPWV [ SEQ ID NO:122]

[0278] Antibody clone: 5H06

[0279] 5H06-VH [SEQ ID NO:15]

[0280] EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVISYDGSDT

[0281] AYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDHSVIGAFDIWGQGTLVT

[0282] VSS

[0283] 5H06-VL[ SEQ ID NO:39]

[0284] QSVLTQPPSASGTPGQRVTISCSGSSSNIGSNTVNWYQQLPGTAPKLLIYDNNKRPSGVP

[0285] DRFSGSKSGTSASLAISGLRSEDEADYYCSSYAGSNNVVFGGGTKLTVLG CDR Area

[0286] CDRH1: SYGMH [ SEQ ID NO:123]

[0287] CDRH2: VISYDGSDTAYADSVKG [ SEQ ID NO:124]

[0288] CDRH3: DHSVIGAFDI [ SEQ ID NO:125]

[0289] CDRL1: SGSSSNIGSNTVN [ SEQ ID NO:126]

[0290] CDRL2: DNNKRPS [ SEQ ID NO:127]

[0291] CDRL3: SSYAGSNNVV [ SEQ ID NO:128]

[0292] Antibody Clone: 6A09

[0293] 6A09-VH [SEQ ID NO:16]

[0294] EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVTSYDGNT

[0295] KYYANSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAREDCGGDCFDYWGQGTL

[0296] VTVSS

[0297] 6A09-VL[ SEQ ID NO:40]

[0298] QSVLTQPPSASGTPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSG

[0299] VPDRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLNEGVFGGGTKLTVLG CDR Area

[0300] CDRH1: SYGMH [ SEQ ID NO:129]

[0301] CDRH2: VTSYDGNTKYYANSVKG [ SEQ ID NO:130]

[0302] CDRH3: EDCGGDCFDY [ SEQ ID NO:131]

[0303] CDRL1: TGSSSNIGAGYDVH [ SEQ ID NO:132]

[0304] CDRL2: GNSNRPS [ SEQ ID NO:133]

[0305] CDRL3: AAWDDSLNEGV [ SEQ ID NO:134]

[0306] Antibody Clone: 6B01

[0307] 6B01-VH [SEQ ID NO:17]

[0308] EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYGMHWVRQAPGKGLEWVAVISYDGSN

[0309] KYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDQLGEAFDIWGQGTLV

[0310] TVSS

[0311] 6B01-VL[ SEQ ID NO:41]

[0312] QSVLTQPPSASGTPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYDNNKRPSG

[0313] VPDRFSGSKSGTSASLAISGLRSEDEADYYCATWDDSLSGPVFGGGTKLTVLG CDR Area

[0314] CDRH1: NYGMH [ SEQ ID NO:135]

[0315] CDRH2: VISYDGSNKYYADSVKG [ SEQ ID NO:136]

[0316] CDRH3: DQLGEAFDI [ SEQ ID NO:137]

[0317] CDRL1: TGSSSNIGAGYDVH [ SEQ ID NO:138]

[0318] CDRL2: DNNKRPS [ SEQ ID NO:139]

[0319] CDRL3: ATWDDSLSGPV [ SEQ ID NO:140]

[0320] Antibody clone: 6C11

[0321] 6C11-VH [SEQ ID NO:18]

[0322] EVQLLESGGGLVQPGGSLRLSCAASGFTFDDYGMSWVRQAPGKGLEWVSAISGSGSST

[0323] YYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAGGDIDYFDYWGQGTLVTV

[0324] SS

[0325] 6C11-VL[ SEQ ID NO:42]

[0326] QSVLTQPPSASGTPGQRVTISCTGSSSNFGAGYDVHWYQQLPGTAPKLLIYENNKRPSG

[0327] VPDRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLNGPVFGGGTKLTVLG CDR Area

[0328] CDRH1: DYGMS [ SEQ ID NO:141]

[0329] CDRH2: AISGSGSSTYYADSVKG [ SEQ ID NO:142]

[0330] CDRH3: GDIDYFDY [ SEQ ID NO:143]

[0331] CDRL1: TGSSSNFGAGYDVH [ SEQ ID NO:144]

[0332] CDRL2: ENNKRPS [ SEQ ID NO:145]

[0333] CDRL3: AAWDDSLNGPV [ SEQ ID NO:146]

[0334] Antibody clone: 6C12

[0335] 6C12-VH [SEQ ID NO:19]

[0336] EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVISYDGSN

[0337] KYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARERRDAFDIWGQGTLVT

[0338] VSS

[0339] 6C12-VL[ SEQ ID NO:43]

[0340] QSVLTQPPSASGTPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYSDNQRPSG

[0341] VPDRFSGSKSGTSASLAISGLRSEDEADYYCATWDSDTPVFGGGTKLTVLG CDR Area

[0342] CDRH1: SYGMH [ SEQ ID NO:147]

[0343] CDRH2: VISYDGSNKYYADSVKG [ SEQ ID NO:148]

[0344] CDRH3: ERRDAFDI [ SEQ ID NO:149]

[0345] CDRL1: TGSSSNIGAGYDVH [ SEQ ID NO:150]

[0346] CDRL2: SDNQRPS [ SEQ ID NO:151]

[0347] CDRL3: ATWDSDTPV [ SEQ ID NO:152]

[0348] Antibody clone: 6D01

[0349] 6D01-VH [SEQ ID NO:20]

[0350] EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVISYDGSN

[0351] KYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAMYYCARDHSAAGYFDYWGQGT

[0352] LVTVSS

[0353] 6D01-VL[ SEQ ID NO:44]

[0354] QSVLTQPPSASGTPGQRVTISCSGSSSNIGSNTVNWYQQLPGTAPKLLIYGNSIRPSGGP

[0355] DRFSGSKSGTSASLAISGLRSEDEADYYCASWDDSLSSPVFGGGTKLTVLG CDR Area

[0356] CDRH1: SYGMH [ SEQ ID NO:153]

[0357] CDRH2: VISYDGSNKYYADSVKG [ SEQ ID NO:154]

[0358] CDRH3: DHSAAGYFDY [ SEQ ID NO:155]

[0359] CDRL1: SGSSSNIGSNTVN [ SEQ ID NO:156]

[0360] CDRL2: GNSIRPS [ SEQ ID NO:157]

[0361] CDRL3: ASWDDSLSSPV [ SEQ ID NO:158]

[0362] Antibody clone: 6G03

[0363] 6G03-VH [SEQ ID NO:21]

[0364] EVQLLESGGGLVQPGGSLRLSCAASGFTFGSYGMHWVRQAPGKGLEWVSGISWDSAII

[0365] DYAGSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDEAAAGAFDIWGQGTLV

[0366] TVSS

[0367] 6G03-VL[SEQ ID NO:45

[0368] QSVLTQPPSASGTPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNTDRPSG

[0369] VPDRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLSGPVVFGGGTKLTVLG CDR Area

[0370] CDRH1: SYGMH [ SEQ ID NO:159

[0371] CDRH2: GISWDSAIIDYAGSVKG [ SEQ ID NO:160

[0372] CDRH3: DEAAAGAFDI [ SEQ ID NO:161

[0373] CDRL1: TGSSSNIGAGYDVH [ SEQ ID NO:162

[0374] CDRL2: GNTDRPS [ SEQ ID NO:163

[0375] CDRL3: AAWDDSLSGPVV [ SEQ ID NO:164

[0376] Antibody clone: 6G08

[0377] 6G08-VH [SEQ ID NO:22]

[0378] EVQLLESGGGLVQPGGSLRLSCAASGFTLSSYGISWVRQAPGKGLEWVSGISGSGGNT

[0379] YYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCASSVGAYANDAFDIWGQGTL

[0380] VTVSS

[0381] 6G08-VL[ SEQ ID NO:46

[0382] QSVLTQPPSASGTPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGDTNRPSG

[0383] VPDRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLNGPVFGGGTKLTVLG CDR Area

[0384] CDRH1: SYGIS [ SEQ ID NO:165]

[0385] CDRH2: GISGSGGNTYYADSVKG [ SEQ ID NO:166]

[0386] CDRH3: SVGAYANDAFDI [ SEQ ID NO:167]

[0387] CDRL1: TGSSSNIGAGYDVH [ SEQ ID NO:168]

[0388] CDRL2: GDTNRPS [ SEQ ID NO:169]

[0389] CDRL3: AAWDDSLNGPV [ SEQ ID NO:170]

[0390] Antibody clone: 6G11

[0391] 6G11-VH [SEQ ID NO:23]

[0392] EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWMAVISYDGSN

[0393] KYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARELYDAFDIWGQGTLVTV

[0394] SS

[0395] 6G11-VL[ SEQ ID NO:47]

[0396] QSVLTQPPSASGTPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYADDHRPSG

[0397] VPDRFSGSKSGTSASLAISGLRSEDEADYYCASWDDSQRAVIFGGGTKLTVLG CDR Area

[0398] CDRH1: SYGMH [ SEQ ID NO:171]

[0399] CDRH2: VISYDGSNKYYADSVKG [ SEQ ID NO:172]

[0400] CDRH3: ELYDAFDI [ SEQ ID NO:173]

[0401] CDRL1: TGSSSNIGAGYDVH [ SEQ ID NO:174]

[0402] CDRL2: ADDHRPS [ SEQ ID NO:175]

[0403] CDRL3: ASWDDSQRAVI [ SEQ ID NO:176]

[0404] Antibody clone: 6H08

[0405] 6H08-VH [SEQ ID NO:24]

[0406] EVQLLESGGGLVQPGGSLRLSCAASGFTFNNYGMHWVRQAPGKGLEWVAVISYDGSN

[0407] KYYADSVKGRFTISKDNSKNTLYLQMNSLRAEDTAVYYCAREYKDAFDIWGQGTLVT

[0408] VSS

[0409] 6H08-VL[ SEQ ID NO:48]

[0410] QSVLTQPPSASGTPGQRVTISCTGSSSNIGSNTVNWYQQLPGTAPKLLIYDNNKRPSGVP

[0411] DRFSGSKSGTSASLAISGLRSEDEADYYCQAWGTGIRVFGGGTKLTVLG CDR Area

[0412] CDRH1: NYGMH [ SEQ ID NO:177]

[0413] CDRH2: VISYDGSNKYYAD SVKG [ SEQ ID NO:178]

[0414] CDRH3: EYKDAFDI [ SEQ ID NO:179]

[0415] CDRL1: TGSSSNIGSNTVN [ SEQ ID NO:180]

[0416] CDRL2: DNNKRPS [ SEQ ID NO:181]

[0417] CDRL3: QAWGTGIRV [ SEQ ID NO:182]

[0418] Antibody clone: 7C07

[0419] 7C07-VH [SEQ ID NO:25]

[0420] EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVISYDGSN

[0421] KYYADSVKGRFTISRDNSQNTLYLQMNSLRAEDTAVYYCAREFGYIILDYWGQGTLVT

[0422] VSS

[0423] 7C07-VL[ SEQ ID NO:49]

[0424] QSVLTQPPSASGTPGQRVTISCSGSSSNIGSNTVNWYQQLPGTAPKLLIYRDYERPSGVP

[0425] DRFSGSKSGTSASLAISGLRSEDEADYYCMAWDDSLSGVVFGGGTKLTVLG CDR Area

[0426] CDRH1: SYGMH [ SEQ ID NO:183]

[0427] CDRH2: VISYDGSNKYYADSVKG [ SEQ ID NO:184]

[0428] CDRH3: EFGYIILDY [ SEQ ID NO:185]

[0429] CDRL1: SGSSSNIGSNTVN [ SEQ ID NO:186]

[0430] CDRL2: RDYERPS [ SEQ ID NO:187]

[0431] CDRL3: MAWDDSLSGVV [ SEQ ID NO:188]

[0432] Antibody clone: 4B02

[0433] 4B02-VH [SEQ ID NO:26]

[0434] EVQLLESGGGLVQPGGSLRLSCAASGFTFSNHGMHWVRQAPGKGLEWVAVISYDGTN

[0435] KYYADSVRGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARETWDAFDVWGQGTLVT

[0436] VSS

[0437] 4B02-VL[ SEQ ID NO:50]

[0438] QSVLTQPPSASGTPGQRVTISCSGSSSNIGSNNANWYQQLPGTAPKLLIYDNNKRPSGVP

[0439] DRFSGSKSGTSASLAISGLRSEDEADYYCQAWDSSTVVFGGGTKLTVLG CDR Area

[0440] CDRH1: NHGMH [ SEQ ID NO:189]

[0441] CDRH2:VISYDGTNKYYADSVRG [ SEQ ID NO: 190]

[0442] CDRH3:ETWDAFDV [ SEQ ID NO: 191]

[0443] CDRL1: SGSSSNIGSNNAN [ SEQ ID NO: 192]

[0444] CDRL2:DNNKRPS [ SEQ ID NO: 193]

[0445] CDRL3:QAWDSSTVV [ SEQ ID NO: 194]

[0446] In some embodiments (which are sometimes preferred embodiments), the antibody molecule that specifically binds to FcγRIIB comprises the following CDR regions: SEQ ID NO: 171 (CDRH1), SEQ ID NO: 172 (CDRH2), SEQ ID NO: 173 (CDRH3), SEQ ID NO: 174 (CDRL1), SEQ ID NO: 175 (CDRL2) and SEQ ID NO: 176 (CDRL3), i.e., the CDR regions of clone 6G11.

[0447] In some embodiments (which are sometimes preferred embodiments), the antibody molecule that specifically binds to FcγRIIB comprises the following constant regions: SEQ ID NO: 1 (CH) and SEQ ID NO: 2 (CL), and the following variable regions: SEQ ID NO: 23 (VL) and SEQ ID NO: 47 (VH), i.e., the constant and variable regions of clone 6G11, which have been further modified to have reduced binding to Fcγ receptors via its Fc region. In some embodiments (which are sometimes preferred embodiments), the antibody molecule that specifically binds to FcγRIIB comprises the following constant regions: SEQ ID NO: 195 (CH) and SEQ ID NO: 2 (CL), and the following variable regions: SEQ ID NO: 23 (VL) and SEQ ID NO: 47 (VH), i.e., the constant and variable regions of clone 6G11 comprising the N297Q mutation.

[0448] In some embodiments, the antibody molecule that specifically binds to the receptor present on the tumor cell is a human antibody molecule or an antibody molecule of human origin. In some such embodiments, the human antibody molecule or the antibody molecule of human origin is an IgG antibody. In some such embodiments, the human antibody molecule or the antibody molecule of human origin is an IgG1 or IgG2 antibody.

[0449] In some embodiments, the antibody molecule that specifically binds to a receptor present on a tumor cell is a humanized antibody molecule.

[0450] In some embodiments, the antibody molecule that specifically binds to a receptor present on a tumor cell is a chimeric antibody.

[0451] As described above, an antibody molecule that specifically binds to a receptor present on tumor cells must have the ability to bind to FcγR.

[0452] The combination of an antibody molecule that specifically binds to FcγRIIB and an antibody molecule that specifically binds to a receptor present on tumor cells can be used to treat cancer.

[0453] As used herein, the term "patient" refers to an animal, including a human, that has been diagnosed with an FcγRIIB-negative cancer, or has been diagnosed with a cancer that is thought to be a FcγRIIB-negative cancer and / or exhibits symptoms of such a cancer.

[0454] We include the following: the patient can be a mammal or a non-mammal. Preferably, the patient is a human or a mammal, such as a horse, or a cow, or a sheep, or a pig, or a camel, or a dog, or a cat. Most preferably, the mammalian patient is a human.

[0455] When referring to "manifesting", we include that the subject exhibits symptoms of cancer and / or cancer diagnostic markers, and / or symptoms of cancer and / or cancer diagnostic markers can be measured, and / or assessed, and / or quantified.

[0456] It will be apparent to those skilled in the medical arts what the symptoms of cancer and the diagnostic markers for cancer would be, and how to measure and / or assess and / or quantify whether there is a decrease or increase in the severity of the symptoms of cancer or whether there is a decrease or increase in the diagnostic markers for cancer; and how those symptoms of cancer and / or the diagnostic markers for cancer can be used to form a prognosis for cancer.

[0457] Cancer treatment is usually administered in the form of a course of treatment, that is, the therapeutic agent is administered over a period of time. The length of the course of treatment will depend on many factors, which may include, among other reasons, the type of therapeutic agent administered, the type of cancer being treated, the severity of the cancer being treated, and the age and health of the patient.

[0458] When referring to "while on treatment", we include that the patient is currently receiving a course of treatment, and / or is receiving a therapeutic agent, and / or is receiving a course of a therapeutic agent.

[0459] In some embodiments, the FcγRIIB-negative cancer to be treated according to the present invention is a solid cancer.

[0460] The clinical definitions of diagnosis, prognosis, and progression of a large number of cancers rely on certain classifications called staging. These staging systems are used to collate many different cancer diagnostic markers and cancer symptoms to provide an overview of the diagnosis and / or prognosis and / or progression of cancer. Those skilled in the art of oncology will know how to use the staging system to assess the diagnosis, and / or prognosis, and / or progression of cancer, and which cancer diagnostic markers and cancer symptoms should be used for assessment.

[0461] “Cancer stage” includes Rai staging, which includes stage 0, stage I, stage II, stage III, and stage IV, and / or Binet staging, which includes stage A, stage B, and stage C, and / or Ann Arbour staging, which includes stage I, stage II, stage III, and stage IV.

[0462] It is known that cancer can cause abnormalities in cell morphology. These abnormalities often occur repeatedly in certain cancers, which means that examining these morphological changes (also known as histological examination) can be used for diagnosis or prognosis of cancer. Techniques for visualizing samples to examine cell morphology and techniques for preparing samples for visualization are well known in the art; for example, optical microscopy or confocal microscopy.

[0463] "Histological examination" includes the presence of small mature lymphocytes, and / or the presence of small mature lymphocytes with narrow cytoplasmic margins, the presence of small mature lymphocytes with pyknotic nuclei lacking discernible nucleoli, and / or the presence of small mature lymphocytes with narrow cytoplasmic margins and pyknotic nuclei lacking discernible nucleoli, and / or the presence of atypical cells, and / or lytic cells, and / or prolymphocytes.

[0464] As is well known, cancer is the result of mutations in cellular DNA, which can cause cells to avoid cell death or proliferate uncontrollably. Therefore, checking these mutations (also referred to as cytogenetic examinations) can be a useful tool for assessing the diagnosis and / or prognosis of cancer. An example is the deletion of chromosome position 13q14.1, which is a characteristic of chronic lymphocytic leukemia. Techniques for checking mutations in cells are well known in the art; for example, fluorescence in situ hybridization (FISH).

[0465] "Cytogenetic examination" includes examination of DNA in cells, especially examination of chromosomes. Cytogenetic examination can be used to identify DNA changes that may be associated with the presence of refractory cancer and / or recurrent cancer. This category may include: deletion of the long arm of chromosome 13, and / or deletion of chromosome position 13q14.1, and / or trisomy of chromosome 12, and / or deletion of the long arm of chromosome 12, and / or deletion of the long arm of chromosome 11, and / or deletion of 11q, and / or deletion of the long arm of chromosome 6, and / or deletion of 6q, and / or deletion of the short arm of chromosome 17, and / or deletion of 17p, and / or t(11:14 ) translocation, and / or (q13:q32) translocation, and / or antigen gene receptor rearrangement, and / or BCL2 rearrangement, and / or BCL6 rearrangement, and / or t(14:18) translocation, and / or t(11:14) translocation, and / or (q13:q32) translocation, and / or (3:v) translocation, and / or (8:14) translocation, and / or (8:v) translocation, and / or t(11:14) and (q13:q32) translocations.

[0466] It is known that patients with cancer show certain physical symptoms, which are usually the result of the burden of cancer on the body. Those symptoms often recur in the same cancer, and therefore can be a feature of the diagnosis, and / or prognosis, and / or progression of the disease. Those skilled in the art of medicine will understand which physical symptoms are relevant to which cancers, and how to assess which body systems can be relevant to the diagnosis, and / or prognosis, and / or progression of the disease." Physical symptoms" include hepatomegaly and / or splenomegaly.

[0467] In some embodiments, the target to which the antibody molecule that specifically binds to a receptor present on tumor cells binds is human epidermal growth factor receptor 2 (HER2). In such embodiments, the FcγRIIB-negative cancer to be treated may be a cancer selected from the group consisting of breast cancer and gastric cancer.

[0468] In this context, breast cancer includes metastatic breast cancer (MBC) and early breast cancer (EBC).

[0469] In this context, gastric cancer may also mean gastric adenocarcinoma or cancer of the stomach, and includes gastroesophageal junction (GEJ) adenocarcinoma. It further includes metastatic gastric cancer (MGC) and metastatic GEJ adenocarcinoma.

[0470] Trastuzumab Currently used alone or in combination with chemotherapy or other drugs to treat breast cancer expressing HER2, and such treatment has significantly improved overall survival. However, many patients are still not cured. Other patients develop trastuzumab resistance, causing recurrence of the disease, and in addition it has been shown that some HER2-positive breast cancers may become HER2-negative or low-expressing HER2 over time. Therefore, in order to cure more patients, it is urgently desired to improve anti-HER2 therapy and the way to overcome resistance.

[0471] In certain embodiments, the FcγRIIB negative cancer to be treated according to the present invention is a cancer with low HER2 expression. Patients suffering from cancer with low HER2 expression generally do not respond or have a good response to standard care treatment (such as treatment with trastuzumab and / or trastuzumab biosimilars). However, by combining with an antibody molecule that specifically binds to FcγRIIB via its Fab region and lacks an Fc region or has a reduced combination with Fcγ receptors via its Fc region, as described herein, the treatment of cancer with low HER2 expression becomes possible.

[0472] To determine if a cancer has low HER2 expression, a standard HER2 assay, such as one using immunohistochemistry (IHC) or fluorescent in situ hybridization (FISH), typically performed on a biopsy taken from the patient can be used.

[0473] IHC test is based on the staining of HER2 protein.When used to determine the amount of HER2 on the surface of the cell in the breast cancer tissue sample, it gives a score of 0 to 3+.If the score is 0 to 1+, it is considered to be HER2 negative.If the score is 2+, it is considered to be critical.The score of 3+ is considered to be HER2 positive.In this context, the IHC score of 0 to 1+ for the patient with breast cancer can classify the patient as suffering from cancer with low HER2 expression.In certain embodiments, the score of 0 to 1+ is considered to represent HER2 low expression cancer.In certain embodiments, the score of 3+ is considered to represent HER2 high expression cancer, that is, not HER2 low expression cancer according to the present invention.In the following examples, a perfect HER2 expression immunoreactive Balb / C TUBO breast cancer tumor model was used.The model is adapted to allow assessment of Fc:FcγR impaired (Fc silence) anti-FcγRIIB antibody enhancement for cancer with low HER2 expression (HER2 low) and (for comparison) also for the ability of anti-HER2 anti-tumor activity of cancer with high HER2 expression (HER2 high). Therefore, in the HER2 high expression tumor model, the animal received a full therapeutic dose of anti-HER2 antibody, which caused a strong occupation of the HER2 expressed by the tumor. In the HER2 low expression model, the animal received a lower dose of antibody, thereby causing about 10 times lower HER2 receptors on the cancer cells targeted by the anti-HER2 antibody, as demonstrated by flow cytometry analysis of tumors harvested from mice treated with anti-HER2 antibodies conjugated with fluorescent dyes. In this way, all other factors except that the antibody targets the HER2 receptor are the same, making this tumor model system ideal for the following: assessing and demonstrating the anti-FcγRIIB-mediated enhancement of the anti-HER2 efficacy for HER2 high expression cancers, and assessing and demonstrating the anti-FcγRIIB-mediated realization of the significant effect of anti-HER2 for HER2 low expression cancers in treatment.

[0474] FISH, based on the HER2 marker, is more accurate than IHC, but it costs more and takes longer to return results. That's why an IHC test is often the first test done to see if a cancer is HER2-positive. With a FISH test, you get a score of either positive or negative (some hospitals refer to a negative test result as a "zero").

[0475] The two tests can be combined, for example, if the IHC test results are borderline, it can be combined with the FISH test to provide a better basis for determining whether the cancer is HER2 positive.

[0476] Generally, only cancers that test IHC 3+ or FISH positive respond to standard of care treatment with drugs that target HER2.

[0477] When the target to which the second antibody molecule binds is HER2, the second antibody molecule may be trastuzumab or trastuzumab biosimilars, such as trastuzumab-ANNS Trastuzumab-qyyp Trastuzumab-PKRB Trastuzumab-dttb or trastuzumab-dkst When referring to a trastuzumab biosimilar, we here mean an antibody molecule that is highly similar to trastuzumab and has no clinically meaningful differences. Alternatively, the second antibody molecule can be an enhanced variant of trastuzumab or a trastuzumab biosimilar conjugated to a toxin, such as fam-trastuzumab-delutec-nxki or T-DM1, or ado-trastuzumab emtansine or other FcγR-engaging anti-HER2 antibody drug conjugates.

[0478] In other cases, the second antibody can be used together with a third antibody, which can be a tumor direct targeting (e.g., anti-HER2 antibody Pertuzumab) or an immunomodulator (e.g., anti-PD-1 / PD-L1 antibody). In addition, the second antibody can be an anti-HER2 antibody used in any anti-HER2-containing treatment regimen.

[0479] In some embodiments, the FcγRIIB negative cancer to be treated according to the present invention is a cancer in a patient who has previously been successfully treated with trastuzumab and / or a trastuzumab biosimilar, but has subsequently developed resistance to trastuzumab or a trastuzumab biosimilar and therefore no longer responds to such treatment. Cancers with low HER2 expression. As described herein, the combination with an antibody molecule that specifically binds to FcγRIIB via its Fab region and lacks an Fc region or has reduced binding to Fcγ receptors via its Fc region makes it possible to overcome such resistance.

[0480] In some embodiments, the target to which the antibody molecule that specifically binds to a receptor present on tumor cells binds is human epidermal growth factor receptor (EGFR). In such embodiments, the FcγRIIB-negative cancer to be treated may be a cancer selected from the group consisting of head and neck cancer and colorectal cancer.

[0481] In this context, head and neck cancer includes locally or regionally advanced squamous cell carcinoma of the head and neck, recurrent locoregional disease or metastatic squamous cell carcinoma of the head and neck, and recurrent or metastatic squamous cell carcinoma of the head and neck.

[0482] In this context, colorectal cancer includes metastatic colorectal cancer that is K-Ras wild-type and expresses EGFR.

[0483] When the target to which the second antibody molecule binds is EGFR, the second antibody molecule may be cetuximab Or cetuximab biosimilars. When referring to cetuximab biosimilars, we mean here an antibody molecule that is highly similar to cetuximab and has no clinically meaningful differences.

[0484] Each of the above cancers is known, and symptoms and cancer diagnostic markers are well described, as are therapeutic agents for treating those cancers. Thus, symptoms, cancer diagnostic markers, and therapeutic agents for treating the above cancer types will be known to those skilled in the medical arts.

[0485] In some embodiments, an antibody molecule that specifically binds to FcγRIIB and an antibody molecule that specifically binds to a receptor present on tumor cells are administered to the patient simultaneously, meaning that they are either administered together at once or separately in very close time to each other.

[0486] In some embodiments, before administering an antibody molecule that specifically binds to a receptor present on a tumor cell, an antibody molecule that specifically binds to FcγRIIB is administered to the patient. Such sequential administration can be achieved by the temporal separation of the two antibodies. Alternatively or in combination with the first option, sequential administration can also be achieved by the spatial separation of the two antibody molecules achieved below: administering an antibody molecule that specifically binds to FcγRIIB in a certain manner (such as intratumorally) so that the antibody molecule that specifically binds to FcγRIIB reaches cancer before the antibody molecule that specifically binds to the receptor present on the tumor cell, and then administering the antibody molecule that specifically binds to the receptor present on the tumor cell in a certain manner (such as systemically) so that the antibody molecule that specifically binds to the receptor present on the tumor cell reaches cancer after the antibody molecule that specifically binds to FcγRIIB.

[0487] In some embodiments, an antibody molecule that specifically binds to a receptor present on tumor cells is administered to the patient prior to administration of an antibody molecule that specifically binds to FcγRIIB.Such sequential administration can be achieved as described above.

[0488] Those skilled in the medical arts will appreciate that drugs can be modified with various additives, for example to alter the rate at which the drug is absorbed by the body; and can be modified in different forms, for example to allow for a particular route of administration to the body.

[0489] We therefore include that the compositions, and / or antibodies, and / or medicaments of the present invention may be combined with excipients, and / or pharmaceutically acceptable carriers, and / or pharmaceutically acceptable diluents and / or adjuvants.

[0490] We also include the following: the compositions, and / or antibodies, and / or medicaments of the invention may be suitable for parenteral administration, including aqueous and / or non-aqueous sterile injection solutions that may contain antioxidants, and / or buffers, and / or bacteriostats, and / or solutes that render the formulation isotonic with the blood of the intended recipient; and / or aqueous and / or non-aqueous sterile suspensions, which may include suspending agents and / or thickening agents. The compositions, and / or antibodies, and / or medicaments, and / or medicaments of the invention may be present in unit-dose or multi-dose containers (e.g., sealed ampoules and vials), and may be stored in a freeze-dried (i.e., lyophilized) condition requiring only the addition of a sterile liquid carrier (e.g., water for injection) immediately prior to use.

[0491] Extemporaneous injection solutions and suspensions may be prepared from sterile powders, and / or granules, and / or tablets of the kind previously described.

[0492] For parenteral administration to human patients, the daily dosage level of antibody molecules that specifically bind to FcγRIIB and / or antibody molecules that specifically bind to receptors present on tumor cells will generally be 1 mg / kg patient body weight to 20 mg / kg, or in some cases even up to 100 mg / kg, administered in single or divided doses. Lower doses may be used in special cases, for example, in combination with long-acting administration. In any case, the doctor will determine the actual dose that will be most suitable for any individual patient, and the actual dose will vary with the age, weight and response of the particular patient. The above dosages are exemplary general situations. Of course, there may be separate situations where a higher or lower dosage range is needed, and this is within the scope of the present invention.

[0493] Generally speaking, in humans, oral or parenteral administration of the composition of the present invention, and / or antibodies, and / or medicaments, and / or drugs is a preferred route, and parenteral administration is the most commonly used for antibodies. For veterinary use, the composition of the present invention, and / or antibodies, and / or medicaments, and / or drugs are administered as suitably acceptable formulations according to normal veterinary practice, and the veterinarian will determine the most appropriate dosage regimen and route of administration for a particular animal. Therefore, the present invention provides a pharmaceutical formulation comprising antibodies and / or medicaments of the present invention in an amount that effectively treats various disorders (as described above and further described below). Preferably, the composition, and / or antibodies, and / or medicaments, and / or drugs are suitable for delivery by a route selected from the group comprising the following items: intravenous (IV); subcutaneous (SC), intramuscular (IM) or intratumoral.

[0494] In some embodiments, the first antibody molecule, or the second antibody, or both can be administered using a plasmid or virus.Then, such plasmids include nucleotide sequences encoding the first antibody molecule, or the second antibody, or both.In some embodiments, the nucleotide sequence encoding the first antibody molecule, or the second antibody, or part or complete sequence of both is integrated into a cell or viral genome or a virus group (viriome) in a virus; Then, such cells or viruses act as delivery vehicles for the first antibody molecule, or the second antibody, or both (or for the delivery vehicles for the nucleotide sequences encoding the first antibody molecule, or the second antibody, or both).For example, in some embodiments, such viruses may be in the form of therapeutic oncolytic viruses, which include nucleotide sequences encoding at least one of the antibody molecules described herein.In some embodiments, such oncolytic viruses include nucleotide sequences encoding full-length human IgG antibodies.Oncolytic viruses are known to those skilled in the art in the field of medicine and virology.

[0495] The present invention also includes compositions, and / or antibodies, and / or medicaments, and / or drugs comprising the pharmaceutically acceptable acid or base addition salts of the polypeptide binding moiety of the present invention. The acid used to prepare the pharmaceutically acceptable acid addition salts of the aforementioned base compounds available in the present invention is to form non-toxic acid addition salts (i.e., salts containing pharmacologically acceptable anions, such as hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, acetate, lactate, citrate, acid citrate, tartrate, bitartrate, succinate, maleate, fumarate, gluconate, sucrose, benzoate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate and pamoate [i.e., 1,1'-methylene-bis-(2-hydroxy-3 naphthoate)] salts, etc.) Those acids. Pharmaceutically acceptable base addition salts can also be used to produce pharmaceutically acceptable salt forms of medicaments according to the present invention. Chemical bases that can be used as reagents to prepare pharmaceutically acceptable alkaline salts of the medicaments of the present invention that are acidic in nature are those chemical bases that form non-toxic alkaline salts with such compounds. Such non-toxic alkaline salts include, but are not limited to, those non-toxic alkaline salts derived from such pharmacologically acceptable cations, such as alkali metal cations (e.g., potassium and sodium) and alkaline earth metal cations (e.g., calcium and magnesium), ammonium or water-soluble amine addition salts (such as N-methylglucamine-(glucamine), and lower alkanolammonium and other alkaline salts of pharmaceutically acceptable organic amines, etc. The medicaments and / or polypeptide binding portions of the present invention can be lyophilized for storage and reconstituted in a suitable carrier before use. Any suitable lyophilization method (e.g., spray drying, filter cake drying) and / or reconstitution technology can be used. It will be understood by those skilled in the art that lyophilization and reconstitution can cause varying degrees of antibody activity loss (e.g., for conventional immunoglobulins, IgM antibodies tend to have greater activity loss than IgG antibodies), and the use level may have to be adjusted upward to compensate. In one embodiment, the lyophilized (freeze-dried) polypeptide binding portion loses no more than about 20%, or no more than about 25%, or no more than about 30%, or no more than about 35%, or no more than about 40%, or no more than about 45%, or no more than about 50% of its activity (before lyophilization) upon rehydration. BRIEF DESCRIPTION OF THE DRAWINGS

[0496] In the following examples, reference is made to the following figures:

[0497] Figure 1 . Figure 1 A to B show the survival curves. 6) were injected subcutaneously into female BalbC mice (n=12). Tumor growth was monitored (measured by calipers), and when tumors reached approximately 7x7 mm, mice were randomized and treated twice a week as indicated. Tumor growth was tracked twice a week until tumors reached a predetermined size (ethical endpoint), at which time mice were euthanized. Figure 1 A. HER2 high cancer model. The therapeutic effect of anti-HER2 (10 mg / kg) in combination with Fc null anti-FcγRIIB-NA (AT130-2NA) was compared to an isotype control antibody (FITC IgG2a) and to anti-HER2 (10 mg / kg) monotherapy. Mice were dosed three times with 2 to 3 days between doses. In this HER2 high cancer model, combined treatment with anti-FcγRIIB-NA (AT130-2NA) delayed tumor growth and increased the number of complete responders compared to anti-HER2 monotherapy. The study was repeated 3 times with comparable results, and results from 1 representative experiment are shown in Figure 1 A. Figure 1 B HER2 low cancer model. This figure shows the therapeutic effect of anti-HER2 (1 mg / kg) in combination with Fc null anti-FcγRIIB-NA (AT130-2NA) compared to an isotype control antibody (FITC IgG2a) and to 1 mg / kg of anti-HER2 as a single treatment. Mice were dosed three times with 2 to 3 days between doses. In this HER2 low cancer model, combination treatment with anti-FcγRIIB-NA (AT130-2NA) delayed tumor growth and increased the number of complete responders compared to anti-HER2 treatment alone, making anti-HER2 treatment of HER2 low cancer as effective as (single agent anti-HER2) treatment of HER2 high cancer. Figure 1 In C, targeted HER2 receptors in HER2 high and HER2 low experimental cancer models. Tumors were established as described in A to B, and mice (n=3) were treated with 1 mg / kg or 10 mg / kg of anti-HER2 labeled with a fluorescent dye (AF647). The mice were dosed twice (with an interval of 2 to 3 days between doses), and then the mice were euthanized and the tumors were collected. Tumors were enzymatically digested, and anti-HER2 labeled with a fluorescent dye (AF647) was quantified by FACS. Tumors from mice in the HER2 high model (injected with 10 mg / kg anti-HER2) showed a 10-fold increase in targeted HER2 receptors compared to tumors from mice in the HER2 low model (injected with 1 mg / kg anti-HER2).

[0498] Figure 2 Survival curve. TUBO cells (1x10 6 ) was injected subcutaneously into female BalbC mice (n=12). Tumor growth was monitored (measured by caliper), and when the tumor reached about 7x7 mm, the mice were randomized and treated with therapeutic mAbs twice a week. Tumor growth was tracked twice a week until the tumor reached a predetermined size (ethical endpoint), at which time the mice were euthanized. The therapeutic effect of anti-HER2 (1 mg / kg) combined with Fc null anti-FcγRIIB-NA (AT130-2NA) was compared with anti-HER2 combined with wild-type anti-FcγRIIB (AT130-2 wt), with isotype control antibody (FITC IgG2a) and with 1 mg / kg of anti-HER2 as a single treatment. Mice were administered three times (with 2 to 3 days between administrations). Compared with anti-HER2 treatment alone, anti-HER2 combined with anti-FcγRIIB-NA (AT130-2NA) showed delayed tumor growth. No delay in tumor growth was observed when anti-HER2 was combined with wild-type anti-FcγRIIB (AT130-2 wt).

[0499] Figure 3 .TUBO cells (1x10 6 ) was injected subcutaneously into female BalbC mice. Tumor growth was monitored (measured by caliper), and when the tumor reached about 7x7 mm, the mice were randomized and treated with therapeutic mAbs twice a week. On the 7th to 8th day after the start of treatment, 24 hours after 3 injections, the mice were killed and the tumors were harvested. The immune cell content of tumor single cell suspensions was analyzed by FACS. Fc null anti-FcγRIIB-NA is named AT-130-2NA in the accompanying drawings. In the group treated with a combination of anti-HER2 and anti-FcγRIIB-NA, bone marrow cells, especially CD11b+F4 / 80+ / MHCII 低 The number has increased significantly.

[0500] Figure 4 . The lung area covered by metastatic tumors. B16 cells (5x10 5) were injected intravenously into female C57 mice. Four days after tumor cell injection, mice were injected with antibodies (10 mg / kg intraperitoneally - isotype control, TA99, AT130-2-NA, and a combination of TA99 and AT130-2-NA). Five treatments were given at intervals of 2 to 3 days. On the 21st day after the start of treatment, mice were killed and the metastasis content in the lungs was quantified. A reduction in lung metastases was observed for TA99 alone, however, the effect of TA99 was greatly increased when combined with anti-FcγRIIB-NA (AT130-2NA). Anti-FcγRIIB-NA had no therapeutic effect as a monotherapy.

[0501] Examples

[0502] Specific non-limiting examples embodying certain aspects of the present invention will now be described. In order to allow examination of the blocking effect of FcγRIIB in complex in vivo systems, two groups of alternative antibodies have been used. The mouse equivalent of the Fc-active antibody 6G11 is called AT130-2. In order to silence the human antibody Fc (so that the binding to FcγR is severely impaired or negligible), we have replaced amino acid position 297 from N to Q. In order to silence the mouse antibody Fc, the same position is replaced from and N to Q. Therefore, in the mouse system, we will refer to AT-130, and this patent application relates to the human counterpart 6G11. In short, human 6G11 corresponds to the mouse substitute AT1302-2 (both are Fc: FcγR proficient, also expressed as Fc activity herein), and 6G11-N297Q corresponds to AT130-3-N297A (both are Fc: FcγR impaired, also expressed as Fc silencing herein). We have previously shown that these human and mouse Fc:FcγR proficient or Fc:FcγR impaired anti-FcγRIIB antibodies are functionally and biochemically equivalent (WO 2019 / 138005 and WO 2021 / 009358).

[0503] A different way for silencing an antibody Fc (and well known to those skilled in the art) would be to remove the Fc part and form a Fab or F(ab')2 fragment.

[0504] The anti-HER2 mAb used below was clone 7.16.4 (mIgG2a) obtained from BioXcell.

[0505] The alternative anti-mouse FcγRIIB mAb AT130-3-N297A improves the in vivo antitumor efficacy of anti-HER2 mAbs. and enables treatment of HER2-low expressing cancers

[0506] Treatment efficacy in the TUBO tumor model (HER2-overexpressing cancer model)

[0507] To assess the in vivo anti-tumor effect of anti-mouse FcγRIIB mAb AT130-3-N297A in combination with anti-HER2 mAb, this combination was studied in vivo in the TUBO tumor model as follows.

[0508] Mice were bred and maintained at the facility in Lund, Sweden, according to applicable rules and guidelines, including those of the facility and the Swedish Board of Agriculture. Six to eight week old female BalbC mice were provided by Taconic (Bomholt, Denmark) and maintained at the local animal facility. TUBO cells (University of Turin) were grown in glutamax-buffered RPMI supplemented with 10% FCS. When the cells were semi-confluent, they were detached with trypsin and 10×10 6 Cells / ml were resuspended in sterile PBS. Glutamax-buffered RPMI (RPMI medium), FCS (fetal calf serum), and PBS (phosphate-buffered saline) were all from Invitrogen and used in the following. 6 Mice were subcutaneously injected with 100 μl cell suspension corresponding to 10 cells / mouse. 12 to 13 days after injection, mice were treated with 10 mg / kg antibody intraperitoneal injection (isotype control, anti-HER2, AT130-3-N297A, and a combination of anti-HER2 and AT130-3-N297A) twice a week, as shown. Tumors were measured twice a week until they reached a diameter of 15 mm, after which mice were killed.

[0509] In this HER2-high experimental cancer model, the anti-mouse FcγRIIB mAb AT130-3-N297A significantly improved anti-HER2-mediated survival compared with single-agent anti-HER2 therapy ( Figure 1 A).

[0510] Anti-FcγRIIB combination treatment achieves anti-HER2 therapeutic effect against HER2-low expressing cancers

[0511] To evaluate the in vivo effect of anti-mouse FcγRIIB mAb AT130-3-N297A in combination with anti-HER2 mAb against HER2 low cancers, the same HER2 high TUBO mouse tumor model described above was used, but with a lower dose of antibody, thereby causing fewer HER2 receptors on the cancer cells to be occupied and targeted by the anti-HER2 antibody. Thus, all other factors except that the antibody targets the HER2 receptor are the same, making this tumor model ideal for evaluating and demonstrating that anti-FcγRIIB achieves anti-HER2 effects against HER2 low expressing cancers.

[0512] Mice were bred and maintained as described above. Six to eight week old female BalbC mice were provided by Taconic (Bomholt, Denmark) and maintained in the local animal facility. TUBO cells (University of Turin) were grown in glutamax-buffered RPMI supplemented with 10% FCS. When cells were semi-confluent, they were detached with trypsin and plated at 10×10 6 Resuspend in sterile PBS. Use 1x10 6 Mice were injected subcutaneously with 100 μl of cell suspension corresponding to 10 cells / mouse. Twelve to 13 days after injection, mice were treated with 1 mg / kg antibody intraperitoneal injection (isotype control, anti-HER2, and a combination of anti-HER2 and AT130-3-N297A) twice a week, as shown. Tumors were measured twice a week until they reached a diameter of 15 mm, after which mice were sacrificed.

[0513] In this experimental HER2 low model, anti-HER2 single agent treatment showed severely impaired therapeutic efficacy, with only 1 / 10 treated animals cured compared to 3 / 10 mice cured in the HER2 high cancer model. After combination with the anti-mouse FcγRIIB mAb AT130-3-N297A, which has no anti-tumor activity on its own, complete therapeutic efficacy was observed in the HER2 low tumor model (3 / 10 mice cured) similar to that observed in the HER2 high model ( Figure 1 B). Next, an experiment was designed to compare the amount of HER2 targeted in the HER2 high model and the HER2 low model. Tumors were established as described in A to B, and mice (n=3) were treated with 1 mg / kg or 10 mg / kg of anti-HER2 labeled with Alexa Flour (AF) 647, so that the antibody targeting HER2 in the HER2 high cancer model and the HER2 low cancer model could be determined. The mice were dosed twice (with an interval of 2 to 3 days between doses). Two days after the second injection, the mice were euthanized and the tumors were collected. The tumors were cut into small pieces and enzymatically digested with a mixture of DNase and Liberase at 37°C. The tumor solution was further filtered through a cell strainer to obtain a single cell solution. Anti-HER2 labeled with a fluorescent dye (AF647) in the tumor was quantified by FACS. Tumor cells from a HER2-high cancer model (mice injected with 10 mg / kg anti-HER2) showed a 10-fold increase in the targeted HER2 receptor compared to tumor cells from a HER2-low cancer model (mice injected with 1 mg / kg anti-HER2).

[0514] No therapeutic effect when Fc-active AT130-2 is combined with anti-HER2 mAb

[0515] To assess whether Fc-active AT130-2 also improves the in vivo anti-tumor effect of anti-HER2 mAbs, the combination was studied in vivo in tumor models as described below.

[0516] Mice were bred and maintained as described above. Six to eight week old female BalbC mice were provided by Taconic (Bomholt, Denmark) and maintained in the local animal facility. TUBO cells (University of Turin) were grown in glutamax-buffered RPMI supplemented with 10% FCS. When cells were semi-confluent, they were detached with trypsin and plated at 10×10 6 Resuspend in sterile PBS. Use 1x10 6 Mice were injected subcutaneously with 100 μl of cell suspension corresponding to 10 cells / mouse. Twelve to 13 days after injection, mice were treated with 10 mg / kg antibody intraperitoneal injection (isotype control, anti-HER2, or a combination of anti-HER2 and AT130-2-N297A or AT130-2 wt) twice a week, as shown. Tumors were measured twice a week until they reached a diameter of 15 mm, after which mice were sacrificed.

[0517] Fc:FcγR-proficient (wt) AT130-2 therefore did not show improved therapeutic anti-tumor effects when combined with anti-HER2 ( Figure 2 ).

[0518] The improved therapeutic effect when anti-HER2 therapy was combined with the anti-mouse FcγRIIB mAb AT130-3-N297A was associated with an increased influx of myeloid cells into the tumor.

[0519] To assess the mode of action of the therapeutic effect of anti-HER2 therapy when combined with the anti-mouse FcγRIIB mAb AT130-2-N297A, the treated tumors were immunoprofiling as described below.

[0520] Mice were bred and maintained as described above. Six to eight week old female BalbC mice were provided by Taconic (Bomholt, Denmark) and maintained in the local animal facility. TUBO cells (University of Turin) were grown in glutamax-buffered RPMI supplemented with 10% FCS. When cells were semi-confluent, they were detached with trypsin and plated at 10×10 6 Resuspend in sterile PBS. Use 1x10 6Mice were injected subcutaneously with 100 μl of cell suspension corresponding to 10 cells / mouse. Once the tumor reached a size of approximately 7x7 mm, the mice were injected with antibodies (10 mg / kg intraperitoneally—isotype control, anti-HER2, AT130-2-N297A, and a combination of anti-HER2 and AT130-2-N297A). Tumors were harvested 24 hours after 3 injections on days 7 to 8 after the start of treatment (when tumors in the combination group significantly regressed).

[0521] The tumor was cut into small pieces and enzymatically digested with a mixture of DNA enzyme and Liberase at 37°C. The tumor solution was filtered through a cell strainer to obtain a single cell solution. The cell solution was blocked with IVIg (human normal immunoglobulin for intravascular administration, Kiovig, Takeda Pharmaceutical (Takeda)) before staining. Immune cells were identified and quantified by FACS using the following markers: CD45, CD3, CD4, CD8, CD25, CD11b, Ly6C, Ly6G, MHCII, F4 / 80, CD49b and NK 1.1 (all from BD Biosciences).

[0522] like Figure 3 As shown, the anti-HER2 / anti-FcγRIIB-NA combination altered the immune cell composition in the tumor. Compared with single treatment, the combination of anti-HER2 and anti-FcγRIIB-NA caused an increase in the CD11b+ / F4 / 80+ population, which is consistent with increased recruitment of effector cells and increased antibody-mediated depletion of HER2-targeted tumor cells. This increase was most significant in the HER2 high model (mice were dosed with 10 mg / kg anti-HER2) ( Figure 3 ).

[0523] B16 lung metastasis model

[0524] To assess whether anti-FcγRIIB-NA could enhance the depletion activity and therapeutic efficacy of other tumor-directed therapeutic antibodies targeting solid tumors, we studied the therapeutic effect of combining anti-FcγRIIB-NA with TA99, an antibody specific for the gp75 melanoma tumor antigen, in the B16 metastatic melanoma model.

[0525] Mice were housed and maintained as described above. Six to eight week old female C57 mice were provided by Taconic (Bomholt, Denmark) and maintained in a local animal facility. B16 cells (ATCC) were grown in glutamax-buffered RPMI supplemented with 10% FCS. When cells were semi-confluent, they were detached with trypsin and plated at 2.5×106 Resuspend in sterile PBS. Use 5x10 5 Mice were injected intravenously with 200 μl of cell suspension corresponding to 10 cells / mouse. Four days after tumor cell injection, mice were injected with antibodies (10 mg / kg intraperitoneally—isotype control, TA99, AT130-2-N297A, and a combination of TA99 and AT130-2-N297A). Five treatments were given at intervals of 2 to 3 days. On day 21 after the start of treatment, mice were killed and the metastasis content in the lungs was quantified. Figure 4

[0526] A modest reduction in lung metastases was observed following treatment with TA99 alone compared to untreated animals ( Figure 4 ). After combined treatment with anti-FcγRIIB-NA (which itself had no effect on metastasis formation), the therapeutic effect of tumor-directed targeting antibody TA99 was greatly enhanced, significantly reducing lung metastases compared to TA99 single agent treatment. Therefore, combined treatment with anti-FcγRIIB-NA enhances the therapeutic efficacy of different tumor-directed targeting antibodies specific for different tumor antigens associated with different FcγRIIB solid cancers.

Claims

1. A first antibody molecule that specifically binds to FcγRIIB via its Fab region and lacks an Fc region or has reduced binding to an Fcγ receptor via its Fc region, the first antibody molecule being used in the treatment of FcγRIIB-negative cancer in a patient in combination with a second antibody molecule that specifically binds to a receptor present on tumor cells, the second antibody molecule having an Fc region that binds to at least one activated Fcγ receptor.

2. A pharmaceutical composition comprising: (i) a first antibody molecule that specifically binds to FcγRIIB via its Fab region and lacks an Fc region or has reduced binding to an Fcγ receptor via its Fc region, and (ii) a second antibody molecule, which specifically binds to a receptor present on tumor cells, and which has an Fc region that binds to at least one activated Fcγ receptor; The pharmaceutical composition is for use in the treatment of FcγRIIB-negative cancer in a patient.

3. A kit for use in the treatment of FcγRIIB-negative cancers, comprising: (i) a first antibody molecule that specifically binds to FcγRIIB via its Fab region and lacks an Fc region or has reduced binding to an Fcγ receptor via its Fc region, and (ii) a second antibody molecule that specifically binds to a receptor present on tumor cells, wherein the second antibody molecule has an Fc region that binds to at least one activated Fcγ receptor.

4. The following items: (i) a first antibody molecule that specifically binds to FcγRIIB via its Fab region and lacks an Fc region or has reduced binding to an Fcγ receptor via its Fc region, and (ii) a second antibody molecule, which specifically binds to a receptor present on tumor cells, and which has an Fc region that binds to at least one activated Fcγ receptor; Use in the manufacture of a medicament for use in the treatment of an FcγRIIB-negative cancer in a patient.

5. A method for treating an FcγRIIB-negative cancer in a patient comprising administering: (i) a first antibody molecule that specifically binds to FcγRIIB via its Fab region and lacks an Fc region or has reduced binding to an Fcγ receptor via its Fc region, and (ii) a second antibody molecule, which specifically binds to a receptor present on tumor cells, and which has an Fc region capable of activating at least one Fcγ receptor.

6. The first antibody molecule for use in combination with a second antibody molecule according to claim 1, the pharmaceutical composition for use according to claim 2, the kit for use according to claim 3, the use according to claim 4 or the method according to claim 5, wherein the FcγRIIB-negative cancer is a solid cancer.

7. A first antibody molecule for use in combination with a second antibody molecule according to claim 1 or 6, a pharmaceutical composition for use according to claim 2 or 6, a kit for use according to claim 3 or 6, a use according to claim 4 or 6, or a method according to claim 5 or 6, wherein the binding of the second antibody molecule to the receptor on the tumor cell causes depletion of the tumor cell.

8. A first antibody molecule for use in combination with a second antibody molecule according to claim 1, 6 or 7, a pharmaceutical composition for use according to claim 2, 6 or 7, a kit for use according to claim 3, 6 or 7, a use according to claim 4, 6 or 7 or a method according to claim 5, 6 or 7, wherein the first antibody lacks an Fc region.

9. A first antibody molecule for use in combination with a second antibody molecule according to any one of claims 1 or 6 to 8, a pharmaceutical composition for use according to any one of claims 21 or 6 to 8, a kit for use according to any one of claims 3 or 6 to 8, a use according to any one of claims 4 or 6 to 8, or a method according to any one of claims 5 or 6 to 8, wherein the second antibody molecule binds to human epidermal growth factor receptor 2 (HER2).

10. The first antibody molecule for use in combination with a second antibody molecule according to claim 9, the pharmaceutical composition for use according to claim 9, the kit for use according to claim 8, the use according to claim 8 or the method according to claim 9, wherein the cancer is selected from the group consisting of breast cancer and gastric cancer.

11. A first antibody molecule for use in combination with a second antibody molecule according to claim 9 or 10, a pharmaceutical composition for use according to claim 9 or 10, a kit for use according to claim 9 or 10, a use according to claim 9 or 10 or a method according to claim 9 or 10, wherein the cancer has low HER2 expression.

12. A first antibody molecule for use in combination with a second antibody molecule according to any one of claims 9 to 11, a pharmaceutical composition for use according to any one of claims 9 to 11, a kit for use according to any one of claims 9 to 11, a use according to any one of claims 9 to 11, or a method according to any one of claims 9 to 11, wherein the cancer is a cancer in a patient who has previously been treated with an antibody molecule that specifically binds to HER2 but has developed resistance to the antibody.

13. A first antibody molecule for use in combination with a second antibody molecule according to any one of claims 9 to 12, a pharmaceutical composition for use according to any one of claims 9 to 12, a kit for use according to any one of claims 9 to 12, a use according to any one of claims 9 to 12, or a method according to any one of claims 9 to 12, wherein the second antibody molecule is trastuzumab or a trastuzumab biosimilar.

14. A first antibody molecule for use in combination with a second antibody molecule according to any one of claims 1 or 6 to 8, a pharmaceutical composition for use according to any one of claims 21 or 6 to 8, a kit for use according to any one of claims 3 or 6 to 8, a use according to any one of claims 4 or 6 to 8, or a method according to any one of claims 5 or 6 to 8, wherein the second antibody molecule binds to human epidermal growth factor receptor (EGFR).

15. A first antibody molecule for use in combination with a second antibody molecule according to claim 14, a pharmaceutical composition for use according to claim 14, a kit for use according to claim 14, a use according to claim 14 or a method according to claim 14, wherein the cancer is selected from the group consisting of head and neck cancer and colorectal cancer.

16. A first antibody molecule for use in combination with a second antibody molecule according to claim 14 or 15, a pharmaceutical composition for use according to claim 14 or 15, a kit for use according to claim 14 or 15, a use according to claim 14 or 15 or a method according to claim 14 or 15, wherein the second antibody molecule is cetuximab or a cetuximab biosimilar.

17. A first antibody molecule for use in combination with a second antibody molecule according to any one of claims 1 or 6 to 16, a pharmaceutical composition for use according to any one of claims 2 or 6 to 16, a kit for use according to any one of claims 3 or 6 to 16, a use according to any one of claims 4 or 6 to 16, or a method according to any one of claims 5 or 6 to 16, wherein the first antibody molecule is selected from the group consisting of a human antibody molecule, a humanized antibody molecule, and an antibody molecule of human origin.

18. A first antibody molecule for use in combination with a second antibody molecule according to any one of claims 1 or 6 to 17, a pharmaceutical composition for use according to any one of claims 2 or 6 to 17, a kit for use according to any one of claims 3 or 6 to 17, a use according to any one of claims 4 or 6 to 17, or a method according to any one of claims 5 or 6 to 17, wherein the first antibody molecule is a monoclonal antibody molecule or an antibody molecule of monoclonal origin.

19. A first antibody molecule for use in combination with a second antibody molecule according to any one of claims 1 or 6 to 18, a pharmaceutical composition for use according to any one of claims 2 or 6 to 18, a kit for use according to any one of claims 3 or 6 to 18, a use according to any one of claims 4 or 6 to 18, or a method according to any one of claims 5 or 6 to 18, wherein the first antibody molecule is selected from the group consisting of a full-length antibody, a chimeric antibody, a single-chain antibody, a Fab fragment, a (Fab')2 fragment, a Fab' fragment, a (Fab')2 fragment, an Fv fragment, and a scFv fragment.

20. A first antibody molecule for use in combination with a second antibody molecule according to any one of claims 1 or 6 to 18, a pharmaceutical composition for use according to any one of claims 2 or 6 to 18, a kit for use according to any one of claims 3 or 6 to 18, a use according to any one of claims 4 or 6 to 18, or a method according to any one of claims 5 or 6 to 18, wherein the first antibody molecule is a human IgG antibody molecule with a deglycosylated Fc region or an IgG antibody molecule of human origin with a deglycosylated Fc region.

21. The first antibody molecule for use in combination with a second antibody molecule according to claim 20, the pharmaceutical composition for use according to claim 20, the kit for use according to claim 20, the use according to claim 20 or the method according to claim 20, wherein the IgG antibody molecule is an IgG1 or IgG2 antibody molecule.

22. A first antibody molecule for use in combination with a second antibody molecule according to claim 21, a pharmaceutical composition for use according to claim 21, a kit for use according to claim 21, a use according to claim 21 or a method according to claim 21, wherein the IgG antibody molecule is a deglycosylated human IgG1, or a deglycosylated humanized mouse antibody, or a deglycosylated humanized alpaca hcIgG antibody or a deglycosylated chimerized mouse IgG.

23. A first antibody molecule for use in combination with a second antibody molecule according to claim 22, a pharmaceutical composition for use according to claim 22, a kit for use according to claim 22, a use according to claim 22 or a method according to claim 22, which has been deglycosylated by amino acid substitution in position 297.

24. A first antibody molecule for use in combination with a second antibody molecule according to claim 23, a pharmaceutical composition for use according to claim 23, a kit for use according to claim 23, a use according to claim 23 or a method according to claim 23, which has been deglycosylated by N297Q substitution.

25. A first antibody molecule for use in combination with a second antibody molecule according to any one of claims 1 or 6 to 24, a pharmaceutical composition for use according to any one of claims 2 or 6 to 24, a kit for use according to any one of claims 3 or 6 to 24, a use according to any one of claims 4 or 6 to 24, or a method according to any one of claims 5 or 6 to 24, wherein the first antibody molecule comprises: (i) a variable heavy chain (VH) comprising the following CDRs: SEQ ID NO: 51 and SEQ ID NO: 52 and SEQ ID NO: 53, and A variable light chain (VL) comprising the following CDRs: SEQ ID NO: 54 and SEQ ID NO: 55 and SEQ ID NO: 56; (ii) a VH comprising the following CDRs: SEQ ID NO:57 and SEQ ID NO:58 and SEQ ID NO:59, and A VL comprising the following CDRs: SEQ ID NO:60 and SEQ ID NO:61 and SEQ ID NO:62; (iii) a VH comprising the following CDRs: SEQ ID NO: 63 and SEQ ID NO: 64 and SEQ ID NO: 65, and a VL comprising the following CDRs: SEQ ID NO:66 and SEQ ID NO:67 and SEQ ID NO:68; (iv) a VH comprising the following CDRs: SEQ ID NO: 69 and SEQ ID NO: 70 and SEQ ID NO: 71, and a VL comprising the following CDRs: SEQ ID NO:72 and SEQ ID NO:73 and SEQ ID NO:74; (v) a VH comprising the following CDRs: SEQ ID NO: 75 and SEQ ID NO: 76 and SEQ ID NO: 77, and a VL comprising the following CDRs: SEQ ID NO:78 and SEQ ID NO:79 and SEQ ID NO:80; (vi) a VH comprising the following CDRs: SEQ ID NO:81 and SEQ ID NO:82 and SEQ ID NO:83, and a VL comprising the following CDRs: SEQ ID NO:84 and SEQ ID NO:85 and SEQ ID NO:86; (vii) a VH comprising the following CDRs: SEQ ID NO: 87 and SEQ ID NO: 88 and SEQ ID NO: 89, and A VL comprising the following CDRs: SEQ ID NO:90 and SEQ ID NO:91 and SEQ ID NO:92; (viii) a VH comprising the following CDRs: SEQ ID NO:93 and SEQ ID NO:94 and SEQ ID NO:95, and A VL comprising the following CDRs: SEQ ID NO:96 and SEQ ID NO:97 and SEQ ID NO:98; (ix) a VH comprising the following CDRs: SEQ ID NO: 99 and SEQ ID NO: 100 and SEQ ID NO: 101, and A VL comprising the following CDRs: SEQ ID NO: 102 and SEQ ID NO: 103 and SEQ ID NO: 104; (x) a VH comprising the following CDRs: SEQ ID NO: 105 and SEQ ID NO: 106 and SEQ ID NO: 107, and A VL comprising the following CDRs: SEQ ID NO: 108 and SEQ ID NO: 109 and SEQ ID NO: 110; (xi) a VH comprising the following CDRs: SEQ ID NO: 111 and SEQ ID NO: 112 and SEQ ID NO: 113, and a VL comprising the following CDRs: SEQ ID NO: 114 and SEQ ID NO: 115 and SEQ ID NO: 116; (xii) a VH comprising the following CDRs: SEQ ID NO: 117 and SEQ ID NO: 118 and SEQ ID NO: 119, and a VL comprising the following CDRs: SEQ ID NO: 120 and SEQ ID NO: 121 and SEQ ID NO: 122; (xiii) a VH comprising the following CDRs: SEQ ID NO: 123 and SEQ ID NO: 124 and SEQ ID NO: 125, and a VL comprising the following CDRs: SEQ ID NO: 126 and SEQ ID NO: 127 and SEQ ID NO: 128; (xiv) a VH comprising the following CDRs: SEQ ID NO: 129 and SEQ ID NO: 130 and SEQ ID NO: 131, and a VL comprising the following CDRs: SEQ ID NO: 132 and SEQ ID NO: 133 and SEQ ID NO: 134; (xv) a VH comprising the following CDRs: SEQ ID NO: 135 and SEQ ID NO: 136 and SEQ ID NO: 137, and a VL comprising the following CDRs: SEQ ID NO: 138 and SEQ ID NO: 139 and SEQ ID NO: 140; (xvi) a VH comprising the following CDRs: SEQ ID NO: 141 and SEQ ID NO: 142 and SEQ ID NO: 143, and a VL comprising the following CDRs: SEQ ID NO: 144 and SEQ ID NO: 145 and SEQ ID NO: 146; (xvii) a VH comprising the following CDRs: SEQ ID NO: 147 and SEQ ID NO: 148 and SEQ ID NO: 149, and A VL comprising the following CDRs: SEQ ID NO: 150 and SEQ ID NO: 151 and SEQ ID NO: 152; (xviii) a VH comprising the following CDRs: SEQ ID NO: 153 and SEQ ID NO: 154 and SEQ ID NO: 155, and a VL comprising the following CDRs: SEQ ID NO: 156 and SEQ ID NO: 157 and SEQ ID NO: 158; (xix) a VH comprising the following CDRs: SEQ ID NO: 159 and SEQ ID NO: 160 and SEQ ID NO: 161, and A VL comprising the following CDRs: SEQ ID NO: 162 and SEQ ID NO: 163 and SEQ ID NO: 164; (xx) a VH comprising the following CDRs: SEQ ID NO: 165 and SEQ ID NO: 166 and SEQ ID NO: 167, and a VL comprising the following CDRs: SEQ ID NO: 168 and SEQ ID NO: 169 and SEQ ID NO: 170; (xxi) a VH comprising the following CDRs: SEQ ID NO: 171 and SEQ ID NO: 172 and SEQ ID NO: 173, and a VL comprising the following CDRs: SEQ ID NO: 174 and SEQ ID NO: 175 and SEQ ID NO: 176; (xxii) a VH comprising the following CDRs: SEQ ID NO: 177 and SEQ ID NO: 178 and SEQ ID NO: 179, and A VL comprising the following CDRs: SEQ ID NO: 180 and SEQ ID NO: 181 and SEQ ID NO: 182; (xxiii) a VH comprising the following CDRs: SEQ ID NO: 183 and SEQ ID NO: 184 and SEQ ID NO: 185, and A VL comprising the following CDRs: SEQ ID NO: 186 and SEQ ID NO: 187 and SEQ ID NO: 188; or (xxiv) a VH comprising the following CDRs: SEQ ID NO: 189 and SEQ ID NO: 190 and SEQ ID NO: 191, and A VL comprising the following CDRs: SEQ ID NO:192 and SEQ ID NO:193 and SEQ ID NO:

194.

26. A first antibody molecule for use in combination with a second antibody molecule according to any one of claims 1 or 6 to 25, a pharmaceutical composition for use according to any one of claims 2 or 6 to 25, a kit for use according to any one of claims 3 or 6 to 25, a use according to any one of claims 4 or 6 to 25, or a method according to any one of claims 5 or 6 to 25, wherein the first antibody molecule comprises: (i) a VH having SEQ ID NO: 3 and a VL having SEQ ID NO: 27; (ii) a VH having SEQ ID NO: 4 and a VL having SEQ ID NO: 28; (iii) a VH having SEQ ID NO: 5 and a VL having SEQ ID NO: 29; (iv) a VH having SEQ ID NO: 6 and a VL having SEQ ID NO: 30; (v) a VH having SEQ ID NO: 7 and a VL having SEQ ID NO: 31; (vi) a VH having SEQ ID NO: 8 and a VL having SEQ ID NO: 32; (vii) a VH having SEQ ID NO: 9 and a VL having SEQ ID NO: 33; (viii) a VH having SEQ ID NO: 10 and a VL having SEQ ID NO: 34; (ix) a VH having SEQ ID NO: 11 and a VL having SEQ ID NO: 35; (x) a VH having SEQ ID NO: 12 and a VL having SEQ ID NO: 36; (xi) a VH having SEQ ID NO: 13 and a VL having SEQ ID NO: 37; (xii) a VH having SEQ ID NO: 14 and a VL having SEQ ID NO: 38; (xiii) a VH having SEQ ID NO: 15 and a VL having SEQ ID NO: 39; (xiv) a VH having SEQ ID NO: 16 and a VL having SEQ ID NO: 40; (xv) a VH having SEQ ID NO: 17 and a VL having SEQ ID NO: 41; (xvi) a VH having SEQ ID NO: 18 and a VL having SEQ ID NO: 42; (xvii) a VH having SEQ ID NO: 19 and a VL having SEQ ID NO: 43; (xviii) a VH having SEQ ID NO: 20 and a VL having SEQ ID NO: 44; (xix) a VH having SEQ ID NO:21 and a VL having SEQ ID NO:45; (xx) a VH having SEQ ID NO: 22 and a VL having SEQ ID NO: 46; (xxi) a VH having SEQ ID NO: 23 and a VL having SEQ ID NO: 47; (xxii) a VH having SEQ ID NO:24 and a VL having SEQ ID NO:48; (xxiii) a VH having SEQ ID NO: 25 and a VL having SEQ ID NO: 49; or (xxiv) a VH having SEQ ID NO: 26 and a VL having SEQ ID NO: 50;.

27. A first antibody molecule for use in combination with a second antibody molecule according to any one of claims 1 or 6 to 26, a pharmaceutical composition for use according to any one of claims 2 or 6 to 26, a kit for use according to any one of claims 3 or 6 to 26, a use according to any one of claims 4 or 6 to 26, or a method according to any one of claims 5 or 6 to 26, The first antibody molecule comprises: a VH comprising the following CDRs: SEQ ID NO: 171, SEQ ID NO: 172 and SEQ ID NO: 173, and a VL comprising the following CDRs: SEQ ID NO: 174, SEQ ID NO: 175 and SEQ ID NO:

176.

28. A first antibody molecule for use in combination with a second antibody molecule according to any one of claims 1 or 6 to 27, a pharmaceutical composition for use according to any one of claims 2 or 6 to 27, a kit for use according to any one of claims 3 or 6 to 27, a use according to any one of claims 4 or 6 to 27, or a method according to any one of claims 5 or 6 to 27, wherein the first antibody molecule comprises: a VH having SEQ ID NO: 23 and a VL having SEQ ID NO:

47.

29. A first antibody molecule for use in combination with a second antibody molecule according to claim 27 or 28, a pharmaceutical composition for use according to claim 27 or 28, a kit for use according to claim 27 or 28, a use according to claim 27 or 28, or a method according to claim 27 or 28, wherein the first antibody molecule has: a constant heavy chain (CH) having SEQ ID NO: 195 and a constant light chain (CL) having SEQ ID NO:

2.

30. An antibody molecule that specifically binds to FcγRIIB for use, a pharmaceutical composition for use, a kit, a use or a method substantially as described herein in the accompanying claims, description, examples and / or drawings.

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