Cancer treatments comprising anti-MSLN / CD137 antibodies and chemotherapeutic agents

Abstract

The present application relates to the use of antibody molecules that bind to MSLN and CD137 and chemotherapeutic agents in the treatment of cancer in patients.

Description

[Technical Field]

[0001] FIELD OF THE INVENTION The present invention relates to the use of bispecific antibody molecules that bind to MSLN and CD137 and chemotherapeutic agents in the treatment of cancer in patients. [Background technology]

[0002] background Costimulatory pathways, such as the CD137 / 4-1BB pathway, are essential for promoting productive anti-cancer immunity, and there is strong genetic evidence supporting their role in mediating anti-cancer immune responses. 1-5 Therefore, an increasing number of studies have aimed to use agonistic antibodies targeting costimulatory molecules to modify the signals and enhance antitumor T cell responses.

[0003] CD137 (also known as 4-1BB or TNFRSF9) is an inducible T cell surface receptor that belongs to the tumor necrosis factor receptor (TNFR) superfamily and activates diverse cellular functions, including the production of type 1 interferon and regulation of antigen-activated T cell survival. 6 CD137 binds activated CD4 + and CD8 + It is expressed on the surface of T cells, monocytes, and B lymphocytes. CD137 expression can be induced via T cell receptor (TCR) stimulation. 7 This is called "signal 1" (TCR / CD3 / MHC interaction between human T cells and target cells). Activation of the CD137 pathway promotes T cell differentiation and survival. 8-10 , providing potent protection against activation-induced T cell death and enhancing cytotoxicity 11-13 .

[0004] The efficacy of anti-CD137 therapy has been demonstrated in multiple preclinical tumor models 14-18 Anti-CD137 agonist antibodies inhibit CD8 +Induce the release of effector molecules from T cells, increase proliferation, and prevent cytotoxic T lymphocyte (CTL) anergy, thereby breaking T cell tolerance to tumor antigens. 19 , which has been shown to increase the persistence of tumor-specific T cells 20 Based on promising preclinical antitumor effects, two first-generation CD137 agonists, utomilumab (PF-05082566) and urelumab (BMS-663513), have been developed and are being clinically investigated. However, clinical trials of both utomilumab and urelumab monotherapy have been discontinued due to the low efficacy of utomilumab and the hepatotoxicity of urelumab. 21,22 Further structural analysis showed that these results are mediated by a recognized epitope on CD137 and Fc gamma receptor (FcγR) ligand-dependent clustering. 23 .

[0005] To overcome either the low antitumor efficacy or hepatotoxicity mediated by FcγR ligand-dependent clustering of first-generation CD137 agonists, strategies are needed to deliver CD137 agonists to the tumor site, enabling clinical administration while reducing systemic toxicity. 24 These second-generation CD137 agonists are either monospecific antibodies that claim to bind to CD137 epitopes that are not associated with hepatotoxicity, or CD137 / tumor-associated antigen (TAA) bispecific antibodies that target the tumor microenvironment (TME) and are linked to antibodies that do not bind FcγRs and target tumor antigens or tumor tissue. 15,25,26 .

[0006] Mesothelin (MSLN) is a 40 kilodalton (kD) membrane-bound protein that is overexpressed in a variety of cancers, including mesothelioma, ovarian cancer, lung cancer, and pancreatic cancer. 27-38 MSLN has limited expression in normal human tissues and is highly expressed in many common cancers, making it an attractive candidate for cancer therapy. Several agents are in various stages of development to treat patients with MSLN-expressing tumors, including monoclonal antibodies, immunotoxins, tumor vaccines, and antibody-drug conjugates.39 .

[0007] M9657 (FS22-172-003-AA / FS28-256-271 in WO 2020 / 011976) is a first-in-class tumor-targeting conditional agonist antibody developed to enhance antitumor immune responses in the TME. The bispecific antibody M9657 is a tetravalent bispecific antibody (mAb) with a Fab portion that binds to the tumor antigen MSLN and a modified CH3 domain as an Fc antigen-binding (Fcab) portion that binds to CD137. 2 M9657 has been engineered into a IgG1-LALA backbone, which blocks Fcγ receptors but retains FcRn binding due to its IgG-like pharmacokinetics (PK). High expression of MSLN on tumor cells should lead to increased binding and cross-linking of antibody molecules and their interaction with CD137 trimers, resulting in increased CD137 agonism. Therefore, clustered M9657 may function as a bridge connecting CD137 trimers to tumor cells. Because M9567 promotes CD137 activation signaling within the TME, which avoids systemic immune activation, M9657 is expected to offer advantages over monospecific CD137 antibodies. In preclinical studies, M9657 demonstrated MSLN target-dependent and dose-dependent antitumor immunity. Nevertheless, there remains a need in the art for additional anti-cancer therapies. Summary of the Invention

[0008] Description of the invention Accumulating evidence indicates that chemotherapeutic agents induce immunogenic cell death of tumor cells, releasing or exposing these immunogenic tumor antigens and allowing their interaction with innate immune cells such as monocytes, macrophages, and dendritic cells (DCs). 40,41This activates and matures these immune cells, which then migrate to the draining lymph nodes loaded with cancer-derived antigen-specific cargo. The cancer antigens are then presented to T cells, enabling a potent anti-cancer adaptive immune response. Traditional chemotherapeutic agents induce immunogenic cell death by directly interfering with DNA or by targeting key proteins required for cell division. 42 Immunogenic dead tumor cells can release tumor-associated antigens (TAAs) and danger-associated molecular patterns (DAMPs), both of which actively recruit immune cells in the TME. 43 Several chemotherapeutic agents have been reported to deplete myeloid-derived suppressor cells (MDSCs), cancer-associated neutrophils, and macrophages. 44, 45 Optimal doses of some chemotherapeutic agents may promote the expansion of effector T cells and the depletion of Tregs. 46 How to optimally integrate immunotherapy with standard of care (SOC) chemotherapy to achieve additive or synergistic clinical activity is being actively investigated in both preclinical and clinical trials.

[0009] As mentioned above, chemotherapy can facilitate the use of immunotherapy in cancer treatment. However, concerns about efficacy and specificity arise because immune cell activation is not restricted to the TME where increased tumor antigen burden occurs. In particular, development of CD137 agonist molecules has been hindered to date due to concerns about liver inflammation and clinical efficacy.

[0010] The present inventors recognized that target-specific antitumor activity could be enhanced by combining chemotherapy with MSLN expression-dependent CD137 costimulation of T cells. Surprisingly, the present inventors were able to demonstrate that the combination of an antibody molecule that binds to MSLN and CD137 with a chemotherapeutic agent resulted in a greater antitumor effect in vivo in a mouse tumor model than the combined increase in antitumor effect observed when mice were treated with either the antibody molecule that binds to MSLN and CD137 or the chemotherapeutic agent alone. In other words, the antitumor effect of the combined treatment was synergistic, not merely additive. This was unexpected. The effect achieved by the combination of two agents is synergistic if it is greater than the sum of the individual effects of the two combined agents. 47 Thus, we found that the combination of antibody molecules that bind to MSLN and CD137 with chemotherapeutic agents synergistically enhanced the antitumor effect in vivo in mouse tumor models. Similar synergistic antitumor effects are expected when human patients are treated with the combination of antibody molecules that bind to MSLN and CD137 with chemotherapeutic agents.

[0011] Due to the lack of cross-reactivity between M9657 (SEQ ID NO: 2 and SEQ ID NO: 10) and mouse MSLN and CD137 proteins, a surrogate antibody to M9657 for in vivo studies in mouse tumor models, anti-mMSLN-mCD137-huIgG1-LALA (FS122m) (SEQ ID NO: 84 and SEQ ID NO: 85), was developed. Similar to M9657, FS122m is a tetravalent bispecific antibody (mAb) in which the Fab portion is targeted to bind to the tumor antigen mouse MSLN and the modified CH3 domain as the Fcab portion is targeted to mouse CD137. 2 ) format. FS122m has a human IgG1 backbone with a LALA mutation that abolishes binding to Fcγ receptors. The binding affinity of FS122m to mouse MSLN / CD137 is similar to that of M9657 to human MSLN / CD137.

[0012] As already summarized above, we have shown that the combination of FS122m with two chemotherapeutic agents, either cisplatin or gemcitabine, can delay tumor growth or reduce tumor volume in ST26 and JC mouse tumor models beyond the combined tumor growth delay or tumor volume reduction observed when mice were treated with either FS122m or cisplatin or gemcitabine alone. We also showed that combined treatment with FS122m and either cisplatin or gemcitabine increased median survival and the percentage of mice with complete tumor regression in the same mouse tumor models compared to the combined increases in median survival and the percentage of mice with complete tumor regression observed when mice were treated with either FS122m or cisplatin or gemcitabine alone. Thus, we have shown that the combination of FS122m with either cisplatin or gemcitabine synergistically enhances antitumor activity by measuring tumor growth delay / tumor volume reduction, median survival, and the percentage of mice exhibiting complete tumor regression in ST26 and JC mouse tumor models.

[0013] These preclinical results using mouse tumor models support the expectation that the combination of antibody molecules that bind to MSLN and CD137 with chemotherapeutic agents will synergistically enhance antitumor activity in human patients. These findings suggest that combination therapy of antibody molecules that bind to MSLN and CD137 with chemotherapeutic agents represents a new therapeutic strategy for improving cancer treatment.

[0014] Thus, the present invention provides an antibody molecule that binds to MSLN and CD137 for use in a method of treating cancer in a patient, the method comprising administering the antibody in combination with a chemotherapeutic agent. The present invention also provides a chemotherapeutic agent for use in a method of treating cancer in a patient, the method comprising administering the chemotherapeutic agent in combination with an antibody molecule that binds to MSLN and CD137.

[0015] The antibody molecule that binds to MSLN and CD137 can be an immunoglobulin or its antigen-binding fragment.For example, the antibody molecule can be an IgG, IgA, IgE or IgM molecule, preferably an IgG molecule, such as an IgG1, IgG2, IgG3 or IgG4 molecule, more preferably an IgG1 or IgG2 molecule, most preferably an IgG1 molecule, or a fragment thereof.In a preferred embodiment, the antibody molecule is a complete immunoglobulin molecule.

[0016] The antibody molecule may comprise at least one, and preferably more than one, complementarity determining region (CDR)-based binding site for MSLN and at least one, and preferably more than one, binding site for CD137 within the constant domain, preferably the CH3 domain, of the bispecific antibody molecule.

[0017] The binding site for CD137 may comprise a first sequence and a second sequence located in the AB and EF structural loops of the CH3 domain of the antibody molecule. Preferably, the first sequence has the sequence set forth in SEQ ID NO: 87. Preferably, the second sequence has the sequence set forth in SEQ ID NO: 88. More preferably, the first sequence has the sequence set forth in SEQ ID NO: 87 and the second sequence has the sequence set forth in SEQ ID NO: 88. According to the IMGT numbering system, the first sequence may be located between positions 14 and 17 of the CH3 domain of the antibody molecule. According to the IMGT numbering system, the second sequence may be located between positions 91 and 99 of the CH3 domain of the antibody molecule. Preferably, the sequence of the CH3 domain of the antibody molecule has the sequence set forth in SEQ ID NO: 86.

[0018] In a preferred embodiment, the bispecific antibody molecule comprises a CH3 domain that comprises, has or consists of the CH3 domain sequence of FS22-172-003 as set forth in SEQ ID NO: 86. The CH3 domain of the bispecific antibody molecule may optionally comprise an additional lysine residue (K) immediately C-terminal to the CH3 domain sequence.

[0019] A number of Fabs that bind to MSLN are known from WO 2020 / 011976. A complementarity-determining region (CDR)-based binding site for MSLN can comprise CDRs 1-6 of any of these Fabs. Thus, an antibody molecule that binds to MSLN and CD137 can comprise CDRs 1-6 as set forth below: SEQ ID NOS: 4, 6, 8, 12, 14, and 16 [FS28-256-271]; SEQ ID NOS: 20, 22, 24, 12, 14, and 28 [FS28-024-052]; SEQ ID NOS: 4, 6, 8, 12, 14, and 34 [FS28-256-021 ... SEQ ID NOs: 4, 6, 8, 12, 14, and 39 [FS28-256-012]; SEQ ID NOs: 43, 6, 45, 12, 14, and 34 [FS28-256-023]; SEQ ID NOs: 4, 6, 8, 12, 14, and 49 [FS28-256-024]; SEQ ID NOs: 43, 6, 45, 12, 14, and 49 [FS28-256-026]; SEQ ID NOs: 4, 6, 8, 12, 14 , and 16 [FS28-256-027]; SEQ ID NOs: 53, 6, 55, 12, 14, and 34 [FS28-256-001]; SEQ ID NOs: 53, 6, 55, 12, 14, and 49 [FS28-256-005]; SEQ ID NOs: 60, 6, 62, 12, 14, and 39 [FS28-256-014]; SEQ ID NOs: 43, 6, 45, 12, 14, and 39 [FS 28-256-018]; SEQ ID NOs: 67, 6, 55, 12, 14, and 39 [FS28-256]; SEQ ID NOs: 21, 23, 72, 12, 14, and 28 [FS28-024-051]; SEQ ID NOs: 21, 23, 77, 12, 14, and 28 [FS28-024-053]; or SEQ ID NOs: 21, 23, 82, 12, 14, and 28 [FS28-024].

[0020] A number of bispecific antibody molecules that bind to MSLN and CD137 are known from WO 2020 / 011976. The antibody M9657 of the present application is identical to the antibody FS22-172-003-AA / FS28-256-271 of WO 2020 / 011976. Any of these antibodies can be used, and they are incorporated herein by reference. Thus, antibody molecules that bind to MSLN and CD137 include SEQ ID NOs: 2 and 10 (FS22-172-003-AA / FS28-256-271), SEQ ID NOs: 18 and 26 (FS22-172-003-AA / FS28-024-052), SEQ ID NOs: 30 and 32 (FS22-172-003-AA / FS28-256-021), SEQ ID NOs: 36 and 37 (FS22-172-003-AA / FS28-256-012), SEQ ID NOs: 41 and 32 (FS22-172-003-AA / FS28-256-023), SEQ ID NOs: 30 and 47 (FS22-172-003-AA / FS28-256-024), SEQ ID NOs: 41 and 47 (FS22-172-003-AA / FS28-256-026), SEQ ID NOs: 30 and 10 (FS22-172-003-AA / FS28-256-0 27), SEQ ID NOs: 51 and 32 (FS22-172-003-AA / FS28-256-001), SEQ ID NOs: 51 and 47 (FS22-172-003-AA / FS28-256-005), SEQ ID NOs: 58 and 37 (FS22-172-003-AA / FS28-256-014), SEQ ID NOs: 41 and 37 (FS22-172-003-AA / FS28-256-018), SEQ ID NOs: 65 and and 37 (FS22-172-003-AA / FS28-256), SEQ ID NOs: 70 and 26 (FS22-172-003-AA / FS28-024-051), SEQ ID NOs: 75 and 26 (FS22-172-003-AA / FS28-024-053), or SEQ ID NOs: 80 and 26 (FS22-172-003-AA / FS28-024). Preferably, the antibody molecule that binds to MSLN and CD137 comprises the heavy chain sequence set forth in SEQ ID NO: 2 and the light chain sequence set forth in SEQ ID NO: 10 (FS22-172-003-AA / FS28-256-271).

[0021] The chemotherapeutic agent can be an alkylating agent or an antimetabolite. Preferably, the antimetabolite is selected from the group consisting of azacitidine, 5-fluorouracil (5-FU), 6-mercaptopurine (6-MP), capecitabine (Xeloda), cladribine, clofarabine, cytarabine (Ara-C), decitabine, floxuridine, fludarabine, gemcitabine (Gemzar), hydroxyurea, methotrexate, nelarabine, pemetrexed (Alimta), pentostatin, pralatrexate, thioguanine, and trifluridine / tipiracil combination. More preferably, the antimetabolite can be gemcitabine.

[0022] The alkylating agent is preferably selected from the group comprising altretamine, bendamustine, busulfan, carboplatin, carmustine, chlorambucil, cisplatin, cyclophosphamide (CPA), dacarbazine, ifosfamide, lomustine, mechlorethamine, melphalan, oxaliplatin, temozolomide, thiotepa, and trabectedin. More preferably, the alkylating agent may be cisplatin.

[0023] The membrane-associated protein mesothelin (MSLN) has been shown to be expressed in several cancers. Ovarian cancer, pancreatic adenocarcinoma, mesothelioma, and non-small cell lung cancer have all been shown to express high levels of MSLN. The inventors have found this to be true in cervical cancer as well. Without wishing to be bound by theory, it is believed that when antibody molecules bind to MSLN, cross-linking of the antibody occurs, leading to binding to CD137 expressed on the surface of immune cells, which then clusters and activates CD137, ultimately activating the immune cells.

[0024] Therefore, the cancer to be treated is preferably a cancer that expresses or is confirmed to express MSLN. More preferably, the cancer is selected from the group including ovarian cancer, pancreatic adenocarcinoma, mesothelioma, cervical cancer, and non-small cell lung cancer.

[0025] Combination therapy of FS122m with cisplatin or gemcitabine resulted in statistically significantly greater antitumor activity in the CT26 and JC mouse tumor models than the combined antitumor activity observed when mice were treated with either FS122m or cisplatin or gemcitabine alone. The effect achieved by the combination of two drugs is synergistic if the effect is greater than the sum of the individual effects of the two combined drugs. 47 Thus, combination therapy of FS122m with cisplatin or gemcitabine synergistically enhanced antitumor activity in CT26 and JC mouse tumor models. Thus, in one embodiment, treatment with an antibody molecule that binds to MSLN and CD137 in combination with a chemotherapeutic agent results in an antitumor effect greater than the combined antitumor effect observed when a patient is treated with either the antibody molecule that binds to MSLN and CD137 or the chemotherapeutic agent alone. Preferably, treatment with an antibody molecule that binds to MSLN and CD137 in combination with a chemotherapeutic agent results in an antitumor effect greater than the combined antitumor effect observed when a patient is treated with either the antibody molecule that binds to MSLN and CD137 or the chemotherapeutic agent alone. The antitumor effect can be tumor growth inhibition or delay, tumor volume reduction, increased median survival, or an increase in the proportion of patients experiencing complete tumor regression. Preferably, the antitumor effect is tumor growth inhibition or delay, or tumor volume reduction. Thus, the antitumor effect can be tumor growth inhibition or delay. The antitumor effect can be tumor volume reduction. An anti-tumor effect can be an increase in the median survival time of patients. An anti-tumor effect can be an increase in the proportion of patients experiencing complete tumor regression, such as clinical complete response or pathological complete response. Determining these anti-tumor effects is within the capabilities of those skilled in the art.

[0026] The bispecific antibody molecule that binds to MSLN and CD137 and the chemotherapeutic agent can be administered to a subject by any suitable means. Thus, in one embodiment, the antibody molecule that binds to MSLN and CD137 and / or the chemotherapeutic agent is administered parenterally. The antibody molecule that binds to MSLN and CD137 and / or the chemotherapeutic agent can be administered intravenously, intramuscularly, subcutaneously, intraperitoneally, or spinally. The antibody molecule that binds to MSLN and CD137 and / or the chemotherapeutic agent can be administered by injection or infusion.

[0027] The antibody molecules and / or chemotherapeutic agents that bind to MSLN and CD137 may be administered non-parenterally. The antibody molecules and / or chemotherapeutic agents that bind to MSLN and CD137 may be administered orally, intranasally, vaginally, rectally, sublingually, or topically.

[0028] The antibody molecule that binds to MSLN and CD137 and the chemotherapeutic agent may be part of the same formulation or part of separate formulations, but are preferably provided as separate formulations. Thus, the antibody molecule that binds to MSLN and CD137 and the chemotherapeutic agent may be administered to a patient simultaneously or sequentially, but are preferably administered sequentially.

[0029] When the antibody molecule that binds to MSLN and CD137 and the chemotherapeutic agent are administered sequentially to the patient, they are preferably administered to the patient within 4 days of each other, more preferably within 3 days of each other, more preferably within 2 days of each other, or sequentially on the same day.

[0030] The present invention also provides a method for treating cancer, comprising administering to an individual in need thereof an antibody molecule that binds to MSLN and CD137 and a chemotherapeutic agent. Preferably, the method for treating cancer comprises administering to an individual in need thereof a therapeutically effective amount of an antibody molecule that binds to MSLN and CD137 and a therapeutically effective amount of a chemotherapeutic agent. In one embodiment, the method may comprise determining whether a patient's cancer expresses MSLN and treating the patient if the cancer is determined to express MSLN. Alternatively, the method may comprise ordering the results of a test to determine whether the patient's cancer expresses MSLN and treating the patient if the test results indicate that the cancer expresses MSLN.

[0031] The present invention also provides use of an antibody molecule that binds to MSLN and CD137 for the manufacture of a medicament for the treatment of cancer, wherein the antibody molecule that binds to MSLN and CD137 is administered in combination with a chemotherapeutic agent.The present invention also provides use of a chemotherapeutic agent for the manufacture of a medicament for the treatment of cancer, wherein the chemotherapeutic agent is administered in combination with an antibody molecule that binds to MSLN and CD137.

[0032] The present invention also provides a kit comprising an antibody molecule that binds to MSLN and CD137 and a pharmaceutically acceptable excipient, and a chemotherapeutic agent and a pharmaceutically acceptable excipient.

[0033] Thus, the present invention provides: [1] An antibody molecule that binds to MSLN and CD137 for use in a method of treating cancer in a patient, the method comprising administering the antibody in combination with a chemotherapeutic agent. [2] A chemotherapeutic agent for use in a method for treating cancer in a patient, the method comprising administering the chemotherapeutic agent in combination with an antibody molecule that binds to MSLN and CD137. [3] A method for treating cancer in an individual, comprising administering to the individual an antibody molecule that binds to MSLN and CD137 and a chemotherapeutic agent.

[0034] [4] Use of an antibody molecule that binds to MSLN and CD137 for the manufacture of a medicament for the treatment of cancer, wherein the antibody molecule that binds to MSLN and CD137 is administered in combination with a chemotherapeutic agent. [5] Use of a chemotherapeutic agent for the manufacture of a medicament for the treatment of cancer, wherein the chemotherapeutic agent is administered in combination with an antibody molecule that binds to MSLN and CD137. [6] Kit containing: (a) an antibody molecule that binds to MSLN and CD137 and a pharmaceutically acceptable excipient; and (b) a chemotherapeutic agent and a pharmaceutically acceptable excipient.

[0035] [7] The antibody molecule or chemotherapeutic agent, method, use, or kit for use according to any one of [1] to [6], wherein the antibody molecule that binds to MSLN and CD137 is selected from the group consisting of: (a) a complementarity-determining region (CDR)-based antigen-binding site for MSLN; and (b) the CD137 antigen-binding site located in the CH3 domain of the antibody molecule; an antibody molecule or chemotherapeutic agent, method, use, or kit for said use, comprising: [8] The antibody molecule or chemotherapeutic agent, method, use, or kit for use according to [7], wherein the antibody molecule that binds to MSLN and CD137 is selected from two or more of the following: (a) a complementarity-determining region (CDR)-based antigen-binding site for MSLN; and (b) the CD137 antigen-binding site located in the CH3 domain of the antibody molecule; an antibody molecule or chemotherapeutic agent, method, use, or kit for said use, comprising: [9] An antibody molecule or chemotherapeutic agent, method, use, or kit for use according to any one of [1] to [8], wherein the antibody molecule that binds to MSLN and CD137 is an IgG molecule, an IgA molecule, an IgE molecule, an IgM molecule, or an antigen-binding fragment thereof.

[0036]

[10] The antibody molecule or chemotherapeutic agent, method, use, or kit for use according to [9], wherein the antibody molecule that binds to MSLN and CD137 is an IgG molecule or an antigen-binding fragment thereof.

[11] The antibody molecule or chemotherapeutic agent, method, use, or kit for use according to

[10] , wherein the antibody molecule that binds to MSLN and CD137 is an IgG1 or IgG2 molecule, or an antigen-binding fragment thereof.

[12] The antibody molecule or chemotherapeutic agent, method, use, or kit for use according to

[11] , wherein the antibody molecule that binds to MSLN and CD137 is an IgG1 molecule or an antigen-binding fragment thereof.

[0037]

[13] The antibody molecule or chemotherapeutic agent, method, use, or kit for use according to any one of [8] to

[12] , wherein the CDR-based antigen-binding site against MSLN comprises CDRs 1 to 6 as follows: (i) SEQ ID NOs: 4, 6, 8, 12, 14, and 16 [FS28-256-271], respectively; (ii) SEQ ID NOs: 20, 22, 24, 12, 14, and 28 [FS28-024-052], respectively; (iii) SEQ ID NOs: 4, 6, 8, 12, 14, and 34 [FS28-256-021], respectively; (iv) SEQ ID NOs: 4, 6, 8, 12, 14, and 39 [FS28-256-012], respectively; (v) SEQ ID NOs: 43, 6, 45, 12, 14, and 34 [FS28-256-023], respectively; (vi) SEQ ID NOs: 4, 6, 8, 12, 14, and 49 [FS28-256-024], respectively; (vii) SEQ ID NOs: 43, 6, 45, 12, 14, and 49 [FS28-256-026], respectively; (viii) SEQ ID NOs: 4, 6, 8, 12, 14, and 16 [FS28-256-027], respectively; (ix) SEQ ID NOs: 53, 6, 55, 12, 14, and 34 [FS28-256-001], respectively; (x) SEQ ID NOs: 53, 6, 55, 12, 14, and 49 [FS28-256-005], respectively; (xi) SEQ ID NOs: 60, 6, 62, 12, 14, and 39 [FS28-256-014], respectively; (xii) SEQ ID NOs: 43, 6, 45, 12, 14, and 39 [FS28-256-018], respectively; (xiii) SEQ ID NOs: 67, 6, 55, 12, 14, and 39 [FS28-256], respectively; (xiv) SEQ ID NOs: 21, 23, 72, 12, 14, and 28 [FS28-024-051], respectively; (xv) SEQ ID NOs: 21, 23, 77, 12, 14, and 28, respectively [FS28-024-053]; or (xvi) SEQ ID NOs: 21, 23, 82, 12, 14, and 28 [FS28-024], respectively; and An antibody molecule or chemotherapeutic agent, method, use, or kit for the above use, wherein the CD137 antigen binding site comprises a first and a second sequence located in the AB and EF structural loops of the CH3 domain, respectively, and wherein the first and second sequences have the sequences set forth in SEQ ID NOs: 87 and 88, respectively.

[0038]

[14] An antibody molecule or chemotherapeutic agent, method, use, or kit for use according to

[13] , wherein: (i) the first sequence is located between positions 14 and 17 of the CH3 domain of the antibody molecule; and / or (ii) wherein the second sequence is located between positions 91 and 99 of the CH3 domain of the antibody molecule; and An antibody molecule or chemotherapeutic agent, method, use, or kit for said use, wherein the numbering of amino acid residues is according to the IMGT numbering system.

[0039]

[15] An antibody molecule or chemotherapeutic agent, method, use, or kit for use according to any one of [1] to

[14] , wherein the antibody molecule that binds to MSLN and CD137 comprises the CH3 domain sequence set forth in SEQ ID NO: 86.

[16] The antibody molecule or chemotherapeutic agent, method, use, or kit for use according to

[15] , wherein the CH3 domain comprises an additional lysine residue (K) adjacent to the C-terminus of the CH3 domain sequence.

[0040]

[17] The antibody molecule or chemotherapeutic agent, method, use, or kit for use according to any one of [1] to

[16] , wherein the antibody molecule that binds to MSLN and CD137 comprises the heavy and light chains of the following antibodies: (i) FS22-172-003-AA / FS28-256-271 set forth in SEQ ID NOs: 2 and 10, respectively; (ii) FS22-172-003-AA / FS28-024-052 set forth in SEQ ID NOs: 18 and 26, respectively; (iii) FS22-172-003-AA / FS28-256-021 set forth in SEQ ID NOs: 30 and 32, respectively; (iv) FS22-172-003-AA / FS28-256-012 set forth in SEQ ID NOs: 36 and 37, respectively; (v) FS22-172-003-AA / FS28-256-023 set forth in SEQ ID NOs: 41 and 32, respectively; (vi) FS22-172-003-AA / FS28-256-024 set forth in SEQ ID NOs: 30 and 47, respectively; (vii) FS22-172-003-AA / FS28-256-026 set forth in SEQ ID NOs: 41 and 47, respectively; (viii) FS22-172-003-AA / FS28-256-027 set forth in SEQ ID NOs: 30 and 10, respectively; (ix) FS22-172-003-AA / FS28-256-001 set forth in SEQ ID NOs: 51 and 32, respectively; (x) FS22-172-003-AA / FS28-256-005 set forth in SEQ ID NOs: 51 and 47, respectively; (xi) FS22-172-003-AA / FS28-256-014 set forth in SEQ ID NOs: 58 and 37, respectively; (xii) FS22-172-003-AA / FS28-256-018 set forth in SEQ ID NOs: 41 and 37, respectively; (xiii) FS22-172-003-AA / FS28-256 set forth in SEQ ID NOs: 65 and 37, respectively; (xiv) FS22-172-003-AA / FS28-024-051 set forth in SEQ ID NOs: 70 and 26, respectively; (xv) FS22-172-003-AA / FS28-024-053 set forth in SEQ ID NOs: 75 and 26, respectively; or (xvi) FS22-172-003-AA / FS28-024 as set forth in SEQ ID NOs: 80 and 26, respectively; An antibody molecule or chemotherapeutic agent, method, use or kit for said use.

[0041]

[18] An antibody molecule or chemotherapeutic agent, method, use, or kit for use according to any one of [1] to

[17] , wherein the antibody molecule that binds to MSLN and CD137 comprises the heavy chain sequence set forth in SEQ ID NO: 2 and the light chain sequence set forth in SEQ ID NO: 10 [FS22-172-003-AA / FS28-256-271].

[19] The antibody molecule or chemotherapeutic agent, method, use, or kit for use according to any one of [1] to

[18] , wherein the chemotherapeutic agent is an alkylating agent or an antimetabolite.

[20] The antibody molecule or chemotherapeutic agent, method, use, or kit for use according to

[19] , wherein the alkylating agent is selected from the group consisting of altretamine, bendamustine, busulfan, carboplatin, carmustine, chlorambucil, cisplatin, cyclophosphamide (CPA), dacarbazine, ifosfamide, lomustine, mechlorethamine, melphalan, oxaliplatin, temozolomide, thiotepa, and trabectedin.

[0042]

[21] The antibody molecule or chemotherapeutic agent, method, use, or kit for use according to

[19] or

[20] , wherein the alkylating agent is cisplatin.

[22] The antibody molecule or chemotherapeutic agent, method, use, or kit for use according to

[18] , wherein the antimetabolite is selected from the group consisting of azacitidine, 5-fluorouracil (5-FU), 6-mercaptopurine (6-MP), capecitabine, cladribine, clofarabine, cytarabine, decitabine, floxuridine, fludarabine, gemcitabine, hydroxyurea, methotrexate, nelarabine, pemetrexed, pentostatin, pralatrexate, thioguanine, and a combination of trifluridine / tipiracil.

[23] The antibody molecule or chemotherapeutic agent, method, use, or kit for use according to

[20] or

[22] , wherein the antimetabolite is gemcitabine.

[0043]

[24] An antibody molecule or chemotherapeutic agent, method, use, or kit for use according to any one of [1] to

[23] , wherein the cancer expresses MSLN or has been determined to express MSLN.

[25] The antibody molecule or chemotherapeutic agent, method, use, or kit for use according to [1] to

[24] , wherein the cancer is selected from the group consisting of ovarian cancer, pancreatic adenocarcinoma, mesothelioma, cervical cancer, and non-small cell lung cancer.

[26] An antibody molecule or chemotherapeutic agent, method, use, or kit for use according to any one of [1] to

[25] , wherein treatment with an antibody molecule that binds to MSLN and CD137 and a chemotherapeutic agent results in greater anti-tumor activity than monotherapy treatment with an antibody molecule that binds to MSLN and CD137 or monotherapy treatment with a chemotherapeutic agent.

[0044]

[27] An antibody molecule or chemotherapeutic agent, method, use, or kit for use according to any one of [1] to

[26] , wherein treatment with an antibody molecule that binds to MSLN and CD137 and a chemotherapeutic agent results in anti-tumor activity greater than the combined anti-tumor activity of monotherapy treatment with the antibody molecule that binds to MSLN and CD137 and monotherapy treatment with the chemotherapeutic agent.

[28] An antibody molecule or chemotherapeutic agent, method, use, or kit for use according to any one of [1] to

[27] , wherein treatment with an antibody molecule that binds to MSLN and CD137 and a chemotherapeutic agent results in greater tumor growth delay, less tumor volume reduction, increased median survival time, and / or more complete tumor regressions than monotherapy treatment with an antibody molecule that binds to MSLN and CD137 or monotherapy treatment with a chemotherapeutic agent.

[29] An antibody molecule or chemotherapeutic agent, method, use, or kit for use according to any of [1] to

[28] , wherein treatment with an antibody molecule that binds to MSLN and CD137 and a chemotherapeutic agent results in a tumor growth delay, tumor volume reduction, increase in median survival time, and / or increase in the number of complete tumor regressions that is greater than the combined tumor growth delay, tumor volume reduction, increase in median survival time, and / or increase in the number of complete tumor regressions of monotherapy treatment with an antibody molecule that binds to MSLN and CD137 and monotherapy treatment with a chemotherapeutic agent.

[0045]

[30] An antibody molecule or chemotherapeutic agent, method, use, or kit for use according to any one of [1] to

[29] , wherein the antibody molecule and / or chemotherapeutic agent that binds to MSLN and CD137 is administered parenterally.

[31] The antibody molecule or chemotherapeutic agent, method, use, or kit for use according to

[30] , wherein the antibody molecule and / or chemotherapeutic agent that binds to MSLN and CD137 is administered intravenously, intramuscularly, subcutaneously, intraperitoneally, or spinally.

[32] The antibody molecule or chemotherapeutic agent, method, use, or kit for use according to

[30] or

[31] , wherein the antibody molecule and / or chemotherapeutic agent that binds to MSLN and CD137 is administered by injection or infusion.

[0046]

[33] An antibody molecule or chemotherapeutic agent, method, use, or kit for use according to any one of [1] to

[29] , wherein the antibody molecule and / or chemotherapeutic agent that binds to MSLN and CD137 is administered non-parenterally.

[34] The antibody molecule or chemotherapeutic agent, method, use, or kit for use according to

[33] , wherein the antibody molecule and / or chemotherapeutic agent that binds to MSLN and CD137 is administered orally, intranasally, vaginally, rectally, sublingually, or topically.

[35] An antibody molecule or chemotherapeutic agent, method, use, or kit for use according to any one of [1] to

[34] , wherein the antibody molecule and chemotherapeutic agent that bind to MSLN and CD137 are administered to a patient simultaneously or sequentially.

[0047]

[36] The antibody molecule or chemotherapeutic agent, method, use, or kit for use according to

[35] , wherein the antibody molecule and / or chemotherapeutic agent that binds to MSLN and CD137 is administered sequentially to a patient.

[37] The antibody molecule or chemotherapeutic agent, method, use, or kit for use according to

[36] , wherein the antibody molecule that binds to MSLN and CD137 and the chemotherapeutic agent are administered to the patient within 4 days of each other.

[38] The antibody molecule or chemotherapeutic agent, method, use, or kit for use according to

[36] , wherein the antibody molecule and the chemotherapeutic agent that bind to MSLN and CD137 are administered to the patient within three days of each other.

[0048]

[39] The antibody molecule or chemotherapeutic agent, method, use, or kit for use according to

[36] , wherein the antibody molecule and chemotherapeutic agent that bind to MSLN and CD137 are administered to the patient within two days of each other.

[40] The antibody molecule or chemotherapeutic agent, method, use, or kit for use according to

[36] , wherein the antibody molecule that binds to MSLN and CD137 and the chemotherapeutic agent are administered to the patient on the same day.

[41] An antibody molecule or chemotherapeutic agent for use, or a method, according to any one of [1] to [3] and [7] to

[40] , wherein the method comprises determining whether the cancer expresses MSLN, and treating the individual if the cancer expresses MSLN. [Brief explanation of the drawings]

[0049] Drawing Overview BRIEF DESCRIPTION OF THE DRAWINGS Embodiments and experiments illustrating the principles of the present invention are described below with reference to the accompanying drawings, in which:

[0050] [Figure 1] Figure 1 shows the efficacy of treatment in a CT26 colon tumor mouse model in BALB / c mice. Tumor progression was measured as mean tumor volume over time (A), median survival (B), % body weight change (C), and change in individual tumor volume over time (D). Mice were treated with either anti-HEL-hIgG1-LALA, FS122m, cisplatin, or FS122m + cisplatin. Treatment with FS122m + cisplatin delayed tumor volume growth to a greater extent than FS122m and cisplatin monotherapy when compared to the anti-HEL-hIgG1-LALA isotype control (A). FS122m plus cisplatin also improved median survival compared with FS122m or cisplatin monotherapy (B) and induced complete tumor regression in 7 of 10 mice, whereas 2 of 10 mice treated with FS122m monotherapy experienced complete tumor regression and none of 10 mice treated with cisplatin monotherapy experienced complete tumor regression (D). Body weight changes were comparable across all treatments, including the anti-HEL-hIgG1-LALA isotype control, demonstrating that all treatments were well tolerated (C). Tumor volume data were log-transformed and subjected to two-way analysis of variance (ANOVA) followed by Tukey's multiple comparison test; ** = P ≤ 0.01, *** = P ≤ 0.001, **** = P ≤ 0.0001. Survival is expressed as median percentage survival, and mean tumor volume and body weight changes are shown as means ± SEM.

[0051] [Figure 2]Figure 2 shows the efficacy of treatment in the JC tumor mouse model in BALB / c mice. Tumor progression was measured as mean tumor volume over time (A), median survival (B), % body weight change (C), and change in individual tumor volume over time (D). Mice were treated with either anti-HEL-hIgG1-LALA, FS122m, cisplatin, or FS122m + cisplatin. Treatment with FS122m + cisplatin delayed tumor volume growth to a greater extent than FS122m and cisplatin monotherapy when compared to the anti-HEL-hIgG1-LALA isotype control (A). FS122m plus cisplatin also improved median survival compared with FS122m or cisplatin monotherapy (B) and induced complete tumor regression in 2 of 10 mice, whereas 2 of 10 mice treated with FS122m monotherapy experienced complete tumor regression and none of 10 mice treated with cisplatin monotherapy experienced complete tumor regression (D). Body weight changes were comparable across all treatments, including the anti-HEL-hIgG1-LALA isotype control, demonstrating that all treatments were well tolerated (C). Tumor volume data were log-transformed and subjected to two-way analysis of variance (ANOVA) followed by Tukey's multiple comparison test; ** = P ≤ 0.01, *** = P ≤ 0.001, **** = P ≤ 0.0001. Survival is expressed as percent median survival, and mean tumor volume and body weight changes are shown as means ± SEM.

[0052] [Figure 3]Figure 3 shows the efficacy of treatment in the JC tumor mouse model in BALB / c mice. Tumor progression was measured as mean tumor volume over time (A), median survival (B), % body weight change (C), and change in individual tumor volume over time (D). Mice were treated with either anti-HEL-hIgG1-LALA, FS122m, gemcitabine, or FS122m + gemcitabine. Treatment with FS122m + gemcitabine delayed tumor volume growth to a greater extent than monotherapy with FS122m and gemcitabine when compared to the anti-HEL-hIgG1-LALA isotype control (A). FS122m plus gemcitabine also improved median survival compared with FS122m or gemcitabine monotherapy (B) and induced complete tumor regression in 5 of 10 mice, whereas 1 of 10 mice treated with FS122m monotherapy experienced complete tumor regression and none of 10 mice treated with gemcitabine monotherapy experienced complete tumor regression (D). Body weight changes were comparable across all treatments, including the anti-HEL-hIgG1-LALA isotype control, demonstrating that all treatments were well tolerated (C). Tumor volume data were log-transformed and subjected to two-way analysis of variance (ANOVA) followed by Tukey's multiple comparison test; ** = P ≤ 0.01, *** = P ≤ 0.001, **** = P ≤ 0.0001. Survival is expressed as percent median survival, and mean tumor volume and body weight changes are shown as mean ± SEM.

[0053] [Figure 4]Figure 4 shows the efficacy of treatment in a CT26 colon tumor mouse model in BALB / c mice. Tumor progression was measured as mean tumor volume over time (A), median survival (B), % body weight change (C), and change in individual tumor volume over time (D). Mice were treated with either anti-HEL-hIgG1-LALA, FS122m, gemcitabine, or FS122m + gemcitabine. Treatment with FS122m + gemcitabine delayed tumor volume growth to a greater extent than monotherapy with FS122m and gemcitabine when compared to the anti-HEL-hIgG1-LALA isotype control (A). FS122m plus gemcitabine also improved median survival compared with FS122m or gemcitabine monotherapy (B) and induced complete tumor regression in one of nine mice, whereas three of nine mice treated with FS122m monotherapy experienced complete tumor regression and none of nine mice treated with gemcitabine monotherapy experienced complete tumor regression (D). Body weight changes were comparable across all treatments, including the anti-HEL-hIgG1-LALA isotype control, demonstrating that all treatments were well tolerated (C). Tumor volume data were log-transformed and subjected to two-way analysis of variance (ANOVA) followed by Tukey's multiple comparison test; ** = P ≤ 0.01, *** = P ≤ 0.001, **** = P ≤ 0.0001. Survival is expressed as percent median survival, and mean tumor volume and body weight changes are shown as means ± SEM.

[0054] Detailed Description of the Invention The present invention relates to an antibody molecule that binds to MSLN and CD137 for use in treating cancer in a patient, said antibody molecule in combination with a chemotherapeutic agent. The present invention also relates to a chemotherapeutic agent for use in treating cancer, said chemotherapeutic agent in combination with an antibody molecule that binds to MSLN and CD137.

[0055] The term "bispecific" refers to a molecule that does not show significant binding to molecules other than its two specific binding partners.This term can also refer to the specific epitopes of the two binding partners that may be carried by other antigens, and in this case, the antibody may also bind to the antigen that carries the specific epitope.In a preferred embodiment, the bispecific antibody molecule does not show significant binding activity to any of OX40, GITR, CD40, CEACAM-5, E-cadherin, thrombomodulin, and EpCAM.

[0056] The term "antibody molecule" refers to an immunoglobulin, whether natural or partially or wholly synthetically produced. The antibody molecule may be human or humanized, preferably human. The antibody molecule may preferably be a monoclonal antibody. Examples of antibody molecules are immunoglobulin isotypes, such as immunoglobulin G, and their isotypic subclasses, such as IgG1, IgG2, IgG3, and IgG4, and fragments thereof. The antibody molecule may be isolated, in the sense of being free from contaminants, such as antibody molecules capable of binding to other polypeptides and / or serum components.

[0057] In the following, the term "bispecific antibody molecule" is used to refer to an antibody molecule that binds to MSLN and CD137. In one embodiment, the bispecific antibody molecule binds independently to MSLN and CD137. In one embodiment, the bispecific antibody binds simultaneously to MSLN and CD137. Bispecific antibody molecules may be naturally occurring or partly or wholly synthetically produced, for example the antibody molecule may be a recombinant antibody molecule.

[0058] The bispecific antibody molecule may comprise at least one, and preferably more than one, complementarity determining region (CDR)-based binding site for MSLN and at least one, and preferably more than one, binding site for CD137 within the constant domains of the bispecific antibody molecule, preferably at least one CH3 domain.

[0059] The bispecific antibody molecule may be an immunoglobulin or an antigen-binding fragment thereof. For example, the bispecific antibody molecule may be an IgG, IgA, IgE, or IgM molecule, preferably an IgG molecule, such as an IgG1, IgG2, IgG3, or IgG4 molecule, more preferably an IgG1 or IgG2 molecule, and most preferably an IgG1 molecule, or a fragment thereof. In a preferred embodiment, the bispecific antibody molecule is a complete immunoglobulin molecule.

[0060] In other embodiments, the bispecific antibody molecule may be an antigen-binding fragment comprising a CDR-based antigen-binding site for MSLN and an antigen-binding site for CD137 located in the constant domain. The antigen-binding fragment may be an scFv, Fab, Fcab, VhH, monovalent IgG, diabody or triabody, IGNAR, V-NAR, hclgG, minibody, or nanobody. For example, the antigen-binding fragment may be an scFv-Fc fusion in which the scFv binds to MSLN and the Fc binds to CD137, or a minibody comprising an scFv linked to a CH3 domain (Hu et al. (1996), Cancer Res., 56(13):3055-61).

[0061] In a preferred embodiment, the bispecific antibody molecule is a mAb 2 (TM) bispecific antibodies. The mAbs referred to herein 2 Bispecific antibodies are IgG immunoglobulins that contain a CDR-based antigen-binding site in each of their variable regions and at least one antigen-binding site in the constant domain of the antibody molecule.

[0062] Antibodies and methods for their construction and use are well known in the art and are described, for example, in Holliger and Hudson, 2005. It is possible to take monoclonal and other antibodies and use recombinant DNA technology techniques to generate other antibodies or chimeric molecules that retain the specificity of the original antibody. Such techniques may involve introducing the CDRs or variable regions of one antibody molecule into another antibody molecule (EP-A-184187, GB 2188638A and EP-A-239400). New antibodies against known targets can be routinely generated by those skilled in the art and can be achieved without undue burden.

[0063] A number of antibody molecules that bind to MSLN and CD137 are known from WO 2020 / 011976. The antibody M9657 of the present application is identical to the antibody FS22-172-003-AA / FS28-256-271 of WO 2020 / 011976. Any of these antibodies can be used. Thus, the CDR-based antigen-binding site of the bispecific antibody molecule can be composed of three VH CDRs or three VL CDRs, preferably three VH CDRs and three VL CDRs, of the following antibodies: The CDRs may include: antibodies FS22-172-003-AA / FS28-256-271, FS22-172-003-AA / FS28-024-052, FS22-172-003-AA / FS28-256-021, FS22-172-003-AA / FS28-256-012, FS22-172-003- AA / FS28-256-023, FS22-172-003-AA / FS28-256-024, FS22-172-003-AA / FS28-256-02 6, FS22-172-003-AA / FS28-256-027, FS22-172-003-AA / FS28-256-001, FS22-172-003- AA / FS28-256-005, FS22-172-003-AA / FS28-256-014, FS22-172-003-AA / FS28-256-01 8, FS22-172-003-AA / FS28-256, FS22-172-003-AA / FS28-024-051, FS22-172-003-AA / F S28-024-053, or FS22-172-003-AA / FS28-024, preferably antibody FS22-172-003-AA / FS28-256-271 or FS22-172-003-AA / FS28-024-052, most preferably antibody FS22-172-003-AA / FS28-256-271.

[0064] The sequences of the CDRs can be readily determined from the VH and VL domain sequences of the antibody molecule using conventional techniques. The following antibodies: FS22-172-003-AA / FS28-256-271, FS22-172-003-AA / FS28-024-052, FS22-172-003-AA / FS28-256-021, FS22-172-003-AA / FS28-256-012, FS22-172-003-AA / FS28-256-023, FS22-172-003-AA / FS28-256-024, FS22-172-003-AA / FS28-256-026, FS22-172-003-AA / FS28-256-027, FS22-172-003-AA / FS28-256-001, FS22- The VH and VL domain sequences of 172-003-AA / FS28-256-005, FS22-172-003-AA / FS28-256-014, FS22-172-003-AA / FS28-256-018, FS22-172-003-AA / FS28-256, FS22-172-003-AA / FS28-024-051, FS22-172-003-AA / FS28-024-053, and FS22-172-003-AA / FS28-024 are set forth herein, and therefore the CDRs of the three VH and three VL domains of the antibodies can be determined from said sequences. CDR sequences can be determined, for example, according to Kabat et al., 1991 or the international ImMunoGeneTics information system (IMGT) (Lefranc et al., 2015).

[0065] Bispecific antibody molecules may or may not have a LALA mutation. The LALA mutation represents a type of mutation intended to disrupt the antibody effector function of an antibody molecule or its fragment. The LALA mutation is associated with several favorable antibody properties, such as reduced toxicity (Lo et al. (2017), The Journal of Biological Chemistry, 292(9):3900-3908). This mutation eliminates Fcγ receptor binding of the antibody molecule or its fragment and is located in the CH2 domain. The VH and VL domains of an antibody containing a LALA mutation, and thus the sequences of the VH domain CDR1, CDR2, and CDR3 and the VL domain CDR1, CDR2, and CDR3, are the same as those of an antibody without a LALA mutation. The LALA mutation involves the substitution of leucine residues at positions 1.3 and 1.2 of the CH2 domain with alanine (L1.3A and L1.2A) according to the IMGT numbering system. According to the Kabat numbering system, the LALA mutation constitutes an L247A L248A substitution. Complement activation (C1q binding) and ADCC are also known to be reduced by mutating the proline at position 114 in the CH2 domain to an alanine or glycine according to the IMGT numbering system (P114A or P114G) (Idusogie et al., 2000; Klein et al., 2016). According to the Kabat numbering system, this mutation constitutes a P348A or P348G substitution. This mutation can also be combined with the LALA mutation to generate antibody molecules with further reduced or no ADCC or CDC activity.

[0066] Thus, a bispecific antibody molecule may comprise a CH2 domain, wherein the CH2 domain comprises an alanine residue at position 1.3 and an alanine residue at position 1.2, where the amino acid numbering is according to the IMGT numbering system. A bispecific antibody molecule may comprise a CH2 domain, wherein the CH2 domain comprises an alanine residue at position 247 and an alanine residue at position 248, where the amino acid numbering is according to the Kabat numbering system. For example, the CH2 domain may have the amino acid sequence set forth in SEQ ID NO: 90. In an alternative embodiment, an antibody molecule may comprise a CH2, wherein the CH2 domain comprises an alanine residue at position 91. An antibody molecule may comprise a CH2, wherein the CH2 domain comprises an alanine residue at position 1.3, an alanine residue at position 1.2, and an alanine residue at position 114. For example, the CH2 domain may have the amino acid sequence set forth in SEQ ID NO: 92.

[0067] The sequences of the VH domain CDR1, CDR2 and CDR3 of the bispecific antibody molecule according to the IMGT numbering may be the sequences located at positions 27-38, 56-65 and 105-117, respectively, of the VH domain of the antibody molecule. The sequences of the VH domain CDR1, CDR2 and CDR3 of the bispecific antibody molecule according to Kabat numbering may be the sequences located at positions 31-35, 50-65 and 95-102 of the VH domain, respectively.

[0068] The sequences of the VL domain CDR1, CDR2 and CDR3 of the bispecific antibody molecule according to the IMGT numbering may be the sequences located at positions 27-38, 56-65 and 105-117 of the VL domain, respectively. The VL domain CDR1, CDR2 and CDR3 sequences of the bispecific antibody molecule according to Kabat numbering may be the sequences located at positions 24-34, 50-56 and 89-97 of the VL domain, respectively.

[0069] For example, the sequences of the VH domains CDR1, CDR2, and CDR3 for the following are as follows: (i) FS22-172-003-AA / FS28-256-271 can be as set forth in SEQ ID NOs: 4, 6, and 8, respectively; (ii) FS22-172-003-AA / FS28-024-052 can be as set forth in SEQ ID NOs: 20, 22, and 24, respectively; (iii) FS22-172-003-AA / FS28-256-021 can be as set forth in SEQ ID NOs: 4, 6, and 8, respectively; (iv) FS22-172-003-AA / FS28-256-012 can be as set forth in SEQ ID NOs: 4, 6, and 8, respectively; (v) FS22-172-003-AA / FS28-256-023 can be as set forth in SEQ ID NOs: 42, 6, and 44, respectively; (vi) FS22-172-003-AA / FS28-256-024 can be as set forth in SEQ ID NOs: 4, 6, and 8, respectively; (vii) FS22-172-003-AA / FS28-256-026 can be as set forth in SEQ ID NOs: 43, 6, and 45, respectively; (viii) FS22-172-003-AA / FS28-256-027 can be as set forth in SEQ ID NOs: 4, 6, and 8, respectively; (ix) FS22-172-003-AA / FS28-256-001 can be as set forth in SEQ ID NOs: 53, 6, and 55, respectively; (x) FS22-172-003-AA / FS28-256-005 can be as set forth in SEQ ID NOs: 53, 6, and 55, respectively; (xi) FS22-172-003-AA / FS28-256-014 can be as set forth in SEQ ID NOs: 60, 6, and 62, respectively; (xii) FS22-172-003-AA / FS28-256-018 can be as set forth in SEQ ID NOs: 43, 6, and 45, respectively; (xiii) FS22-172-003-AA / FS28-256 can be as set forth in SEQ ID NOs: 67, 6, and 55, respectively; (xiv) FS22-172-003-AA / FS28-024-051 can be as set forth in SEQ ID NOs: 21, 23, and 72, respectively; (xv) FS22-172-003-AA / FS28-024-053 can be as set forth in SEQ ID NOs: 21, 23, and 77, respectively; and (xvi) FS22-172-003-AA / FS28-024 can be as set forth in SEQ ID NOs: 21, 23, and 82, respectively; The CDR sequences herein are defined according to the IMGT numbering system.

[0070] The sequences of the VL domains CDR1, CDR2, and CDR3 for: (i) FS22-172-003-AA / FS28-256-271 can be as set forth in SEQ ID NOs: 12, 14, and 16, respectively; (ii) FS22-172-003-AA / FS28-024-052 can be as set forth in SEQ ID NOs: 12, 14, and 18, respectively; (iii) FS22-172-003-AA / FS28-256-021 can be as set forth in SEQ ID NOs: 12, 14, and 34, respectively; (iv) FS22-172-003-AA / FS28-256-012 can be as set forth in SEQ ID NOs: 12, 14, and 39, respectively; (v) FS22-172-003-AA / FS28-256-023 can be as set forth in SEQ ID NOs: 12, 14, and 34, respectively; (vi) FS22-172-003-AA / FS28-256-024 can be as set forth in SEQ ID NOs: 12, 14, and 49, respectively; (vii) FS22-172-003-AA / FS28-256-026 can be as set forth in SEQ ID NOs: 12, 14, and 49, respectively; (viii) FS22-172-003-AA / FS28-256-027 can be as set forth in SEQ ID NOs: 12, 14, and 16, respectively; (ix) FS22-172-003-AA / FS28-256-001 can be as set forth in SEQ ID NOs: 12, 14, and 34, respectively; (x) FS22-172-003-AA / FS28-256-005 can be as set forth in SEQ ID NOs: 12, 14, and 49, respectively; (xi) FS22-172-003-AA / FS28-256-014 can be as set forth in SEQ ID NOs: 12, 14, and 39, respectively; (xii) FS22-172-003-AA / FS28-256-018 can be as set forth in SEQ ID NOs: 12, 14, and 39, respectively; (xiii) FS22-172-003-AA / FS28-256 can be as set forth in SEQ ID NOs: 12, 14, and 39, respectively; (xiv) FS22-172-003-AA / FS28-024-051 can be as set forth in SEQ ID NOs: 12, 14, and 28, respectively; (xv) FS22-172-003-AA / FS28-024-053 can be as set forth in SEQ ID NOs: 12, 14, and 28, respectively; and (xvi) FS22-172-003-AA / FS28-024 can be as set forth in SEQ ID NOs: 12, 14, and 28, respectively; The CDR sequences herein are defined according to the IMGT numbering system.

[0071] For example, the sequences of the VH domains CDR1, CDR2, and CDR3 for the following are as follows: (i) FS22-172-003-AA / FS28-256-271 can be as set forth in SEQ ID NOs: 5, 7, and 9, respectively; (ii) FS22-172-003-AA / FS28-024-052 can be as set forth in SEQ ID NOs: 21, 23, and 25, respectively; (iii) FS22-172-003-AA / FS28-256-021 can be as set forth in SEQ ID NOs: 5, 31, and 9, respectively; (iv) FS22-172-003-AA / FS28-256-012 can be as set forth in SEQ ID NOs: 5, 31, and 9, respectively; (v) FS22-172-003-AA / FS28-256-023 can be as set forth in SEQ ID NOs: 44, 31, and 46, respectively; (vi) FS22-172-003-AA / FS28-256-024 can be as set forth in SEQ ID NOs: 5, 31, and 9, respectively; (vii) FS22-172-003-AA / FS28-256-026 can be as set forth in SEQ ID NOs: 44, 31, and 46, respectively; (viii) FS22-172-003-AA / FS28-256-027 can be as set forth in SEQ ID NOs: 5, 31, and 9, respectively; (ix) FS22-172-003-AA / FS28-256-001 can be as set forth in SEQ ID NOs: 54, 31, and 56, respectively; (x) FS22-172-003-AA / FS28-256-005 can be as set forth in SEQ ID NOs: 54, 31, and 56, respectively; (xi) FS22-172-003-AA / FS28-256-014 can be as set forth in SEQ ID NOs: 61, 31, and 63, respectively; (xii) FS22-172-003-AA / FS28-256-018 can be as set forth in SEQ ID NOs: 44, 31, and 46, respectively; (xiii) FS22-172-003-AA / FS28-256 can be as set forth in SEQ ID NOs: 68, 31, and 56, respectively; (xiv) FS22-172-003-AA / FS28-024-051 can be as set forth in SEQ ID NOs: 22, 24, and 73, respectively; (xv) FS22-172-003-AA / FS28-024-053 can be as set forth in SEQ ID NOs: 22, 24, and 78, respectively; and (xvi) FS22-172-003-AA / FS28-024 can be as set forth in SEQ ID NOs: 22, 24, and 83, respectively; CDR sequences herein are defined according to the Kabat numbering system.

[0072] The sequences of the VL domains CDR1, CDR2, and CDR3 for: (i) FS22-172-003-AA / FS28-256-271 can be as set forth in SEQ ID NOs: 13, 15, and 16, respectively; (ii) FS22-172-003-AA / FS28-024-052 can be as set forth in SEQ ID NOs: 13, 15, and 28, respectively; (iii) FS22-172-003-AA / FS28-256-021 can be as set forth in SEQ ID NOs: 13, 15, and 34, respectively; (iv) FS22-172-003-AA / FS28-256-012 can be as set forth in SEQ ID NOs: 13, 15, and 39, respectively; (v) FS22-172-003-AA / FS28-256-023 can be as set forth in SEQ ID NOs: 13, 15, and 34, respectively; (vi) FS22-172-003-AA / FS28-256-024 can be as set forth in SEQ ID NOs: 13, 15, and 49, respectively; (vii) FS22-172-003-AA / FS28-256-026 can be as set forth in SEQ ID NOs: 13, 15, and 49, respectively; (viii) FS22-172-003-AA / FS28-256-027 can be as set forth in SEQ ID NOs: 13, 15, and 16, respectively; (ix) FS22-172-003-AA / FS28-256-001 can be as set forth in SEQ ID NOs: 13, 15, and 34, respectively; (x) FS22-172-003-AA / FS28-256-005 can be as set forth in SEQ ID NOs: 13, 15, and 49, respectively; (xi) FS22-172-003-AA / FS28-256-014 can be as set forth in SEQ ID NOs: 13, 15, and 39, respectively; (xii) FS22-172-003-AA / FS28-256-018 can be as set forth in SEQ ID NOs: 13, 15, and 39, respectively; (xiii) FS22-172-003-AA / FS28-256 can be as set forth in SEQ ID NOs: 13, 15, and 39, respectively; (xiv) FS22-172-003-AA / FS28-024-051 can be as set forth in SEQ ID NOs: 13, 15, and 28, respectively; (xv) FS22-172-003-AA / FS28-024-053 can be as set forth in SEQ ID NOs: 13, 15, and 28, respectively; and (xvi) FS22-172-003-AA / FS28-024 can be as set forth in SEQ ID NOs: 13, 15, and 28, respectively; CDR sequences herein are defined according to the Kabat numbering system.

[0073] The CDR-based antigen-binding site may comprise the VH or VL domains, preferably the VH and VL domains, of the following antibodies: antibodies FS22-172-003-AA / FS28-256-271, FS22-172-003-AA / FS28-024-052, FS22-172-003-AA / FS28-256-021 ... 2-003-AA / FS28-256-012, FS22-172-003-AA / FS28-256-023, FS22-172-003-AA / FS28-256-02 4, FS22-172-003-AA / FS28-256-026, FS22-172-003-AA / FS28-256-027, FS22-172-003-AA / FS2 8-256-001, FS22-172-003-AA / FS28-256-005, FS22-172-003-AA / FS28-256-014, FS22-172-0 03-AA / FS28-256-018, FS22-172-003-AA / FS28-256, FS22-172-003-AA / FS28-024-051, FS22- 172-003-AA / FS28-024-053, or FS22-172-003-AA / FS28-024, preferably antibody FS22-172-003-AA / FS28-256-271 or FS22-172-003-AA / FS28-024-052, most preferably antibody FS22-172-003-AA / FS28-256-271.

[0074] Antibodies FS22-172-003-AA / FS28-256-271, FS22-172-003-AA / FS28-024-052, FS22-172-003-AA / FS28-256-021, FS22-172-003-AA / FS28-256-012, FS22-172-003-AA / FS28-256-023, FS22-172-003-AA / FS28-256-024, FS22-172-003-AA / FS28-256-026, FS2 2-172-003-AA / FS28-256-027, FS22-172-003-AA / FS28-256-001, FS22-172-003-AA / FS28-256-0 05, FS22-172-003-AA / FS28-256-014, FS22-172-003-AA / FS28-256-018, FS22-172-003-AA / FS28 The VH domains of FS22-172-003-AA / FS28-024-051, FS22-172-003-AA / FS28-024-053, and FS22-172-003-AA / FS28-024 may have the sequences set forth in SEQ ID NOs: 3, 19, 3, 3, 42, 3, 42, 3, 52, 52, 59, 42, 66, 71, 76, and 81, respectively.

[0075] Antibodies FS22-172-003-AA / FS28-256-271, FS22-172-003-AA / FS28-024-052, FS22-17 2-003-AA / FS28-256-021, FS22-172-003-AA / FS28-256-012, FS22-172-003-AA / FS28-256-023, FS22-172-003-AA / FS28-256-024, FS22-172-003-AA / FS28-256- 026, FS22-172-003-AA / FS28-256-027, FS22-172-003-AA / FS28-256-001, FS22- The VL domains of 172-003-AA / FS28-256-005, FS22-172-003-AA / FS28-256-014, FS22-172-003-AA / FS28-256-018, FS22-172-003-AA / FS28-256, FS22-172-003-AA / FS28-024-051, FS22-172-003-AA / FS28-024-053, and FS22-172-003-AA / FS28-024 may have the sequences set forth in SEQ ID NOs: 11, 27, 33, 38, 33, 48, 48, 11, 33, 48, 38, 38, 38, 27, 27, and 27, respectively.

[0076] In one embodiment, the bispecific antibody molecule of the present invention comprises a CD137 antigen-binding site. The CD137 antigen-binding site may be located in the constant domain of the antibody molecule, preferably in the CH3 domain. The CD137 antigen-binding site may comprise one or more modified structural loops in the constant domain of the antibody molecule. Engineering of antibody constant domain structural loops to create antigen-binding sites for target antigens is known in the art and is described, for example, in Wozniak-Knopp G et al. (2010) Protein Eng Des. 23 (4): 289-297; WO2006 / 072620 and WO2009 / 132876. The CD137 constant domain antigen-binding site contained in the antibody molecule of the present invention was identified through extensive selection and affinity maturation programs and preferentially binds to dimeric human CD137 over monomeric human CD137.

[0077] The CD137 antigen-binding site of the bispecific antibody molecule can comprise a first sequence and a second sequence, wherein the first sequence and the second sequence are located within the AB and EF structural loops, respectively, of the constant domain of the bispecific antibody molecule, preferably the CH3 domain. The first and second sequences are preferably the first and second sequences of FS22-172-003 set forth in SEQ ID NOs: 87 and 88, respectively. The first and second sequences are preferably located between positions 14 and 17 and between positions 91 and 99, respectively, of the CH3 domain of the bispecific antibody molecule, where residue numbering is according to the IMGT numbering system. The CD loop sequences of the bispecific antibody molecule are preferably unmodified, i.e., wild-type.

[0078] Thus, the CD loop sequence preferably has the sequence set forth in SEQ ID NO: 89. The CD loop sequence is preferably located at positions 43 to 78 of the CH3 domain of the bispecific antibody molecule, where residue numbering is according to IMGT numbering. In a preferred embodiment, the bispecific antibody molecule comprises a CH3 domain that comprises, has or consists of the CH3 domain sequence of FS22-172-003 as set forth in SEQ ID NO: 86. The CH3 domain of the bispecific antibody molecule may optionally comprise an additional lysine residue (K) immediately C-terminal to the CH3 domain sequence.

[0079] In a preferred embodiment, the antibody molecule comprises the heavy and / or light chains, preferably the heavy and light chains, of the following antibodies: (i) FS22-172-003-AA / FS28-256-271 set forth in SEQ ID NOs: 2 and 10, respectively; (ii) FS22-172-003-AA / FS28-024-052 set forth in SEQ ID NOs: 18 and 26, respectively; (iii) FS22-172-003-AA / FS28-256-021 set forth in SEQ ID NOs: 30 and 32, respectively; (iv) FS22-172-003-AA / FS28-256-012 set forth in SEQ ID NOs: 36 and 37, respectively; (v) FS22-172-003-AA / FS28-256-023 set forth in SEQ ID NOs: 41 and 32, respectively; (vi) FS22-172-003-AA / FS28-256-024 set forth in SEQ ID NOs: 30 and 47, respectively; (vii) FS22-172-003-AA / FS28-256-026 set forth in SEQ ID NOs: 41 and 47, respectively; (viii) FS22-172-003-AA / FS28-256-027 set forth in SEQ ID NOs: 30 and 10, respectively; (ix) FS22-172-003-AA / FS28-256-001 set forth in SEQ ID NOs: 51 and 32, respectively; (x) FS22-172-003-AA / FS28-256-005 set forth in SEQ ID NOs: 51 and 47, respectively; (xi) FS22-172-003-AA / FS28-256-014 set forth in SEQ ID NOs: 58 and 37, respectively; (xii) FS22-172-003-AA / FS28-256-018 set forth in SEQ ID NOs: 41 and 37, respectively; (xiii) FS22-172-003-AA / FS28-256 set forth in SEQ ID NOs: 65 and 37, respectively; (xiv) FS22-172-003-AA / FS28-024-051 set forth in SEQ ID NOs: 70 and 26, respectively; (xv) FS22-172-003-AA / FS28-024-053 set forth in SEQ ID NOs: 75 and 26, respectively; or (xvi) FS22-172-003-AA / FS28-024 as set forth in SEQ ID NOs: 80 and 26, respectively.

[0080] In a more preferred embodiment, the bispecific antibody molecule comprises the heavy and / or light chain, preferably the heavy and light chain, of antibody FS22-172-003-AA / FS28-256-271 or FS22-172-003-AA / FS28-024-052, most preferably antibody FS22-172-003-AA / FS28-256-271, wherein the heavy and light chain sequences of these antibodies are as described above.

[0081] The bispecific antibody molecules of the present invention may also comprise variants of the first, second, or third sequences, AB, CD, or EF structural loop sequences, CH3 domains, CH2 domains, CDRs, VH domains, VL domains, light chain and / or heavy chain sequences disclosed herein. Suitable variants can be obtained by sequence alteration or mutation and screening methods. In preferred embodiments, antibody molecules comprising one or more variant sequences retain one or more functional properties of the parent antibody molecule, e.g., binding specificity and / or binding affinity to MSLN and CD137. For example, antibody molecules comprising one or more variant sequences preferably bind to MSLN and / or CD137 with affinity comparable to or higher than that of the (parent) antibody molecule. The parent antibody molecule is an antibody molecule that does not contain the amino acid substitution(s), deletion(s), and / or insertion(s) incorporated into the variant antibody molecule.

[0082] Antibody molecules, comprising the following antibodies: FS22-172-003-AA / FS28-256-271, FS22-172-003-AA / FS28-024-052, FS22-172-003-AA / FS28-256-021, FS22-172-003-AA / FS28-256-012, FS22-172-003 -AA / FS28-256-023, FS22-172-003-AA / FS28-256-024, FS22-172-003-AA / FS28-256- 026, FS22-172-003-AA / FS28-256-027, FS22-172-003-AA / FS28-256-001, FS22-172- The antibody molecules comprising CDRs 1-6, the VH domain, and / or the heavy chain of FS22-172-003-AA / FS28-256-005, FS22-172-003-AA / FS28-256-014, FS22-172-003-AA / FS28-256-018, FS22-172-003-AA / FS28-256, FS22-172-003-AA / FS28-024-051, FS22-172-003-AA / FS28-024-053, or FS22-172-003-AA / FS28-024 may comprise an amino acid substitution at position 55 or 57 of the VH domain, wherein the numbering of amino acid residues is according to the IMGT numbering system.

[0083] For example, an antibody molecule may comprise CDRs 1-6, a VH domain, and / or a heavy chain of antibody FS22-172-003-AA / FS28-256-027, wherein the antibody molecule comprises an amino acid substitution at position 55 of the VH domain, where the numbering of the amino acid residues is according to the IMGT numbering system. For example, an antibody molecule of the present invention can comprise a first, second or third sequence, an AB, CD or EF structural loop sequence, a CH3 domain, a CH2 domain, a CDR, a VH domain, a VL domain, a light chain and / or a heavy chain sequence that has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.1%, at least 99.2%, at least 99.3%, at least 99.4%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, or at least 99.9% sequence identity to a structural loop, CH3 domain, CH2 domain, CDR, VH domain, VL domain, light chain or heavy chain sequence disclosed herein.

[0084] In preferred embodiments, a bispecific antibody molecule of the invention comprises a CH3 domain sequence that has at least 97%, at least 98%, at least 99%, at least 99.1%, at least 99.2%, at least 99.3%, at least 99.4%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, or at least 99.9% sequence identity to a CH3 domain disclosed herein.

[0085] In further preferred embodiments, the bispecific antibody molecule has or comprises a CH2 domain sequence that has at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.1%, at least 99.2%, at least 99.3%, at least 99.4%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, or at least 99.9% sequence identity to a CH2 domain disclosed herein.

[0086] Sequence identity is generally defined with reference to the GAP algorithm (Wisconsin GCG package, Accelerys Inc, San Diego, USA). GAP uses the Needleman and Wunsch algorithm to align two complete sequences, maximizing the number of matches and minimizing the number of gaps. Generally, default parameters are used, with a gap creation penalty of 12 and a gap extension penalty of 4. While the use of GAP may be preferred, other algorithms, such as BLAST (using the method of Altschul et al., 1990), FASTA (using the method of Pearson and Lipman, 1988), or the Smith-Waterman algorithm (Smith and Waterman, 1981), or the TBLASTN program (Altschul et al., 1990, supra), can be used, generally with default parameters. In particular, the psi-Blast algorithm (Altschul et al., 1997) may also be used.

[0087] Bispecific antibody molecules of the invention may also comprise a first, second or third sequence, AB, CD or EF structural loop sequence, CH3 domain, CH2 domain, VH domain, VL domain, light chain and / or heavy chain that has one or more amino acid sequence changes (addition, deletion, substitution and / or insertion of amino acid residues), preferably no more than 20 changes, no more than 15 changes, no more than 10 changes, no more than 5 changes, no more than 4 changes, no more than 3 changes, no more than 2 changes, or no more than 1 change, compared to the first, second or third sequence, AB, CD or EF structural loop sequence, CH3 domain, CH2 domain, Fcab, CDR, VH domain, VL domain, light chain or heavy chain sequence disclosed herein. In particular, changes may be made to one or more framework regions of the antibody molecule outside the VH and VL domain sequences and / or to one or more framework regions of the CH3 domain. For example, variations can occur in the CH3 domain outside of the sequences described herein, as first, second and third sequences, or as AB, CD or EF structural loop sequences.

[0088] The bispecific antibody molecule may comprise a VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and / or VL CDR3 that has one or more amino acid sequence changes (addition, deletion, substitution and / or insertion of amino acid residues) compared to the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and / or VL CDR3 disclosed herein, preferably no more than three, no more than two, or one change.

[0089] In a preferred embodiment, a bispecific antibody molecule of the invention comprises a CH3 domain sequence which has one or more amino acid sequence changes (addition, deletion, substitution and / or insertion of amino acid residues) compared to the CH3 domains disclosed herein, preferably no more than 20 changes, no more than 15 changes, no more than 10 changes, no more than 5 changes, no more than 4 changes, no more than 3 changes, no more than 2 changes, or no more than 1 change.

[0090] In preferred embodiments, where one or more amino acids are substituted with another amino acid, the substitution is a conservative substitution, for example, according to the following table: In some embodiments, amino acids in the same category in the middle column are substituted for each other, i.e., a non-polar amino acid is substituted for another non-polar amino acid, for example. In some embodiments, amino acids in the same row in the rightmost column are substituted for each other. [Table 1]

[0091] In some embodiments, the substitution(s) are functionally conservative, i.e., in some embodiments, the substitution does not affect (or does not substantially affect) one or more functional properties (e.g., binding affinity) of an antibody molecule comprising the substitution, compared to a comparable unsubstituted antibody molecule.

[0092] The term "chemotherapeutic agent" refers to a broad range of drugs used in the treatment of cancer. Chemotherapeutic agents may be naturally occurring compounds or may be partially or wholly synthetically produced.

[0093] For example, the chemotherapeutic agent may be an alkylating agent, a nitrosourea, an antimetabolite, a cytotoxic antibiotic, a topoisomerase inhibitor, a mitotic inhibitor, a corticosteroid, or any of the drug groups including trans-retinoic acid, arsenic trioxide, asparaginase, eribulin, hydroxyurea, ixabepilone, mitotane, omacetaxine, pegaspargase, procarbazine, romidepsin, or vorinostat. The topoisomerase inhibitor may be a type I topoisomerase inhibitor or a type II topoisomerase inhibitor. The mitotic inhibitor may be a taxane or a vinca alkaloid.

[0094] Preferably, the chemotherapeutic agent is an alkylating agent or an antimetabolite. Preferably, the antimetabolite is selected from the group consisting of azacitidine, 5-fluorouracil (5-FU), 6-mercaptopurine (6-MP), capecitabine, cladribine, clofarabine, cytarabine, decitabine, floxuridine, fludarabine, gemcitabine, hydroxyurea, methotrexate, nelarabine, pemetrexed, pentostatin, pralatrexate, thioguanine, and the combination of trifluridine / tipiracil. More preferably, the antimetabolite is gemcitabine. Preferably, the alkylating agent is selected from the group comprising altretamine, bendamustine, busulfan, carboplatin, carmustine, chlorambucil, cisplatin, cyclophosphamide (CPA), dacarbazine, ifosfamide, lomustine, mechlorethamine, melphalan, oxaliplatin, temozolomide, thiotepa, and trabectedin. More preferably, the alkylating agent is cisplatin.

[0095] Preferably, the nitrosourea is selected from the group consisting of camstine, lomustine, and streptozocin. Preferably, the cytotoxic antibiotic is selected from the group consisting of daunorubicin, doxorubicin, liposomal doxorubicin, epirubicin, idarubicin, valrubicin, bleomycin, dactinomycin, and mitomycin C. Preferably, the type I topoisomerase inhibitor is selected from the group consisting of irinotecan, liposomal irinotecan, and topotecan. Preferably, the type II topoisomerase inhibitor is selected from the group consisting of etoposide, mitoxantrone, and teniposide. Preferably, the taxane is selected from the group consisting of cabazitaxel, docetaxel, nab-paclitaxel, and paclitaxel. Preferably, the vinca alkaloid is selected from the group consisting of vinblastine, vincristine, liposomal vincristine, and vinorelbine. Preferably, the corticosteroid is selected from the group comprising prednisone, methylprednisolone, and dexamethasone.

[0096] The term "tumor" refers to a mass of cells of abnormal size and / or composition resulting from increased proliferation and / or prolonged survival of the cells. Tumors can be benign or malignant; in the latter case, they are called "cancer." Thus, a "tumor" cell is one that has an abnormally increased ability to divide and / or resist cell death compared to other cells of the same cell type.

[0097] Cancer is characterized by the abnormal proliferation of malignant tumor cells. When a specific type of cancer, such as ovarian cancer, is mentioned, this refers to the abnormal proliferation of malignant cells in related tissues, such as breast tissue. A secondary cancer that is located in the breast but is the result of the abnormal proliferation of malignant cells in another tissue, such as ovarian tissue, is not breast cancer as used herein, but is ovarian cancer.

[0098] MSLN is expressed on the surface of some tumor cells, and high expression levels of soluble MSLN correlate with poor prognosis in several cancers. Anti-MSLN antibodies are being investigated as anticancer therapies. These anti-MSLN antibodies either directly induce cell killing through their ADCC activity or are used in the form of ADCs.

[0099] Thus, cancers to be treated with a bispecific antibody molecule that binds to MSLN and CD137 in combination with a chemotherapeutic agent preferably express or are determined to express MSLN. More preferably, the cells of the treated cancer contain or are determined to contain MSLN on their cell surface, i.e., are determined to contain cell surface-bound MSLN.

[0100] The cancer preferably comprises or is determined to comprise tumor-infiltrating lymphocytes (TILs) that express CD137. Specifically, the TILs preferably comprise or are determined to comprise CD137 on their cell surface.

[0101] Cancer can be primary cancer or secondary cancer.Accordingly, the antibody molecule that binds to MSLN and CD137 described herein can be used in combination with a chemotherapeutic agent in a method for treating cancer in an individual, wherein the cancer is a primary tumor and / or tumor metastasis. The cancer to be treated may be a solid tumor.

[0102] As described above, the cancer to be treated can be a cancer that expresses MSLN or a cancer that has been confirmed to express MSLN. Preferably, the cancer is selected from the group including ovarian cancer, pancreatic adenocarcinoma, mesothelioma, cervical cancer, and non-small cell lung cancer.

[0103] The patient to be treated can be selected for treatment when cancer expresses MSLN.The patient can be selected when cancer is confirmed to express MSLN.Preferably, the patient is selected for treatment when cancer is ovarian cancer, pancreatic adenocarcinoma, mesothelioma, cervical cancer or non-small cell lung cancer and expresses MSLN.

[0104] The present inventors have shown that the combined treatment of FS122m (SEQ ID NO: 84 and SEQ ID NO: 85) with either cisplatin or gemcitabine resulted in tumor growth delay, tumor volume reduction, increased median survival time, and increased percentage of mice with complete tumor regression in ST26 and JC mouse tumor models, which were greater than the combined tumor growth delay, tumor volume reduction, increased median survival time, and increased percentage of mice with complete tumor regression observed when mice were treated with either FS122m or cisplatin or gemcitabine alone.Therefore, the present inventors have shown that the combined treatment of FS122m with either cisplatin or gemcitabine synergistically delayed tumor growth, reduced tumor volume, increased median survival time, and increased percentage of mice with complete tumor regression in ST26 and JC mouse tumor models.

[0105] In one embodiment, the antibody molecule that binds to MSLN and CD137, when combined with a chemotherapeutic agent, therefore, reduces tumor volume, delays tumor growth, increases survival, and / or increases the proportion of patients with complete tumor regression. Preferably, the antibody molecule that binds to MSLN and CD137, when combined with a chemotherapeutic agent, reduces tumor volume, delays tumor growth, increases survival, and / or increases the proportion of patients with complete tumor regression to a statistically significantly greater extent than monotherapy with the bispecific antibody molecule that binds to MSLN and CD137 or monotherapy with the chemotherapeutic agent. More preferably, the bispecific antibody molecule that binds to MSLN and CD137, when combined with a chemotherapeutic agent, synergistically reduces tumor volume, delays tumor growth, increases survival, and / or increases the proportion of patients with complete tumor regression, i.e., compared to the combined tumor growth delay, tumor volume reduction, increased median survival, and / or increased proportion of patients with complete tumor regression when patients are treated with the antibody molecule that binds MSLN and CD137 or the chemotherapeutic agent alone.

[0106] In one embodiment, the antibody molecule that binds to MSLN and CD137 and the chemotherapeutic agent are administered as a first ("front") line treatment (e.g., initial or first treatment). In another embodiment, the antibody molecule that binds to MSLN and CD137 and the chemotherapeutic agent are administered as a second line treatment (e.g., after initial treatment with the same or different therapeutic agent, including after relapse and / or if first treatment has failed).

[0107] As used herein, "administering" refers to the physical introduction of a therapeutic agent into a subject using any of a variety of methods and delivery systems known to those skilled in the art. Exemplary routes of administration for antibody molecules and / or chemotherapeutic agents that bind to MSLN and CD137 include intravenous, intramuscular, subcutaneous, intraperitoneal, spinal, or other parenteral routes, such as injection or infusion. The phrase "parenteral administration" as used herein refers to modes of administration other than enteral and topical administration, typically by injection, including, but not limited to, intravenous, intramuscular, intraarterial, intrathecal, intralymphatic, intralesional, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injection and infusion, and in vivo electroporation. Therapeutic agents may also be administered via non-parenteral routes. Non-parenteral routes include topical, epidermal, or mucosal routes of administration, such as oral, intranasal, vaginal, rectal, sublingual, or topical.

[0108] Therefore, in some embodiments, the antibody molecule and / or chemotherapeutic agent that binds to MSLN and CD137 is administered parenterally. The antibody molecule and / or chemotherapeutic agent that binds to MSLN and CD137 may be administered intravenously, intramuscularly, subcutaneously, intraperitoneally, or spinally. Alternatively, the antibody molecule and / or chemotherapeutic agent that binds to MSLN and CD137 may be administered by injection or infusion.

[0109] In other embodiments, the antibody molecule and / or chemotherapeutic agent that binds to MSLN and CD137 is administered non-parenterally. The antibody molecule and / or chemotherapeutic agent that binds to MSLN and CD137 may be administered orally, intranasally, vaginally, rectally, sublingually, or topically.

[0110] "Concurrent administration" refers to simultaneous administration of the antibody molecule that binds to MSLN and CD137 and the chemotherapeutic agent in the same or separate formulations. "Sequential administration" refers to separate timely administration of the bispecific antibody molecule and the chemotherapeutic agent in separate formulations.

[0111] Thus, the antibody molecule that binds to MSLN and CD137 and the chemotherapeutic agent may be part of the same formulation or may be part of separate formulations. Preferably, the antibody molecule that binds to MSLN and CD137 and the chemotherapeutic agent are provided as separate formulations.

[0112] In one embodiment of the present invention, the antibody molecule that binds to MSLN and CD137 and the chemotherapeutic agent are administered simultaneously. For example, the antibody molecule that binds to MSLN and CD137 may be administered together with the chemotherapeutic agent in the same formulation. Alternatively, the antibody molecule that binds to MSLN and CD137 and the chemotherapeutic agent may be administered in separate formulations immediately before or after each other.

[0113] Preferably, the antibody molecule that binds to MSLN and CD137 and the chemotherapeutic agent are administered to the patient sequentially. More preferably, the antibody molecule that binds to MSLN and CD137 and the chemotherapeutic agent are administered to the patient within 4 days of each other, preferably within 3 days of each other, more preferably within 2 days of each other, or sequentially on the same day. The present invention also relates to a method of treating cancer, comprising administering to an individual in need thereof an antibody molecule that binds to MSLN and CD137 and a chemotherapeutic agent.

[0114] The administration may be a "therapeutically effective amount," which is sufficient to provide benefit to the individual. The actual amount administered, as well as the rate and time course of administration, will vary depending on the nature and severity of what is being treated, the particular individual being treated, the individual's clinical condition, the cause of the disorder, the site of delivery of the composition, the type of antibody molecule, the method of administration, the schedule of administration, and other factors known to physicians. Prescribing treatment, such as determining dosage, is within the responsibility of general practitioners and other physicians and may vary depending on the severity of the symptoms and / or the progression of the disease being treated. Appropriate dosages of antibody molecules are well known in the art (Ledermann et al., 1991; Bagshawe et al., 1991). Therapeutically effective or suitable dosages of antibody molecules can be determined by comparing in vitro and in vivo activity in animal models. Methods for extrapolating effective dosages in mice and other test animals to humans are known. The exact dosage will depend on many factors, including the size and location of the area to be treated and the precise nature of the antibody molecule.

[0115] Thus, the present invention may relate to a method for treating cancer, comprising administering to an individual in need thereof a therapeutically effective amount of an antibody molecule that binds to MSLN and CD137 and a therapeutically effective amount of a chemotherapeutic agent.

[0116] Also provided is a kit comprising an antibody molecule that binds to MSLN and CD137 and a chemotherapeutic agent. Preferably, the kit comprises an antibody molecule that binds to MSLN and CD137 and a pharmaceutically acceptable excipient, and a chemotherapeutic agent and a pharmaceutically acceptable excipient.

[0117] The kit may be a package including a first container containing an antibody molecule that binds to MSLN and CD137 and a second container containing a chemotherapeutic agent, and the package may include instructions for using the antibody molecule that binds to MSLN and CD137 in combination with the chemotherapeutic agent to treat cancer in an individual.

[0118] In one embodiment, the kit may be a package containing at least one dose of a pharmaceutical comprising an antibody molecule that binds to MSLN and CD137 and one dose of a pharmaceutical comprising a chemotherapeutic agent. Preferably, the kit contains at least one dose of a pharmaceutical comprising an antibody molecule that binds to MSLN and CD137 and a pharmaceutically acceptable excipient, and one dose of a chemotherapeutic agent and a pharmaceutically acceptable excipient. More preferably, the kit contains a package insert containing instructions for using the pharmaceutical to treat cancer in an individual.

[0119] In another embodiment, the kit may be a package including a first container and a second container, wherein the first container contains an antibody molecule that binds to MSLN and CD137, and the second container contains a chemotherapeutic agent. The first container may contain at least one dose of a medicament comprising an antibody molecule that binds to MSLN and CD137 and a pharmaceutically acceptable excipient, and the second container may contain at least one dose of a medicament comprising a chemotherapeutic agent. The package may further include an insert containing instructions for using the medicament to treat cancer in an individual.

[0120] The features disclosed in the foregoing description, or the following claims, or the accompanying drawings, are expressed in their specific form, or in terms of means for performing a disclosed function or a method or process for obtaining a disclosed result, and can, where appropriate, be utilized separately or in any combination of such features to realize the invention in various of its forms.

[0121] The features disclosed in the foregoing description, or the following claims, or the accompanying drawings, are expressed in their specific form, or in terms of means for performing a disclosed function or a method or process for obtaining a disclosed result, and can, where appropriate, be utilized separately or in any combination of such features to realize the invention in various of its forms.

[0122] While the invention has been described in conjunction with the exemplary embodiments set forth above, many equivalent modifications and variations will become apparent to those skilled in the art upon reading this disclosure. Accordingly, the exemplary embodiments of the invention set forth above are considered to be illustrative and not limiting. Various modifications to the described embodiments can be made without departing from the spirit and scope of the invention.

[0123] For the avoidance of doubt, the theoretical explanations provided herein are provided for the purpose of enhancing the understanding of the reader, and the inventors do not wish to be bound by any of these theoretical explanations. Any section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.

[0124] Throughout this specification, including the claims that follow, unless the context requires otherwise, the words "comprise" and "include," and variations such as "comprises," "comprising," and "including," are understood to imply the inclusion of a stated integer or step or group of integers or steps, but not the exclusion of any other integer or step or group of integers or steps.

[0125] It should be noted that, as used in the specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Ranges may be expressed herein as from "about" one particular value and / or to "about" another particular value. When such a range is expressed, another embodiment includes from the one particular value and / or to the other particular value. Similarly, when values ​​are expressed as approximations by use of the antecedent "about," it will be understood that the particular value forms another embodiment. The term "about" with respect to numerical values ​​is optional and means, for example, + / - 10%. [Example]

[0126] material and method The efficacy of the combination of an antibody molecule that binds to MSLN and CD137 and a chemotherapeutic agent in enhancing anti-cancer immune responses, compared with either the antibody molecule that binds to MSLN and CD137 or the chemotherapeutic agent alone, was evaluated by examining mean tumor volume over time and survival time in mouse tumor models, which are described in detail below.

[0127] CT26 cells were obtained from ATCC and cultured in RPMI medium containing 2 mM L-glutamine, 10 mM HEPES, 1 mM sodium pyruvate, 4500 mg / L glucose, 1500 mg / L sodium bicarbonate, and 10% fetal bovine serum (FBS). JC cells were obtained from ATCC and maintained in vitro as monolayer cultures in RPMI-1640 supplemented with 10% heat-inactivated FBS at WuXi. These tumor cell lines were individually maintained in vitro as monolayer cultures and grown at 37°C in a 5% O2 atmosphere. Cells were typically subcultured twice weekly with TrypLE treatment. Cells growing during the exponential growth phase were harvested and counted for tumor inoculation. Wild-type BALB / c female mice were purchased from Charles River Laboratories or Lingchang Biological Technology Co., Ltd. All mice were 8–12 weeks old at the start of the study and were bred and maintained as described below under conditions consistent with The Guide for the Care and Use of Laboratory Animals, 8th Edition. All animal experiments were performed in accordance with the guidelines of the EMD Serono Research Institutional (protocols 17-008, 20-005) and the Wuxi AppTec Animal Care and Use Committee (IACUC).

[0128] Upon arrival at the research institution's animal facility, all animals underwent a detailed physical examination, including weight measurement, by research staff. All animals were confirmed to be in satisfactory health. Animals were housed in EMD Serono's specific pathogen-free barrier animal facility. Mice were maintained in individual ventilated cages at constant temperature and humidity, with five mice per cage. Identification labels on each cage included information such as the animal number, sex, strain, date of receipt, treatment, study number, group number, and treatment start date. Animals were marked with ear notches or ear tags. The animal room was maintained at 20–26°C and humidity at 40–70%. Lighting was on a 12-hour light / dark cycle. Animals were fed standard, certified, commercially available laboratory chow ad libitum. Maximum allowable concentrations of contaminants in the chow were controlled by the manufacturer and regularly analyzed. Animals were provided with ad libitum access to autoclaved municipal tap water suitable for human consumption. The dietary materials were believed to be free of known contaminants that could affect tumor growth. Approximately one week was allowed between the animals' arrival and tumor inoculation to allow them to acclimate to the laboratory environment. All procedures related to the handling, care, and treatment of animals in this study were performed in accordance with guidelines approved by the institutional animal care and use committees (IACUC) at EMD Serono and in accordance with the guidance of the Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC).

[0129] Using the CT26 and JC tumor models, the antitumor efficacy of a combination of an antibody molecule that binds to MSLN and CD137 and a chemotherapeutic agent was investigated compared with monotherapy with either an antibody molecule that binds to MSLN and CD137 or a chemotherapeutic agent. Mice were treated with the anti-mMSLN-mCD137-huIgG1-LALA antibody molecule FS122m in combination with either of two chemotherapeutic agents, cisplatin or gemcitabine. In the CT26 model, female BALB / c mice were inoculated with 0.3 × 10 IgG1-binding IgG1-hMSLN antibody molecules in 0.1 mL of PBS into the right flank. 6CT26 cells were inoculated by subcutaneous (sc) injection. Mice developed tumors with an average tumor volume of approximately 50–100 mm. 3 When the mice reached 100 mg / kg, they were randomly assigned to treatment groups (n=10 mice / group). For the JC model, female BALB / c mice were inoculated with JC tumor cells (5×10) in 0.1 mL of PBS into the right upper flank. 6 ) was inoculated by sc injection. Treatment was initiated 14 days after tumor inoculation when the average tumor size reached approximately 80 mm 3 Treatment was initiated when the FS122m and anti-HEL hIgG1 LALA isotype control were administered at 5 mg / kg by intraperitoneal (ip) injection on days 1, 3, and 5 of treatment (days 0, 2, and 4). Cisplatin was administered at 10 mg / kg by ip injection on day 0, and gemcitabine was administered at 120 mg / kg by ip injection on day 0.

[0130] Mortality checks were performed daily. Animals were checked daily for the effects of tumor growth and treatment on normal behavior, such as motor ability, food and water consumption (observation only), eye / hair matting, and any other abnormal effects described in the protocol. Body weight was recorded twice weekly, and mice that lost 20% of their initial weight were humanely sacrificed. Mice were kept at 4°C until the subcutaneous tumors reached 2500 mm 3 When the volume reached 100 mg / kg, the animals were humanely euthanized.

[0131] If tumor ulceration occurred, animals with ulcerated tumors were monitored at least three times per week, increasing to once daily depending on clinical signs. Ulcerated tumors that did not form scabs were cleaned with an appropriate wound cleanser (e.g., Novalsan). Antibiotic cream was applied to ulcerations / lesions only if directed by veterinary staff.

[0132] Criteria for euthanasia included lesions that did not heal or form scabs within 1 week, lesions exceeding 5 mm in diameter, lesions that cavitated or showed signs of infection (e.g., presence of pus) or bleeding, or animals showing signs of discomfort (e.g., excessive licking or chewing at the site) or systemic signs of illness (lethargy, decreased activity, decreased food intake, poor body condition, or weight loss). Veterinary staff were contacted to discuss possible exceptions.

[0133] Animals were euthanized if found to be moribund. Examples of clinical morbidity may include hunched posture, persistent recumbency, and unresponsiveness to handling or other stimuli, signs of severe organ or systemic failure, weakness, hypothermia, central nervous system defects (convulsions), respiratory signs (increased respiratory rate, labored breathing, coughing, rales), and gastrointestinal signs (diarrhea lasting more than 2 days, jaundice). Animals exhibiting the above clinical problems were humanely sacrificed with CO2.

[0134] Body weight was measured and recorded twice weekly until the endpoint was reached. Tumor size was measured twice weekly in three dimensions using calipers, and volume was calculated in mm using the formula: width x length x height x 0.5236. 3 It is expressed as:

[0135] Differences in tumor growth between treatment groups were determined using two-way analysis of variance (ANOVA) and Tukey's multiple comparison test. The significance of survival was determined using the Log-rank (Mantel-Cox) test. All analyses were performed using the GraphPad Prism software package (Prism 5 for Windows, Version 8.0, GraphPad Software Inc., San Diego, CA), and statistical significance was accepted at the p≦0.05 level.

[0136] Example 1: In vivo proof of concept To test whether the combination of an antibody molecule that binds to MSLN and CD137 and a chemotherapeutic agent exhibits improved antitumor activity compared with monotherapy with a bispecific antibody molecule or a chemotherapeutic agent, an in vivo mouse tumor model was treated with the combination of an antibody molecule that binds to MSLN and CD137 and a chemotherapeutic agent. The results of tumor growth delay, tumor volume reduction, and survival rate were then compared with monotherapy with an antibody molecule that binds to MSLN and CD137 and a chemotherapeutic agent.

[0137] Because there is no cross-reactivity between the bispecific anti-MSLN / CD137 antibody molecule M9657 and mouse MSLN and mouse CD137, we developed the anti-mMSLN-mCD137-huIgG1-LALA (FS112m) bispecific antibody as an alternative antibody (SEQ ID NOs: 84 and 85), which has similar binding affinity for mouse CD137 as M9657 for human CD137. Cisplatin and gemcitabine are two chemotherapeutic agents that belong to the groups of alkylating agents and antimetabolites, respectively. FS112m was combined with either cisplatin or gemcitabine, and the tumor efficacy of this combination was compared with FS112m monotherapy and cisplatin or gemcitabine monotherapy in two different mouse tumor models. To this end, female mice were inoculated with either CT26 colon cancer cells or JC breast cancer cells, and tumors with an average tumor volume of approximately 50–100 mm were grown. 3 Mice were inoculated, treated, and terminated according to the methods described in Materials and Methods.

[0138] 1.1 Combination of FS221m and cisplatin The antitumor efficacy of the combination of FS122m and cisplatin was first evaluated in a CT26 colon tumor model in BALB / c mice. FS122m and cisplatin monotherapy induced significant tumor growth inhibition (TGI) (61.3% and 77.8%, respectively) compared with the isotype control (both P ≤ 0.0001, day 17) and prolonged median survival (24 and 36.5 days, respectively) compared with the isotype control (17 days) (Figure 1A, B, and D). The combination of FS122m and cisplatin resulted in a TGI (96.3%) that was greater than that observed with FS122m monotherapy (P ≤ 0.0001, day 17) and cisplatin monotherapy (P ≤ 0.0001, day 17) (Figure 1A and D). The combination of FS122m and cisplatin also prolonged median survival (Fig. 1B) and induced complete tumor regression in 7 of 10 mice, compared with 2 of 10 mice treated with FS122m monotherapy and none of 10 mice treated with cisplatin monotherapy (Fig. 1D).

[0139] In JC tumor-bearing mice, treatment with cisplatin induced a marginal but statistically significant TGI (44.9%, P ≤ 0.0001) compared with the isotype control at day 22 after treatment initiation, and FS122m monotherapy induced a moderate TGI (60.1%, P ≤ 0.0001) at day 22 (Figure 2A and D). Cisplatin and FS122m monotherapy extended survival compared with the isotype control (22 days vs. 29 and 33 days, respectively) (Figure 2B). The combination of FS122m and cisplatin resulted in a TGI (83.3%) greater than that observed with cisplatin monotherapy (P ≤ 0.0001) and FS122m monotherapy (P ≤ 0.001) (Figure 2A and D). The combination of FS122m and cisplatin also prolonged median survival (36.5 days) (Fig. 2B) and induced complete tumor regression in 2 of 10 mice, whereas 2 of 10 mice treated with FS122m monotherapy experienced complete tumor regression and none of 10 mice treated with cisplatin monotherapy experienced complete tumor regression (Fig. 2D).

[0140] In both CT26 colon tumor- and JC tumor-bearing mice, the combination of FS122m and cisplatin resulted in a greater TGI than that observed when mice were treated with either FS122m or cisplatin monotherapy. The difference compared to FS122m and cisplatin monotherapy was particularly striking in the CT26 mouse tumor model, where the combination of FS122m and cisplatin resulted in complete tumor regression in 7 of 10 mice, compared with 2 of 10 mice treated with FS122m monotherapy and none in the cisplatin monotherapy group (Figure 1D). No significant weight loss was observed in either mouse tumor model after treatment with the combination of FS122m and cisplatin, suggesting that the combined treatment was well tolerated by the animals (Figures 1C and 2C).

[0141] 1.2 Combination of FS221m and gemcitabine The antitumor efficacy of the combination of FS122m and gemcitabine was evaluated for the first time in JC tumor-bearing mice. Treatment with gemcitabine or FS122m monotherapy induced marginal but statistically significant TGI (47.8% and 45.8%, respectively) compared with isotype control (both P ≤ 0.0001, day 25) (Figure 3A and D). Gemcitabine and FS122m monotherapy extended survival compared with isotype control (36 days and 34 days, respectively, compared with 27 days) (Figure 3B). The combination of FS122m and gemcitabine resulted in a greater TGI (80.2%) than gemcitabine monotherapy (P ≤ 0.0001) and FS122m monotherapy (P ≤ 0.001) (Figure 3A). The combination of FS122m and gemcitabine also prolonged median survival (74.5 days) (Fig. 3B) and induced complete tumor regression in 5 of 10 mice, whereas 1 of 10 mice treated with FS122m monotherapy experienced complete tumor regression and none of 10 mice treated with cisplatin monotherapy experienced complete tumor regression (Fig. 3D).

[0142] In a CT26 colon tumor model in BALB / c mice, FS122m and gemcitabine monotherapy induced moderate TGI (61.4% and 58.2%, respectively) compared with isotype control (P ≤ 0.0001, day 14) and prolonged median survival (35 and 22 days, respectively) compared with isotype control (17 days) (Figure 4A, B, D). The combination of FS122m and gemcitabine resulted in a greater TGI (86.1%) than gemcitabine monotherapy (P ≤ 0.0001, day 18) and FS122m monotherapy (P = 0.0113, day 18) (Figure 4A). The combination therapy also prolonged median survival (35 days) (Figure 4B) and induced complete tumor regression in 1 of 9 mice, whereas complete tumor regression was observed in 3 of 9 mice treated with FS122m monotherapy and in none of the 9 mice treated with cisplatin monotherapy (Figure 4D).

[0143] In both JC tumor-bearing mice and CT26 colon tumor-bearing mice, treatment with the combination of FS122m and gemcitabine resulted in a greater TGI than that observed when mice were treated with either FS122m or gemcitabine monotherapy. The difference compared to FS122m and gemcitabine monotherapy was particularly striking in the JC mouse tumor model, where the combination of FS122m and cisplatin resulted in complete tumor regression in 5 of 10 mice, compared with 1 of 10 mice treated with FS122m monotherapy and none in the gemcitabine monotherapy group (Figure 3D). The combination of FS122m and gemcitabine did not cause significant weight loss in either mouse tumor model, suggesting that the combination treatment was well tolerated in the animals (Figures 3C and 4C).

[0144] Sequence Listing Heavy chain annotation i.mAb 2 In the amino acid sequence of the heavy chain of Figure 1, the variable domain is shown in italics, the CDRs according to IMGT are shown in bold italics, the CDRs according to Kabat are shown in italic and underlined (thus the overlapping IMGT and Kabat CDR sequences are in bold italic and underlined), the CH1 domain is underlined, the hinge region is double underlined, the CH2 domain is shown in bold (and, if applicable, the position of the LALA mutation is shown in bold and underlined), the CH3 domain is in plain font, and the modified region of the CH3 structural loop is underlined (or not underlined if the loop is unchanged). ii. In the amino acid sequences of the variable domains, the CDRs according to IMGT are shown in bold italics and the CDRs according to Kabat are shown in italic and underlined (thus the overlapping IMGT and Kabat CDR sequences are shown in bold italic and underlined). iii. CDR amino acid sequences according to both IMGT and Kabat are provided.

[0145] Light chain annotation i.mAb2 In the amino acid sequence of the light chain of , the variable domain is shown in italics, the CDRs according to IMGT are shown in bold italics, and the CDRs according to Kabat are shown in italics and underlined (thus the overlapping sequences of the IMGT and Kabat CDRs are shown in bold italics and underlined). ii. In the amino acid sequences of the variable domains, the CDRs according to IMGT are shown in bold italics and the CDRs according to Kabat are shown in italic and underlined (thus the overlapping IMGT and Kabat CDR sequences are shown in bold italic and underlined). iii. CDR amino acid sequences according to both IMGT and Kabat are provided.

[0146] FS22-172-003-AA / FS28-256-271 mAb 2 The amino acid sequence of SEQ ID NO: 1 Heavy chain AA (no LALA) [ka] SEQ ID NO: 2 Heavy chain AA (with LALA) [ka] SEQ ID NO: 3 VH domain AA [ka] SEQ ID NO: 4 HCDR1 (AA) (IMGT) GFTFTHTY SEQ ID NO: 5 HCDR1 (AA) (Kabat) HTYMS SEQ ID NO: 6 HCDR2 (AA) (IMGT) ISPTYSTT SEQ ID NO: 7 HCDR2 (AA) Kabat) AISPTYSTTNYADSVKG SEQ ID NO: 8 HCDR3 (AA) (IMGT) ARYNAYHAALDY SEQ ID NO: 9 HCDR3 (AA) (Kabat) YNAYHAALDY SEQ ID NO: 10 Light chain AA [ka] SEQ ID NO: 11 VL domain AA [ka] SEQ ID NO: 12 LCDR1 (AA) (IMGT) QSVSSSY SEQ ID NO: 13 LCDR1 (AA) (Kabat) RASQSVSSSYLA SEQ ID NO: 14 LCDR2 (AA) (IMGT) GAS SEQ ID NO: 15 LCDR2 (AA) (Kabat) GASSRAT SEQ ID NO: 16 LCDR3 (AA) (IMGT) QQTVPYPYT SEQ ID NO: 16 LCDR3 (AA) (Kabat) QQTVPYPYT

[0147] FS22-172-003-AA / FS28-024-052 mAb 2 The amino acid sequence of SEQ ID NO: 17 Heavy chain AA (no LALA) [ka] SEQ ID NO: 18 Heavy chain AA (with LALA) [ka] SEQ ID NO: 19 VH domain AA [ka] SEQ ID NO: 20 HCDR1 (AA) (IMGT) GFTLSYSS SEQ ID NO: 21 HCDR1 (AA) (Kabat) YSSMS SEQ ID NO: 22 HCDR2 (AA) (IMGT) ITPSTGYT SEQ ID NO: 23 HCDR2 (AA) Kabat) FITPSTGYTHYADSVKG SEQ ID NO: 24 HCDR3 (AA) (IMGT) ARRALLFDY SEQ ID NO: 25 HCDR3 (AA) (Kabat) RALLFDY SEQ ID NO: 26 Light chain AA [ka] SEQ ID NO: 27 VL domain AA [ka] SEQ ID NO: 12 LCDR1 (AA) (IMGT) QSVSSSY SEQ ID NO: 13 LCDR1 (AA) (Kabat) RASQSVSSSYLA SEQ ID NO: 14 LCDR2 (AA) (IMGT) GAS SEQ ID NO: 15 LCDR2 (AA) (Kabat) GASSRAT SEQ ID NO: 28 LCDR3 (AA) (IMGT) QQASSYPLT SEQ ID NO: 28 LCDR3 (AA) (Kabat) QQASSYPLT

[0148] FS22-172-003-AA / FS28-256-021 mAb 2 The amino acid sequence of SEQ ID NO: 29 Heavy chain AA (no LALA) [ka] SEQ ID NO: 30 Heavy chain AA (with LALA) [ka] SEQ ID NO: 3 VH domain AA [ka] SEQ ID NO: 4 HCDR1 (AA) (IMGT) GFTFTHTY SEQ ID NO: 5 HCDR1 (AA) (Kabat) HTYMS SEQ ID NO: 6 HCDR2 (AA) (IMGT) ISPTYSTT SEQ ID NO: 31 HCDR2 (AA) Kabat) NISPTYSTTNYADSVKG SEQ ID NO: 8 HCDR3 (AA) (IMGT) ARYNAYHAALDY SEQ ID NO: 9 HCDR3 (AA) (Kabat) YNAYHAALDY SEQ ID NO: 32 Light chain AA [ka] SEQ ID NO: 33 VL domain AA [ka] SEQ ID NO: 12 LCDR1 (AA) (IMGT) QSVSSSY SEQ ID NO: 13 LCDR1 (AA) (Kabat) RASQSVSSSYLA SEQ ID NO: 14 LCDR2 (AA) (IMGT) GAS SEQ ID NO: 15 LCDR2 (AA) (Kabat) GASSRAT SEQ ID NO: 34 LCDR3 (AA) (IMGT) QQHNQYPNT SEQ ID NO: 34 LCDR3 (AA) (Kabat) QQHNQYPNT

[0149] FS22-172-003-AA / FS28-256-012 mAb 2 The amino acid sequence of SEQ ID NO: 35 Heavy chain AA (no LALA) [ka] SEQ ID NO: 36 Heavy chain AA (with LALA) [ka] SEQ ID NO: 3 VH domain AA [ka] SEQ ID NO: 4 HCDR1 (AA) (IMGT) GFTFTHTY SEQ ID NO: 5 HCDR1 (AA) (Kabat) HTYMS SEQ ID NO: 6 HCDR2 (AA) (IMGT) ISPTYSTT SEQ ID NO: 31 HCDR2 (AA) Kabat) NISPTYSTTNYADSVKG SEQ ID NO: 8 HCDR3 (AA) (IMGT) ARYNAYHAALDY SEQ ID NO: 9 HCDR3 (AA) (Kabat) YNAYHAALDY SEQ ID NO: 37 Light chain AA [ka] SEQ ID NO: 38 VL domain AA [ka] SEQ ID NO: 12 LCDR1 (AA) (IMGT) QSVSSSY SEQ ID NO: 13 LCDR1 (AA) (Kabat) RASQSVSSSYLA SEQ ID NO: 14 LCDR2 (AA) (IMGT) GAS SEQ ID NO: 15 LCDR2 (AA) (Kabat) GASSRAT SEQ ID NO: 39 LCDR3 (AA) (IMGT) QQSYYYPIT SEQ ID NO: 39 LCDR3 (AA) (Kabat) QQSYYYPIT

[0150] FS22-172-003-AA / FS28-256-023 mAb 2 The amino acid sequence of SEQ ID NO: 40 Heavy chain AA (no LALA) [ka] SEQ ID NO: 41 Heavy chain AA (with LALA) [ka] SEQ ID NO: 42 VH domain AA [ka] SEQ ID NO: 43 HCDR1 (AA) (IMGT) GFTFTQTY SEQ ID NO: 44 HCDR1 (AA) (Kabat) QTYMS SEQ ID NO: 6 HCDR2 (AA) (IMGT) ISPTYSTT SEQ ID NO: 31 HCDR2 (AA) Kabat) NISPTYSTTNYADSVKG SEQ ID NO: 45 HCDR3 (AA) (IMGT) ARYNAYQIGLDY SEQ ID NO: 46 HCDR3 (AA) (Kabat) YNAYQIGLDY SEQ ID NO: 32 Light chain AA [ka] SEQ ID NO: 33 VL domain AA [ka] SEQ ID NO: 12 LCDR1 (AA) (IMGT) QSVSSSY SEQ ID NO: 13 LCDR1 (AA) (Kabat) RASQSVSSSYLA SEQ ID NO: 14 LCDR2 (AA) (IMGT) GAS SEQ ID NO: 15 LCDR2 (AA) (Kabat) GASSRAT SEQ ID NO: 34 LCDR3 (AA) (IMGT) QQHNQYPNT SEQ ID NO: 34 LCDR3 (AA) (Kabat) QQHNQYPNT

[0151] FS22-172-003-AA / FS28-256-024 mAb 2 The amino acid sequence of SEQ ID NO: 29 Heavy chain AA (no LALA) [ka] SEQ ID NO: 30 Heavy chain AA (with LALA) [ka] SEQ ID NO: 3 VH domain AA [ka] SEQ ID NO: 4 HCDR1 (AA) (IMGT) GFTFTHTY SEQ ID NO: 5 HCDR1 (AA) (Kabat) HTYMS SEQ ID NO: 6 HCDR2 (AA) (IMGT) ISPTYSTT SEQ ID NO: 31 HCDR2 (AA) Kabat) NISPTYSTTNYADSVKG SEQ ID NO: 8 HCDR3 (AA) (IMGT) ARYNAYHAALDY SEQ ID NO: 9 HCDR3 (AA) (Kabat) YNAYHAALDY SEQ ID NO: 47 Light chain AA [ka] SEQ ID NO: 48 VL domain AA [ka] SEQ ID NO: 12 LCDR1 (AA) (IMGT) QSVSSSY SEQ ID NO: 13 LCDR1 (AA) (Kabat) RASQSVSSSYLA SEQ ID NO: 14 LCDR2 (AA) (IMGT) GAS SEQ ID NO: 15 LCDR2 (AA) (Kabat) GASSRAT SEQ ID NO: 49 LCDR3 (AA) (IMGT) QQALGYPHT SEQ ID NO: 49 LCDR3 (AA) (Kabat) QQALGYPHT

[0152] FS22-172-003-AA / FS28-256-026 mAb 2 The amino acid sequence of SEQ ID NO: 40 Heavy chain AA (no LALA) [ka] SEQ ID NO: 41 Heavy chain AA (with LALA) [ka] SEQ ID NO: 42 VH domain AA [ka] SEQ ID NO: 43 HCDR1 (AA) (IMGT) GFTFTQTY SEQ ID NO: 44 HCDR1 (AA) (Kabat) QTYMS SEQ ID NO: 6 HCDR2 (AA) (IMGT) ISPTYSTT SEQ ID NO: 31 HCDR2 (AA) Kabat) NISPTYSTTNYADSVKG SEQ ID NO: 45 HCDR3 (AA) (IMGT) ARYNAYQIGLDY SEQ ID NO: 46 HCDR3 (AA) (Kabat) YNAYQIGLDY SEQ ID NO: 47 Light chain AA [ka] SEQ ID NO: 48 VL domain AA [ka] SEQ ID NO: 12 LCDR1 (AA) (IMGT) QSVSSSY SEQ ID NO: 13 LCDR1 (AA) (Kabat) RASQSVSSSYLA SEQ ID NO: 14 LCDR2 (AA) (IMGT) GAS SEQ ID NO: 15 LCDR2 (AA) (Kabat) GASSRAT SEQ ID NO: 49 LCDR3 (AA) (IMGT) QQALGYPHT SEQ ID NO: 49 LCDR3 (AA) (Kabat) QQALGYPHT

[0153] FS22-172-003-AA / FS28-256-027 mAb 2 The amino acid sequence of SEQ ID NO: 29 Heavy chain AA (no LALA) [ka] SEQ ID NO: 30 Heavy chain AA (with LALA) [ka] SEQ ID NO: 3 VH domain AA [ka] SEQ ID NO: 4 HCDR1 (AA) (IMGT) GFTFTHTY SEQ ID NO: 5 HCDR1 (AA) (Kabat) HTYMS SEQ ID NO: 6 HCDR2 (AA) (IMGT) ISPTYSTT SEQ ID NO: 31 HCDR2 (AA) Kabat) NISPTYSTTNYADSVKG SEQ ID NO: 8 HCDR3 (AA) (IMGT) ARYNAYHAALDY SEQ ID NO: 9 HCDR3 (AA) (Kabat) YNAYHAALDY SEQ ID NO: 10 Light chain AA [ka] SEQ ID NO: 11 VL domain AA [ka] SEQ ID NO: 12 LCDR1 (AA) (IMGT) QSVSSSY SEQ ID NO: 13 LCDR1 (AA) (Kabat) RASQSVSSSYLA SEQ ID NO: 14 LCDR2 (AA) (IMGT) GAS SEQ ID NO: 15 LCDR2 (AA) (Kabat) GASSRAT SEQ ID NO: 16 LCDR3 (AA) (IMGT) QQTVPYPYT SEQ ID NO: 16 LCDR3 (AA) (Kabat) QQTVPYPYT

[0154] FS22-172-003-AA / FS28-256-001 mAb 2 The amino acid sequence of SEQ ID NO: 50 Heavy chain AA (no LALA) [ka] SEQ ID NO: 51 Heavy chain AA (with LALA) [ka] SEQ ID NO: 52 VH domain AA [ka] SEQ ID NO: 53 HCDR1 (AA) (IMGT) GFTFTETY SEQ ID NO: 54 HCDR1 (AA) (Kabat) ETYMS SEQ ID NO: 6 HCDR2 (AA) (IMGT) ISPTYSTT SEQ ID NO: 31 HCDR2 (AA) Kabat) NISPTYSTTNYADSVKG SEQ ID NO: 55 HCDR3 (AA) (IMGT) ARYNSYQGGLDY SEQ ID NO: 56 HCDR3 (AA) (Kabat) YNSYQGGLDY SEQ ID NO: 32 Light chain AA [ka] SEQ ID NO: 33 VL domain AA [ka] SEQ ID NO: 12 LCDR1 (AA) (IMGT) QSVSSSY SEQ ID NO: 13 LCDR1 (AA) (Kabat) RASQSVSSSYLA SEQ ID NO: 14 LCDR2 (AA) (IMGT) GAS SEQ ID NO: 15 LCDR2 (AA) (Kabat) GASSRAT SEQ ID NO: 34 LCDR3 (AA) (IMGT) QQHNQYPNT SEQ ID NO: 34 LCDR3 (AA) (Kabat) QQHNQYPNT

[0155] FS22-172-003-AA / FS28-256-005 mAb 2 The amino acid sequence of SEQ ID NO: 50 Heavy chain AA (no LALA) [ka] SEQ ID NO: 51 Heavy chain AA (with LALA) [ka] SEQ ID NO: 52 VH domain AA [ka] SEQ ID NO: 53 HCDR1 (AA) (IMGT) GFTFTETY SEQ ID NO: 54 HCDR1 (AA) (Kabat) ETYMS SEQ ID NO: 6 HCDR2 (AA) (IMGT) ISPTYSTT SEQ ID NO: 31 HCDR2 (AA) Kabat) NISPTYSTTNYADSVKG SEQ ID NO: 55 HCDR3 (AA) (IMGT) ARYNSYQGGLDY SEQ ID NO: 56 HCDR3 (AA) (Kabat) YNSYQGGLDY SEQ ID NO: 47 Light chain AA [ka] SEQ ID NO: 48 VL domain AA [ka] SEQ ID NO: 12 LCDR1 (AA) (IMGT) QSVSSSY SEQ ID NO: 13 LCDR1 (AA) (Kabat) RASQSVSSSYLA SEQ ID NO: 14 LCDR2 (AA) (IMGT) GAS SEQ ID NO: 15 LCDR2 (AA) (Kabat) GASSRAT SEQ ID NO: 49 LCDR3 (AA) (IMGT) QQALGYPHT SEQ ID NO: 49 LCDR3 (AA) (Kabat) QQALGYPHT

[0156] FS22-172-003-AA / FS28-256-014 mAb 2 The amino acid sequence of SEQ ID NO: 57 Heavy chain AA (no LALA) [ka] SEQ ID NO: 58 Heavy chain AA (with LALA) [ka] SEQ ID NO: 59 VH domain AA [ka] SEQ ID NO: 60 HCDR1 (AA) (IMGT) GFTFTDTY SEQ ID NO: 61 HCDR1 (AA) (Kabat) DTYMS SEQ ID NO: 6 HCDR2 (AA) (IMGT) ISPTYSTT SEQ ID NO: 31 HCDR2 (AA) Kabat) NISPTYSTTNYADSVKG SEQ ID NO: 62 HCDR3 (AA) (IMGT) ARYNAYAAGLDY SEQ ID NO: 63 HCDR3 (AA) (Kabat) YNAYAAGLDY SEQ ID NO: 37 Light chain AA [ka] SEQ ID NO: 38 VL domain AA [ka] SEQ ID NO: 12 LCDR1 (AA) (IMGT) QSVSSSY SEQ ID NO: 13 LCDR1 (AA) (Kabat) RASQSVSSSYLA SEQ ID NO: 14 LCDR2 (AA) (IMGT) GAS SEQ ID NO: 15 LCDR2 (AA) (Kabat) GASSRAT SEQ ID NO: 39 LCDR3 (AA) (IMGT) QQSYYYPIT SEQ ID NO: 39 LCDR3 (AA) (Kabat) QQSYYYPIT

[0157] FS22-172-003-AA / FS28-256-018 mAb 2 The amino acid sequence of SEQ ID NO: 40 Heavy chain AA (no LALA) [ka] SEQ ID NO: 41 Heavy chain AA (with LALA) [ka] SEQ ID NO: 42 VH domain AA [ka] SEQ ID NO: 43 HCDR1 (AA) (IMGT) GFTFTQTY SEQ ID NO: 44 HCDR1 (AA) (Kabat) QTYMS SEQ ID NO: 6 HCDR2 (AA) (IMGT) ISPTYSTT SEQ ID NO: 31 HCDR2 (AA) Kabat) NISPTYSTTNYADSVKG SEQ ID NO: 45 HCDR3 (AA) (IMGT) ARYNAYQIGLDY SEQ ID NO: 46 HCDR3 (AA) (Kabat) YNAYQIGLDY SEQ ID NO: 37 Light chain AA [ka] SEQ ID NO: 38 VL domain AA [ka] SEQ ID NO: 12 LCDR1 (AA) (IMGT) QSVSSSY SEQ ID NO: 13 LCDR1 (AA) (Kabat) RASQSVSSSYLA SEQ ID NO: 14 LCDR2 (AA) (IMGT) GAS SEQ ID NO: 15 LCDR2 (AA) (Kabat) GASSRAT SEQ ID NO: 39 LCDR3 (AA) (IMGT) QQSYYYPIT SEQ ID NO: 39 LCDR3 (AA) (Kabat) QQSYYYPIT

[0158] FS22-172-003-AA / FS28-256 mAb 2 The amino acid sequence of SEQ ID NO: 64 Heavy chain AA (no LALA) [ka] SEQ ID NO: 65 Heavy chain AA (with LALA) [ka] SEQ ID NO: 66 VH domain AA [ka] SEQ ID NO: 67 HCDR1 (AA) (IMGT) GFTFTNTY SEQ ID NO: 68 HCDR1 (AA) (Kabat) NTYMS SEQ ID NO: 6 HCDR2 (AA) (IMGT) ISPTYSTT SEQ ID NO: 31 HCDR2 (AA) Kabat) NISPTYSTTNYADSVKG SEQ ID NO: 55 HCDR3 (AA) (IMGT) ARYNSYQGGLDY SEQ ID NO: 56 HCDR3 (AA) (Kabat) YNSYQGGLDY SEQ ID NO: 37 Light chain AA [ka] SEQ ID NO: 38 VL domain AA [ka] SEQ ID NO: 12 LCDR1 (AA) (IMGT) QSVSSSY SEQ ID NO: 13 LCDR1 (AA) (Kabat) RASQSVSSSYLA SEQ ID NO: 14 LCDR2 (AA) (IMGT) GAS SEQ ID NO: 15 LCDR2 (AA) (Kabat) GASSRAT SEQ ID NO: 39 LCDR3 (AA) (IMGT) QQSYYYPIT SEQ ID NO: 39 LCDR3 (AA) (Kabat) QQSYYYPIT

[0159] FS22-172-003-AA / FS28-024-051 mAb 2 The amino acid sequence of SEQ ID NO: 69 Heavy chain AA (no LARA) [ka] SEQ ID NO: 70 Heavy chain AA (with LALA) [ka] SEQ ID NO: 71 VH domain AA [ka] SEQ ID NO: 21 HCDR1 (AA) (IMGT) GFTLSYSS SEQ ID NO: 22 HCDR1 (AA) (Kabat) YSSMS SEQ ID NO: 23 HCDR2 (AA) (IMGT) ITPSTGYT SEQ ID NO: 24 HCDR2 (AA) Kabat) FITPSTGYTHYADSVKG SEQ ID NO: 72 HCDR3 (AA) (IMGT) ARRALIFDY SEQ ID NO: 73 HCDR3 (AA) (Kabat) RALIFDY SEQ ID NO: 26 Light chain AA [ka] SEQ ID NO: 27 VL domain AA [ka] SEQ ID NO: 12 LCDR1 (AA) (IMGT) QSVSSSY SEQ ID NO: 13 LCDR1 (AA) (Kabat) RASQSVSSSYLA SEQ ID NO: 14 LCDR2 (AA) (IMGT) GAS SEQ ID NO: 15 LCDR2 (AA) (Kabat) GASSRAT SEQ ID NO: 28 LCDR3 (AA) (IMGT) QQASSYPLT SEQ ID NO: 28 LCDR3 (AA) (Kabat) QQASSYPLT

[0160] FS22-172-003-AA / FS28-024-053 mAb 2 The amino acid sequence of SEQ ID NO: 74 Heavy chain AA (no LALA) [ka] SEQ ID NO: 75 Heavy chain AA (with LALA) [ka] SEQ ID NO: 76 VH domain AA [ka] SEQ ID NO: 21 HCDR1 (AA) (IMGT) GFTLSYSS SEQ ID NO: 22 HCDR1 (AA) (Kabat) YSSMS SEQ ID NO: 23 HCDR2 (AA) (IMGT) ITPSTGYT SEQ ID NO: 24 HCDR2 (AA) Kabat) FITPSTGYTHYADSVKG SEQ ID NO: 77 HCDR3 (AA) (IMGT) ARRALVFDY SEQ ID NO: 78 HCDR3 (AA) (Kabat) RALVFDY SEQ ID NO: 26 Light chain AA [ka] SEQ ID NO: 27 VL domain AA [ka] SEQ ID NO: 12 LCDR1 (AA) (IMGT) QSVSSSY SEQ ID NO: 13 LCDR1 (AA) (Kabat) RASQSVSSSYLA SEQ ID NO: 14 LCDR2 (AA) (IMGT) GAS SEQ ID NO: 15 LCDR2 (AA) (Kabat) GASSRAT SEQ ID NO: 28 LCDR3 (AA) (IMGT) QQASSYPLT SEQ ID NO: 28 LCDR3 (AA) (Kabat) QQASSYPLT

[0161] FS22-172-003-AA / FS28-024 mAb 2 The amino acid sequence of SEQ ID NO: 79 Heavy chain AA (no LALA) [ka] SEQ ID NO: 80 Heavy chain AA (with LALA) [ka] SEQ ID NO: 81 VH domain AA [ka] SEQ ID NO: 21 HCDR1 (AA) (IMGT) GFTLSYSS SEQ ID NO: 22 HCDR1 (AA) (Kabat) YSSMS SEQ ID NO: 23 HCDR2 (AA) (IMGT) ITPSTGYT SEQ ID NO: 24 HCDR2 (AA) Kabat) FITPSTGYTHYADSVKG SEQ ID NO: 82 HCDR3 (AA) (IMGT) ARRALTFDY SEQ ID NO: 83 HCDR3 (AA) (Kabat) RALTFDY SEQ ID NO: 26 Light chain AA [ka] SEQ ID NO: 27 VL domain AA [ka] SEQ ID NO: 12 LCDR1 (AA) (IMGT) QSVSSSY SEQ ID NO: 13 LCDR1 (AA) (Kabat) RASQSVSSSYLA SEQ ID NO: 14 LCDR2 (AA) (IMGT) GAS SEQ ID NO: 15 LCDR2 (AA) (Kabat) GASSRAT SEQ ID NO: 28 LCDR3 (AA) (IMGT) QQASSYPLT SEQ ID NO: 28 LCDR3 (AA) (Kabat) QQASSYPLT

[0162] Amino acid sequence of FS122m (alternative anti-mMSLN mCD137 Fcab G1-AA): SEQ ID NO: 84 Heavy chain AA (with LALA) [ka] SEQ ID NO: 85 Light chain AA [ka]

[0163] All FS22-172-003 Fcab-containing mAbs 2 Amino acid sequences of the CH3 domain and modified regions of the CH3 AB and EF structural loops of the clones and FS22-172-003 Fcab SEQ ID NO: 86 CH3 domain [ka] SEQ ID NO: 87 AB Loop PYIIPPY SEQ ID NO: 88 EF Loop GADRWLE SEQ ID NO: 89 CD Loop SNGQPENNY

[0164] Amino acid sequences of the CH2 domain containing the LALA mutation, PA mutation, and LALA-PA mutation (mutations are bold and underlined) SEQ ID NO: 90 CH2 (LALA) [ka] SEQ ID NO: 91 CH2(PA) [ka] SEQ ID NO: 92 CH2 (LALA-PA) [ka]

[0165] References Numerous publications have been cited above in order to more fully describe and disclose the present invention and the state of the art to which it pertains. Full citations for these references are set forth below. Each of these references is incorporated herein in its entirety.

[0166] [Table 2-1] [Table 2-2] [Table 2-3] [Table 2-4] [Table 2-5]

[0167] For standard molecular biology techniques, see Sambrook, J., Russell, D.W. Molecular Cloning, A Laboratory Manual. 3rd ed. 2001, Cold Spring Harbor, New York: Cold Spring Harbor Laboratory Press.

Claims

1. 1. An antibody molecule that binds to MSLN and CD137 for use in a method of treating cancer in a patient, the method comprising administering the antibody in combination with a chemotherapeutic agent.

2. A method for treating cancer in an individual, comprising administering to the individual a therapeutically effective amount of an antibody molecule that binds to MSLN and CD137 and a chemotherapeutic agent.

3. The kit includes: (a) an antibody molecule that binds to MSLN and CD137 and a pharmaceutically acceptable excipient; and (b) a chemotherapeutic agent and a pharmaceutically acceptable excipient.

4. 4. The antibody molecule for use, method of treating cancer, or kit according to any one of claims 1 to 3, wherein the antibody molecule that binds to MSLN and CD137 is selected from the group consisting of: (a) a complementarity-determining region (CDR)-based antigen-binding site for MSLN; and (b) the CD137 antigen-binding site located in the CH3 domain of the antibody molecule; The antibody molecule, method, or kit comprising:

5. 5. The antibody molecule for use, method of treating cancer, or kit according to any one of claims 1 to 4, wherein the CDR-based antigen binding site against MSLN comprises CDRs 1 to 6 as set out below: (i) SEQ ID NOs: 4, 6, 8, 12, 14, and 16 [FS28-256-271], respectively; (ii) SEQ ID NOs: 20, 22, 24, 12, 14, and 28 [FS28-024-052], respectively; (iii) SEQ ID NOs: 4, 6, 8, 12, 14, and 34 [FS28-256-021], respectively; (iv) SEQ ID NOs: 4, 6, 8, 12, 14, and 39 [FS28-256-012], respectively; (v) SEQ ID NOs: 43, 6, 45, 12, 14, and 34 [FS28-256-023], respectively; (vi) SEQ ID NOs: 4, 6, 8, 12, 14, and 49 [FS28-256-024], respectively; (vii) SEQ ID NOs: 43, 6, 45, 12, 14, and 49 [FS28-256-026], respectively; (viii) SEQ ID NOs: 4, 6, 8, 12, 14, and 16 [FS28-256-027], respectively; (ix) SEQ ID NOs: 53, 6, 55, 12, 14, and 34 [FS28-256-001], respectively; (x) SEQ ID NOs: 53, 6, 55, 12, 14, and 49 [FS28-256-005], respectively; (xi) SEQ ID NOs: 60, 6, 62, 12, 14, and 39 [FS28-256-014], respectively; (xii) SEQ ID NOs: 43, 6, 45, 12, 14, and 39 [FS28-256-018], respectively; (xiii) SEQ ID NOs: 67, 6, 55, 12, 14, and 39 [FS28-256], respectively; (xiv) SEQ ID NOs: 21, 23, 72, 12, 14, and 28 [FS28-024-051], respectively; (xv) SEQ ID NOs: 21, 23, 77, 12, 14, and 28, respectively [FS28-024-053]; or (xvi) SEQ ID NOs: 21, 23, 82, 12, 14, and 28 [FS28-024], respectively; and wherein the CD137 antigen binding site comprises a first sequence and a second sequence located in the AB and EF structural loops of the CH3 domain, respectively, and wherein the first and second sequences have the sequences set forth in SEQ ID NOs: 87 and 88 [FS22-172-003], respectively. The antibody molecule, method, or kit.

6. An antibody molecule for use, a method for treating cancer or a kit according to any one of claims 1 to 5, wherein (i) the first sequence is located between positions 14 and 17 of the CH3 domain of the antibody molecule; and / or (ii) wherein the second sequence is located between positions 91 and 99 of the CH3 domain of the antibody molecule; and Here, the numbering of amino acid residues is according to the IMGT numbering system: The antibody molecule, method, or kit.

7. 7. The antibody molecule for use, method of treating cancer, or kit according to any one of claims 1 to 6, wherein the antibody molecule comprises a CH3 domain sequence set forth in SEQ ID NO: 86 [FS22-172-003].

8. 8. The antibody molecule for use, method of treating cancer or kit according to any one of claims 1 to 7, wherein the antibody molecule comprises the heavy and light chains of the following antibodies: (i) FS22-172-003-AA / FS28-256-271 set forth in SEQ ID NOs: 2 and 10, respectively; (ii) FS22-172-003-AA / FS28-024-052 set forth in SEQ ID NOs: 18 and 26, respectively; (iii) FS22-172-003-AA / FS28-256-021 set forth in SEQ ID NOs: 30 and 32, respectively; (iv) FS22-172-003-AA / FS28-256-012 set forth in SEQ ID NOs: 36 and 37, respectively; (v) FS22-172-003-AA / FS28-256-023 set forth in SEQ ID NOs: 41 and 32, respectively; (vi) FS22-172-003-AA / FS28-256-024 set forth in SEQ ID NOs: 30 and 47, respectively; (vii) FS22-172-003-AA / FS28-256-026 set forth in SEQ ID NOs: 41 and 47, respectively; (viii) FS22-172-003-AA / FS28-256-027 set forth in SEQ ID NOs: 30 and 10, respectively; (ix) FS22-172-003-AA / FS28-256-001 set forth in SEQ ID NOs: 51 and 32, respectively; (x) FS22-172-003-AA / FS28-256-005 set forth in SEQ ID NOs: 51 and 47, respectively; (xi) FS22-172-003-AA / FS28-256-014 set forth in SEQ ID NOs: 58 and 37, respectively; (xii) FS22-172-003-AA / FS28-256-018 set forth in SEQ ID NOs: 41 and 37, respectively; (xiii) FS22-172-003-AA / FS28-256 set forth in SEQ ID NOs: 65 and 37, respectively; (xiv) FS22-172-003-AA / FS28-024-051 set forth in SEQ ID NOs: 70 and 26, respectively; (xv) FS22-172-003-AA / FS28-024-053 set forth in SEQ ID NOs: 75 and 26, respectively; or (xvi) FS22-172-003-AA / FS28-024 set forth in SEQ ID NOs: 80 and 26, respectively; The antibody molecule, use, or kit comprising:

9. 9. The antibody molecule for use, method of treating cancer, or kit according to claim 8, wherein the antibody molecule comprises the heavy chain sequence set forth in SEQ ID NO: 2 and the light chain sequence set forth in SEQ ID NO: 10 [FS22-172-003-AA / FS28-256-271].

10. 10. The antibody molecule for use, method of treating cancer, or kit according to any one of the preceding claims, wherein the chemotherapeutic agent is an alkylating agent or an antimetabolite.

11. 11. The antibody molecule for use, method of treating cancer, or kit of claim 10, wherein the alkylating agent is selected from the group consisting of altretamine, bendamustine, busulfan, carboplatin, carmustine, chlorambucil, cisplatin, cyclophosphamide (CPA), dacarbazine, ifosfamide, lomustine, mechlorethamine, melphalan, oxaliplatin, temozolomide, thiotepa, and trabectedin.

12. 11. The antibody molecule for use, method of treating cancer, or kit of claim 10, wherein the antimetabolite is selected from the group consisting of azacitidine, 5-fluorouracil (5-FU), 6-mercaptopurine (6-MP), capecitabine, cladribine, clofarabine, cytarabine, decitabine, floxuridine, fludarabine, gemcitabine, hydroxyurea, methotrexate, nelarabine, pemetrexed, pentostatin, pralatrexate, thioguanine, and a trifluridine / tipiracil combination.

13. 13. The antibody molecule for use or method of treating cancer according to any one of claims 1, 2, and 4-12, wherein the cancer expresses or is determined to express MSLN and is selected from the group consisting of ovarian cancer, pancreatic adenocarcinoma, mesothelioma, cervical cancer, and non-small cell lung cancer.

14. 14. The antibody molecule for use or method of treating cancer according to any one of claims 1, 2 and 4 to 13, wherein treatment with the antibody molecule that binds to MSLN and CD137 and the chemotherapeutic agent results in an anti-tumor effect that is greater than the combined anti-tumor effect of treating a patient with the antibody molecule that binds to MSLN and CD137 alone or the chemotherapeutic agent alone.

15. 15. The antibody molecule for use or method of treating cancer according to claim 14, wherein the anti-tumor effect is tumor growth inhibition, reduction in tumor volume, increased median survival, and / or an increase in the proportion of patients experiencing complete tumor regression.

16. 16. The antibody molecule for use or method of treating cancer according to any one of claims 1, 2, and 4 to 15, wherein the antibody molecule that binds to MSLN and CD137 and the chemotherapeutic agent are administered to the patient simultaneously or sequentially.

17. 17. The antibody molecule for use, method of treating cancer, or kit of any one of claims 1, 2, and 4 to 16, wherein the method comprises determining whether the cancer expresses MSLN, and treating the individual if the cancer expresses MSLN.

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