Combination cancer treatments including anti-MSLN / CD137 antibodies and PD-1 / PD-L1 inhibitors

Abstract

The present application relates to the use of antibody molecules that bind to MSLN and CD137 and PD-1 / PD-L1 inhibitors in the treatment of cancer in patients.

Description

[Technical Field]

[0001] FIELD OF THE INVENTION The present invention relates to the use of a bispecific antibody molecule that binds to MSLN and CD137 and a PD-1 / PD-L1 inhibitor in the treatment of cancer in a patient. [Background technology]

[0002] background Antagonist antibodies targeting immune checkpoint co-inhibitory receptors can reverse immune resistance in some tumors, but the majority of patients do not respond to treatment or eventually experience resistance. 1 Costimulatory pathways, such as the CD137 / 4-1BB pathway, are also important in promoting productive anti-cancer immunity, and there is strong genetic evidence supporting their role in mediating anti-cancer immune responses. 2-6 Therefore, an increasing number of studies are aimed at using agonistic antibodies targeting costimulatory molecules to modify signaling 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. 7 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. 8 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. 9-11 , providing potent protection against activation-induced T cell death and enhancing cytotoxicity 12-14 .

[0004] The efficacy of anti-CD137 therapy has been demonstrated in multiple preclinical tumor models 15-19 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. 20 , which has been shown to increase the persistence of tumor-specific T cells 21 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. 22,23 Further structural analysis showed that these results are mediated by a recognized epitope on CD137 and Fc gamma receptor (FcγR) ligand-dependent clustering. 24 .

[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. 25 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. 16,26,27 .

[0006] Mesothelin (MSLN) is a 40 kD membrane-bound protein that is overexpressed in a variety of cancers, including mesothelioma, ovarian cancer, lung cancer, and pancreatic cancer. 28-39MSLN 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. 40 .

[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 anti-tumor 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 that binds to CD137. 2 M9657 has a human IgG1-LALA backbone, which does not bind to Fcγ receptors but retains FcRn binding and IgG-like pharmacokinetics (PK). High expression of MSLN on tumor cells increases antibody binding to tumor cells, cross-linking of antibody molecules, and interaction of antibody molecules with CD137 trimers, resulting in increased CD137 agonism. Therefore, clustered M9657 may function as a bridge connecting CD137 trimers to tumor cells. M9657 promotes CD137 activation signaling within the TME, which avoids systemic immune activation, and is therefore expected to offer advantages over monospecific CD137 antibodies. In preclinical studies, M9657 demonstrated MSLN target-dependent and dose-dependent antitumor immunity.

[0008] Monoclonal antibodies that block the interaction between programmed cell death protein 1 (PD-1) and its ligand, programmed cell death ligand 1 (PD-L1), such as anti-PD-1 / PD-L1 antibodies, are promising immune checkpoint antagonists that can enhance the immune response against cancer. Currently, the FDA has approved eight checkpoint inhibitors, including six anti-PD-1 / PD-L1 antibodies, one anti-CTLA-4 antibody, and one LAG3 monoclonal antibody, for the treatment of more than 12 major cancer types. 41,42 The successful application of anti-PD-1 / PD-L1 monoclonal antibodies in various clinical trials has demonstrated their remarkable potential in cancer immunotherapy. However, clinical results have not always been satisfactory, with large inter-patient variability often observed, and typically only a minority of patients responding to treatment.

[0009] Therefore, combination therapy has become a new research focus in the development of PD-1 / PD-L1 blockade-based therapies. Anti-PD-1 / PD-L1 antibodies and CD137 agonists have different but complementary mechanisms of action in cancer immunotherapy. In particular, anti-PD-1 / PD-L1 antibodies suppress negative signals that reduce T cell activation during the priming process or inhibit effector T cell activity at the tumor site, while CD137 agonists promote T cell activation through antigen presentation and cytokine secretion. The combination of PD-1 / PD-L1 inhibitors and CD137 agonists has been shown to lead to improved T cell activation. 43,44 . Summary of the Invention

[0010] Description of the invention As discussed in the background section, the combination of PD-1 / PD-L1 inhibitors and CD137 agonists has been shown to improve T cell activation. However, the effects of such combinations in treating cancer patients are not limited to the tumor microenvironment, raising concerns about the efficacy and specificity of such treatments. The development of CD137 agonist molecules as anticancer therapies has also been hindered by concerns about liver inflammation and clinical efficacy, and anti-PD-1 / PD-L1 monoclonal antibody therapy has shown wide variations in outcomes among patients.

[0011] The present inventors recognized a need for enhanced target-specific T cell activation and antitumor activity, which could be achieved by combining MSLN expression-dependent CD137 costimulation with PD-1 / PD-L1 blockade. Surprisingly, the present inventors were able to demonstrate that the combination of an antibody molecule that binds to MSLN and CD137 with a PD-1 / PD-L1 inhibitor resulted in an antitumor effect in a mouse tumor model that was greater than the combined antitumor effect observed when mice were treated with either the antibody molecule that binds to MSLN and CD137 or the PD-1 / PD-L1 inhibitor 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. 45 Thus, we found that the combination of antibody molecules that bind to MSLN and CD137 with PD-1 / PD-L1 inhibitors synergistically enhanced antitumor efficacy in mouse tumor models. Similar synergistic antitumor efficacy is expected when human patients are treated with the combination of antibody molecules that bind to MSLN and CD137 with PD-1 / PD-L1 inhibitors.

[0012] 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 mouse MSLN and the modified CH3 domain 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 and mouse CD137 is similar to that of M9657 to human MSLN and human CD137.

[0013] As already summarized above, the inventors have shown that the combination of FS122m and anti-mPD-1 can delay tumor growth or reduce tumor volume in the E0771, JC, and Eph4-1424 mouse tumor models to a greater extent than the combined tumor growth delay or tumor volume reduction observed when mice were treated with either FS122m or anti-mPD-1 alone. The inventors have also shown that combined treatment with FS122m and anti-mPD-1 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 or the percentage of mice with complete tumor regression observed when mice were treated with either FS122m or anti-mPD-1 alone. Thus, we demonstrated that the combination of FS122m and anti-mPD-1 synergistically enhanced anti-tumor activity by measuring tumor growth delay / tumor volume reduction, median survival, and the percentage of mice exhibiting complete tumor regression in E0771, JC, and Eph4-1424 mouse tumor models.

[0014] These preclinical results using mouse tumor models support the expectation that the combination of an antibody molecule that binds to MSLN and CD137 with a PD-1 / PD-L1 inhibitor synergistically enhances antitumor activity. These findings suggest that combination therapy of an antibody molecule that binds to MSLN and CD137 with a PD-1 / PD-L1 inhibitor represents a new therapeutic strategy for improving cancer treatment.

[0015] We were also able to show that the combination of M9657 with the anti-PD-1 antibody pembrolizumab synergistically increased T cell activation, tumor cell killing, and cytokine release in vitro, supporting the expectation that the combination of a bispecific anti-MSLN / CD137 antibody molecule with a PD-1 / PD-L1 inhibitor in the treatment of cancer in human patients will also show synergistic anti-tumor effects.

[0016] Accordingly, the present invention provides an antibody molecule that binds MSLN and CD137 for use in a method of treating cancer in a patient, the method comprising administering an antibody molecule that binds MSLN and CD137 in combination with a PD-1 / PD-L1 inhibitor.The present invention also relates to a PD-1 / PD-L1 inhibitor for use in a method of treating cancer in a patient, the method comprising administering a PD-1 / PD-L1 inhibitor in combination with an antibody molecule that binds MSLN and CD137.

[0017] 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.

[0018] 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.

[0019] 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. The second sequence may be located between positions 91 and 99 of the CH3 domain of the antibody molecule according to the IMGT numbering system. Preferably, the CH3 domain of the antibody molecule has the sequence set forth in SEQ ID NO: 86.

[0020] 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.

[0021] Numerous Fab regions 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].

[0022] A number of bispecific antibody molecules that bind to MSLN and CD137 are also 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).

[0023] PD-1 / PD-L1 inhibitors inhibit the PD-1 immune checkpoint. PD-1 / PD-L1 inhibitors can inhibit PD-1 activation directly or indirectly, but preferably directly inhibit PD-1 activation. PD-1 activation can be directly inhibited by inhibiting the binding of PD-L1 to PD-1 and / or inhibiting PD-L1-mediated PD-1 activation. PD-1 activation can also be indirectly inhibited, for example, through reducing PD-L1 expression.

[0024] PD-1 / PD-L1 inhibitors include small molecules, peptides, and antibody molecules. In this context, antibody molecules are immunoglobulins such as IgG1, chimeric antibody molecules, antibody fusion proteins, or antigen-binding antibody fragments, such as scFv, Fab, Fcab, VhH, monovalent IgG, diabodies, triabodies, immunoglobulin novel antigen receptors (IGNAR), single-domain shark variable domain novel antigen receptors (V-NAR), human chimeric IgG (hcIgG), minibodies, or nanobodies. In a preferred embodiment, the PD-1 / PD-L1 inhibitor is an antibody molecule or antigen-binding fragment thereof that binds to PD-1 or PD-L1.

[0025] The antibody molecule or antigen-binding fragment thereof that binds to PD-1 may be selected from the group consisting of nivolumab, pembrolizumab, cemiplimab, dostallimab, sintilimab, camrelizumab, toripalimab, tislelizumab, spartalizumab, zimberelimab, penprimab, and candonilimab. In a preferred embodiment, the antibody molecule or antigen-binding fragment thereof that binds to PD-1 may be selected from the group consisting of nivolumab, pembrolizumab, and cemiplimab. More preferably, the antibody molecule that binds to PD-1 is pembrolizumab. The heavy and light chain sequences of pembrolizumab are known in the art and are set forth in SEQ ID NO:93 and SEQ ID NO:94, respectively.

[0026] The antibody molecule or antigen-binding fragment thereof that binds to PD-L1 may be selected from the group consisting of avelumab, atezolizumab, durvalumab, sugemalimab, and emvafolimab. Preferably, the antibody molecule or antigen-binding fragment thereof that binds to PD-L1 is selected from the group consisting of avelumab, atezolizumab, and durvalumab. More preferably, the antibody molecule that binds to PD-L1 is avelumab.

[0027] The small molecule inhibitor of PD-1 / PD-L1 may be selected from the group consisting of BMS202, CA-170, fraxinerone, BMS-1166, N-deacetylated BMS-202, BMS-1001 hydrochloride, INCB086550, tomivosertib, PD-1 / PD-L1-IN-9, PROTAC PD-1 / PD-L1 degrader-1, PD-L1-IN-1, sulfamethoxypyridazine, PD-1-IN-17, PD-1 / PD-L1-IN-10, PD-1-IN-24, BMS-8, ebixapodrin, PD-1-IN-18, PD-1-IN-17 TFA, BMS-1166 hydrochloride, ARB272572, PD-1, PD-L1-IN-13, sulindac sodium, PD-1 / PD-L1-IN 5, PD-1-IN-22, PD-1 / PD-L1-IN-NP19, PD-1 / PD-L1-IN-27, PD-1-IN-20, PD-1 / PD-L1-IN-23, PD-1 / PD-L1-IN 5 TFA, PD-1 / PD-L1-IN 6, PD-1 / PD-L1-IN-14, PD-1 / PD-L1-IN-26, PD1-PDL1-IN 1, [D-Leu-4]-OB3, PD-1 / PD-L1-IN-16, PD-1 / PD-L1-IN-20, PD-1 / PD-L1-IN-19, PD-1 / PD-L1-IN-17, BMSpep-57, PD-1 / PD-L1-IN-24, PD-1 / PD-L1-IN-21, HE-S2, PD-1 / PD-L1-IN-22, PD-1 / PD-L1-IN-18, and PD-1 / PD-L1-IN-15.

[0028] The membrane-associated protein mesothelin (MSLN) has been shown to be expressed in several cancers. Specifically, high levels of MSLN have been shown to be expressed in ovarian cancer, pancreatic adenocarcinoma, mesothelioma, and non-small cell lung cancer. The present inventors have found that this is also the case in cervical cancer. Without wishing to be bound by theory, it is believed that when antibody molecules bind to MSLN, the antibody crosslinks and binds to CD137 expressed on the surface of immune cells, which then clusters and activates CD137, ultimately activating the immune cells.

[0029] Therefore, the cancer to be treated is preferably a cancer that expresses or is confirmed to express MSLN.Preferably, the cancer is selected from the group consisting of ovarian cancer, pancreatic adenocarcinoma, mesothelioma, cervical cancer, and non-small cell lung cancer.

[0030] The combination of FS122m and anti-mPD-1 resulted in anti-tumor activity in the E0771, JC, and Eph4-1424 mouse tumor models that was greater than the combined anti-tumor activity observed when mice were treated with either FS122m or anti-PD-1 alone, as described above. In other words, the anti-tumor effect of the combination was synergistic.

[0031] Thus, in one embodiment, treatment with an antibody molecule that binds to MSLN and CD137 in combination with a PD-1 / PD-L1 inhibitor results in an anti-tumor effect that is greater than the anti-tumor effect observed when a patient is treated with either the antibody molecule that binds to MSLN and CD137 or the PD-1 / PD-L1 inhibitor alone. Preferably, treatment with an antibody molecule that binds to MSLN and CD137 in combination with a PD-1 / PD-L1 inhibitor results in an anti-tumor effect that is greater than the combined anti-tumor effect observed when a patient is treated with either the antibody molecule that binds to MSLN and CD137 or the PD-1 / PD-L1 inhibitor alone. An anti-tumor effect can be the inhibition or delay of tumor growth. Thus, an anti-tumor effect can be a reduction in tumor volume. An anti-tumor effect can be an increase in the median survival time of a patient. 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 one skilled in the art.

[0032] The bispecific antibody molecule that binds to MSLN and CD137 and the PD-1 / PD-L1 inhibitor 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 PD-1 / PD-L1 inhibitor is administered parenterally. The antibody molecule that binds to MSLN and CD137 and / or the PD-1 / PD-L1 inhibitor may be administered intravenously, intramuscularly, subcutaneously, intraperitoneally, or spinally. Alternatively, the antibody molecule that binds to MSLN and CD137 and / or the PD-1 / PD-L1 inhibitor may be administered by injection or infusion.

[0033] The antibody molecule that binds to MSLN and CD137 and / or the PD-1 / PD-L1 inhibitor may be administered non-parenterally. The antibody molecule that binds to MSLN and CD137 and / or the PD-1 / PD-L1 inhibitor may be administered orally, intranasally, vaginally, rectally, sublingually, or topically.

[0034] The antibody molecule that binds to MSLN and CD137 and the PD-1 / PD-L1 inhibitor 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 PD-1 / PD-L1 inhibitor may be administered to a patient simultaneously or sequentially, but are preferably administered sequentially.

[0035] When the antibody molecule that binds to MSLN and CD137 and the PD-1 / PD-L1 inhibitor are administered to a patient sequentially, 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.

[0036] 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 PD-1 / PD-L1 inhibitor. 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 PD-1 / PD-L1 inhibitor. 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.

[0037] The present invention also provides the 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 PD-1 / PD-L1 inhibitor.The present invention also provides the use of a PD-1 / PD-L1 inhibitor for the manufacture of a medicament for the treatment of cancer, wherein the PD-1 / PD-L1 inhibitor is administered in combination with an antibody molecule that binds MSLN and CD137.

[0038] The present invention also provides a kit comprising an antibody molecule that binds to MSLN and CD137 and a pharmaceutically acceptable excipient, and a PD-1 / PD-L1 inhibitor and a pharmaceutically acceptable excipient.

[0039] 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 PD-1 / PD-L1 inhibitor. [2] A PD-1 / PD-L1 inhibitor for use in a method of treating cancer in a patient, the method comprising administering the PD-1 / PD-L1 inhibitor in combination with an antibody molecule that binds 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 PD-1 / PD-L1 inhibitor.

[0040] [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 PD-1 / PD-L1 inhibitor. [5] Use of a PD-1 / PD-L1 inhibitor for the manufacture of a medicament for the treatment of cancer, wherein the PD-1 / PD-L1 inhibitor 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 PD-1 / PD-L1 inhibitor and a pharmaceutically acceptable excipient.

[0041] [7] The antibody molecule or PD-1 / PD-L1 inhibitor, method, use, or kit for use according to [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 PD-1 / PD-L1 inhibitor, method, use, or kit for said use, comprising: [8] The antibody molecule or PD-1 / PD-L1 inhibitor for use, method, use, or kit 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 PD-1 / PD-L1 inhibitor, method, use, or kit for said use, comprising: [9] An antibody molecule or PD-1 / PD-L1 inhibitor, 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.

[0042]

[10] The antibody molecule or PD-1 / PD-L1 inhibitor, 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 PD-1 / PD-L1 inhibitor, 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 PD-1 / PD-L1 inhibitor, 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.

[0043]

[13] The antibody molecule or PD-1 / PD-L1 inhibitor, method, use, or kit for use according to any one of [7] 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 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.

[0044]

[14] The antibody molecule or PD-1 / PD-L1 inhibitor, 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 wherein the numbering of amino acid residues is according to the IMGT numbering system; an antibody molecule or PD-1 / PD-L1 inhibitor, method, use, or kit for said use.

[15] An antibody molecule or PD-1 / PD-L1 inhibitor, 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 PD-1 / PD-L1 inhibitor, 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.

[0045]

[17] The antibody molecule or PD-1 / PD-L1 inhibitor, 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 set forth in SEQ ID NOs: 80 and 26, respectively; an antibody molecule or PD-1 / PD-L1 inhibitor, method, use, or kit for said use, comprising:

[0046]

[18] An antibody molecule or PD-1 / PD-L1 inhibitor, 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 PD-1 / PD-L1 inhibitor, method, use, or kit for use according to [1] to

[18] , wherein the PD-1 / PD-L1 inhibitor is selected from the group consisting of small molecule inhibitors, peptides, antibodies, chimeric antibodies, antibody fusion proteins, and antigen-binding fragments thereof.

[20] The antibody molecule or PD-1 / PD-L1 inhibitor, method, use, or kit for use according to

[19] , wherein the antigen-binding fragment is selected from the group consisting of scFv, Fab, Fcab, VhH, monovalent IgG, diabody or triabody, immunoglobulin novel antigen receptor (IGNAR), single domain shark variable domain of novel antigen receptor (V-NAR), hIgG, minibody, or nanobody.

[0047]

[21] An antibody molecule or PD-1 / PD-L1 inhibitor, method, use, or kit for use according to any one of [1] to

[20] , wherein the PD-1 / PD-L1 inhibitor is an antibody or an antigen-binding fragment thereof that binds to PD-1 or PD-L1.

[22] The antibody molecule or PD-1 / PD-L1 inhibitor, method, use, or kit for use according to

[21] , wherein the antibody molecule or antigen-binding fragment thereof that binds to PD-1 or PD-L1 is an IgG molecule, an IgA molecule, an IgE molecule, an IgM molecule, or a fragment thereof.

[23] The antibody molecule or PD-1 / PD-L1 inhibitor, method, use, or kit for use according to

[21] or

[22] , wherein the antibody or antigen-binding fragment thereof that binds to PD-1 is selected from the group consisting of nivolumab, pembrolizumab, and cemiplimab.

[0048]

[24] The antibody molecule or PD-1 / PD-L1 inhibitor, method, use, or kit for use according to

[23] , wherein the antibody or antigen-binding fragment thereof that binds to PD-1 is pembrolizumab.

[25] The antibody molecule or PD-1 / PD-L1 inhibitor, method, use, or kit for use according to

[21] or

[22] , wherein the antibody or antigen-binding fragment thereof that binds to PD-L1 is selected from the group consisting of avelumab, atezolizumab, and durvalumab.

[26] The antibody molecule or PD-1 / PD-L1 inhibitor, method, use, or kit for use according to

[25] , wherein the antibody or antigen-binding fragment thereof that binds to PD-L1 is avelumab.

[0049]

[27] An antibody molecule or PD-1 / PD-L1 inhibitor, method, use, or kit for use according to any one of [1] to

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

[28] The antibody molecule or PD-1 / PD-L1 inhibitor, method, use, or kit for use according to [1] to

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

[29] An antibody molecule or PD-1 / PD-L1 inhibitor for use, method, use, or kit according to any one of [1] to

[28] , wherein treatment with an antibody molecule that binds to MSLN and CD137 and a PD-1 / PD-L1 inhibitor results in greater anti-tumor activity than monotherapy treatment with an antibody molecule that binds to MSLN and CD137 or monotherapy treatment with a PD-1 / PD-L1 inhibitor.

[0050]

[30] An antibody molecule or PD-1 / PD-L1 inhibitor for use, method, use, or kit according to any one of [1] to

[29] , wherein treatment with an antibody molecule that binds to MSLN and CD137 and a PD-1 / PD-L1 inhibitor results in anti-tumor activity that is 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 PD-1 / PD-L1 inhibitor.

[31] An antibody molecule or PD-1 / PD-L1 inhibitor for use according to any one of [1] to

[30] , a method, a use, or a kit, wherein treatment with an antibody molecule that binds to MSLN and CD137 and a PD-1 / PD-L1 inhibitor results in greater tumor growth delay, tumor volume reduction, median survival time, and / or number of complete tumor regressions than monotherapy treatment with an antibody molecule that binds to MSLN and CD137 or monotherapy treatment with a PD-1 / PD-L1 inhibitor.

[32] An antibody molecule or PD-1 / PD-L1 inhibitor for use, method, use, or kit according to any one of [1] to

[31] , wherein treatment with an antibody molecule that binds to MSLN and CD137 and a PD-1 / PD-L1 inhibitor results in a tumor growth delay, tumor volume reduction, median survival time, and / or 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 number of complete tumor regressions of monotherapy treatment with an antibody molecule that binds to MSLN and CD137 and monotherapy treatment with a PD-1 / PD-L1 inhibitor.

[0051]

[33] The antibody molecule or PD-1 / PD-L1 inhibitor, method, use, or kit for use according to [1] to

[32] , wherein the antibody molecule and / or PD-1 / PD-L1 inhibitor that binds to MSLN and CD137 is administered parenterally.

[34] The antibody molecule or PD-1 / PD-L1 inhibitor, method, use, or kit for use according to

[33] , wherein the antibody molecule and / or PD-1 / PD-L1 inhibitor that binds to MSLN and CD137 is administered intravenously, intramuscularly, subcutaneously, intraperitoneally, or spinally.

[35] The antibody molecule or PD-1 / PD-L1 inhibitor, method, use, or kit for use according to

[33] or

[34] , wherein the antibody molecule and / or PD-1 / PD-L1 inhibitor that binds to MSLN and CD137 is administered by injection or infusion.

[0052]

[36] An antibody molecule or PD-1 / PD-L1 inhibitor, method, use, or kit for use according to any one of [1] to

[32] , wherein the antibody molecule and / or PD-1 / PD-L1 inhibitor that binds to MSLN and CD137 is administered non-parenterally.

[37] The antibody molecule or PD-1 / PD-L1 inhibitor, method, use, or kit for use according to

[36] , wherein the antibody molecule and / or PD-1 / PD-L1 inhibitor that binds to MSLN and CD137 is administered orally, intranasally, vaginally, rectally, sublingually, or topically.

[38] An antibody molecule or a PD-1 / PD-L1 inhibitor, method, use, or kit for use according to any one of [1] to

[37] , wherein the antibody molecule that binds to MSLN and CD137 and the PD-1 / PD-L1 inhibitor are administered to a patient simultaneously or sequentially.

[0053]

[39] An antibody molecule or PD-1 / PD-L1 inhibitor, method, use, or kit for use according to

[38] , wherein the antibody molecule that binds to MSLN and CD137 and the PD-1 / PD-L1 inhibitor are administered sequentially to a patient.

[40] The antibody molecule or PD-1 / PD-L1 inhibitor, method, use, or kit for use according to

[39] , wherein the antibody molecule that binds to MSLN and CD137 and the PD-1 / PD-L1 inhibitor are administered to the patient within 4 days of each other.

[41] The antibody molecule or PD-1 / PD-L1 inhibitor, method, use, or kit for use according to

[39] or

[40] , wherein the antibody molecule that binds to MSLN and CD137 and the PD-1 / PD-L1 inhibitor are administered to the patient within 3 days of each other.

[0054]

[42] An antibody molecule or a PD-1 / PD-L1 inhibitor, method, use, or kit for use according to

[39] to

[41] , wherein the antibody molecule that binds to MSLN and CD137 and the PD-1 / PD-L1 inhibitor are administered to a patient within two days of each other.

[43] An antibody molecule or PD-1 / PD-L1 inhibitor, method, use, or kit for use according to

[39] to

[42] , wherein the antibody molecule that binds to MSLN and CD137 and the PD-1 / PD-L1 inhibitor are administered to the patient on the same day.

[44] An antibody molecule or PD-1 / PD-L1 inhibitor for use, or a method according to any one of [1] to [3] and [7] to

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

[0055] 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:

[0056] [Figure 1] Figure 1 shows the results of a bioluminescence report assay, in which bioluminescence signals were measured as a function of NF-κB expression in PD-1+CD137 effector cells cocultured with PD-L1 aAPC / CHO-K1-expressing and MSLN-expressing CHO tumor cells. Cells were treated with various concentrations of either M9657, M9657+pembrolizumab, M9657+anti-HEL-hIgG1-LALA antibody (isotype control), CD137L+pembrolizumab, or CD137L+anti-HEL-hIgG1-LALA antibody. Treatment with M9657+pembrolizumab resulted in significantly higher bioluminescence signals than treatment with M9657, M9657+anti-HEL-hIgG1-LALA antibody, CD137L+pembrolizumab, or CD137L+anti-HEL-hIgG1-LALA antibody. Mean values ​​± standard error of the mean (SEM) are shown and data represent three replicate experiments.

[0057] [Figure 2]Figure 2 shows the results of an assay measuring target cell cytotoxicity and cytokine release of CD8+ T cells cocultured with NCI-H226 tumor target cells. Cytotoxicity was measured as the % kill of target cells (A) and the normalized area under the curve (AUC) (B) of the results in (A). Cytokine release was measured as the INFγ level in the supernatant (C) and the normalized area under the curve (AUC) (D) of the results in (C). Cells were treated with various concentrations of either M9657 + pembrolizumab, M9657, anti-HEL-hIgG1-LALA antibody (isotype control), or BiTE (anti-CD3 x anti-EGFR BiTE). Treatment with M9657 + pembrolizumab increased CD8+ T cell cytotoxicity compared to monotherapy with M9657, anti-HEL-hIgG1-LALA antibody, or BiTE (anti-CD3 x anti-EGFR BiTE). % Kill (A) and AUC (B) were measured in cells from five different donors and are shown as mean ± SEM. IFNγ concentrations (C) and AUC (D) were measured in cells from six different donors and are shown as mean ± SEM.

[0058] [Figure 3]Figure 3 shows the efficacy of treatment in the E0771 orthotopic mouse breast cancer model in C57BL / 6 mice. Progression of the mouse cancer model 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 antibody (isotype control), FS122m, anti-mPD-1, or FS122m + anti-mPD-1. Treatment with FS122m + anti-mPD-1 demonstrated a reduction in mean tumor volume over time, whereas FS122m and anti-mPD-1 monotherapy only slowed tumor growth compared to the anti-HEL-hIgG1-LALA isotype control over the study period (A). Combination treatment with FS122m and anti-mPD-1 also improved median survival compared with FS122m and anti-mPD-1 monotherapy (B), inducing complete tumor regression in 7 of 9 mice, compared with none of the 9 mice treated with FS122m monotherapy and only 1 of the 9 mice treated with anti-mPD-1 monotherapy. 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 change are shown as means ± SEM.

[0059] [Figure 4]Figure 4 shows the efficacy of treatment in the JC subcutaneous breast tumor model in BALB / c mice. Progression of the mouse cancer model 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, anti-mPD-1, or FS122m + anti-mPD-1. Treatment with FS122m + anti-mPD-1 delayed tumor growth to a greater extent than FS122m and anti-mPD-1 monotherapy when compared to the anti-HEL-hIgG1-LALA isotype control (A). Combination treatment with FS122m and anti-mPD-1 also improved median survival compared with FS122m and anti-mPD-1 monotherapy (B), inducing complete tumor regression in 3 of 10 mice, whereas no mice achieved complete tumor regression when treated with FS122m or anti-mPD-1 monotherapy. 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.

[0060] [Figure 5]Figure 5 shows the efficacy of treatment in an Eph4-1424 subcutaneous breast tumor model in BALB / c mice. Progression of the mouse cancer model 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, anti-mPD-1, or FS122m + anti-mPD-1. Treatment with FS122m + anti-mPD-1 demonstrated a reduction in mean tumor volume over time, whereas monotherapy with FS122m and anti-mPD-1 only delayed tumor growth over the study period compared to treatment with the anti-HEL-hIgG1-LALA isotype control (A). Combination therapy with FS122m and anti-mPD-1 also improved median survival compared with FS122m and anti-mPD-1 monotherapy (B), inducing complete tumor regression in 10 of 10 mice, but only 1 of 10 mice treated with FS122m monotherapy and 6 of 10 mice treated with anti-mPD-1 monotherapy. 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.

[0061] Detailed Description of the Invention The present invention relates to an antibody molecule that binds to MSLN and CD137 for use in the treatment of cancer, said antibody molecule in combination with a PD-1 / PD-L1 inhibitor. The present invention also relates to a PD-1 / PD-L1 inhibitor for use in the treatment of cancer, said inhibitor in combination with an antibody molecule that binds to MSLN and CD137.

[0062] The term "PD-1 / PD-L1 inhibitor" refers to a molecule that inhibits the PD-1 immune checkpoint. A PD-1 / PD-L1 inhibitor can inhibit PD-1 activation directly or indirectly, but preferably directly inhibits PD-1 activation. PD-1 activation can be inhibited directly by inhibiting PD-L1 binding to PD-1 and / or inhibiting PD-L1-mediated activation of PD-1. PD-1 activation can be inhibited indirectly, for example, through reducing PD-L1 expression. A PD-1 / PD-L1 inhibitor can be a small molecule inhibitor, peptide, antibody, chimeric antibody, antibody fusion protein, or antibody fragment, such as scFv, Fab, Fcab, VhH, monovalent IgG, diabody or triabody, IGNAR, V-NAR, hclgG, minibody, or nanobody. For example, a PD-1 / PD-L1 inhibitor can be an antibody or antigen-binding fragment thereof that binds to PD-1 or PD-L1.

[0063] 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, M, A, E, and D, and their isotypic subclasses, such as IgG1, IgG2, IgG3, and IgG4, and antigen-binding 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.

[0064] A PD-1 / PD-L1 inhibitor may be naturally occurring or partly or wholly synthetically produced, for example, a PD-1 / PD-L1 inhibitor may be a recombinant antibody molecule.

[0065] The antibody or antigen-binding fragment thereof that binds to PD-1 or PD-L1 may be an anti-PD-1 antibody or antigen-binding fragment thereof. Preferably, the anti-PD-1 antibody is selected from the list consisting of nivolumab, pembrolizumab, and cemiplimab. More preferably, the anti-PD-1 antibody is pembrolizumab.

[0066] The antibody or antigen-binding fragment thereof that binds to PD-1 or PD-L1 may be an anti-PD-L1 antibody or antigen-binding fragment thereof. Preferably, the anti-PD-1 antibody is selected from the list consisting of avelumab, atezolizumab, and durvalumab. More preferably, the anti-PD-L1 antibody is avelumab.

[0067] 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.

[0068] Thus, the PD-1 / PD-L1 inhibitor may be any anti-PD-1 / PD-L1 antibody not listed above that is within the state of the art or that is not yet within the state of the art and can be arrived at using commonly available techniques known in the art.

[0069] In the following, the term "bispecific antibody molecule" is used to refer to an antibody molecule that binds to MSLN and CD137.

[0070] The bispecific antibody molecule may be an immunoglobulin or an antigen-binding fragment thereof. In one embodiment, the bispecific antibody molecule binds independently to MSLN and CD137. In one embodiment, the bispecific antibody simultaneously binds to MSLN and CD137.

[0071] 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.

[0072] Bispecific antibody molecules may be naturally occurring or partly or wholly synthetically produced, for example the antibody molecule may be a recombinant antibody molecule.

[0073] 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.

[0074] 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 an antigen-binding fragment thereof. In a preferred embodiment, the bispecific antibody molecule is a complete immunoglobulin molecule.

[0075] 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. 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).

[0076] 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.

[0077] 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.

[0078] 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 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, 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).

[0079] A bispecific antibody molecule may or may not have a LALA mutation. In a preferred embodiment, the bispecific antibody molecule has a LALA mutation. The LALA mutation represents a type of mutation for disrupting 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 an 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. Alternatively, complement activation (C1q binding) and ADCC are 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. These two mutations can also be combined to generate antibody molecules with further reduced or no ADCC or CDC activity.

[0080] 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, wherein 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, wherein 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 another embodiment, an antibody molecule may comprise a CH2, wherein the CH2 domain comprises an alanine residue at position 114. For example, the CH2 domain may have the amino acid sequence set forth in SEQ ID NO:91. In another embodiment, 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.

[0081] 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.

[0082] 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.

[0083] 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.

[0084] 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.

[0085] 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.

[0086] 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 28, 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.

[0087] 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.

[0088] 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.

[0089] The CDR-based antigen-binding site may comprise the VH or VL domain, preferably the VH and VL domain, 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, 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 / 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.

[0090] Antibodies FS22-172-003-AA / FS28-256-271, FS22-172-003-AA / FS28-024-052, FS22-1 72-003-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 / FS28-25 6-026, FS22-172-003-AA / FS28-256-027, FS22-172-003-AA / FS28-256-001, FS2 The VH domains 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, 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.

[0091] 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.

[0092] The bispecific antibody molecule of the present invention comprises a CD137 antigen-binding site. The CD137 antigen-binding site is located in the constant domain of the antibody molecule, preferably in the CH3 domain. The CD137 antigen-binding site comprises 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.

[0093] The CD137 antigen-binding site of the bispecific antibody molecule comprises 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.

[0094] 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.

[0095] 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.

[0096] In a preferred embodiment, the bispecific 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.

[0097] 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.

[0098] 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, such as binding specificity and / or 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.

[0099] 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.

[0100] 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.

[0101] 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.

[0102] 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.

[0103] 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.

[0104] 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 having 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.

[0105] 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.

[0106] In one embodiment, a bispecific antibody molecule of the invention comprises a CH3 domain sequence that has one or more amino acid sequence changes (addition, deletion, substitution and / or insertion of amino acid residues) compared to a CH3 domain 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.

[0107] In embodiments in which one or more amino acids are substituted with another amino acid, the substitution may be 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, In some embodiments, amino acids in the same row in the right-most column are substituted for each other. [Table 1]

[0108] In some embodiments, the substitution(s) may be functionally conservative, i.e., the substitution may not affect (or may 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.

[0109] 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.

[0110] 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. Secondary or metastatic 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.

[0111] 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.

[0112] Thus, cancers to be treated with a bispecific antibody molecule that binds MSLN and CD137 in combination with a PD-1 / PD-L1 inhibitor 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.

[0113] 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.

[0114] The cancer may be a primary cancer or a secondary cancer. Thus, the antibody molecules that bind to MSLN and CD137 described herein can be for use in combination with a PD-1 / PD-L1 inhibitor in a method of treating cancer in an individual, wherein the cancer is a primary tumor and / or a secondary or metastatic tumor. The cancer to be treated may be a solid tumor.

[0115] 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 consisting of ovarian cancer, pancreatic adenocarcinoma, mesothelioma, cervical cancer, and non-small cell lung cancer.

[0116] The patient to be treated can be selected for treatment if the cancer expresses MSLN.The patient to be treated can be selected if it is determined that the cancer expresses MSLN.Preferably, the patient is selected for treatment if the cancer is ovarian cancer, pancreatic adenocarcinoma, mesothelioma, cervical cancer or non-small cell lung cancer and expresses MSLN.

[0117] The inventors have demonstrated that combination treatment with M9657 (SEQ ID NO: 2 and SEQ ID NO: 10) and pembrolizumab (SEQ ID NO: 93 and SEQ ID NO: 94) significantly reduces T cell activation, CD8 + T cell activation, CD8 + The inventors also demonstrated that combination therapy with FS122m and anti-mPD-1 resulted in tumor target cell killing and increased cytokine release by FS122m and anti-mPD-1 cells. The inventors also demonstrated that combination therapy with FS122m and anti-mPD-1 resulted in tumor growth delay, tumor volume reduction, increased median survival, and / or increased number of complete tumor regressions that were greater than the tumor growth delay, tumor volume reduction, increased median survival, and / or increased number of complete tumor regressions observed when mice were treated with either FS122m or anti-mPD-1 alone. Thus, the inventors demonstrated that combination treatment with M9657 and pembrolizumab enhanced T cell activation, CD8 + The combination of F122m (SEQ ID NO: 84 and SEQ ID NO: 85) and anti-mPD-1 was found to synergistically increase tumor target cell killing and cytokine release by the cells. Similarly, combined treatment with F122m (SEQ ID NO: 84 and SEQ ID NO: 85) and anti-mPD-1 synergistically slowed tumor growth, reduced tumor volume, increased median survival, and increased the number of complete tumor regressions.

[0118] In one embodiment, combining an antibody molecule that binds to MSLN and CD137 with a PD-1 / PD-L1 inhibitor results in tumor growth delay, reduction in tumor volume, increased median survival, and / or an increase in the number of complete tumor regressions. Preferably, combining an antibody molecule that binds to MSLN and CD137 with a PD-1 / PD-L1 inhibitor results in tumor growth delay, reduction in tumor volume, increased median survival, and / or an increase in the number of complete tumor regressions that is statistically significantly greater than monotherapy treatment with an antibody molecule that binds to MSLN and CD137 or monotherapy treatment with a PD-1 / PD-L1 inhibitor. More preferably, the combination of an antibody molecule that binds MSLN and CD137 with a PD-1 / PD-L1 inhibitor results in a tumor growth delay, tumor volume reduction, increase in median survival, 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, and / or increase in the number of complete tumor regressions of monotherapy treatment with an antibody molecule that binds MSLN and CD137 and monotherapy treatment with a PD-1 / PD-L1 inhibitor.

[0119] In another embodiment, an antibody molecule that binds to MSLN and CD137 in combination with a PD-1 / PD-L1 inhibitor enhances T cell activation, CD8 + Preferably, the combination of an antibody molecule that binds to MSLN and CD137 with a PD-1 / PD-L1 inhibitor increases T cell activation, CD8 T cell activation, and cytokine release that are statistically significantly greater than monotherapy treatment with either the bispecific antibody molecule that binds to MSLN and CD137 or the PD-1 / PD-L1 inhibitor. + More preferably, combining an antibody molecule that binds to MSLN and CD137 with a PD-1 / PD-L1 inhibitor results in the combined T cell activation, CD8 + T cell activation, CD8 cells killing tumor target cells, and cytokine release +This results in the cells killing tumor target cells and releasing cytokines.

[0120] The ability of antibody molecules that bind MSLN and CD137 in combination with a PD-1 / PD-L1 inhibitor to activate T cells can be determined by measuring the maximum bioluminescent signal transmitted by T cells in the presence of antibody molecules that bind MSLN and CD137 in combination with a PD-1 / PD-L1 inhibitor using a Bio-Glo-NL™ luciferase assay.

[0121] The ability of antibody molecules that bind MSLN and CD137 to activate T cells in combination with a PD-1 / PD-L1 inhibitor can also be determined by measuring IFNγ release in a cytokine release assay in the presence of the bispecific antibody molecule in combination with a PD-1 / PD-L1 inhibitor.

[0122] In one embodiment, the antibody molecule that binds to MSLN and CD137 and the PD-1 / PD-L1 inhibitor 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 PD-1 / PD-L1 inhibitor are administered as a second line treatment (e.g., after initial treatment with the same or a different therapeutic agent, including after relapse and / or if first treatment has failed).

[0123] 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 that bind to MSLN and CD137 and / or PD-1 / PD-L1 inhibitors 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.

[0124] Thus, in some embodiments, the antibody molecule that binds to MSLN and CD137 and / or the PD-1 / PD-L1 inhibitor is administered parenterally. The antibody molecule that binds to MSLN and CD137 and / or the PD-1 / PD-L1 inhibitor may be administered intravenously, intramuscularly, subcutaneously, intraperitoneally, or spinally. Alternatively, the antibody molecule that binds to MSLN and CD137 and / or the PD-1 / PD-L1 inhibitor may be administered by injection or infusion.

[0125] In other embodiments, the antibody molecule that binds to MSLN and CD137 and / or the PD-1 / PD-L1 inhibitor is administered non-parenterally. The antibody molecule that binds to MSLN and CD137 and / or the PD-1 / PD-L1 inhibitor may be administered orally, intranasally, vaginally, rectally, sublingually, or topically.

[0126] "Concurrent administration" refers to the simultaneous administration of the antibody molecule that binds to MSLN and CD137 and the PD-1 / PD-L1 inhibitor in the same or separate formulations. "Sequential administration" refers to the separate, timely administration of the bispecific antibody molecule and the PD-1 / PD-L1 inhibitor in separate formulations.

[0127] Thus, the antibody molecule that binds to MSLN and CD137 and the PD-1 / PD-L1 inhibitor may be part of the same formulation or part of separate formulations. Preferably, the antibody molecule that binds to MSLN and CD137 and the PD-1 / PD-L1 inhibitor are provided as separate formulations.

[0128] In one embodiment of the present invention, the antibody molecule that binds to MSLN and CD137 and the PD-1 / PD-L1 inhibitor are administered simultaneously. For example, the bispecific antibody molecule that binds to MSLN and CD137 may be administered together with the PD-1 / PD-L1 inhibitor in the same formulation. Alternatively, the antibody molecule that binds to MSLN and CD137 and the PD-1 / PD-L1 inhibitor may be administered immediately before or after each other in separate formulations.

[0129] Preferably, the antibody molecule that binds to MSLN and CD137 and the PD-1 / PD-L1 inhibitor are administered to the patient sequentially. More preferably, the antibody molecule that binds to MSLN and CD137 and the PD-1 / PD-L1 inhibitor are administered to the patient sequentially within 4 days of each other, preferably within 3 days of each other, more preferably within 2 days of each other, or on the same day.

[0130] 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 PD-1 / PD-L1 inhibitor.

[0131] 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.

[0132] 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 PD-1 / PD-L1 inhibitor.

[0133] Also provided is a kit comprising an antibody molecule that binds to MSLN and CD137 and a PD-1 / PD-L1 inhibitor. Preferably, the kit may comprise an antibody molecule that binds to MSLN and CD137 and a pharmaceutically acceptable excipient, and a PD-1 / PD-L1 inhibitor and a pharmaceutically acceptable excipient.

[0134] 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 PD-1 / PD-L1 inhibitor, and the package may include instructions for using the antibody molecule that binds to MSLN and CD137 in combination with the PD-1 / PD-L1 inhibitor to treat cancer in an individual.

[0135] In one embodiment, the kit may be a package containing at least one dose of a medicament comprising an antibody molecule that binds to MSLN and CD137 and one dose of a medicament comprising a PD-1 / PD-L1 inhibitor. Preferably, the kit may include at least one dose of a medicament comprising an antibody molecule that binds to MSLN and CD137 and a pharmaceutically acceptable excipient, and one dose of a PD-1 / PD-L1 inhibitor and a pharmaceutically acceptable excipient. More preferably, the kit may further include a package insert containing instructions for using the medicament to treat cancer in an individual.

[0136] 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 PD-1 / PD-L1 inhibitor. 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 PD-1 / PD-L1 inhibitor and a pharmaceutically acceptable excipient. The package may further include an insert containing instructions for using the medicament to treat cancer in an individual.

[0137] 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.

[0138] 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.

[0139] 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.

[0140] Any section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.

[0141] 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.

[0142] 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]

[0143] material and method The efficacy of the combination of MSLN with a CD137-binding antibody molecule and a PD-1 / PD-L1 inhibitor in enhancing anti-cancer immune responses compared with either MSLN with a CD137-binding antibody molecule or a PD-1 / PD-L1 inhibitor monotherapy was assessed by examining various parameters, including in vitro T cell activation, target cell cytotoxicity, and cytokine release in human cells, and in vivo mean tumor volume and survival time in mouse tumor models. These methods are described in detail below.

[0144] Bio-Glo-NL™ Luciferase Assay To investigate the combined effects of PD-1 blocking and CD137 stimulation, combined treatment with M9657 and pembrolizumab inhibited PD-1 + The effect on NF-κB expression in CD137 effector cells was measured using a Bio-Glo™-NL luciferase assay. The assay was performed according to the manufacturer's instructions. One day before the assay, PD-L1 aAPC / CHO-K1 cells expressing PD-L1 and T cell activators were thawed and collected in 14.5 mL of cell collection medium (10% FBS / F-12). The cells were then seeded into white 96-well plates and incubated overnight at 37°C in a 5% CO2 incubator. On the day of the assay, serial dilutions of M9657 (250 nM, serially diluted 4-fold to 0.003815 nM), pembrolizumab (167.875 nM), CD137L (250 nM, serially diluted 4-fold to 0.003815 nM), and anti-HIS-tagged antibody (125 nM, serially diluted 4-fold to 0.0019 nM) were prepared, the latter of which was added to CD137L for crosslinking. CD137 ligand (CD137L) interacts with CD137 to induce T cell activation. Here, CD137L was used as a positive control for the reporting assay. An antibody against an unrelated target (anti-HEL-hIgG1-LALA antibody) was used as a negative control. The medium was aspirated from the PD-L1 aAPC / CHO-K1 cells, and a total of 40 μL of the prepared antibody solution was added to each well. +CD137 effector cells were thawed and harvested, and 40 μL of cell suspension was added to each well. MSLN-expressing CHO cells were added to the wells at a 1:1 ratio of PD-L1 aAPC / CHO-K1 (75,000 cells):CHO-MSLN cells (75,000 cells). The plate was incubated for 6 hours. At the end of the incubation period, Bio-Glo-NL luciferase reagent was prepared by combining 1 volume of Bio-Glo-NL luciferase assay substrate with 50 volumes of Bio-Glo-NL luciferase assay buffer. The assay plate was removed from the incubator, and 80 μL of Bio-Glo-NL reagent was added to each well. The plate was then incubated at room temperature for 5 minutes. After incubation, the plate was read using a Tecan plate reader with the luminescence setting. GraphPad Prism V9 software was used for statistical analysis and graph plotting. To calculate the efficacy of the combination treatment of antibody molecules that bind to MSLN and CD137 and PD-1 / PD-L1 inhibitors in inducing luciferase activity compared with monotherapy with antibody molecules that bind to MSLN and CD137 or PD-1 / PD-L1 inhibitors, the half-maximal effective concentration (EC 50 The antibody concentration at which ΔΨ ≈ 0.05 was achieved was calculated using a nonlinear regression fit.

[0145] Target-dependent cytotoxicity and cytokine release assays Combination of PD-1 blocking and CD137 stimulation + The efficacy of T cell-mediated killing of tumor target cells and cytokine release was also investigated. To this end, CD8 T cells were expressed against co-cultured NCI-H226 target cancer cells. + T cell toxicity and IFNγ release were monitored ex vivo. NCI-H226 cells were cultured in RPMI-1640 medium containing 10% FBS. Human CD8 +T cells were purchased from Hemacare. All cells were cultured at 37°C, 5% CO2, and 95% relative humidity. NCI-H226 cells were treated with 10 ng / mL human IFNγ for 48 hours, washed, labeled with IncuCyte Cytolight Rapid Red dye one day before the experiment, and then plated onto assay plates. Human CD8 + T cells were thawed and recovered in 250 U / ml IL-2 for 24 hours before being added to assay plates on the day of the experiment. NCI-H226 cells express high levels of MSLN and EGFR in the presence or absence of 2 pM anti-CD3 × anti-EGFR BiTE. Cells were then incubated for 72 hours with 2 pM anti-CD3 × anti-EGFR BiTE, 10 μg / ml pembrolizumab, a fixed concentration of 1 μM Cytolight Rapid Dye, and serial dilutions of M9657 or control antibody. Cell proliferation was detected using an Incucyte® Live Imaging System. Target cell cytotoxicity was determined by calculating the % killing rate relative to the BiTE-only group using an Incucyte® Live-Cell Analysis System. Both total and labeled cells were monitored using phase contrast and red fluorescence channels, respectively. IFNγ release was detected using an AlphaLisa according to the manufacturer's instructions.

[0146] Mouse tumor model Wild-type BALB / c and C57BL / 6 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 housed and maintained under conditions consistent with The Guide for the Care and Use of Laboratory Animals, 8th Edition, as described below. All animal experiments were performed in accordance with the guidelines of the EMD Serono Research Institution (protocols 17-008, 20-005) and the Wuxi AppTec Animal Care and Use Committee (IACUC).

[0147] 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).

[0148] Using the E0771, JC, and Eph4-1424 tumor models, the antitumor efficacy of a combination of an antibody molecule binding to MSLN and CD137 with a PD-1 / PD-L1 inhibitor was investigated compared with monotherapy with an antibody molecule binding to MSLN and CD137 or a PD-1 / PD-L1 inhibitor. Because M9657 does not show cross-reactivity to mouse MSLN and mouse CD137, anti-mMSLN-mCD137-huIgG1-LALA(FS122m) (SEQ ID NOs: 84 and 85) was developed as a surrogate antibody to M9657 for in vivo pharmacology studies. In the E0771 model, 0.5 x 106 E0771 tumor cells in 0.1 mL of PBS were orthotopically inoculated into the right mammary fat pad of female C57BL / 6 mice. Mice were cultured for a mean tumor volume of approximately 50-100 mm. 3 When they reached maturity, they were randomly assigned to treatment groups (n = 9 mice / group). For the JC model, female BALB / c mice were inoculated subcutaneously (sc) into the right upper flank with JC tumor cells (5 × 106) in 0.1 mL of PBS. Mice were cultured until a mean tumor volume of approximately 50–100 mm2 was achieved. 3 When they reached maturity, they were randomly assigned to treatment groups (n = 10 mice / group). For the Eph4-1424 model, female BALB / c mice were inoculated subcutaneously (sc) into the right upper flank with Eph4-1424 tumor cells (1 × 10) in 0.1 mL of PBS. Mice were cultured until a mean tumor volume of approximately 50–100 mm was achieved. 3 When they reached 10 days, they were randomly assigned to treatment groups (n=10 mice / group).

[0149] 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.

[0150] If tumor ulceration occurred, animals with ulcerated tumors were monitored at least three times per week, increasing frequency 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 ulcers / lesions only when directed by veterinary staff.

[0151] 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). Potential exceptions were consulted and discussed with veterinary staff.

[0152] Animals were euthanized if they were 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 system failure, weakness, hypothermia, central nervous system disorders (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.

[0153] 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:

[0154] Differences in tumor growth between treatment groups were determined using two-way analysis of variance (ANOVA) and Tukey's multiple comparison test. 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 recognized at the p ≤ 0.05 level.

[0155] Example 1: NF-κB expression Activation of NF-κB expression is an indicator of T cell activation. CD137 clustering activates the NF-κB signaling pathway upon interaction with its cognate ligand, CD137L. Agonist antibody molecules mimic the ligand by promoting CD137 clustering and activation, thereby activating the NF-κB signaling pathway. Anti-PD-1 / PD-L1 antibodies, on the other hand, limit the negative effects of PD-1 signaling on T cell activation by blocking the interaction between PD-L1 and PD-1. NF-κB expression is measured as a function of luciferase activity, resulting in a bioluminescent signal in the luciferase reporter assay described in Materials and Methods. Luciferase expression in the assay is controlled by an NF-κB-sensitive promoter and therefore directly correlates with NF-κB expression.

[0156] To investigate the combined effects of PD-1 blocking and CD137 stimulation, the effect of combined treatment with M9657 and pembrolizumab was measured using the Bio-Glo-NL™ luciferase assay. +CD137 effector cells were cocultured with PD-L1aAPC / CHO K1 cells and treated with either M9657 + pembrolizumab, M9657, M9657 + anti-HEL-hIgG1-LALA, CD137L + anti-HEL-hIgG1-LALA, CD137L + pembrolizumab, or CD137L + anti-HEL-hIgG1-LALA. Because the bispecific antibody molecule M9657 requires binding to MSLN for its bioactivity and PD-L1aAPC / CHO-K1 cells do not express MSLN, MSLN-expressing CHO cells were added to the PD-L1aAPC / CHO-K1 cells and CHO-MSLN cells. Luciferase activity was measured as a function of bioluminescence using a Tecan device and analyzed using GraphPad Prism V9 software. Treatment with M9657 induced a concentration-dependent bioluminescence signal with an EC of 0.08024 nM. 50 Combination treatment with M9567 + pembrolizumab enhanced T cell activation. Combination treatment with M9567 + pembrolizumab increased bioluminescence signal more than treatment with M9657, M9657 + anti-HEL-hIgG1-LALA, CD137L + anti-HEL-hIgG1-LALA, CD137L + pembrolizumab, and CD137L + anti-HEL-hIgG1-LALA. Pembrolizumab had an EC 50 (Figure 1).

[0157] Example 2: Cytotoxicity and cytokine release Two additional hallmarks of T cell activation are human CD8 + T cell-mediated killing of tumor target cells and cytokine release. To this end, NCI-H226 cancer cells were transfected with CD8 + The effect of combining CD137 stimulation with PD-1 blocking was investigated by ex vivo co-culture with T cells (Materials and Methods). Tumor target cell killing was measured as the % killing of NCI-H226 cancer cells, while CD8 + Cytokine release by T cells was measured as INFγ levels in T cell culture supernatants.

[0158] To calculate the effect of treating co-cultured cells with M9657 + pembrolizumab, M9657, an isotype control (anti-HEL-hIgG1-LALA antibody), or a BiTE (anti-CD3xanti-EGFR BiTE), cytotoxicity and INFγ release were measured. Treatment with M9657 resulted in concentration-dependent killing of tumor target cells and INFγ release, which was further increased in cells treated with the combination of M9657 and pembrolizumab (Figure 2). Pembrolizumab increased CD8 expression of target cells. + T cell killing was increased compared to single-agent treatment with M9657 and isotype control at all concentrations of M9657 tested, with particularly rapid increases at concentrations between 10 and 100 pM (Fig. 2A).

[0159] Example 3: In vivo proof of concept CD137 / MSLN mAb 2 Because the combination of PD-1 / PD-L1 antibodies with anti-PD-1 / PD-L1 antibodies was shown to improve T cell activation, target cell killing, and T cell cytokine release, the effects of this combination were also tested in an in vivo mouse tumor model.

[0160] Because there is no cross-reactivity between M9657 and mouse CD137, we developed the anti-mMSLN-mCD137-huIgG1-LALA (FS112m) bispecific antibody, whose binding affinity for mouse CD137 is similar to that of M9657 for human CD137, as a surrogate antibody for use in mouse tumor models. FS112m was combined with anti-mPD-1 to test the anti-tumor efficacy of the combination of both antibodies compared to monotherapy with FS112m and anti-mPD-1 in three different mouse breast tumor models. To this end, female mice were inoculated with either E0771, JC, or Eph4-1424 breast cancer cells, and tumors with a mean 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.

[0161] The combination of FS112m and anti-mPD-1 was compared with monotherapy with FS112m and anti-mPD-1 and with monotherapy with an anti-HEL-hIgG1-LALA isotype control antibody in all three mouse breast tumor models. The results are summarized below.

[0162] 3.1 E0771 Mouse Breast Tumor Model In the E0771 mouse breast tumor model, FS122m and anti-mPD-1 monotherapy induced marginal or moderate tumor growth inhibition (TGI) (26.6% and 55.5%, respectively) compared with isotype control (P<0.01, P<0.0001, day 14, respectively) and prolonged median survival (18 and 24 days, respectively) compared with isotype control (16 days) (Figure 3A, B, D). TGI was further enhanced when mice were treated with the combination of FS122m and anti-mPD1 (102%) compared with FS122m (P<0.0001, day 14) and anti-mPD1 (P<0.0001, day 14) monotherapy (Figure 3A). Combination treatment with FS122m and anti-mPD-1 also prolonged median survival (Figure 3B) and induced complete tumor regression in 7 of 9 mice, compared with none of the 9 mice treated with FS122m monotherapy and only 1 of the 9 mice treated with anti-mPD-1 monotherapy (Figure 3D). Mice in the monotherapy and combination treatment groups experienced no additional mortality or clinical signs, and changes in body weight (BW) were comparable to those in the isotype control group, indicating that all treatments were well tolerated (Figure 3C).

[0163] 3.2 JC mouse breast tumor model In the JC mouse breast tumor model, anti-mPD1 monotherapy showed no antitumor effect (TGI = 7%), whereas FS122m monotherapy showed marginal TGI (47.7%) compared to the isotype control (P < 0.0001, day 22) and slightly prolonged median survival compared to the isotype control (26 days vs. 33 days) (Figure 4A, B, D). Combination treatment of FS122m and anti-mPD1 increased TGI (85.1%) compared to FS122m (P < 0.0001, day 22) and anti-mPD1 (P < 0.0001, day 22) monotherapy (Figure 4A). Combination treatment with FS122m and anti-mPD-1 also extended median survival (50 days) (Figure 4B) and induced complete tumor regression in 3 of 10 mice, whereas neither FS112 nor anti-mPD-1 monotherapy achieved complete tumor regression in any of the 10 mice (Figure 4D). There were no relevant changes in body weight between treatment groups compared to isotype control, demonstrating that all treatments were well tolerated (Figure 4C).

[0164] 3.3 Eph-1424 mouse mammary tumor model The antitumor efficacy of the combination of FS122m and anti-mPD1 was also evaluated in an Eph4-1424 subcutaneous breast tumor model in BALB / c mice. Both anti-mPD1 and FS122m monotherapy demonstrated significant antitumor efficacy (TGI = 68.8% and 51.3%, respectively, P < 0.0001), significantly extending median survival compared with isotype control (43 days vs. 56 days vs. undefined) (Figure 5A, B, D). Treatment with the combination of FS122m and anti-mPD1 increased TGI (105.6%) compared with FS122m (P < 0.0001, day 28) and anti-mPD1 (P < 0.0001, day 28) monotherapy (Figure 5A). Combination treatment with FS122m and anti-mPD-1 also prolonged median survival (Figure 5B) and induced complete tumor regression in 10 of 10 mice, compared with only 1 of 10 mice treated with FS122m monotherapy and 6 of 10 mice treated with anti-mPD-1 monotherapy (Figure 5D). There were no relevant changes in body weight between treatment groups compared to isotype control, suggesting that all treatments were well tolerated (Figure 5C).

[0165] Overall, combined treatment with FS122m and anti-mPD-1 resulted in statistically significantly higher TGI than either FS122m or anti-mPD-1 monotherapy. The combination of FS122m and anti-mPD-1 achieved complete tumor regression in the majority of mice in two of three mouse breast tumor models (Figures 3D and 5D). The difference compared to FS122m and anti-PD-1 monotherapy was particularly striking in the E0771 mouse tumor model, where the combination of FS122m and anti-mPD-1 resulted in complete tumor regression in seven of nine mice, compared with one of nine mice in the anti-mPD1 monotherapy group and none in the FS122m monotherapy group (Figure 3D). Complete tumor regression was also achieved in some mice in the JC mouse tumor model as a result of combination therapy with FS122m and anti-mPD-1, whereas monotherapy with both FS112m and anti-mPD-1 failed to result in complete tumor regression in any of the treated mice (Figure 4D).In all three mouse tumor models, combination therapy resulted in an extended median survival compared with monotherapy (Figures 3B, 4B, and 5B).

[0166] 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.

[0167] Light chain annotation i.mAb 2 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.

[0168] 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

[0169] 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

[0170] 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

[0171] 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

[0172] 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

[0173] 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

[0174] 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

[0175] 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

[0176] 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

[0177] 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

[0178] 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

[0179] 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

[0180] 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

[0181] 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

[0182] 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

[0183] 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

[0184] 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]

[0185] 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

[0186] Amino acid sequences of the CH2 domain containing the LALA, PA, and LALA-PA mutations (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]

[0187] Pembrolizumab amino acid sequence SEQ ID NO: 93 Heavy chain AA [ka] SEQ ID NO: 94 Light chain AA [ka]

[0188] 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.

[0189] [Table 2-1] [Table 2-2] [Table 2-3] [Table 2-4] [Table 2-5]

[0190] 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 molecule in combination with a PD-1 / PD-L1 inhibitor.

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 PD-1 / PD-L1 inhibitor.

3. The kit includes: (a) an antibody molecule that binds to MSLN and CD137 and a pharmaceutically acceptable excipient; and (b) a PD-1 / PD-L1 inhibitor 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, each located in the AB and EF structural loops of the CH3 domain, and wherein the first and second sequences have the sequences set forth in SEQ ID NOs: 87 and 88, respectively [FS22-172-003].

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 claims 1 to 9, wherein the PD-1 / PD-L1 inhibitor is an antibody that binds to PD-1 or PD-L1.

11. 11. The antibody molecule for use, method of treating cancer, or kit according to claim 10, wherein the antibody molecule that binds to PD-1 is selected from the group consisting of nivolumab, pembrolizumab, and cemiplimab.

12. 11. The antibody molecule for use, method of treating cancer, or kit according to claim 10, wherein the antibody molecule that binds to PD-L1 is selected from the group consisting of avelumab, atezolizumab, and durvalumab.

13. 13. The antibody molecule for use or method of treating cancer according to any one of claims 1, 2 and 4 to 12, wherein the cancer 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 PD-1 / PD-L1 inhibitor results in an anti-tumor effect that is greater than the combined anti-tumor effect resulting from treatment of the patient with either the antibody molecule that binds to MSLN and CD137 or the PD-1 / PD-L1 inhibitor 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 PD-1 / PD-L1 inhibitor are administered to a patient simultaneously or sequentially.

17. 17. The antibody molecule for use or method of treating cancer according to 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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