Antibody, method of production and use thereof, isolated nucleic acid, vector, host cell, immunoconjugate and pharmaceutical formulation.

Description

Antibody, method of production and use thereof, isolated nucleic acid, vector, host cell, immunoconjugate and pharmaceutical formulation. Separated from BR112021002037-3, filed on August 9, 2019. Technical Field

[001] This disclosure relates to anti-CD137 antigen-binding molecules and methods of using them. Previous Technique

[002] Cancer is a fatal disease that is difficult to cure completely, except in a few cases. The outcome of treatment with chemotherapy, which is the main therapeutic method, is not good. It has been suggested that not only the heterogeneity of the cancer cells themselves, but the tumor microenvironment plays a significant role as a factor that hinders cancer treatment (NPL 1). Recently, unresectable malignant melanoma and others have shown themselves to be potentially curable with an anti-CTLA-4 antibody, which suppresses the immunosuppressive function of CTLA-4 and therefore promotes T cell activation (NPL 2). In 2011, an anti-human CTLA4 monoclonal antibody (ipilimumab) was approved by the US Food and Drug Administration (FDA) as the world's first immune-activating antibody drug.In addition, inhibitory antibodies against PD-1 and PD-L1, other immune checkpoint molecules besides CTLA-4, have also been reported as possessing therapeutic effects (NPL 3) and have been approved by the FDA.

[003] It is understood that T cells, which have important roles in tumor immunity, are activated by two signals: 1) binding of a T cell receptor (TCR) to an antigenic peptide presented by major histocompatibility complex (MHC) class I molecules and TCR activation; and 2) binding of a co-stimulatory molecule on the surface of T cells to its ligands. Petition 870260042208, dated 05 / 05 / 2026, page 18 / 1364 2 / 448 in antigen-presenting cells and activation of the co-stimulatory molecule. In addition, the activation of co-stimulatory molecules belonging to the tumor necrosis factor receptor (TNFRSF) superfamily, including CD137 (4-1BB), on the surface of T cells has been described as important for T cell activation (NPL 4).

[004] TNFRSF includes CD137, CD40, OX40, RANK, GITR and such molecules. CD137 is expressed not only on the surface of T cells, but also on the surface of other immune cells, such as dendritic cells (DCs), B cells, NK cells, macrophages and neutrophils (NPL 5).

[005] The CD137 agonist antibody has already demonstrated an antitumor effect in a mouse model, and this was shown to result primarily from the activation of CD8-positive T cells and NK cells by mouse model experiments (NPL 6). However, side effects due to nonspecific hepatotoxicity of the CD137 agonist antibody have become clinical and non-clinical problems, hindering the desired progress in drug development (NPL 7, NPL 8). It is suggested that the side effects are mainly caused by the activation of immune cells in non-tumor and non-immune tissues, such as the liver, which involves the binding of the antibody to the Fcy receptor through the antibody's constant region (NPL 9).On the other hand, it has been reported that, for agonistic antibodies of the anti-TNF receptor superfamily member to exhibit agonistic activity in vivo, the antibody needs to be cross-linked by cells expressing the Fcy receptor (cells expressing FcyRII) (NPL 10). That is, the binding of the CD137 agonist antibody to the Fcy receptor is involved both in the drug's efficacy—the antibody's antitumor effect—and in its side effects, such as hepatotoxicity. Thus, increased binding between the antibody and the Fcy receptor should increase the drug's efficacy, but it may also... Petition 870260042208, dated 05 / 05 / 2026, p. 19 / 1364 3 / 448 Increasing hepatotoxic side effects and reducing antibody-Fcy receptor binding may reduce side effects, but also reduce drug efficacy. There have been no reports to date of a CD137 antibody agonist whose drug efficacy and side effects are separate. Furthermore, the antitumor effect of the CD137 antibody agonist itself is by no means clinically potent, and further enhancement of drug efficacy along with prevention of toxicity is desired. Consequently, the development of a novel drug capable of inducing antitumor immune responses while reducing these side effects is desirable.

[006] When a therapeutic antibody is administered to a living body, it is desirable that its target antigen be specifically expressed at the site of injury only. However, in many cases, the same antigen is also expressed at sites without injury, i.e., normal tissues, and this can be a cause of undesirable side effects from a treatment standpoint. For example, although antibodies against tumor antigens may exhibit cytotoxic activity in tumor cells by ADCC, etc., they may also damage normal cells if the same antigen is expressed in normal cells. In order to address the problems mentioned above, a focus has been placed on the phenomenon where certain compounds are abundantly present in target tissues (e.g., tumor tissues) and a technology to search for antigen-binding molecules with variable antigen-binding activity depending on the concentration of such compounds (e.g., PTL 1). List of Citations Patent Literature

[007] [PTL 1] WO2013 / 180200 Non-Patent Literature

[008] [NPL 1] Hanahan, Cell, 2011, 144, 646-74 Petition 870260042208, dated 05 / 05 / 2026, p. 20 / 1364 4 / 448

[009] [NPL 2] Prieto, Clin Cancer Res. 2012, 18, 2039-47

[0010] [NPL 3] Hamid, Expert Opin. Biol. Ther., 2013, 6, 847-61

[0011] [NPL 4] Summers, Nat Rev Immunol, 2012, 12, 339-51

[0012] [NPL 5] Vinay, Cellular and Molecular Immunology, 2011, 8, 281-284

[0013] [NPL 6] Houot, Blood, 2009, 114, 3431-8

[0014] [NPL 7] Ascierto, Semin Oncol, 2010, 37, 508-16

[0015] [NPL 8] Dubrot, Cancer Immunol Immunother, 2010, 59, 1223-33

[0016] [NPL 9] Schabowsky, Vaccine, 2009, 28, 512-22

[0017] [NPL 10] Li, Proc Natl Acad Sci US A. 2013, 110 (48), 195016 Summary of the Invention Technical Problem

[0018] This disclosure relates to anti-CD137 antigen-binding molecules and methods of using them. Solution to the Problem

[0019] In order to provide anti-CD137 antigen-binding molecules that have immunocyte-activating, cytotoxic, or antitumor activity, yet have reduced effects on non-tumor tissues such as normal tissues and have fewer side effects, and to provide methods of use thereof, the present disclosure provides anti-CD137 antigen-binding molecules characterized by their CD137-binding activity varying depending on various compounds (e.g., small molecule compounds) in target tissues (e.g., tumor tissues), and provides methods of use thereof, pharmaceutical formulations, and others. In one embodiment, the anti-CD137 antigen-binding molecules of the present disclosure have low side effects, and thus the dosage can be increased without concern. Petition 870260042208, dated 05 / 05 / 2026, page 21 / 1364 5 / 448 regarding side effects and, as a result, they may exhibit the efficacy of the stronger drug (cytotoxic activity or antitumor activity).

[0020] Specifically, this disclosure provides anti-CD137 antigen-binding molecules, methods of use thereof, pharmaceutical formulations, and such, as described exemplarily below.

[0021] [1] An anti-CD137 antigen-binding molecule that has CD137-dependent binding activity of a small molecule compound.

[0022] [2] The anti-CD137 antigen-binding molecule of [1], in which the CD137 binding activity in the presence of 10 μM, 50 μM, 100 μM, 150 μM, 200 μM or 250 pM of the small molecule compound is two or more times greater than the CD137 binding activity in the absence of the small molecule compound.

[0023] [2.1] The anti-CD137 antigen-binding molecule of [1] or [2], where the CD137 binding activity in the presence of 10 μM or more of the small molecule compound is two or more times greater than the CD137 binding activity in the absence of the small molecule compound.

[0024] [2.2] The anti-CD137 antigen-binding molecule of any of [1] to [2.1], wherein the KD value for CD137 in the presence of 10 μM or more of the small molecule compound is 5 x 10-7M or less.

[0025] [2.3] Anti-CD137 antigen-binding molecule of any of [1] to [2.2], where the KD value for CD137 in the absence of the small molecule compound is 1 x 10-6M or more.

[0026] [2.4] The anti-CD137 antigen-binding molecule of [1], wherein the KD value for CD137 in a solution is prepared such that the concentration of the small molecule compound is 10 μM Petition 870260042208, dated 05 / 05 / 2026, p. 22 / 1364 6 / 448 or more, that is, 5 x 10-7M or less, and the KD value for CD137 in a solution to which the small molecule compound is not added is 1 x 10-6M or more.

[0027] [2.5] The anti-CD137 antigen-binding molecule of [1], wherein the KD value for CD137 in a solution that is prepared so that the concentration of the small molecule compound is 10 μM or more, and the KD value for CD137 in a solution to which the small molecule compound is not added, are measured by a Biacore assay within 24 hours after CD137 and the anti-CD137 antigen-binding molecule are contacted in the solution.

[0028] [2.6] The anti-CD137 antigen-binding molecule of any of [1] to [2.5], which forms a trimolecular complex with the small molecule compound and CD137.

[0029] [2.7] Anti-CD137 antigen-binding molecule of any of [1] to [2.6], which binds to human and monkey-derived CD137.

[0030] [2.8] Anti-CD137 antigen-binding molecule of any of [1] to [2.7], wherein the small molecule compound is an adenosine-containing compound.

[0031] [2.9] The anti-CD137 antigen-binding molecule of any of [1] to [2.8], where the small molecule compound is ATP.

[0032] [3] The anti-CD137 antigen-binding molecule of any one of [1] to [2.9], comprising any combination of HVR-H1, HVR-H2 and HVR-H3 selected from (a) to (k) below: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7, HVR-H2 comprising the amino acid sequence of SEQ ID NO: 8 and HVR-H3 comprising the amino acid sequence of SEQ ID NO: 17; (b) HVR-H1 comprising the amino acid sequence Petition 870260042208, dated 05 / 05 / 2026, p. 23 / 1364 7 / 448 of SEQ ID NO: 7, HVR-H2 comprising the amino acid sequence of SEQ ID NO: 9, and HVR-H3 comprising the amino acid sequence of SEQ ID NO: 17; (c) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7, HVR-H2 comprising the amino acid sequence of SEQ ID NO: 10, and HVR-H3 comprising the amino acid sequence of SEQ ID NO: 17; (d) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7, HVR-H2 comprising the amino acid sequence of SEQ ID NO: 11, and HVR-H3 comprising the amino acid sequence of SEQ ID NO: 18; (e) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7, HVR-H2 comprising the amino acid sequence of SEQ ID NO: 8, and HVR-H3 comprising the amino acid sequence of SEQ ID NO: 18; (f) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7, HVR-H2 comprising the amino acid sequence of SEQ ID NO: 12 and HVR-H3 comprising the amino acid sequence of SEQ ID NO: 18; (g) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7, HVR-H2 comprising the amino acid sequence of SEQ ID NO: 13, and HVR-H3 comprising the amino acid sequence of SEQ ID NO: 18; (h) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7, HVR-H2 comprising the amino acid sequence of SEQ ID NO: 14 and HVR-H3 comprising the amino acid sequence of SEQ ID NO: 19; (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7, HVR-H2 comprising the amino acid sequence of SEQ ID NO: 15, and HVR-H3 comprising the Petition 870260042208, dated 05 / 05 / 2026, p. 24 / 1364 8 / 448 amino acid sequence of SEQ ID NO: 20; (j) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7, HVR-H2 comprising the amino acid sequence of SEQ ID NO: 16, and HVR-H3 comprising the amino acid sequence of SEQ ID NO: 20; and (k) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7, HVR-H2 comprising the amino acid sequence of SEQ ID NO: 14, and HVR-H3 comprising the amino acid sequence of SEQ ID NO: 17.

[0033] [3.1] The anti-CD137 antigen-binding molecule of any one of [1] to [3], comprising any combination of HVR-L1, HVR-L2 and HVR-L3 selected from (a) to (g) below: (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 21, HVR-L2 comprising the amino acid sequence of SEQ ID NO: 26, and HVR-L3 comprising the amino acid sequence of SEQ ID NO: 27; (b) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 22, HVR-L2 comprising the amino acid sequence of SEQ ID NO: 26, and HVR-L3 comprising the amino acid sequence of SEQ ID NO: 27; (c) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 21, HVR-L2 comprising the amino acid sequence of SEQ ID NO: 26, and HVR-L3 comprising the amino acid sequence of SEQ ID NO: 28; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 21, HVR-L2 comprising the amino acid sequence of SEQ ID NO: 26, and HVR-L3 comprising the amino acid sequence of SEQ ID NO: 29; (e) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 23, HVR-L2 comprising the sequence of Petition 870260042208, dated 05 / 05 / 2026, p. 25 / 1364 9 / 448 amino acids of SEQ ID NO: 26, and HVR-L3 comprising the amino acid sequence of SEQ ID NO: 27; (f) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 24, HVR-L2 comprising the amino acid sequence of SEQ ID NO: 26, and HVR-L3 comprising the amino acid sequence of SEQ ID NO: 27; and (g) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 25, HVR-L2 comprising the amino acid sequence of SEQ ID NO: 26, and HVR-L3 comprising the amino acid sequence of SEQ ID NO: 27.

[0034] [4] An anti-CD137 antigen-binding molecule comprising any combination of HVR-H1, HVR-H2, HVR-H3, HVR-L1, HVR-L2 and HVR-L3 selected from (a) to (m) below: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7, HVR-H2 comprising the amino acid sequence of SEQ ID NO: 8, HVR-H3 comprising the amino acid sequence of SEQ ID NO: 17, HVR-L1 comprising the amino acid sequence of SEQ ID NO: 21, HVR-L2 comprising the amino acid sequence of SEQ ID NO: 26 and HVR-L3 comprising the amino acid sequence of SEQ ID NO: 27; (b) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7, HVR-H2 comprising the amino acid sequence of SEQ ID NO: 9, HVR-H3 comprising the amino acid sequence of SEQ ID NO: 17, HVR-L1 comprising the amino acid sequence of SEQ ID NO: 22, HVR-L2 comprising the amino acid sequence of SEQ ID NO: 26 and HVR-L3 comprising the amino acid sequence of SEQ ID NO: 27; (c) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7, HVR-H2 comprising the amino acid sequence of SEQ ID NO: 10, HVR-H3 comprising the sequence Petition 870260042208, dated 05 / 05 / 2026, p. 26 / 1364 10 / 448 amino acids of SEQ ID NO: 17, HVR-L1 comprising the amino acid sequence of SEQ ID NO: 22, HVR-L2 comprising the amino acid sequence of SEQ ID NO: 26 and HVR-L3 comprising the amino acid sequence of SEQ ID NO: 27; (d) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7, HVR-H2 comprising the amino acid sequence of SEQ ID NO: 11, HVR-H3 comprising the amino acid sequence of SEQ ID NO: 18, HVR-L1 comprising the amino acid sequence of SEQ ID NO: 21, HVR-L2 comprising the amino acid sequence of SEQ ID NO: 26 and HVR-L3 comprising the amino acid sequence of SEQ ID NO: 27; (e) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7, HVR-H2 comprising the amino acid sequence of SEQ ID NO: 8, HVR-H3 comprising the amino acid sequence of SEQ ID NO: 18, HVR-L1 comprising the amino acid sequence of SEQ ID NO: 21, HVR-L2 comprising the amino acid sequence of SEQ ID NO: 26 and HVR-L3 comprising the amino acid sequence of SEQ ID NO: 27; (f) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7, HVR-H2 comprising the amino acid sequence of SEQ ID NO: 12, HVR-H3 comprising the amino acid sequence of SEQ ID NO: 18, HVR-L1 comprising the amino acid sequence of SEQ ID NO: 21, HVR-L2 comprising the amino acid sequence of SEQ ID NO: 26, and HVR-L3 comprising the amino acid sequence of SEQ ID NO: 28; (g) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7, HVR-H2 comprising the amino acid sequence of SEQ ID NO: 13, HVR-H3 comprising the amino acid sequence of SEQ ID NO: 18, HVR-L1 comprising the amino acid sequence of SEQ ID NO: 21, HVR-L2 comprising Petition 870260042208, dated 05 / 05 / 2026, p. 27 / 1364 11 / 448 the amino acid sequence of SEQ ID NO: 26, and HVR-L3 comprising the amino acid sequence of SEQ ID NO: 29; (h) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7, HVR-H2 comprising the amino acid sequence of SEQ ID NO: 14, HVR-H3 comprising the amino acid sequence of SEQ ID NO: 19, HVR-L1 comprising the amino acid sequence of SEQ ID NO: 23, HVR-L2 comprising the amino acid sequence of SEQ ID NO: 26, and HVR-L3 comprising the amino acid sequence of SEQ ID NO: 27; (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7, HVR-H2 comprising the amino acid sequence of SEQ ID NO: 15, HVR-H3 comprising the amino acid sequence of SEQ ID NO: 20, HVR-L1 comprising the amino acid sequence of SEQ ID NO: 24, HVR-L2 comprising the amino acid sequence of SEQ ID NO: 26 and HVR-L3 comprising the amino acid sequence of SEQ ID NO: 27; (j) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7, HVR-H2 comprising the amino acid sequence of SEQ ID NO: 15, HVR-H3 comprising the amino acid sequence of SEQ ID NO: 20, HVR-L1 comprising the amino acid sequence of SEQ ID NO: 25, HVR-L2 comprising the amino acid sequence of SEQ ID NO: 26 and HVR-L3 comprising the amino acid sequence of SEQ ID NO: 27; (k) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7, HVR-H2 comprising the amino acid sequence of SEQ ID NO: 16, HVR-H3 comprising the amino acid sequence of SEQ ID NO: 20, HVR-L1 comprising the amino acid sequence of SEQ ID NO: 25, HVR-L2 comprising the amino acid sequence of SEQ ID NO: 26 and HVR-L3 comprising the amino acid sequence of SEQ ID NO: 27; Petition 870260042208, dated 05 / 05 / 2026, p. 28 / 1364 12 / 448 (l) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7, HVR-H2 comprising the amino acid sequence of SEQ ID NO: 14, HVR-H3 comprising the amino acid sequence of SEQ ID NO: 19, HVR-L1 comprising the amino acid sequence of SEQ ID NO: 24, HVR-L2 comprising the amino acid sequence of SEQ ID NO: 26 and HVR-L3 comprising the amino acid sequence of SEQ ID NO: 27; and (m) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7, HVR-H2 comprising the amino acid sequence of SEQ ID NO: 14, HVR-H3 comprising the amino acid sequence of SEQ ID NO: 17, HVR-L1 comprising the amino acid sequence of SEQ ID NO: 21, HVR-L2 comprising the amino acid sequence of SEQ ID NO: 26 and HVR-L3 comprising the amino acid sequence of SEQ ID NO: 27.

[0035] [5] An anti-CD137 antigen-binding molecule comprising: (a) a VH possessing at least 95% sequence identity with any of the amino acid sequences of SEQ ID NOs: 43 to 53; or (b) a VL with at least 95% sequence identity with any of the amino acid sequences of SEQ ID NOs: 54 to 60.

[0036] [5.1] An anti-CD137 antigen-binding molecule comprising any combination of VH and VL selected from (a) to (m) below: (a) a VH having at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 43, and a VL having at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 54; (b) a VH possessing at least 95% identity of Petition 870260042208, dated 05 / 05 / 2026, page 29 / 1364 13 / 448 sequence with the amino acid sequence of SEQ ID NO: 44, and a VL possessing at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 55; (c) a VH having at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 45, and a VL having at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 55; (d) a VH having at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 46, and a VL having at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 54; (e) a VH possessing at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 47, and a VL possessing at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 54; (f) a VH having at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 48, and a VL having at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 56; (g) a VH having at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 49, and a VL having at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 57; (h) a VH having at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 50, and a VL having at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 58; (i) a VH possessing at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 51, and a VL possessing at least 95% sequence identity with the Petition 870260042208, dated 05 / 05 / 2026, page 30 / 1364 14 / 448 amino acid sequence of SEQ ID NO: 59; (j) a VH having at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 51, and a VL having at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 60; (k) a VH having at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 52, and a VL having at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 60; (l) a VH having at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 50, and a VL having at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 59; and (m) a VH having at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 53, and a VL having at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 54.

[0037] [5.2] An anti-CD137 antigen-binding molecule comprising any combination of VH and VL selected from (a) to (m) below: (a) a VH comprising the amino acid sequence of SEQ ID NO: 43, and a VL comprising the amino acid sequence of SEQ ID NO: 54; (b) a VH comprising the amino acid sequence of SEQ ID NO: 44, and a VL comprising the amino acid sequence of SEQ ID NO: 55; (c) a VH comprising the amino acid sequence of SEQ ID NO: 45, and a VL comprising the amino acid sequence of SEQ ID NO: 55; (d) a VH comprising the amino acid sequence Petition 870260042208, dated 05 / 05 / 2026, page 31 / 1364 15 / 448 of SEQ ID NO: 46, and a VL comprising the amino acid sequence of SEQ ID NO: 54; (e) a VH comprising the amino acid sequence of SEQ ID NO: 47, and a VL comprising the amino acid sequence of SEQ ID NO: 54; (f) a VH comprising the amino acid sequence of SEQ ID NO: 48, and a VL comprising the amino acid sequence of SEQ ID NO: 56; (g) a VH comprising the amino acid sequence of SEQ ID NO: 49, and a VL comprising the amino acid sequence of SEQ ID NO: 57; (h) a VH comprising the amino acid sequence of SEQ ID NO: 50, and a VL comprising the amino acid sequence of SEQ ID NO: 58; (i) a VH comprising the amino acid sequence of SEQ ID NO: 51 and a VL comprising the amino acid sequence of SEQ ID NO: 59; (j) a VH comprising the amino acid sequence of SEQ ID NO: 51 and a VL comprising the amino acid sequence of SEQ ID NO: 60; (k) a VH comprising the amino acid sequence of SEQ ID NO: 52, and a VL comprising the amino acid sequence of SEQ ID NO: 60; (l) a VH comprising the amino acid sequence of SEQ ID NO: 50, and a VL comprising the amino acid sequence of SEQ ID NO: 59; and (m) a VH comprising the amino acid sequence of SEQ ID NO: 53, and a VL comprising the amino acid sequence of SEQ ID NO: 54.

[0038] [5.3] An anti-CD137 antigen-binding molecule whose Petition 870260042208, dated 05 / 05 / 2026, p. 32 / 1364 16 / 448 value of [binding activity (amount of binding) to CD137 in the presence of 10 μM or more of a small molecule compound] / [binding activity (amount of binding) to CD137 in the absence of the molecule compound] is equal to or greater than that of a reference antigen-binding molecule, wherein the reference antigen-binding molecule is an anti-CD137 antigen-binding molecule comprising a combination of HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7, HVR-H2 comprising the amino acid sequence of SEQ ID NO: 8, HVR-H3 comprising the amino acid sequence of SEQ ID NO: 17, HVR-L1 comprising the amino acid sequence of SEQ ID NO: 21, HVR-L2 comprising the amino acid sequence of SEQ ID NO: 26, and HVR-L3 comprising the amino acid sequence of SEQ ID NO: 27.

[0039] [5.4] The anti-CD137 antigen-binding molecule of [5.3], wherein the reference antigen-binding molecule is an anti-CD137 antigen-binding molecule comprising a combination of a VH comprising the amino acid sequence of SEQ ID NO: 43 and a VL comprising the amino acid sequence of SEQ ID NO: 54.

[0040] [5.5] An anti-CD137 antigen-binding molecule whose value of [CD137 binding activity (KD) in the presence of 1 μM of a small molecule compound] / [CD137 binding activity (KD) in the presence of 10 μM or more of the small molecule compound] is equal to or greater than that of a reference antigen-binding molecule, wherein the reference antigen-binding molecule is an anti-CD137 antigen-binding molecule comprising a combination of HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7, HVR-H2 comprising the sequence of Petition 870260042208, dated 05 / 05 / 2026, p. 33 / 1364 17 / 448 amino acids of SEQ ID NO: 8, HVR-H3 comprising the amino acid sequence of SEQ ID NO: 17, HVR-L1 comprising the amino acid sequence of SEQ ID NO: 21, HVR-L2 comprising the amino acid sequence of SEQ ID NO: 26 and HVR-L3 comprising the amino acid sequence of SEQ ID NO: 27.

[0041] [5.6] The anti-CD137 antigen-binding molecule of [5.5], wherein the reference antigen-binding molecule is an anti-CD137 antigen-binding molecule comprising a combination of a VH comprising the amino acid sequence of SEQ ID NO: 43 and a VL comprising the amino acid sequence of SEQ ID NO: 54.

[0042] [5.7] An anti-CD137 antigen-binding molecule that has CD137-binding activity dependent on a small molecule compound, wherein the anti-CD137 antigen-binding molecule competes with the antigen-binding molecule of any of [3] to [5.2] to bind to CD137 in the presence of 10 μM or more, 50 μM or more, 100 μM or more, 150 μM or more, 200 μM or more, or 250 μM or more of the small molecule compound.

[0043] [5.8] An anti-CD137 antigen-binding molecule that has CD137-dependent small molecule compound binding activity, wherein the anti-CD137 antigen-binding molecule binds to the same CD137 epitope bound by the antigen-binding molecule of any of [3] to [5.2] in the presence of 10 μM or more, 50 μM or more, 100 μM or more, 150 μM or more, 200 μM or more, or 250 μM or more of the small molecule compound.

[0044] [5.8A] Anti-CD137 antigen-binding molecule of any of [5.3] to [5.8], wherein the small molecule compound is an adenosine-containing compound.

[0045] [5.8B] The anti-CD137 antigen-binding molecule of any of [5.3] to [5.8A], wherein the small molecule compound Petition 870260042208, dated 05 / 05 / 2026, p. 34 / 1364 18 / 448 is ATP.

[0046] [5.9] The anti-CD137 antigen-binding molecule of any of [1] to [5.8B], which is a monoclonal antibody or an antigen-binding fragment thereof.

[0047] [5.10] The anti-CD137 antigen-binding molecule of any one of [1] to [5.9], which is a human antibody, humanized antibody or chimeric antibody, or an antigen-binding fragment thereof.

[0048] [5.11] The anti-CD137 antigen-binding molecule of any of [1] to [5.10], which is a full-length IgG1 antibody.

[0049] [5.12] The anti-CD137 antigen-binding molecule of any of [1] to [5.11], comprising an altered Fc region in which at least one amino acid is altered, wherein the altered Fc region increased FcyRIIb binding activity compared with a parental Fc region that does not comprise the amino acid alteration.

[0050] [5.13] The anti-CD137 antigen-binding molecule of [5.12], where the altered Fc binding activity to FcyRIIb is equal to or greater than that of a reference Fc region, where the reference Fc is a human IgG1 Fc region comprising a combination of G236N / H268D / A330K amino acid substitutions according to EU numbering.

[0051] [5.14] The anti-CD137 antigen-binding molecule of [5.12] or [5.13], wherein the reference Fc region comprises the amino acid sequence of SEQ ID NO: 153.

[0052] [5.15] The anti-CD137 antigen-binding molecule of [5.12], where at least one amino acid change is at least one amino acid substitution selected from the group consisting of G236N, H268D and A330K according to EU numbering. Petition 870260042208, dated 05 / 05 / 2026, p. 35 / 1364 19 / 448

[0053] [5.16] The anti-CD137 antigen-binding molecule of [5.12] or [5.15], wherein at least one amino acid change is a combination of G236N / H268D / A330K amino acid substitutions according to EU numbering.

[0054] [5.17] The anti-CD137 antigen-binding molecule of any of [5.12] to [5.16], wherein the parental Fc region is derived from a human IgG1 Fc region.

[0055] [5.18] The anti-CD137 antigen-binding molecule of any one of [1] to [5.17], comprising an altered Fc region in which at least one amino acid is altered, wherein the anti-CD137 antigen-binding molecule has an increased isoelectric point (pI) compared with that of a parental anti-CD137 antigen-binding molecule comprising a parental Fc region that does not comprise the amino acid alteration.

[0056] [5.19] The anti-CD137 antigen-binding molecule of [5.18], where at least one amino acid change is a change of an amino acid residue that can be exposed on the surface of the parental Fc region.

[0057] [5.20] The anti-CD137 antigen-binding molecule of [5.18] or [5.19], in which at least one amino acid change is: (i) replacement of at least one amino acid residue with a negative side chain charge in the parental Fc region by an amino acid residue with no side chain charge, (ii) replacement of at least one amino acid residue with no side chain charge in the parental Fc region by an amino acid residue with a positive side chain charge, and / or (iii) replacement of at least one amino acid residue with a negative side chain charge in the parental Fc region by an amino acid residue with a positive side chain charge. Petition 870260042208, dated 05 / 05 / 2026, p. 36 / 1364 20 / 448

[0058] [5.21] The anti-CD137 antigen-binding molecule of any one of [5.18] to [5.20], wherein at least one amino acid change is a combination of amino acid substitutions and wherein the amino acid substitutions are located in positions that are conformationally close to each other.

[0059] [5.22] The anti-CD137 antigen-binding molecule of any of [5.18] to [5.21], in which the binding activity of the altered Fc region to an Fcy receptor (FcyR) is not substantially reduced compared with that of the parental Fc region.

[0060] [5.23] The anti-CD137 antigen-binding molecule of [5.2 2], where the Fcy receptor (FcyR) is FcyRIIb.

[0061] [5.24] The anti-CD137 antigen-binding molecule of any one of [5.18] to [5.23], in which at least one amino acid change is at least one amino acid substitution selected from the group consisting of Q311R, P343R and D413K according to EU numbering.

[0062] [5.25] The anti-CD137 antigen-binding molecule of any one of [5.18] to [5.24], in which at least one amino acid change is (i) a P343R amino acid substitution, (ii) a combination of Q311R / P343R amino acid substitutions, or (iii) a combination of Q311R / D413K amino acid substitutions, according to EU numbering.

[0063] [6] The anti-CD137 antigen-binding molecule of any one of [1] to [5,25], comprising an altered Fc region, wherein the altered Fc region comprises any combination of amino acid changes selected from the following: L235W / G236N / H268D / Q295L / K326T / A330K / P343R / D413K; K214R / L235W / G236N / H268D / Q295L / K326T / A330K / P343R / D413K; L234Y / P238D / T250V / V264I / T307P / A330K / P343R / D413K; L234Y / P238D / V264I / A330K / P343R / D413K; L234Y / G237D / P238D / T250V / T307P / A330K / P343R / D413K; Petition 870260042208, dated 05 / 05 / 2026, p. 37 / 1364 21 / 448 L234Y / G237D / P238D / A330K / P343R / D413K; L235W / G236N / H268D / Q295L / K326T / A330K / Q311R / P343R; L234Y / P238D / T250V / V264I / T307P / A330K / Q311R / P343R; L234Y / P238D / V264I / A330K / Q311R / P343R; L234Y / G237D / P238D / T250V / T307P / A330K / Q311R / P343R; L234Y / G237D / P238D / A330K / Q311R / P343R; L235W / G236N / H268D / Q295L / K326T / A330K / P343R; K214R / L235W / G236N / H268D / Q295L / K326T / A330K / P343R; L235W / G236N / H268D / Q295L / K326T / A330K / D413K; K214R / G236N / H268D / A330K / P343R; K214R / L235W / G236N / H268D / A330K / P343R; K214R / G236N / H268D / A330K / D413K; K214R / G236N / H268D / A330K / P343R / D413K; K214R / L235W / G236N / H268D / A330K / P343R / D413K; K214R / G236N / H268D / A330K / Q311R; K214R / L235W / G236N / H268D / A330K / Q311R; K214R / G236N / H268D / A330K / Q311R / P343R; K214R / L235W / G236N / H268D / A330K / Q311R / P343R; K214R / G236N / H268D / A330K / Q311R / D413K; K214R / L235W / G236N / H268D / A330K / Q311R / D413K; and K214R / L235W / G236N / H268D / Q295L / K326T / A330K / Q311R, according to EU numbering.

[0064] [6.1] The anti-CD137 antigen-binding molecule of any of [1] to [6], wherein the altered Fc region is derived from a human IgG1 Fc region.

[0065] [6.2] The anti-CD137 antigen-binding molecule of any of [1] to [6.1], wherein the altered Fc region additionally comprises deletions at positions 446 and 447 according to EU numbering.

[0066] [7] The anti-CD137 antigen-binding molecule of any one of [1] to [6.2], comprising a heavy chain constant region comprising any one of the amino acid sequences of SEQ ID NOs: 64 to 85.

[0067] [7.1] Anti-CD137 antigen-binding molecule, which Petition 870260042208, dated 05 / 05 / 2026, page 38 / 1364 22 / 448 comprises any combination of VH, VL, CH and CL selected from (i) to (xxxviii) below: (i) VH comprising the amino acid sequence with SEQ ID NO: 43, CH comprising the amino acid sequence with SEQ ID NO: 64, VL comprising the amino acid sequence with SEQ ID NO: 54, and CL comprising the amino acid sequence with SEQ ID NO: 63; (ii) VH comprising the amino acid sequence of SEQ ID NO: 43, CH comprising the amino acid sequence of SEQ ID NO: 66, VL comprising the amino acid sequence of SEQ ID NO: 54, and CL comprising the amino acid sequence of SEQ ID NO: 63; (iii) VH comprising the amino acid sequence of SEQ ID NO: 43, CH comprising the amino acid sequence of SEQ ID NO: 67, VL comprising the amino acid sequence of SEQ ID NO: 54, and CL comprising the amino acid sequence of SEQ ID NO: 63; (iv) VH comprising the amino acid sequence of SEQ ID NO: 43, CH comprising the amino acid sequence of SEQ ID NO: 68, VL comprising the amino acid sequence of SEQ ID NO: 54, and CL comprising the amino acid sequence of SEQ ID NO: 63; (v) VH comprising the amino acid sequence of SEQ ID NO: 43, CH comprising the amino acid sequence of SEQ ID NO: 69, VL comprising the amino acid sequence of SEQ ID NO: 54, and CL comprising the amino acid sequence of SEQ ID NO: 63; (vi) VH comprising the amino acid sequence of SEQ ID NO: 43, CH comprising the amino acid sequence of SEQ ID NO: 70, VL comprising the amino acid sequence of Petition 870260042208, dated 05 / 05 / 2026, page 39 / 1364 23 / 448 SEQ ID NO: 54, and CL comprising the amino acid sequence of SEQ ID NO: 63; (vii) VH comprising the amino acid sequence of SEQ ID NO: 43, CH comprising the amino acid sequence of SEQ ID NO: 71, VL comprising the amino acid sequence of SEQ ID NO: 54, and CL comprising the amino acid sequence of SEQ ID NO: 63; (viii) VH comprising the amino acid sequence of SEQ ID NO: 43, CH comprising the amino acid sequence of SEQ ID NO: 73, VL comprising the amino acid sequence of SEQ ID NO: 54, and CL comprising the amino acid sequence of SEQ ID NO: 63; (ix) VH comprising the amino acid sequence of SEQ ID NO: 43, CH comprising the amino acid sequence of SEQ ID NO: 75, VL comprising the amino acid sequence of SEQ ID NO: 54, and CL comprising the amino acid sequence of SEQ ID NO: 63; (x) VH comprising the amino acid sequence of SEQ ID NO: 43, CH comprising the amino acid sequence of SEQ ID NO: 78, VL comprising the amino acid sequence of SEQ ID NO: 54, and CL comprising the amino acid sequence of SEQ ID NO: 63; (xi) VH comprising the amino acid sequence of SEQ ID NO: 43, CH comprising the amino acid sequence of SEQ ID NO: 80, VL comprising the amino acid sequence of SEQ ID NO: 54, and CL comprising the amino acid sequence of SEQ ID NO: 63; (xii) VH comprising the amino acid sequence of SEQ ID NO: 43, CH comprising the amino acid sequence of SEQ ID NO: 82, VL comprising the amino acid sequence of Petition 870260042208, dated 05 / 05 / 2026, page 40 / 1364 24 / 448 SEQ ID NO: 54, and CL comprising the amino acid sequence of SEQ ID NO: 63; (xiii) VH comprising the amino acid sequence of SEQ ID NO: 43, CH comprising the amino acid sequence of SEQ ID NO: 84, VL comprising the amino acid sequence of SEQ ID NO: 54, and CL comprising the amino acid sequence of SEQ ID NO: 63; (xiv) VH comprising the amino acid sequence of SEQ ID NO: 43, CH comprising the amino acid sequence of SEQ ID NO: 85, VL comprising the amino acid sequence of SEQ ID NO: 54, and CL comprising the amino acid sequence of SEQ ID NO: 63; (xv) VH comprising the amino acid sequence of SEQ ID NO: 51, CH comprising the amino acid sequence of SEQ ID NO: 65, VL comprising the amino acid sequence of SEQ ID NO: 59, and CL comprising the amino acid sequence of SEQ ID NO: 63; (xvi) VH comprising the amino acid sequence of SEQ ID NO: 51, CH comprising the amino acid sequence of SEQ ID NO: 72, VL comprising the amino acid sequence of SEQ ID NO: 59, and CL comprising the amino acid sequence of SEQ ID NO: 63; (xvii) VH comprising the amino acid sequence of SEQ ID NO: 51, CH comprising the amino acid sequence of SEQ ID NO: 74, VL comprising the amino acid sequence of SEQ ID NO: 59, and CL comprising the amino acid sequence of SEQ ID NO: 63; (xviii) VH comprising the amino acid sequence of SEQ ID NO: 51, CH comprising the amino acid sequence of SEQ ID NO: 75, VL comprising the amino acid sequence of Petition 870260042208, dated 05 / 05 / 2026, page 41 / 1364 25 / 448 SEQ ID NO: 59, and CL comprising the amino acid sequence of SEQ ID NO: 63; (xix) VH comprising the amino acid sequence of SEQ ID NO: 51, CH comprising the amino acid sequence of SEQ ID NO: 77, VL comprising the amino acid sequence of SEQ ID NO: 59, and CL comprising the amino acid sequence of SEQ ID NO: 63; (xx) VH comprising the amino acid sequence of SEQ ID NO: 51, CH comprising the amino acid sequence of SEQ ID NO: 78, VL comprising the amino acid sequence of SEQ ID NO: 59, and CL comprising the amino acid sequence of SEQ ID NO: 63; (xxi) VH comprising the amino acid sequence of SEQ ID NO: 51, CH comprising the amino acid sequence of SEQ ID NO: 79, VL comprising the amino acid sequence of SEQ ID NO: 59, and CL comprising the amino acid sequence of SEQ ID NO: 63; (xxii) VH comprising the amino acid sequence of SEQ ID NO: 51, CH comprising the amino acid sequence of SEQ ID NO: 80, VL comprising the amino acid sequence of SEQ ID NO: 59, and CL comprising the amino acid sequence of SEQ ID NO: 63; (xxiii) VH comprising the amino acid sequence of SEQ ID NO: 51, CH comprising the amino acid sequence of SEQ ID NO: 81, VL comprising the amino acid sequence of SEQ ID NO: 59, and CL comprising the amino acid sequence of SEQ ID NO: 63; (xxiv) VH comprising the amino acid sequence of SEQ ID NO: 51, CH comprising the amino acid sequence of SEQ ID NO: 82, VL comprising the amino acid sequence of Petition 870260042208, dated 05 / 05 / 2026, page 42 / 1364 26 / 448 SEQ ID NO: 59, and CL comprising the amino acid sequence of SEQ ID NO: 63; (xxv) VH comprising the amino acid sequence of SEQ ID NO: 51, CH comprising the amino acid sequence of SEQ ID NO: 83, VL comprising the amino acid sequence of SEQ ID NO: 59, and CL comprising the amino acid sequence of SEQ ID NO: 63; (xxvi) VH comprising the amino acid sequence of SEQ ID NO: 51, CH comprising the amino acid sequence of SEQ ID NO: 84, VL comprising the amino acid sequence of SEQ ID NO: 59, and CL comprising the amino acid sequence of SEQ ID NO: 63; (xxvii) VH comprising the amino acid sequence of SEQ ID NO: 51, CH comprising the amino acid sequence of SEQ ID NO: 72, VL comprising the amino acid sequence of SEQ ID NO: 60, and CL comprising the amino acid sequence of SEQ ID NO: 63; (xxviii) VH comprising the amino acid sequence of SEQ ID NO: 51, CH comprising the amino acid sequence of SEQ ID NO: 74, VL comprising the amino acid sequence of SEQ ID NO: 60, and CL comprising the amino acid sequence of SEQ ID NO: 63; (xxix) VH comprising the amino acid sequence of SEQ ID NO: 51, CH comprising the amino acid sequence of SEQ ID NO: 75, VL comprising the amino acid sequence of SEQ ID NO: 60, and CL comprising the amino acid sequence of SEQ ID NO: 63; (xxx) VH comprising the amino acid sequence of SEQ ID NO: 51, CH comprising the amino acid sequence of SEQ ID NO: 77, VL comprising the amino acid sequence of Petition 870260042208, dated 05 / 05 / 2026, page 43 / 1364 27 / 448 SEQ ID NO: 60, and CL comprising the amino acid sequence of SEQ ID NO: 63; (xxxi) VH comprising the amino acid sequence of SEQ ID NO: 51, CH comprising the amino acid sequence of SEQ ID NO: 78, VL comprising the amino acid sequence of SEQ ID NO: 60, and CL comprising the amino acid sequence of SEQ ID NO: 63; (xxxii) VH comprising the amino acid sequence of SEQ ID NO: 51, CH comprising the amino acid sequence of SEQ ID NO: 79, VL comprising the amino acid sequence of SEQ ID NO: 60, and CL comprising the amino acid sequence of SEQ ID NO: 63; (xxxiii) VH comprising the amino acid sequence of SEQ ID NO: 51, CH comprising the amino acid sequence of SEQ ID NO: 80, VL comprising the amino acid sequence of SEQ ID NO: 60, and CL comprising the amino acid sequence of SEQ ID NO: 63; (xxxiv) VH comprising the amino acid sequence of SEQ ID NO: 51, CH comprising the amino acid sequence of SEQ ID NO: 81, VL comprising the amino acid sequence of SEQ ID NO: 60, and CL comprising the amino acid sequence of SEQ ID NO: 63; (xxxv) VH comprising the amino acid sequence of SEQ ID NO: 51, CH comprising the amino acid sequence of SEQ ID NO: 82, VL comprising the amino acid sequence of SEQ ID NO: 60, and CL comprising the amino acid sequence of SEQ ID NO: 63; (xxxvi) VH comprising the amino acid sequence of SEQ ID NO: 51, CH comprising the amino acid sequence of SEQ ID NO: 83, VL comprising the amino acid sequence of Petition 870260042208, dated 05 / 05 / 2026, p. 44 / 1364 28 / 448 SEQ ID NO: 60, and CL comprising the amino acid sequence of SEQ ID NO: 63; (xxxvii) VH comprising the amino acid sequence of SEQ ID NO: 51, CH comprising the amino acid sequence of SEQ ID NO: 84, VL comprising the amino acid sequence of SEQ ID NO: 60, and CL comprising the amino acid sequence of SEQ ID NO: 63; and (xxxviii) VH comprising the amino acid sequence of SEQ ID NO: 51, CH comprising the amino acid sequence of SEQ ID NO: 85, VL comprising the amino acid sequence of SEQ ID NO: 60, and CL comprising the amino acid sequence of SEQ ID NO: 63.

[0068] [8] An isolated nucleic acid encoding the anti-CD137 antigen-binding molecule from any of [1] to [7.1].

[0069] [9] A vector comprising the nucleic acid of [8].

[0070]

[10] A host cell comprising the nucleic acid of [8] or the vector of [9].

[0071]

[11] Method for producing an anti-CD137 antigen-binding molecule, comprising culturing the host cell of

[10] such that the anti-CD137 antigen-binding molecule is produced.

[0072]

[12] Immunoconjugate comprising the anti-CD137 antigen-binding molecule of any one of [1] to [7.1] and a cytotoxic agent.

[0073]

[13] A pharmaceutical formulation comprising the anti-CD137 antigen-binding molecule of any of [1] to [7.1] or the immunoconjugate of

[12] ; and a pharmaceutically acceptable carrier.

[0074]

[14] The anti-CD137 antigen-binding molecule of any of [1] to [7.1] or the immunoconjugate of

[12] , which is for use Petition 870260042208, dated 05 / 05 / 2026, page 45 / 1364 29 / 448 as a pharmaceutical product.

[0075] [14.1] The anti-CD137 antigen-binding molecule of any of [1] to [7.1], the immunoconjugate of

[12] or the pharmaceutical formulation of

[13] , which is for use in the treatment of a tumor.

[0076] [14.2] The anti-CD137 antigen-binding molecule, immunoconjugate or pharmaceutical formulation of [14.1], wherein the tumor is a solid tumor infiltrated by a B cell, a dendritic cell, a natural killer cell, a macrophage and / or a CD8-positive T cell.

[0077] [14.3] The anti-CD137 antigen-binding molecule, immunoconjugate or pharmaceutical formulation of [14.1], wherein the tumor is a solid tumor infiltrated by a regulatory T cell (Treg).

[0078]

[15] The anti-CD137 antigen-binding molecule of any of [1] to [7.1], the immunoconjugate of

[12] or the pharmaceutical formulation of

[13] , which is for use in activating an immune cell.

[0079] [15.1] The anti-CD137 antigen-binding molecule, immunoconjugate or pharmaceutical formulation of

[15] , wherein the immune cell is a B cell, a dendritic cell, a natural killer cell, a macrophage and / or a T cell.

[0080] [15.2] The anti-CD137 antigen-binding molecule of any of [1] to [7.1], or the pharmaceutical formulation of

[13] , which is to activate an immune cell in a tumor tissue.

[0081] [15.3] The anti-CD137 antigen-binding molecule or pharmaceutical formulation of [15.2], wherein the immune cell is a B cell, a dendritic cell, a natural killer cell, a macrophage and / or a T cell.

[0082] [15.4] The anti-CD137 antigen-binding molecule of any of [1] to [7.1], the immunoconjugate of

[12] or the pharmaceutical formulation of

[13] , which is for use in cell injury.

[0083]

[16] The anti-CD137 antigen-binding molecule of any of [1] to [7.1], the immunoconjugate of

[12] or the formulation Petition 870260042208, dated 05 / 05 / 2026, page 46 / 1364 30 / 448 pharmaceutical

[13] , whose level of immune activation in a non-tumor tissue is lower than that of an anti-CD137 antigen-binding molecule that does not have CD137-dependent small molecule compound binding activity.

[0084] [16.1] The anti-CD137 antigen-binding molecule, immunoconjugate or pharmaceutical formulation of

[16] , wherein the non-tumor tissue is a lymph node, spleen and / or liver.

[0085] [16.2] The anti-CD137 antigen-binding molecule of any of [1] to [7.1] or the immunoconjugate of

[12] , which does not bind substantially to CD137 expressed in a non-tumor tissue.

[0086] [16.3] The anti-CD137 antigen-binding molecule of any of [1] to [7.1] or the immunoconjugate of

[12] , which has a longer half-life in blood than that of an anti-CD137 antigen-binding molecule that does not have CD137-dependent small molecule compound binding activity.

[0087]

[17] The anti-CD137 antigen-binding molecule of any of [1] to [7.1], the immunoconjugate of

[12] or the pharmaceutical formulation of

[13] , which has a lower level of side effect than an anti-CD137 antigen-binding molecule that does not have CD137-dependent binding activity of a small molecule compound.

[0088] [17.1] Anti-CD137 antigen-binding molecule, immunoconjugate or pharmaceutical formulation of

[17] , wherein the side effect is increased AST, increased ALT, fever, nausea, acute hepatitis, hepatopathy, splenomegaly, enteritis, purulent skin inflammation, reduced neutrophils, reduced lymphocytes, reduced platelets, transaminase expression and / or hyperbilirubinemia.

[0089]

[18] An anti-CD137 antigen-binding molecule that has CD137-dependent agonist activity based on a small molecule compound. Petition 870260042208, dated 05 / 05 / 2026, page 47 / 1364 31 / 448

[0090] [18.1] The anti-CD137 antigen-binding molecule of

[18] , in which the agonist activity for CD137 in the presence of 10 μM, 50 μM, 100 μM, 150 μM, 200 μM or 250 μM of the small molecule compound is two or more times greater than the agonist activity for CD137 in the absence of the small molecule compound.

[0091] [18.2] The anti-CD137 antigen-binding molecule of

[18] or [18.1], wherein the agonist activity for CD137 in the presence of 10 μM or more of the small molecule compound is two or more times greater than the agonist activity for CD137 in the absence of the small molecule compound.

[0092] [18.3] The anti-CD137 antigen-binding molecule of

[18] or [18.1], wherein the agonist activity for CD137 in the presence of 50 μM or more of the small molecule compound is two or more times greater than the agonist activity for CD137 in the absence of the small molecule compound.

[0093] [18.4] The anti-CD137 antigen-binding molecule of

[18] or [18.1], wherein the agonist activity for CD137 in the presence of 250 μM or more of the small molecule compound is two or more times greater than the agonist activity for CD137 in the absence of the small molecule compound.

[0094] [18.5] Anti-CD137 antigen-binding molecule of any of

[18] to [18.4], wherein agonist activity for CD137 is assessed with the amount of IL-2 and / or IFN-γ produced by a cell expressing CD137.

[0095] [18.6] The anti-CD137 antigen-binding molecule of [18.5], where the cell expressing CD137 is an isolated human peripheral blood mononuclear cell (PBMC) or a human PBMC-derived T cell.

[0096] [18.7] Anti-CD137 antigen-binding molecule of any of

[18] to [18.4], wherein agonist activity for CD137 Petition 870260042208, dated 05 / 05 / 2026, page 48 / 1364 32 / 448 is evaluated by a reporter gene assay.

[0097] [18.8] The anti-CD137 antigen-binding molecule of

[18] , which exhibits agonist activity for CD137 in a solution that is prepared such that the final concentration of a small molecule compound is 50 μM or more, and does not exhibit substantially agonist activity for CD137 in a solution to which the small molecule compound is not added.

[0098] [18.9] The anti-CD137 antigen-binding molecule of [18.8], in which the agonist activity for CD137 in a solution that is prepared so that the final concentration of a small molecule compound is 50 pM or more, and the agonist activity for CD137 in a solution to which the small molecule compound is not added, are each evaluated with the amount of IL-2, IFN-γ and / or IL-6 measured within 72 hours after a cell expressing CD137 and the anti-CD137 antigen-binding molecule are contacted in the solution.

[0099] [18.10] The anti-CD137 antigen-binding molecule of [18.8], in which the agonist activity for CD137 in a solution that is prepared so that the final concentration of a small molecule compound is 50 μM or more, and the agonist activity for CD137 in a solution to which the small molecule compound is not added, are each evaluated with a luciferase luminescence signal that is measured within 6 hours after a T cell expressing an NF-kappaB-luciferase and CD137 reporter construct is contacted with the anti-CD137 antigen-binding molecule.

[00100] [18.11] Anti-CD137 antigen-binding molecule of any of

[18] to [18.10], wherein the small molecule compound is an adenosine-containing compound.

[00101] [18.12] The anti-CD137 antigen-binding molecule of any of

[18] to [18.11], wherein the small molecule compound Petition 870260042208, dated 05 / 05 / 2026, page 49 / 1364 33 / 448 is ATP.

[00102]

[19] Anti-CD137 antigen-binding molecule of any of [1] to [7.1], which has CD137-dependent agonist activity of a small molecule compound.

[00103] [19.1] The anti-CD137 antigen-binding molecule of

[19] , in which the agonist activity for CD137 in the presence of 10 μM, 50 μM, 100 μM, 150 μM, 200 μM or 250 μM of the small molecule compound is two or more times greater than the agonist activity for CD137 in the absence of the small molecule compound.

[00104] [19.2] The anti-CD137 antigen-binding molecule of

[19] or [19.1], wherein the agonist activity for CD137 in the presence of 10 μM or more of the small molecule compound is two or more times greater than the agonist activity for CD137 in the absence of the small molecule compound.

[00105] [19.3] The anti-CD137 antigen-binding molecule of

[19] or [19.1], wherein the agonist activity for CD137 in the presence of 50 μM or more of the small molecule compound is two or more times greater than the agonist activity for CD137 in the absence of the small molecule compound.

[00106] [19.4] The anti-CD137 antigen-binding molecule of

[19] or [19.1], wherein the agonist activity for CD137 in the presence of 250 μM or more of the small molecule compound is two or more times greater than the agonist activity for CD137 in the absence of the small molecule compound.

[00107] [19.5] Anti-CD137 antigen-binding molecule of any of

[19] to [19.4], wherein agonist activity for CD137 is assessed with the amount of IL-2 and / or IFN-γ produced by a cell expressing CD137.

[00108] [19.6] The anti-CD137 antigen-binding molecule of [19.5], in which the cell expressing CD137 is a mononuclear cell Petition 870260042208, dated 05 / 05 / 2026, p. 50 / 1364 34 / 448 of human peripheral blood (PBMC) alone or a T cell derived from human PBMC.

[00109] [19.7] Anti-CD137 antigen-binding molecule of any of

[19] to [19.4], wherein agonist activity for CD137 is assessed by a reporter gene assay.

[00110] [19.8] The anti-CD137 antigen-binding molecule of

[19] , which exhibits agonist activity for CD137 in a solution prepared such that the final concentration of a small molecule compound is 50 μM or more, and does not exhibit substantially agonist activity for CD137 in a solution to which the small molecule compound is not added.

[00111] [19.9] The anti-CD137 antigen-binding molecule of [19.8], in which the agonist activity for CD137 in a solution that is prepared so that the final concentration of a small molecule compound is 50 pM or more, and the agonist activity for CD137 in a solution to which the small molecule compound is not added, are each evaluated with the amount of IL-2, IFN-γ and / or IL-6 measured within 72 hours after a cell expressing CD137 and the anti-CD137 antigen-binding molecule are contacted in the solution.

[00112] [19.10] The anti-CD137 antigen-binding molecule of [19.8], in which the agonist activity for CD137 in a solution that is prepared so that the final concentration of a small molecule compound is 50 μM or more, and the agonist activity for CD137 in a solution to which the small molecule compound is not added, are each evaluated with a luciferase luminescence signal that is measured within 6 hours after a T cell expressing an NF-kappaB-luciferase and CD137 reporter construct is contacted with the anti-CD137 antigen-binding molecule.

[00113] [19.11] The anti-CD137 antigen-binding molecule of Petition 870260042208, dated 05 / 05 / 2026, page 51 / 1364 35 / 448 any of

[19] to [19.10], wherein the small molecule compound is an adenosine-containing compound.

[00114] [19.12] The anti-CD137 antigen-binding molecule of any of

[19] to [19.11], wherein the small molecule compound is ATP.

[00115]

[20] An agonist antigen-binding molecule comprising an altered Fc region, wherein the altered Fc region comprises at least one amino acid change leading to an increased isoelectric point (pI) compared with that of a parental agonist antigen-binding molecule comprising a parental Fc region, and wherein the agonist antigen-binding molecule has increased agonist activity compared with that of the parental agonist antigen-binding molecule.

[00116] [20.1] The antigen-binding agonist molecule of

[20] , in which at least one amino acid change is a change of an amino acid residue that can be exposed on the surface of the parental Fc region.

[00117] [20.2] The antigen-binding antibody agonist of

[20] or [20.1], in which at least one amino acid change is: (i) replacement of at least one amino acid residue with a negative side chain charge in the parental Fc region by an amino acid residue with no side chain charge, (ii) replacement of at least one amino acid residue with no side chain charge in the parental Fc region by an amino acid residue with a positive side chain charge, and / or (iii) replacement of at least one amino acid residue with a negative side chain charge in the parental Fc region by an amino acid residue with a positive side chain charge.

[00118] [20.3] The antigen-binding molecule of any agonist Petition 870260042208, dated 05 / 05 / 2026, p. 52 / 1364 36 / 448 one of

[20] to [20.2], where at least one amino acid change is a combination of amino acid substitutions and where the amino acid substitutions are located in positions that are conformationally close to each other.

[00119] [20.4] The antigen-binding agonist molecule of any of

[20] to [20.3], in which the binding activity of the altered Fc region to an Fcy receptor is not substantially reduced compared with that of the parental Fc region.

[00120] [20.5] The antigen-binding agonist molecule of [20.4], where the Fcy receptor is FcyRIIb.

[00121] [20.6] The antigen-binding agonist molecule of any one of

[20] to [20.4], in which at least one amino acid change is at least one amino acid substitution selected from the group consisting of Q311R, P343R and D413K, according to EU numbering.

[00122] [20.7] The agonist antigen-binding molecule of any one of

[20] to [20.6], wherein at least one amino acid change is the amino acid change of (i) P343R / D413K, (ii) Q311R / P343R, (iii) P343R, (iv) D413K, (v) Q311R, or (vi) Q311R / D413K or a combination thereof, according to EU numbering.

[00123] [20.8] The antigen-binding molecule agonist of any of

[20] to [20.7], which is an antiCD137 antigen-binding molecule.

[00124] [20.9] The antigen-binding agonist molecule of any of

[20] to [20.8], which is an anti-CD137 antibody.

[00125]

[21] A method for producing an antigen-binding agonist molecule, comprising an altered Fc region, wherein the method comprises: Introduce into a parental Fc at least one amino acid change that leads to an increased isoelectric point (pI) in Petition 870260042208, dated 05 / 05 / 2026, page 53 / 1364 37 / 448 comparison with that of a parental agonist antigen-binding molecule comprising the parental Fc region, wherein the agonist activity of the agonist antigen-binding molecule comprising the altered Fc region is increased compared with that of the parental agonist antigen-binding molecule.

[00126] [21.1] The method of

[21] , in which the agonist activity of the antigen-binding molecule is agonist for the antigen in the presence of 10 μM, 50 μM, 100 μM, 150 μM, 200 μM or 250 μM of a small molecule compound is two or more times greater than the agonist activity for the antigen in the absence of the small molecule compound.

[00127] [21.2] The method of

[21] or [21.1], in which the agonist activity of the antigen-binding molecule is agonist for the antigen in the presence of 10 μM or more of a small molecule compound, which is two or more times greater than the agonist activity for the antigen in the absence of the small molecule compound.

[00128] [21.3] The method of

[21] or [21.1], in which the agonist activity of the antigen-binding molecule is agonist for the antigen in the presence of 50 μM or more of a small molecule compound, which is two or more times greater than the agonist activity for the antigen in the absence of the small molecule compound.

[00129] [21.4] The method of

[21] or [21.1], in which the agonist activity of the antigen-binding molecule is agonist for the antigen in the presence of 250 μM or more of a small molecule compound, which is two or more times greater than the agonist activity for the antigen in the absence of the small molecule compound.

[00130] [21.5] The method of any of

[21] to [21.4], in which agonist activity for the antigen is assessed with the amount of IL2 and / or IFN-γ produced by a cell expressing the antigen. Petition 870260042208, dated 05 / 05 / 2026, page 54 / 1364 38 / 448

[00131] [21.6] The method of [21.5], in which the cell expressing the antigen is an isolated human peripheral blood mononuclear cell (PBMC) or a human PBMC-derived T cell.

[00132] [21.7] The method of any of

[21] to [21.4], in which agonist activity for the antigen is evaluated by a reporter gene assay.

[00133] [21.8] The method of any of

[21] to [21.7], additionally comprising: (i) obtain an expression vector comprising an appropriate promoter operationally linked to a gene encoding the agonist antigen-binding molecule produced by the method of any of

[21] to [21.7], (ii) introduce the vector into a host cell and culture the host cell to produce the agonist antigen-binding molecule, and (iii) collect the agonist antigen-binding molecule from the host cell culture.

[00134] [21.9] The method of any of

[21] to [21.8], which is an anti-CD137 antigen-binding molecule.

[00135] [21.10] The method of any of

[21] to [21.9], which is an anti-CD137 antibody.

[00136] [21.11] The method of any one of [21.1] to [21.10], wherein the small molecule compound is an adenosine-containing compound.

[00137] [21.12] The method of any one of [21.1] to [21.11], wherein the small molecule compound is ATP.

[00138]

[22] A method for increasing the agonist activity of an antigen-binding molecule comprising an Fc region, wherein the method comprises introducing into the Fc region at least one amino acid change leading to an increased isoelectric point (pI) compared with that of a molecule of Petition 870260042208, dated 05 / 05 / 2026, page 55 / 1364 39 / 448 binding to parental agonist antigen comprising a parental Fc region.

[00139] [22.1] The method of

[22] , in which the agonist activity of the antigen-binding molecule is agonist for the antigen in the presence of 10 μM, 50 μM, 100 μM, 150 μM, 200 μM or 250 μM of a small molecule compound is two or more times greater than the agonist activity for the antigen in the absence of the small molecule compound.

[00140] [22.2] The method of

[22] or [22.1], in which the agonist activity of the antigen-binding molecule is agonist for the antigen in the presence of 10 μM or more of a small molecule compound, which is two or more times greater than the agonist activity for the antigen in the absence of the small molecule compound.

[00141] [22.3] The method of

[22] or [22.1], in which the agonist activity of the antigen-binding molecule is two or more times greater than the agonist activity for the antigen in the presence of 50 μM or more of a small molecule compound than the agonist activity for the antigen in the absence of the small molecule compound.

[00142] [22.4] The method of

[22] or [22.1], in which the agonist activity of the antigen-binding molecule for the antigen in the presence of 250μM or more of the small molecule compound is two or more times greater than the agonist activity for the antigen in the absence of the small molecule compound.

[00143] [22.5] The method of any of

[22] to [22.4], in which agonist activity for the antigen is assessed with the amount of IL2 and / or IFN-γ produced by a cell expressing the antigen.

[00144] [22.6] The method of [22.5], in which the cell expressing the antigen is an isolated human peripheral blood mononuclear cell (PBMC) or a human PBMC-derived T cell.

[00145] [22.7] The method of any of

[22] to [22.4], where the Petition 870260042208, dated 05 / 05 / 2026, page 56 / 1364 40 / 448 Agonist activity for the antigen is evaluated by a reporter gene assay.

[00146] [22.8] The method of any of

[22] to [22.7], which is an anti-CD137 antigen-binding molecule.

[00147] [22.9] The method of any of

[22] to [22.8], which is an anti-CD137 antibody.

[00148] [22.10] The method of any one of [22.1] to [22.9], wherein the small molecule compound is an adenosine-containing compound.

[00149] [21.11] The method of any one of [22.1] to [22.10], wherein the small molecule compound is ATP.

[00150]

[23] A method of using at least one amino acid change to increase the agonist activity of an agonist antigen-binding molecule comprising an Fc region, wherein the amino acid change leads to an increased isoelectric point (pI) compared with that of a parental agonist antigen-binding molecule comprising a parental Fc region.

[00151] [23.1] The method of

[23] , in which the agonist activity of the antigen-binding molecule is agonist for the antigen in the presence of 10 μM, 50 μM, 100 μM, 150 μM, 200 μM or 250 μM of a small molecule compound is two or more times greater than the agonist activity for the antigen in the absence of the small molecule compound.

[00152] [23.2] The method of

[23] or [23.1], in which the agonist activity of the antigen-binding molecule is agonist for the antigen in the presence of 10 μM or more of a small molecule compound, which is two or more times greater than the agonist activity for the antigen in the absence of the small molecule compound.

[00153] [23.3] The method of

[23] or [23.1], in which the agonist activity of the antigen-binding molecule is agonist for the antigen in the presence of 50 μM or more of a small molecule compound is Petition 870260042208, dated 05 / 05 / 2026, page 57 / 1364 41 / 448 two or more times greater than the agonist activity for the antigen in the absence of the small molecule compound.

[00154] [23.4] The method of

[23] or [23.1], in which the agonist activity of the antigen-binding molecule is two or more times greater than the agonist activity for the antigen in the presence of 250 μM or more of a small molecule compound than the agonist activity for the antigen in the absence of the small molecule compound.

[00155] [23.5] The method of any of

[23] to [23.4], in which agonist activity for the antigen is assessed with the amount of IL2 and / or IFN-γ produced by a cell expressing the antigen.

[00156] [23.6] The method of [23.5], in which the cell expressing the antigen is an isolated human peripheral blood mononuclear cell (PBMC) or a human PBMC-derived T cell.

[00157] [23.7] The method of any of

[23] to [23.4], in which agonist activity for the antigen is evaluated by a reporter gene assay.

[00158] [23.8] The method of any of

[23] to [23.7], which is an anti-CD137 antigen-binding molecule.

[00159] [23.9] The method of any of

[23] to [23.8], which is an anti-CD137 antibody.

[00160] [23.10] The method of any one of [23.1] to [23.9], wherein the small molecule compound is an adenosine-containing compound.

[00161] [23.11] The method of any one of [23.1] to [23.10], wherein the small molecule compound is ATP.

[00162]

[24] A screening method for an antigen-binding domain or antigen-binding molecule that has small molecule compound-dependent antigen-binding activity, wherein the method comprises: (a) contact an antigen-binding domain or antigen-binding molecule or a library of antigen-binding domains Petition 870260042208, dated 05 / 05 / 2026, page 58 / 1364 42 / 448 antigen or antigen-binding molecules with a fusion molecule in the presence of a small molecule compound, wherein in the fusion molecule, two or more units of an antigen are fused to one unit of a fusion partner, (b) placing an antigen-binding domain or antigen-binding molecule bound to the antigen within the fusion molecule in step (a) in the absence or in the presence of a low concentration of the small molecule compound, and (c) isolating an antigen-binding domain or antigen-binding molecule dissociated in step (b).

[00163] [24.1] The method of

[24] , in which the fusion partner molecule is an Fc dimer region.

[00164] [24.2] The method of [24.1], in which the Fc region comprises a first Fc subunit and a second Fc subunit, and in which one unit of the antigen is fused to each of the first and second Fc subunits.

[00165] [24.3] The method of [24.1] or [24.2], in which one antigen unit is fused to the N-terminal of each of the first and second Fc subunits.

[00166] [24.4] The method of any of

[24] to [24.3], wherein the library of antigen-binding domains or antigen-binding molecules is a phage library.

[00167] [24.5] The method of any of

[24] to [24.4], wherein the phages included in the phage library are phages that have on their surface two or more antigen-binding domains or antigen-binding molecules.

[00168] [24.6] The method of any of

[24] to [24.5], wherein the phages included in the phage library are phages with a defect in the pIII gene derived from the helper phage.

[00169]

[25] Screening method for a linkage domain to Petition 870260042208, dated 05 / 05 / 2026, page 59 / 1364 43 / 448 antigen or antigen-binding molecule having antigen-binding activity dependent on two or more different small molecule compounds, wherein the method comprises: (a) contacting an antigen-binding domain or antigen-binding molecule or a library of antigen-binding domains or antigen-binding molecules with an antigen in the presence of a first small molecule compound, (b) placing an antigen-binding domain or antigen-binding molecule bound to the antigen in step (a) in the absence of, or in the presence of a low concentration of, the first small molecule compound, (c) isolating an antigen-binding domain or antigen-binding molecule dissociated in step (b), (d) contacting an antigen-binding domain or antigen-binding molecule isolated in step (c) with the antigen in the presence of a second small molecule compound, (e) placing an antigen-binding domain or antigen-binding molecule bound to the antigen in step (d) in the absence of, or in the presence of a low concentration of, the second small molecule compound,(e) and (f) isolate an antigen-binding domain or antigen-binding molecule dissociated in step (e), wherein the method does not comprise, between steps (c) and (d), amplifying a gene encoding the antigen-binding domain or antigen-binding molecule isolated in step (c).

[00170] [25.1] The method of

[25] , in which the library of antigen-binding domains or antigen-binding molecules is a phage library.

[00171]

[26] A screening method for an antigen-binding domain or antigen-binding molecule that has activity of Petition 870260042208, dated 05 / 05 / 2026, page 60 / 1364 44 / 448 antigen-dependent binding of a small molecule compound, wherein the method comprises: (a) contacting a virgin library of antigen-binding domains or antigen-binding molecules with an antigen in the presence of a small molecule compound, (b) placing an antigen-binding domain or antigen-binding molecule bound to the antigen in step (a) in the absence or in the presence of a low concentration of the small molecule compound, and (c) isolating an antigen-binding domain or antigen-binding molecule dissociated in step (b), wherein the virgin library is a library of phages including phages that display on their surface two or more antigen-binding domains or antigen-binding molecules.

[00172]

[27] A screening method for an antigen-binding domain or antigen-binding molecule having small molecule compound-dependent antigen-binding activity, characterized in that the method comprises: (a) contacting a library of antigen-binding domains or antigen-binding molecules with an antigen in the presence of a small molecule compound, (b) placing an antigen-binding domain or antigen-binding molecule bound to the antigen in step (a) in the absence or in the presence of a low concentration of the small molecule compound, and (c) isolating an antigen-binding domain or antigen-binding molecule dissociated in step (b), wherein the library is a library including phages with a defect in the pIII gene derived from the helper phage.

[00173]

[28] Screening method for a linkage domain to Petition 870260042208, dated 05 / 05 / 2026, page 61 / 1364 45 / 448 antigen or antigen-binding molecule having antigen-binding activity dependent on a small molecule compound, wherein the method comprises: (a) contacting a library of antigen-binding domains or antigen-binding molecules with an antigen in the presence of a small molecule compound, (b) placing an antigen-binding domain or antigen-binding molecule bound to the antigen in step (a) in the absence or presence of a low concentration of the small molecule compound, and (c) isolating an antigen-binding domain or antigen-binding molecule dissociated in step (b), wherein the library is a library including phages prepared increasing the expression of the antigen-binding domain or antigen-binding molecule with a small molecule additive increasing the expression level of the promoter that regulates the expression of the antigen-binding domain or antigen-binding molecule.

[00174] [28.1] The screening method of

[28] , wherein the small molecule additive is isopropyl-e-thiogalactopyranoside or arabinose.

[00175] [28.2] The method of any of

[24] to [28.[1], wherein the small molecule compound is an adenosine-containing compound.

[00176] [28.3] The method of any of

[24] to [28.2], wherein the small molecule compound is ATP.

[00177]

[29] An antigen-binding molecule that has antigen-binding activity dependent on the concentration of a specific tumor tissue compound, wherein the antigen-binding activity in the presence of 100 μM of the compound is two or more times greater than the antigen-binding activity in the absence of the compound.

[00178] [29.1] The antigen-binding molecule of

[29] , wherein the KD value in the presence of 100 pM of the compound is 5 x 10-7M or less. Petition 870260042208, dated 05 / 05 / 2026, page 62 / 1364 46 / 448

[00179] [29.2] The antigen-binding molecule of

[29] or [29.1], wherein the KD value in the absence of the compound is 1 x 10-6M or more.

[00180] [29.3] The antigen-binding molecule of any of

[29] to [29.2], which has neutralizing activity against the antigen.

[00181] [29.4] The antigen-binding molecule of any of

[29] a [29.3], which has cytotoxic activity against a cell expressing the antigen.

[00182] [29.5] The antigen-binding molecule of any of

[29] to [29.4], wherein the antigen is an antigen expressed or secreted by any tumor cell, immune cell and stromal cell in a tumor tissue.

[00183] [29.6] The antigen-binding molecule of any of

[29] to [29.5], wherein the compound is an adenosine-containing compound.

[00184] [29.7] The antigen-binding molecule of any of

[29] to [29.6], which comprises an Fc region.

[00185] [29.8] The antigen-binding molecule of [29.7], wherein the Fc region is a mutated Fc region comprising an amino acid change, wherein the mutated Fc region has improved binding activity to at least one Fcy receptor selected from the group consisting of FcyRIa, FcyRIIa, FcyRIIb and FcyRIIIa, compared with a wild-type Fc region.

[00186] [29.9] The antigen-binding molecule of any of

[29] to [29.8], wherein the antigen-binding molecule is an antibody or an antibody fragment.

[00187]

[30] A pharmaceutical formulation comprising the antigen-binding molecule of any one of

[29] to [29.9] and a pharmaceutically acceptable carrier.

[00188] [30.1] The pharmaceutical formulation of

[30] , which is for use in the treatment of a tumor.

[00189] [30.2] The pharmaceutical formulation of [30.1], which has Petition 870260042208, dated 05 / 05 / 2026, page 63 / 1364 47 / 448 less cytotoxic activity in a non-tumor tissue than a pharmaceutical formulation comprising a control antigen-binding molecule.

[00190] [30.3] The pharmaceutical formulation of [30.1] or [30.2], which has a lower level of side effects than a pharmaceutical formulation comprising a control antigen-binding molecule.

[00191] [30.4] The pharmaceutical formulation of [30.2] or [30.3], wherein the control antigen-binding molecule is an antigen-binding molecule that does not have concentration-dependent antigen-binding activity of a specific tumor tissue compound.

[00192]

[31] A method for producing an antigen-binding molecule for use in the treatment of a tumor, wherein the method comprises the step of selecting an antigen-binding molecule whose antigen-binding activity in the presence of 100 μM of a specific tumor tissue compound is two or more times greater than its antigen-binding activity in the absence of the compound.

[00193]

[32] A method for producing a pharmaceutical formulation for use in the treatment of a tumor, wherein the method comprises the step of mixing the antigen-binding molecule from any of

[29] to [29.9] with a pharmaceutically acceptable carrier.

[00194]

[33] An antigen-binding molecule that has antigen-binding activity dependent on the concentration of a specific target tissue compound, wherein the antigen-binding activity in the presence of 1 μM of the compound is two or more times lower than the antigen-binding activity in the presence of a sufficient amount of the compound.

[00195] [33.1] The antigen-binding molecule of

[33] , where the KD value in the presence of 1 pM of the compound is 2 x 10-7M or more. Petition 870260042208, dated 05 / 05 / 2026, p. 64 / 1364 48 / 448

[00196] [33.2] The antigen-binding molecule of

[33] or [33.1], where the KD value in the presence of a sufficient amount of the compound is 1 x 10-7 M or less.

[00197] [33.3] The antigen-binding molecule of any of

[33] to [33.2], where the compound is a tumor tissue-specific compound.

[00198] [33.4] The antigen-binding molecule of [33.3], wherein the compound is an adenosine-containing compound.

[00199] [33.5] The antigen-binding molecule of any of

[33] a [33.4], which has greater plasma retention capacity and / or lower antigen accumulation capacity in plasma than a control antigen-binding molecule.

[00200] [33.6] The antigen-binding molecule of [33.5], wherein the control antigen-binding molecule is an antigen-binding molecule that does not have antigen-binding activity dependent on the concentration of a specific target tissue compound.

[00201] [33.7] The antigen-binding molecule of any of

[33] to [33.6], wherein the antigen-binding molecule is an antibody or an antibody fragment.

[00202]

[34] A pharmaceutical formulation comprising the antigen-binding molecule of any one of

[33] to [33.7] and a pharmaceutically acceptable carrier.

[00203]

[35] A method for producing an antigen-binding molecule that has a higher plasma retention property and / or lower antigen accumulation capacity in plasma than a control antigen-binding molecule, wherein the method comprises the steps of (a) producing an antigen-binding molecule whose antigen-binding activity increases as the concentration of a specific target tissue compound increases, and (b) measuring the plasma retention property and / or the capacity to Petition 870260042208, dated 05 / 05 / 2026, page 65 / 1364 49 / 448 accumulation of antigen in plasma from antigen-binding molecule produced in (a).

[00204] [35.1] The method of

[35] , which comprises the step of selecting an antigen-binding molecule whose antigen-binding activity in the presence of 1 μM of a specific target tissue compound is two or more times lower than the antigen-binding activity in the presence of a sufficient amount of the compound.

[00205] [35.2] The method of

[35] or [35.1], wherein the control antigen-binding molecule is an antigen-binding molecule that does not have antigen-binding activity dependent on the concentration of a specific target tissue compound.

[00206]

[36] A method for producing a pharmaceutical formulation, comprising the step of mixing the antigen-binding molecule from any of

[33] to [33.7] with a pharmaceutically acceptable carrier.

[00207]

[37] A method for measuring ATP concentration in a solution, comprising the steps of (i) contacting a split Luc / HEK293 cell expressing P2Y11 with the solution and (ii) measuring luciferase activity in the cell.

[00208] [37.1] The method of

[37] , which additionally comprises the step of contacting a solution containing a luciferase substrate with the cell.

[00209] [37.2] The method of

[37] or [37.1], in which the solution is the intercellular fluid within a tissue in vivo.

[00210] [37.3] The method of [37.2], in which the tissue is a tumor tissue.

[00211] [37.4] The method of [37.2] or [37.3], wherein step (i) is the transplantation step of a divided Luc / HEK293 cell expressing P2Y11 into the tissue in vivo. Brief Description of the Drawings Petition 870260042208, dated 05 / 05 / 2026, page 66 / 1364 50 / 448

[00212] Fig. 1 is a diagram showing the agonist activity of various anti-CD137 antibodies tested using Jurkat cells in the presence or absence of ATP.

[00213] The X-axis shows the antibody concentration (pg / mL) and the Y-axis shows the relative light unit.

[00214] Fig. 2 is a diagram showing the agonist activity of various anti-CD137 antibodies tested using Jurkat cells in the presence or absence of ADP.

[00215] The X-axis shows the antibody concentration (pg / mL) and the Y-axis shows the relative light unit.

[00216] Fig. 3 is a diagram showing the agonist activity of various anti-CD137 antibodies tested using human T cells in the presence or absence of ADPbetaS.

[00217] Fig. 4 is a diagram showing the agonist activity of dBBAT119-P253 / dBBAT119L-LamLib (small molecule exchange anti-CD137 antibody) or NS1-P253 (non-exchange anti-CD137 antibody) tested using human T cells in the presence or absence of ADPbetaS.

[00218] The X-axis shows the antibody concentration (pg / mL) and the Y-axis shows the amount of IFN-γ produced (ng / mL).

[00219] Fig. 5 is a diagram showing the ATP-dependent antigen-binding activity of various anti-CD137 antibodies (anti-CD137 antibody exchange with enhanced binding activity) tested with phage ELISA.

[00220] The Y-axis shows the S / N ratio of absorption in the presence / absence of ATP, and the X-axis shows the S / N ratio in the presence / absence of the antigen.

[00221] Fig. 6 is a diagram showing the binding activity of various variants of the anti-CD137 antibody (dBBAT119HP253 / dBBAT119L-LamLib) to human CD137 in the presence or absence of CD137. Petition 870260042208, dated 05 / 05 / 2026, page 67 / 1364 51 / 448 of ATP.

[00222] The top row shows the binding activity to human CD137 in the absence of ATP and the bottom row shows the binding activity to human CD137 in the presence of ATP.

[00223] Fig. 7 is a diagram showing the agonist activity of dBBAT119H-P253 / dBBAT119L-LamLib, dBBATk119H024P253 / dBBATk119L020-LamLib, IC17HdK-hIgG1 / IC17L-k0 (control), or NS1-P253 (or non-exchangeable anti-CD137 antibody) tested using human T cells in the presence or absence of ADPbetaS.

[00224] Subfigure (A) shows the test results in the absence of ADPbetaS and subfigure (B) shows the test results in the presence of ADPbetaS.

[00225] The X-axis shows the antibody concentration (pg / mL) and the Y-axis shows the amount of IFN-γ produced (ng / mL).

[00226] Fig. 8 is a diagram showing the agonist activity of various exchangeable anti-CD137 antibodies tested using the Jurkat 4-1BB reporter gene assay in the presence or absence of ATP.

[00227] Subfigure (A) shows the test results in the absence of ATP and subfigure (B) shows the test results in the presence of ATP.

[00228] Fig. 9 is a diagram showing the increase in agonist activity of various exchangeable anti-CD137 antibodies in the presence of ATP due to increased binding activity of heavy chain constant regions to Fcy receptors, tested using human peripheral blood mononuclear cells.

[00229] Subfigure (A) shows the agonist activity determined using the amount of IL-2 production as an index, and subfigure (B) shows the agonist activity determined using the amount of IFN-γ production as an index.

[00230] Fig. 10 is a diagram showing the increase in activity Petition 870260042208, dated 05 / 05 / 2026, page 68 / 1364 52 / 448 agonist of several anti-CD137 antibodies in the presence of ATP due to increased binding activity of heavy chain constant regions to Fcy receptors or increased pI of heavy chain constant regions, tested using human peripheral blood mononuclear cells.

[00231] Subfigure (A) shows the agonist activity determined using the amount of IL-2 production as an index, and subfigure (B) shows the agonist activity determined using the amount of IFN-γ production as an index.

[00232] Fig. 11 is a diagram showing the increase in agonist activity of various exchangeable anti-CD137 antibodies in the presence or absence of ATP due to increased binding activity of heavy chain constant regions to Fcy receptors, tested using human peripheral blood mononuclear cells.

[00233] Subfigure (A) shows the agonist activity determined using the amount of IL-2 production as an index, and subfigure (B) shows the agonist activity determined using the amount of IFN-γ production as an index.

[00234] Fig. 12 is a diagram showing the increase in agonist activity of various exchangeable anti-CD137 antibodies in the presence or absence of ATP due to increased binding activity of heavy chain constant regions to Fcy receptors, tested using human peripheral blood mononuclear cells.

[00235] Subfigure (A) shows the agonist activity determined using the amount of IL-2 production as an index, and subfigure (B) shows the agonist activity determined using the amount of IFN-γ production as an index.

[00236] Fig. 13 is a diagram showing the enhancement in agonist activity of various anti-CD137 exchange antibodies in the presence or absence of ATP due to increased binding activity of the regions Petition 870260042208, dated 05 / 05 / 2026, page 69 / 1364 53 / 448 heavy chain constants for Fcy receptors, tested using human peripheral blood mononuclear cells.

[00237] Subfigure (A) shows the agonist activity determined using the amount of IL-2 production as an index, and subfigure (B) shows the agonist activity determined using the amount of IFN-γ production as an index.

[00238] Fig. 14 is a diagram showing the increase in agonist activity of various exchangeable anti-CD137 antibodies in the presence or absence of ATP due to the increased binding activity of the heavy chain constant regions to Fcy receptors, tested using human peripheral blood mononuclear cells.

[00239] Subfigure (A) shows the agonist activity determined using the amount of IL-2 production as an index, and subfigure (B) shows the agonist activity determined using the amount of IFN-γ production as an index.

[00240] Fig. 15 is a diagram showing the increase in agonist activity of various exchangeable anti-CD137 antibodies in the presence or absence of ATP due to increased binding activity of heavy chain constant regions to Fcy receptors, tested using human peripheral blood mononuclear cells.

[00241] Subfigure (A) shows the agonist activity determined using the amount of IL-2 production as an index, and subfigure (B) shows the agonist activity determined using the amount of IFN-γ production as an index.

[00242] Fig. 16 is a diagram showing the increase in agonist activity of various exchangeable anti-CD137 antibodies in the presence or absence of ATP due to the increase in pI of the heavy chain constant regions, tested using human peripheral blood mononuclear cells.

[00243] Subfigure (A) shows the determined agonist activity Petition 870260042208, dated 05 / 05 / 2026, page 70 / 1364 54 / 448 using the amount of IL-2 production as an index, and subfigure (B) shows the agonist activity determined using the amount of IFN-γ production as an index.

[00244] Fig. 17 is a diagram showing the increase in agonist activity of various exchangeable anti-CD137 antibodies in the presence or absence of ATP due to the increase in pI of the heavy chain constant regions, tested using human peripheral blood mononuclear cells.

[00245] Subfigure (A) shows the agonist activity determined using the amount of IL-2 production as an index, and subfigure (B) shows the agonist activity determined using the amount of IFN-γ production as an index.

[00246] Fig. 18 is a diagram showing the increase in agonist activity of various exchangeable anti-CD137 antibodies in the presence or absence of ATP due to the increase in pI of the heavy chain constant regions, tested using human peripheral blood mononuclear cells.

[00247] Subfigure (A) shows the agonist activity determined using the amount of IL-2 production as an index, and subfigure (B) shows the agonist activity determined using the amount of IFN-γ production as an index.

[00248] Fig. 19 is a diagram showing the increase in agonist activity of various exchangeable anti-CD137 antibodies in the presence or absence of ATP due to the increase in pI of the heavy chain constant regions, tested using human peripheral blood mononuclear cells.

[00249] Subfigure (A) shows the agonist activity determined using the amount of IL-2 production as an index, and subfigure (B) shows the agonist activity determined using the amount of IFN-γ production as an index. Petition 870260042208, dated 05 / 05 / 2026, page 71 / 1364 55 / 448

[00250] Fig. 20 is a diagram showing the increase in agonist activity of various exchangeable anti-CD137 antibodies in the presence or absence of ATP due to the increase in pI of the heavy chain constant regions, tested using human peripheral blood mononuclear cells.

[00251] Subfigure (A) shows the agonist activity determined using the amount of IL-2 production as an index, and subfigure (B) shows the agonist activity determined using the amount of IFN-γ production as an index.

[00252] Fig. 21 is a diagram showing the increase in agonist activity of various exchangeable anti-CD137 antibodies in the presence or absence of ATP due to the increase in pI of the heavy chain constant regions, tested using human peripheral blood mononuclear cells.

[00253] Subfigure (A) shows the agonist activity determined using the amount of IL-2 production as an index, and subfigure (B) shows the agonist activity determined using the amount of IFN-γ production as an index.

[00254] Fig. 22 is a diagram showing the increase in agonist activity of various exchangeable anti-CD137 antibodies in the presence or absence of ATP due to the increase in pI of the heavy chain constant regions, tested using human peripheral blood mononuclear cells.

[00255] Subfigure (A) shows the agonist activity determined using the amount of IL-2 production as an index, and subfigure (B) shows the agonist activity determined using the amount of IFN-γ production as an index.

[00256] Fig. 23 is a diagram showing the increase in agonist activity of various anti-CD137 exchange antibodies in the presence or absence of ATP due to increased binding activity of the regions Petition 870260042208, dated 05 / 05 / 2026, page 72 / 1364 56 / 448 heavy chain constants for Fcy receptors, tested using human peripheral blood mononuclear cells.

[00257] Subfigure (A) shows the agonist activity determined using the amount of IL-2 production as an index and subfigure (B) shows the agonist activity determined using the amount of IFN-γ production as an index.

[00258] Fig. 24 is a diagram showing the plasma concentration of various exchangeable and non-exchangeable anti-CD137 antibodies tested using human CD137 knock-in mice.

[00259] The FCs are all mIgG1.

[00260] Fig. 25 is a diagram showing the plasma concentration of various exchangeable and non-exchangeable anti-CD137 antibodies tested using human CD137 knock-in mice.

[00261] The FCs are all MB110.

[00262] Fig. 26 is a diagram showing the plasma concentration of various exchangeable and non-exchangeable anti-CD137 antibodies tested using human CD137 knock-in mice.

[00263] The FCs are all MB492.

[00264] Fig. 27 is a diagram showing the antitumor effect of A375-mIgG1 / B167-ml0r in a mouse model transplanted with MC38 cells.

[00265] Each point shows the average value of a group (n = 5) of tumor volumes.

[00266] Fig. 28 is a diagram showing the weight of organs in a mouse model transplanted with MC38 cells after administration of antibodies (NO1-mIgG1 or A375-mIgG1 / B167-ml0r).

[00267] Subfigure (A) shows the weight of the lymph node and subfigure (B) shows the weight of the spleen.

[00268] Fig. 29 is a diagram showing the degree of T cell activation in lymph nodes of a transplanted mouse model. Petition 870260042208, dated 05 / 05 / 2026, p. 73 / 1364 57 / 448 with MC38 cells after administration of NO1-mIgG1 or A375mIgG1 / B167-ml0r.

[00269] Subfigure (A) shows the percentage of PD-1 positive T cells in CD8 positive T cells, subfigure (B) shows the percentage of ICOS positive T cells in CD8 positive T cells and subfigure (C) shows the percentage of Granzyme B positive T cells in CD8 positive T cells.

[00270] Fig. 30 is a diagram showing the degree of T cell activation in the spleen of a mouse model transplanted with the MC38 cell line after administration of NO1-mIgG1 or A375mIgG1 / B167-ml0r.

[00271] Subfigure (A) shows the percentage of PD-1 positive T cells in CD8 positive T cells, subfigure (B) shows the percentage of ICOS positive T cells in CD8 positive T cells and subfigure (C) shows the percentage of Granzyme B positive T cells in CD8 positive T cells.

[00272] Fig. 31 is a diagram showing the degree of T cell activation in the liver of a mouse model transplanted with the MC38 cell line after administration of NO1-mIgG1 or A375mIgG1 / B167-ml0r.

[00273] Subfigure (A) shows the percentage of PD-1 positive T cells in CD8 positive T cells and subfigure (B) shows the percentage of Granzyme B positive T cells in CD8 positive T cells.

[00274] Fig. 32 is a diagram showing the antitumor effect of A356-MB110 / B040-ml0r in a mouse model transplanted with the MC38 cell line.

[00275] Each point shows the average value of a group (n = 5) of tumor volumes.

[00276] Fig. 33 is a diagram showing the weight of the organs in Petition 870260042208, dated 05 / 05 / 2026, page 74 / 1364 58 / 448 a mouse model transplanted with the MC38 cell line after administration of NS2-MB110 or A356-MB110 / B040ml0r.

[00277] Subfigure (A) shows the weight of the lymph node and subfigure (B) shows the weight of the spleen.

[00278] Fig. 34 is a diagram showing the degree of T cell activation in the liver of a mouse model transplanted with the MC38 cell line after administration of NS2-MB110 or A356MB110 / B040-ml0r.

[00279] Subfigure (A) shows the percentage of PD-1 positive T cells in CD8 positive T cells and subfigure (B) shows the percentage of ICOS positive T cells in CD8 positive T cells.

[00280] Fig. 35 is a diagram showing the antitumor effect of A372-mIgG1 / B040-ml0r in a mouse model transplanted with the MC38 cell line.

[00281] Each point shows the average value of a group (n = 5) of tumor volumes.

[00282] Fig. 36 shows the number of lymph node cells (subfigure (A)) and spleen weight (subfigure (B)) in a mouse model transplanted with the MC38 cell line after administration of A372-mIgG1 / B040-ml0r.

[00283] Fig. 37 is a diagram showing the degree of T cell activation in the liver of a mouse model transplanted with the MC38 cell line after administration of A372-mIgG1 / B040ml0r (percentage of Granzyme B-positive T cells in CD8-positive T cells).

[00284] Fig. 38 is a diagram showing the antitumor effect of A372-MB110 / B040-ml0r in a mouse model transplanted with the MC38 cell line. Petition 870260042208, dated 05 / 05 / 2026, page 75 / 1364 59 / 448

[00285] Each point shows the average value of a group (n = 5) of tumor volumes.

[00286] Fig. 39 is a diagram showing the weight of organs in a mouse model transplanted with the MC38 cell line after administration of NS2-MB110 or A372-MB110 / B040ml0r.

[00287] Subfigure (A) shows the weight of the lymph node and subfigure (B) shows the weight of the spleen.

[00288] Fig. 40 is a diagram showing the degree of T cell activation in the liver of a mouse model transplanted with the MC38 cell line after administration of NS2-MB110 or A372MB110 / B040-ml0r (percentage of PD-1 positive T cells in CD8 positive T cells).

[00289] Fig. 41 is a diagram showing the antitumor effect of A372-MB492 / B040-ml0r in a mouse model transplanted with the MC38 cell line.

[00290] Each point shows the average value of a group (n = 5) of tumor volumes.

[00291] Fig. 42 is a diagram showing the number of lymph node cells and the organ weight of the spleen in a mouse model transplanted with the MC38 cell line after administration of NS1-MB492 or A372-MB492 / B040-ml0r.

[00292] Subfigure (A) shows the number of cells in the lymph node and subfigure (B) shows the weight of the spleen organ.

[00293] Fig. 43 is a diagram showing the degree of T cell activation in the liver of a mouse model transplanted with the MC38 cell line after administration of NS1-MB492 or A372MB492 / B040-ml0r (percentage of Granzyme B-positive T cells in CD8-positive T cells).

[00294] Fig. 44 is a diagram showing the antitumor effect of Petition 870260042208, dated 05 / 05 / 2026, page 76 / 1364 60 / 448 A486-MB492 / B167-ml0r or A488-MB492 / B226-ml0r in a mouse model transplanted with the MC38 cell line.

[00295] Each point shows the average value of a group (n = 5) of tumor volumes.

[00296] Fig. 45 is a diagram showing the number of cells per lymph node and the weight of the spleen in a mouse model transplanted with the MC38 cell line after administration of NS1-MB492, A486-MB492 / B167-ml0r or A488-MB492 / B226-ml0r.

[00297] Subfigure (A) shows the number of cells per lymph node and subfigure (B) shows the weight of the spleen.

[00298] Fig. 46 is a diagram showing the level of effector cell infiltration in the liver of a mouse model transplanted with the MC38 cell line after administration of NS1-MB492, A486-MB492 / B167-ml0r or A488-MB492 / B226-ml0r (percentage of CD3-positive and CD8-positive T cells in CD45-positive T cells).

[00299] Fig. 47 is a diagram showing the antitumor effect of A489-MB492 / B223-ml0r in a mouse model transplanted with the MC38 cell line.

[00300] Each point shows the average value of a group (n = 5) of tumor volumes.

[00301] Fig. 48 is a diagram showing the number of lymph node cells and the number of cells in a splenic lymphocyte fraction in a mouse model transplanted with the MC38 cell line after administration of NS1-MB492 or A489-MB492 / B223ml0r.

[00302] Subfigure (A) shows the number of cells in the lymph node and subfigure (B) shows the number of cells in a splenic lymphocyte fraction.

[00303] Fig. 49 is a diagram showing the degree of activation of Petition 870260042208, dated 05 / 05 / 2026, page 77 / 1364 61 / 448 T cells in the liver of a transplanted mouse model with the MC38 cell line after administration of NS1-MB492 or A489MB492 / B223-ml0r (percentage of CD8-positive T cells in CD45-positive T cells).

[00304] Fig. 50 is a diagram showing the antitumor effect of A548-mIgG1 / B256-ml0r and A551-mIgG1 / B256-ml0r in a mouse model transplanted with the MC38 cell line.

[00305] Subfigure (A) shows the antitumor effect of A548mIgG1 / B256-ml0r and subfigure (B) shows the antitumor effect of A551mIgG1 / B256-ml0r.

[00306] Fig. 51 is a diagram showing the weight of organs in a mouse model transplanted with the MC38 cell line after administration of NS1-mIgG1, A548-mIgG1 / B256-ml0r or A551-mIgG1 / B256-ml0r.

[00307] Subfigure (A) shows the weight of the lymph node and subfigure (B) shows the weight of the spleen.

[00308] Fig. 52 is a diagram showing the degree of T cell activation in the liver of a mouse model transplanted with the MC38 cell line after administration of NS1-mIgG1, A548mIgG1 / B256-ml0r or A551-mIgG1 / B256-ml0r.

[00309] Subfigure (A) shows the percentage of PD-1 positive T cells in CD8 positive T cells and subfigure (B) shows the percentage of Granzyme B positive T cells in CD8 positive T cells.

[00310] Fig. 53 is a diagram showing the antitumor effect of A551-MB110 / B379-ml0r in a mouse model transplanted with the MC38 cell line.

[00311] Fig. 54 is a diagram showing the weight of organs in a mouse model transplanted with the MC38 cell line after administration of NS1-mIgG1 or A551-MB110 / B379-ml0r. Petition 870260042208, dated 05 / 05 / 2026, page 78 / 1364 62 / 448

[00312] Subfigure (A) shows the weight of the lymph node and subfigure (B) shows the weight of the spleen.

[00313] Fig. 55 is a diagram showing the degree of T cell activation in the spleen of a mouse model transplanted with the MC38 cell line after administration of NS1-mIgG1 or A551MB110 / B379-ml0r.

[00314] Subfigure (A) shows the percentage of PD-1 positive T cells in CD8 positive T cells, subfigure (B) shows the percentage of ICOS positive T cells in CD8 positive T cells and subfigure (C) shows the percentage of Granzyme B positive T cells in CD8 positive T cells.

[00315] Fig. 56 is a diagram showing the degree of T cell activation in the liver of a mouse model transplanted with the MC38 cell line after administration of NS1-mIgG1 or A551MB110 / B379-ml0r.

[00316] Subfigure (A) shows the percentage of PD-1 positive T cells in CD8 positive T cells, subfigure (B) shows the percentage of ICOS positive T cells in CD8 positive T cells and subfigure (C) shows the percentage of Granzyme B positive T cells in CD8 positive T cells.

[00317] Fig. 57 is a diagram showing the agonist activity of various anti-CD137 antibodies tested using Jurkat cells in the presence or absence of L-kynurenine.

[00318] The X-axis shows the antibody concentration (pg / mL) and the Y-axis shows the relative light unit.

[00319] Fig. 58 is a diagram showing the agonist activity of various anti-CD137 antibodies tested using Jurkat 4-1BB cells in the presence or absence of the small molecule compound (ATP or ADP).

[00320] The X-axis shows the antibody concentration (pg / mL) and the Petition 870260042208, dated 05 / 05 / 2026, page 79 / 1364 63 / 448 Y-axis shows the relative light unit.

[00321] Fig. 59 is a diagram showing the ATP responsiveness (ATP concentration-dependent luciferin luminescence) of P2Y11-divided Luc / HEK293 cells produced to determine elevated extracellular ATP levels.

[00322] Fig. 60 is a diagram showing the in vivo ATP responsiveness (ATP concentration-dependent luciferin luminescence) of P2Y11 split Luc / HEK293 cells when transplanted subcutaneously into a mouse.

[00323] Fig. 61 is a diagram showing the luminescence imaging results of mice subcutaneously transplanted with P2Y11 split Luc / HEK293 cells and predetermined ATP concentrations and FM3A tumor-bearing mice subcutaneously transplanted with P2Y11 split Luc / HEK293 cells. Marks on the ventral portion of the mice indicate the detected luminescence.

[00324] Fig. 62 is a diagram showing the ATP concentration-dependent binding activity (KD value) of antihIL6R antibodies MRAH-G4T1 / MRAL-k0 (control antibody) and H0002G4T1 / L1058-lam1, H0041-G4T1 / L1088-lam1 and H0052-G4T1 / L1083lam1 (all are exchange antibodies) against hIL6R.

[00325] Fig. 63 is a diagram showing the ADP concentration-dependent binding activity (KD value) of antihIL6R antibodies MRAH-G4T1 / MRAL-k0 (control antibody) and H0002G4T1 / L1058-lam1, H0041-G4T1 / L1088-lam1 and H0052-G4T1 / L1083lam1 (all are exchange antibodies) against hIL6R.

[00326] Fig. 64 is a diagram showing the AMP concentration-dependent binding activity (KD value) of antihIL6R antibodies MRAH-G4T1 / MRAL-k0 (control antibody) and H0002G4T1 / L1058-lam1, H0041-G4T1 / L1088-lam1 and H0052-G4T1 / L1083. Petition 870260042208, dated 05 / 05 / 2026, page 80 / 1364 64 / 448 laml (all are exchange antibodies) against hIL6R.

[00327] Fig. 65 is a diagram showing the ATP concentration-dependent ADCC activity of anti-hIL6R antibodies MRAHmFa55 / MRAL-mk0 (control antibody) and H0002-mFa55 / L1058-ml0, H0041-mFa55 / L1088-ml0 and H0052-mFa55 / L1083-ml0 (all are exchange antibodies).

[00328] Fig. 66 is a diagram showing the in vivo antitumor activity of anti-hIL6R antibodies MRAH-mFa55 / MRAL-mk0 (control antibody) and H0002-mFa55 / L1058-ml0, H0041-mFa55 / L1088-ml0 and H0052-mFa55 / L1083-ml0 (all are exchange antibodies). IC17HdkmFa55 / IC17L-mk1 is the negative control antibody.

[00329] Fig. 67 is a diagram showing a comparison of the plasma kinetics of an anti-hIL6R antibody MRAH-mFa55 / MRALmk0 (control antibody) in normal mice and hIL6R transgenic mice. The vertical axis of the graph shows the plasma concentration of the antibody.

[00330] Fig. 68 is a diagram showing a comparison of the plasma kinetics of an anti-hIL6R antibody H0002-mFa55 / L1058ml0 (antibody exchange) in normal mice and hIL6R transgenic mice. The vertical axis of the graph shows the plasma concentration of the antibody.

[00331] Fig. 69 is a diagram showing a comparison of the plasma kinetics of an anti-hIL6R antibody H0041-mFa55 / L1088ml0 (antibody exchange) in normal mice and hIL6R transgenic mice. The vertical axis of the graph shows the plasma concentration of the antibody.

[00332] Fig. 70 is a diagram showing a comparison of the plasma kinetics of an anti-hIL6R antibody H0052-mFa55 / L1083ml0 (antibody exchange) in normal mice and hIL6R transgenic mice. The vertical axis of the graph shows the concentration Petition 870260042208, dated 05 / 05 / 2026, page 81 / 1364 65 / 448 plasma antibody.

[00333] Fig. 71 is a diagram showing the accumulation of antigens in hIL6R transgenic mice after administration of each of a non-exchange anti-hIL6R antibody MRAH-mFa55 / MRAL-mk0 (control antibody) and exchange anti-hIL6R antibodies H0002mFa55 / L1058-ml0, H0041-mFa55 / L1088-ml0 and H0052-mFa55 / L1083ml0 (all are exchange antibodies). The vertical axis of the graph shows the plasma concentration of soluble hIL6R. IC17Hdk-mFa55 / IC17L-mk1 (indicated as KLH-mFa55 in the figure) was used as the negative control antibody.

[00334] Fig. 72 is a diagram showing the in vivo antitumor activity of a non-exchangeable anti-hIL6R antibody MRAH-mFa55 / MRALmk0 (control antibody) and exchangeable anti-hIL6R antibodies H0002mFa55 / L1058-ml0 and H0041-mFa55 / L1088-ml0 (both are exchangeable antibodies). IC17Hdk-mFa55 / IC17L-mk1 is the negative control antibody.

[00335] Fig. 73 is a diagram showing a comparison of the plasma kinetics of a non-exchangeable anti-hIL6R antibody MRAHmFa55 / MRAL-mk0 (control antibody) and exchangeable anti-hIL6R antibodies H0002-mFa55 / L1058-ml0 and H0041-mFa55 / L1088-ml0 (both are exchangeable antibodies). The vertical axis of the graph shows the plasma concentration of the antibody.

[00336] Fig. 74 is a diagram showing the accumulation of antigens after administration of each of a non-exchangeable anti-hIL6R antibody MRAH-mFa55 / MRAL-mk0 (control antibody) and exchangeable anti-hIL6R antibodies H0002-mFa55 / L1058-ml0 and H0041-mFa55 / L1088ml0 (both are exchangeable antibodies). The vertical axis of the graph shows the plasma concentration of soluble hIL6R. IC17Hdk-mFa55 / IC17Lmk1 (indicated as KLH-mFa55 in the figure) was used as the negative control antibody. Petition 870260042208, dated 05 / 05 / 2026, page 82 / 1364 66 / 448

[00337] Fig. 75 is a diagram showing the in vivo antitumor activity of a non-exchangeable anti-hIL6R antibody MRAH-mFa55 / MRAL-mk0 (control antibody) and exchangeable anti-hIL6R antibodies H0041mFa55 / L1088-ml0 and H0052-mFa55 / L1083-ml0 (both are exchangeable antibodies). IC17Hdk-mFa55 / IC17L-mk1 is the negative control antibody.

[00338] Fig. 76 is a diagram showing a comparison of the plasma kinetics of a non-exchangeable anti-hIL6R antibody MRAHmFa55 / MRAL-mk0 (control antibody) and an exchangeable anti-hIL6R antibody H0052-mFa55 / L1083-ml0 (exchange antibody). The vertical axis of the graph shows the plasma concentration of the antibody.

[00339] Fig. 77 is a diagram showing the accumulation of antigens after administration of each of a non-exchangeable anti-hIL6R antibody MRAH-mFa55 / MRAL-mk0 (control antibody) and an exchangeable anti-hIL6R antibody H0052-mFa55 / L1083-ml0 (exchange antibody). The vertical axis of the graph shows the plasma concentration of soluble hIL6R. IC17Hdk-mFa55 / IC17L-mk1 (indicated as KLH-mFa55 in the figure) was used as the negative control antibody.

[00340] Fig. 78 is a diagram showing the ATP concentration-dependent activity of anti-PD1 antibodies mPD1F2VHmF18 / mPD1F2VL-mk1 (control antibody) and H5029-mFa31 / L3021ml0 (antibody switch) in inhibiting obligate PD-1 / PDL-1.

[00341] Fig. 79 is a diagram showing the ATP concentration-dependent activity of anti-PD1 antibodies mPD1F2VHmF18 / mPD1F2VL-mk1 (control antibody) and H5041-mFa31 / L3021ml0 (antibody switch) in inhibiting obligate PD-1 / PDL-1.

[00342] Fig. 80 is a diagram showing the in vitro AMP concentration-dependent neutralization activity of anti-PD1 antibodies mPD1F2VH-mF18 / mPD1F2VL-mk1 (control antibody) and H5029-mFa31 / L3021-ml0 and H5041-mFa31 / L3021-ml0 Petition 870260042208, dated 05 / 05 / 2026, page 83 / 1364 67 / 448 (both are alternating antibodies).

[00343] Fig. 81 is a diagram showing the ATP concentration-dependent in vitro neutralization activity of anti-PD1 antibodies mPD1F2VH-mF18 / mPD1F2VL-mk1 (control antibody) and H5029-mFa31 / L3021-ml0 and H5041-mFa31 / L3021-ml0 (both are alternate antibodies).

[00344] Fig. 82 is a diagram showing the in vivo antitumor activity of anti-PD1 antibodies mPD1F2VH-mFa55 / mPD1F2VL-mk1 (control antibody) and H5041-mFa55 / L3023-ml0 (antibody exchange). IC17Hdk-mFa55 / IC17L-mk1 is the negative control antibody.

[00345] Fig. 83 is a diagram showing the activity of anti-PD1 antibodies mPD1F2VH-mFa55 / mPD1F2VL-mk1 (control antibody) and H5041-mFa55 / L3023-ml0 (antibody switch) in clearing PD-1 expressing cells (A) from the tumor and (B) the spleen. In the figure, isotype represents the negative control antibody (IC17HdkmFa55 / IC17L-mk1).

[00346] Fig. 84 is a diagram showing the binding mode between ATP and the Fab H0041L1088 fragment of the anti-hIL6R exchange antibody. In the figure, ATP is shown with the ball-and-stick model and the amino acid residues that interact with ATP are shown with the stick model. The dashed lines indicate the hydrogen bonds between the antibody and ATP.

[00347] Fig. 85 is a diagram showing the amino acid sequence of the extracellular domain hIL6R (shIL6R) mapped to the epitope of the anti-hIL6R exchange antibody H0041L1088. In the figure, the amino acid residues shaded in gray are those (epitope residues) of shIL6R comprising one or more non-hydrogen atoms positioned at a distance of 4.2 Angstroms or less from ATP or Fab H0041L1088 in a crystal structure.

[00348] Fig. 86 is a diagram showing the connection details. Petition 870260042208, dated 05 / 05 / 2026, page 84 / 1364 68 / 448 between shIL6R and the Fab fragment H0041L1088 bound to ATP. In the figure, the antibody heavy chain is represented in black, the light chain is represented in gray, and shIL6R is represented in white. In the figure, ATP is shown with the ball model, and the shIL6R epitope residue within 4.2 Angstroms of the antibody or ATP and the antibody paratope residue within 4.2 Angstroms of the epitope residue are shown with the stick model. Dashed lines indicate hydrogen bonds between the antibody and shIL6R. To clarify the interaction with ATP, only the F298 of shIL6R is shown with the ball model.

[00349] Fig. 87 is a diagram showing a structure in which the structure of Fig. 86 is rotated 180 degrees (rear view).

[00350] Fig. 88 is a diagram showing the agonist activity of various exchangeable anti-CD137 antibodies tested using the Jurkat 4-1BB reporter gene assay in the presence of ATP.

[00351] Fig. 89 is a diagram showing a comparison of the plasma kinetics of each of the anti-CD137 exchange antibodies A375-SCF041aPh / B167-Lamlib and A375-MY201aPh / B167-Lamlib. The vertical axis of the graph shows the plasma concentration of each antibody.

[00352] Fig. 90 is a diagram showing the antitumor effect of each of A375 / B167-SCF041aPh and A375 / B167-MY201aPh in a mouse model prepared by transplantation of the LLC1 / OVA / GPC3 cell line into hCD137KI / mFcYR2bKO / hFcYR2bTg#90 mice.

[00353] Each point shows the average value of a group (n = 5) of tumor volumes.

[00354] Fig. 91 is a diagram showing the agonist activity of various exchangeable anti-CD3 antibodies tested by a reporter gene assay using T-cell activation bioassay (NFAT) in the presence Petition 870260042208, dated 05 / 05 / 2026, page 85 / 1364 69 / 448 ATP. Method for Carrying Out the Invention I. Definitions

[00355] The term binding activity refers to the strength of the total sum of non-covalent interactions between one or more binding sites of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). Here, binding activity is not strictly limited to a 1:1 interaction between members of a binding pair (e.g., antibody and antigen). For example, when the members of a binding pair reflect a monovalent 1:1 interaction, the binding activity is specifically called intrinsic binding affinity (affinity). When a member of a binding pair is capable of both monovalent and multivalent binding, the binding activity is the sum of each binding strength. The binding activity of a molecule X to its partner Y can generally be represented by the dissociation constant (KD) or the amount of analyte binding per unit amount of ligand (hereinafter referred to as the amount of binding).Those skilled in the art would understand that, generally, a lower dissociation constant (KD) value means higher binding activity, and a higher value of analyte binding per unit amount of ligand or amount of binding means higher binding activity. Binding activity can be measured by common methods known in the art, including those described herein. Illustrative embodiments and specific examples for measuring binding activity are described below.

[00356] An antigen-binding molecule or antibody with matured binding activity, or an antigen-binding antibody or molecule with enhanced (improved) binding activity, refers to an antibody with one or more alterations in one or more hypervariable regions (HVRs), compared to a binding molecule. Petition 870260042208, dated 05 / 05 / 2026, page 86 / 1364 70 / 448 parental antigen or a parental antibody that does not carry such alterations, such alterations resulting in an improvement in the binding activity of the antigen-binding molecule or antibody to the antigen.

[00357] The terms anti-CD137 antigen-binding molecule or anti-CD137 antibody and an antigen-binding molecule that binds to CD137 or an antibody that binds to CD137 refer to an antigen-binding molecule or antibody that is capable of binding to CD137 with sufficient binding activity, such that the antigen-binding molecule or antibody is useful as a diagnostic and / or therapeutic agent in targeting CD137. In certain embodiments, an anti-CD137 antibody binds to a CD137 epitope that is conserved among CD137 of different species.

[00358] The term an anti-CD137 antigen-binding molecule or an anti-CD137 antibody having CD137-binding activity dependent on a small molecule compound means an antigen-binding molecule or an antibody that shows greater CD137-binding activity in the presence of the small molecule compound compared to CD137-binding activity in the absence of the small molecule compound. In one embodiment, the presence of a small molecule compound refers to the condition where the small molecule compound is present at a concentration of 10 micromolar or more, 50 micromolar or more, 100 micromolar or more, 150 micromolar or more, 200 micromolar or more, or 250 micromolar or more.In one embodiment, the extent of binding activity of an anti-CD137 antigen-binding molecule or antibody to an unrelated non-CD137 protein in the presence of a small molecule compound is less than about 10% of the binding of the antigen-binding molecule or antibody to CD137 as measured, for example, by radioimmunoassay (RIA) or surface plasmon resonance (SPR). In certain embodiments, Petition 870260042208, dated 05 / 05 / 2026, page 87 / 1364 71 / 448 In the presence of a low molecular weight compound, an anti-CD137 antigen-binding molecule or antibody has a dissociation constant (KD) of 1 micromolar or less, 100 nM or less, 10 nM or less, 1 nM or less, 0.1 nM or less, 0.01 nM or less, or 0.001 nM or less (e.g., 10⁻⁶M or less, 10⁻⁷M or less, 10⁻⁸M or less, 10⁻⁹M or less, 10⁻¹⁰M or less, e.g., from 10⁻⁶M to 10⁻¹⁰M, from 10⁻⁷M to 10⁻⁹M, e.g., from 10⁻⁷M to 10⁻⁸M).

[00359] Here, the term antigen-binding molecule is used in its broadest sense and refers to a molecule that binds specifically to an antigenic determinant. In one embodiment, the antigen-binding molecule is an antibody, antibody fragment, or antibody derivative.

[00360] An agonistic antigen-binding molecule or agonistic antibody, as used in this document, is an antigen-binding molecule or antibody that significantly induces or enhances a biological activity of the antigen to which it binds (e.g., CD137 and CD3).

[00361] Therefore, if the antigen is, for example, CD137, such an antigen-binding molecule or antibody with agonistic action is called a CD137 agonistic antigen-binding molecule or a CD137 agonistic antibody, respectively. Similarly, if the antigen is, for example, CD3, such an antigen-binding molecule or antibody with agonistic action is called a CD3 agonistic antigen-binding molecule or a CD3 agonistic antibody, respectively.

[00362] The term antibody here is used in the broadest sense and encompasses various antibody structures, including but not limited to monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments, provided they exhibit antigen-binding activity. Petition 870260042208, dated 05 / 05 / 2026, p. 88 / 1364 72 / 448 desired.

[00363] An antibody fragment refers to a molecule other than an intact antibody comprising a portion of an intact antibody that binds to the antigen to which the intact antibody binds. Examples of antibody fragments include, but are not limited to, Fv, Fab, Fab', Fab'-SH, F(ab')2; diabodies; linear antibodies; single-chain antibody molecules (e.g., scFv); and multispecific antibodies formed from antibody fragments.

[00364] An antigen-binding molecule that binds to the same epitope or antibody that binds to the same epitope as a reference antigen-binding molecule or reference antibody refers to an antibody or antigen-binding molecule that blocks the binding of the reference antibody or reference antigen-binding molecule to its antigen in a competitive assay by 50% or more, and conversely, the reference antibody blocks the binding of the antibody to its antigen in a competitive assay by 50% or more. An exemplary competitive assay is provided here. In one embodiment, in the case where the reference antigen-binding molecule or the reference antibody shows antigen-binding activity in a low molecular weight compound-dependent manner, the competitive assay is performed in the presence of the low molecular weight compound.

[00365] The term chimeric antibody refers to an antibody in which a portion of the heavy and / or light chain is derived from a particular source or species, while the remainder of the heavy and / or light chain is derived from a different source or species.

[00366] The class of an antibody refers to the type of constant domain or constant region possessed by its heavy chain. There are five main classes of antibodies: IgA, IgD, IgE, IgG, and IgM. Petition 870260042208, dated 05 / 05 / 2026, p. 89 / 1364 73 / 448 and several of them can be divided into subclasses (isotypes), for example, IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2. The constant heavy chain domains that correspond to the different classes of immunoglobulins are called alpha, delta, epsilon, gamma and mu, respectively.

[00367] Effector functions refer to the biological activities attributable to the Fc region of an antibody, which vary with the antibody isotype. Examples of antibody effector functions include: C1q binding and complement-dependent cytotoxicity (CDC); Fc receptor binding; antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis; downregulation of cell surface receptors (e.g., B cell receptor); and B cell activation.

[00368] Cytotoxicity refers to activity that inhibits or prevents cellular function and / or causes cell death or destruction. Cytotoxicity can be, for example, antibody-dependent cell-mediated cytotoxicity (ADCC) activity, complement-dependent cytotoxicity (CDC) activity, and T-cell cytotoxicity; and it can be cytotoxicity caused by cytotoxic agents (e.g., radioisotopes and chemotherapeutic agents), such as immunoconjugates.

[00369] The term Fc region here is used to define a C-terminal region of an immunoglobulin heavy chain that contains at least a portion of the constant region. The term includes native sequence Fc regions and variant Fc regions. In one embodiment, a human IgG heavy chain Fc region extends from Cys226, or Pro230, to the carboxyl terminus of the heavy chain. However, the C-terminal lysine (Lys447) or glycine-lysine (residues 446-447) of the Fc region may or may not be present.Unless otherwise specified herein, the numbering of amino acid residues in the Fc region or constant region is in accordance with the EU numbering system. Petition 870260042208, dated 05 / 05 / 2026, page 90 / 1364 74 / 448, also called the EU index, as described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD, 1991.

[00370] The term variant Fc region herein comprises an amino acid sequence that differs from that of a native sequence Fc region by virtue of at least one amino acid modification, preferably one or more amino acid substitutions. Preferably, the variant Fc region has at least one amino acid substitution compared to a native sequence Fc region or to the Fc region of a parental polypeptide, for example, from about one to about ten amino acid substitutions, and preferably from about one to about five amino acid substitutions in a native sequence Fc region or in the Fc region of the parental polypeptide. The variant Fc region herein will preferably have at least about 80% homology with a native sequence Fc region and / or with an Fc region of a parental polypeptide, and more preferably at least about 90% homology with the same, most preferably at least about 95% homology with the same.

[00371] Here, amino acid changes or substitutions within an Fc region or a constant region can be represented by combining the EU numbering system and amino acids. For example, S424N represents a substitution at position 424 in the EU numbering of serine (Ser) to asparagine (Asn). EU424N means a substitution at position 424 in the EU numbering of an amino acid (any type) to asparagine (Asn).

[00372] The term antibody comprising the Fc region here refers to an antibody that comprises an Fc region. The C-terminal lysine (residue 447 according to the EU numbering system) or C-terminal glycinelysine (residues 446-447) of the Fc region may be removed, Petition 870260042208, dated 05 / 05 / 2026, page 91 / 1364 75 / 448, for example, during antibody purification or by recombinant engineering of the nucleic acid encoding the antibody. Consequently, a composition comprising an antibody possessing an Fc region according to the present disclosure may comprise an antibody with G446-K447, with G446 and without K447, with all G446-K447 removed, or a mixture of three types of antibodies described above.

[00373] The terms full-length antibody, intact antibody, and whole antibody are used herein interchangeably to refer to an antibody having a structure substantially similar to a native antibody structure or having heavy chains containing an Fc region or a variant Fc region as defined herein.

[00374] A human antibody is one that has an amino acid sequence that corresponds to that of an antibody produced by a human cell or derived from a non-human source using human antibody repertoires or other human antibody-coding sequences. This definition of a human antibody specifically excludes a humanized antibody comprising non-human antigen-binding residues.

[00375] Structure or FR refers to variable domain residues other than hypervariable region (HVR) residues. A variable domain FR generally consists of four FR domains: FR1, FR2, FR3, and FR4. Consequently, HVR and FR sequences generally appear in the following sequence in VH (or VL): FR1-H1(L1)-FR2-H2(L2)-FR3-H3(L3)-FR4.

[00376] An acceptable human structure for the purposes of this document is a structure comprising the amino acid sequence of a variable light chain (VL) domain structure or a variable heavy chain (VH) domain structure derived Petition 870260042208, dated 05 / 05 / 2026, page 92 / 1364 76 / 448 of a human immunoglobulin structure or a human consensus structure, as defined below. A human acceptor structure derived from a human immunoglobulin structure or a human consensus structure may comprise the same amino acid sequence as the same, or may contain changes in the amino acid sequence. In some embodiments, the number of amino acid changes is 10 or less, 9 or less, 8 or less, 7 or less, 6 or less, 5 or less, 4 or less, 3 or less, or 2 or less. In some embodiments, the VL acceptor structure is sequence-identical to the VL human immunoglobulin structure sequence or human consensus structure sequence.

[00377] A human consensus structure is a structure that represents the most commonly occurring amino acid residues in a selection of human immunoglobulin VL or VH structure sequences. Generally, the selection of human immunoglobulin VL or VH sequences is from a variable domain sequence subgroup. Generally, the sequence subgroup is a subgroup as in Kabat et al., Sequences of Proteins of Immunological Interest, Fifth Edition, NIH Publication 91-3242, Bethesda MD (1991), vols. 1-3. In one embodiment, for VL, the subgroup is kappa subgroup I as in Kabat et al., Supra. In one embodiment, for VH, the subgroup is subgroup III as in Kabat et al., Supra.

[00378] A humanized antibody refers to a chimeric antibody comprising amino acid residues from non-human HVRs and amino acid residues from human FRs. In certain embodiments, a humanized antibody will comprise substantially all of at least one, and typically two, variable domains, where all or substantially all HVRs (e.g., CDRs) correspond to those of a non-human antibody, and all or substantially all FRs correspond to those of a human antibody. Petition 870260042208, dated 05 / 05 / 2026, p. 93 / 1364 77 / 448 human antibody. A humanized antibody may optionally comprise at least a portion of a constant antibody region derived from a human antibody. A humanized form of an antibody, for example, a non-human antibody, refers to an antibody that has undergone humanization.

[00379] The term variable region or variable domain refers to the domain of an antibody heavy or light chain that is involved in antibody binding to the antigen. The variable domains of the heavy chain and light chain (VH and VL, respectively) of a native antibody generally have similar structures, with each domain comprising four conserved structural regions (FRs) and three hypervariable regions (HVRs). (See, for example, Kindt et al. Kuby Immunology, 6th ed., WH Freeman and Co., page 91 (2007).) A single VH or VL domain may be sufficient to confer antigen-binding specificity. Furthermore, antibodies that bind to a given antigen can be isolated using a VH or VL domain from an antibody that binds to the antigen to screen a library of complementary VL or VH domains, respectively. See, for example, Portolano et al., J. Immunol. 150: 880-887 (1993); Clarkson et al., Nature 352: 624-628 (1991).

[00380] The term hypervariable region or HVR, as used herein, refers to each of the regions of an antibody variable domain that are hypervariable in sequence (complementarity-determining regions or CDRs) and / or form structurally defined loops (hypervariable loops) and / or contain antigen-contact residues (antigen contacts). Generally, antibodies comprise six HVRs: three in the VH (H1, H2, H3) and three in the VL (L1, L2, L3). Exemplary HVRs herein include: (a) Hypervariable loops that occur at amino acid residues 26-32 (L1), 50-52 (L2), 91-96 (L3), 26-32 (H1), 53-55 (H2) Petition 870260042208, dated 05 / 05 / 2026, page 94 / 1364 78 / 448 and 96-101 (H3) (Chothia and Lesk, J. Mol. Biol. 196: 901-917 (1987)); (b) CDRs occurring at amino acid residues 24-34 (L1), 50-56 (L2), 89-97 (L3), 31-35b (H1), 50-65 (H2), and 95-102 (H3) (Kabat et al., Sequences of Proteins of Immunological Interest, 5thEd. Public Health Service, National Institutes of Health, Bethesda, MD (1991)); (c) Antigen contacts that occur at amino acid residues 27c-36 (L1), 46-55 (L2), 89-96 (L3), 30-35b (H1), 47-58 (H2), and 93-101 (H3) (MacCallum et al. J. Mol. Biol. 262: 732-745 (1996)); and (d) Combinations of (a), (b), and / or (c), including HVR amino acid residues 46-56 (L2), 47-56 (L2), 48-56 (L2), 49-56 (L2), 26-35 (H1), 26-35b (H1), 49-65 (H2), 93-102 (H3), and 94-102 (H3).

[00381] Unless otherwise indicated, HVR residues and other residues in the variable domain (e.g., FR residues) are numbered here according to Kabat et al., Supra. Here, HVR residues or other residues within a variable domain (e.g., FR residues) and amino acid changes or substitutions at such residues may be represented by combining the Kabat numbering system and amino acids. For example, N99 represents asparagine (Asn) at position 99 in the Kabat numbering, and N99A represents a substitution at position 99 in the Kabat numbering of asparagine (Asn) to alanine (Ala).

[00382] An immunoconjugate is an antibody conjugated to one or more heterologous molecules, including, but not limited to, a cytotoxic agent.

[00383] The term cytotoxic agent, as used herein, refers to a substance that inhibits or prevents a cellular function and / or causes cell death or destruction. Cytotoxic agents include, but are not limited to, radioactive isotopes (e.g., 211At, 131I, 125I, Petition 870260042208, dated 05 / 05 / 2026, page 95 / 1364 79 / 448 90Y, 186Re, 188Re, 153Sm, 212Bi, 32P, 212Pb and radioactive isotopes of Lu); chemotherapeutic agents or drugs (for example, methotrexate, adriamycin, vinca alkaloids (vincristine, vinblastine, etoposide), doxorubicin, melphalan, mitomycin C, chlorambucil, daunorubicin or other intercalating agents); growth-inhibiting agents; enzymes and their fragments, such as nucleolytic enzymes; antibiotics; toxins, such as small molecule toxins or enzymatically active toxins of bacterial, fungal, plant or animal origin, including fragments and / or variants thereof; and the various antitumor or anticancer agents disclosed below.

[00384] An isolated antibody is one that has been separated from a component of its natural environment. In some embodiments, an antibody is purified to more than 95% or 99% purity as determined, for example, by electrophoresis (e.g., SDS-PAGE, isoelectric focusing (IEF), capillary electrophoresis) or chromatography (e.g., ion exchange or reverse phase HPLC). For a review of methods for assessing antibody purity, see, for example, Flatman et al., J. Chromatogr. B 848: 79-87 (2007).

[00385] An isolated nucleic acid refers to a nucleic acid molecule that has been separated from a component of its natural environment. An isolated nucleic acid includes a nucleic acid molecule contained in cells that normally contain the nucleic acid molecule, but the nucleic acid molecule is present extrachromosomally or in a chromosomal location that is different from its natural chromosomal location.

[00386] The term vector, as used herein, refers to a nucleic acid molecule capable of propagating another nucleic acid to which it is attached. The term includes the vector as a self-replicating nucleic acid structure, as well as the vector incorporated into the genome of Petition 870260042208, dated 05 / 05 / 2026, page 96 / 1364 80 / 448 a host cell into which it was introduced. Certain vectors are capable of directing the expression of nucleic acids to which they are operationally linked. These vectors are referred to here as expression vectors.

[00387] Nucleic acid encoding for anti-CD137 antigen-binding molecule refers to one or more nucleic acid molecules encoding polypeptide(s) that constitute the antigen-binding molecule. Isolated nucleic acid encoding an anti-CD137 antibody refers to one or more nucleic acid molecules encoding the heavy and light chains of the antibody (or fragments thereof), including such nucleic acid molecule(s) in a single vector or separate vectors, and such nucleic acid molecule(s) present in one or more locations on a host cell.

[00388] The terms host cell, host cell line, and host cell culture are used interchangeably and refer to cells into which exogenous nucleic acid has been introduced, including the progeny of such cells. Host cells include transformants and transformed cells, which include the primary transformed cell and the progeny derived from it regardless of the number of passages. The progeny may not be completely identical in nucleic acid content to a parent cell, but may contain mutations. Mutant progeny that possesses the same biological function or activity as screened for or selected in the originally transformed cell is included here.

[00389] The term monoclonal antibody, as used herein, refers to an antibody obtained from a population of substantially homogeneous antibodies, that is, the individual antibodies that make up the population are identical and / or bind to the same epitope, except for possible variant antibodies, for example, containing naturally occurring mutations or arising during the production of a Petition 870260042208, dated 05 / 05 / 2026, page 97 / 1364 81 / 448 monoclonal antibody preparation, such variants generally being present in smaller quantities. In contrast to polyclonal antibody preparations, which typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody in a monoclonal antibody preparation is directed against a single determinant on an antigen. Thus, the monoclonal modifier indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies and should not be interpreted as requiring the production of the antibody by any particular method.For example, the monoclonal antibodies to be used in accordance with this disclosure can be made by a variety of techniques, including, but not limited to, the hybridoma method, recombinant DNA methods, phage display methods, and methods using transgenic animals containing all or part of the human immunoglobulin loci, such methods and other exemplary methods for the production of monoclonal antibodies being described herein.

[00390] A nude antibody refers to an antibody that is not conjugated to a heterologous moiety (e.g., a cytotoxic moiety) or radiolabel. Nude antibodies may be present in a pharmaceutical formulation.

[00391] Native antibodies refer to naturally occurring immunoglobulin molecules with variable structures. For example, native IgG antibodies are heterotetrameric glycoproteins of approximately 150,000 daltons, composed of two identical light chains and two identical heavy chains that are disulfide-linked. From the N- to the C-terminal, each heavy chain has a variable region (VH), also called a variable heavy domain or variable heavy chain domain, followed by three constant domains (CH1, CH2, and CH3). Similarly, from the N- to the C-terminal, Petition 870260042208, dated 05 / 05 / 2026, page 98 / 1364 82 / 448 Each light chain has a variable region (VL), also called a variable light domain or variable light chain domain, followed by a constant light domain (CL). The light chain of an antibody can be assigned to one of two types, called kappa (κ) and lambda (λ), based on the amino acid sequence of its constant domain.

[00392] Percentage (%) of amino acid sequence identity with respect to a reference polypeptide sequence is defined as the percentage of amino acid residues in a candidate sequence that is identical to the amino acid residues in the reference polypeptide sequence, after sequence alignment and introduction of gaps, if necessary, to achieve maximum sequence identity, and not considering any conservative substitutions as part of the sequence identity. Alignment for the purpose of determining percentage of amino acid sequence identity can be achieved in several ways that are within the expertise of the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN, Megalign software (DNASTAR) or GENETYX (registered trademark) (Genetyx Co., Ltd.).Those skilled in the technique can determine the appropriate parameters for aligning sequences, including any algorithms needed to achieve maximum alignment across the entire length of the sequences being compared.

[00393] The ALIGN-2 sequence comparison computer program was authored by Genentech, Inc., and the source code was deposited with the user documentation at the U.S. Copyright Office, Washington DC, 20559, where it is registered under U.S. Copyright Registration No. TXU510087. The ALIGN-2 program is publicly available from Genentech, Inc., South San Francisco, California, or can be compiled from the source code. The ALIGN-2 program should Petition 870260042208, dated 05 / 05 / 2026, page 99 / 1364 83 / 448 to be compiled for use on a UNIX operating system, including digital UNIX V4.0D. All sequence comparison parameters are defined by the ALIGN-2 program and do not vary. In situations where ALIGN-2 is employed for amino acid sequence comparisons, the % amino acid sequence identity of a given amino acid sequence A to, with, or against a given amino acid sequence B (which may alternatively be expressed as a given amino acid sequence A having or comprising a certain % amino acid sequence identity to, with, or against a given amino acid sequence B) is calculated as follows: 100 times the fraction X / Y where X is the number of amino acid residues marked as identical matches by the ALIGN-2 sequence alignment program in that program's alignment of A and B, and where Y is the total number of amino acid residues in B. It will be appreciated that where the length of amino acid sequence A is not equal to the length of amino acid sequence B, the % amino acid sequence identity from A to B will not be equal to the % amino acid sequence identity from B to A. Unless specifically indicated otherwise, all % amino acid sequence identity values ​​used herein are obtained as described in the immediately preceding paragraph using the ALIGN-2 computer program.

[00394] The term pharmaceutical formulation refers to a preparation that is in a form that allows the biological activity of an active ingredient contained therein to be effective, and that does not contain additional components that are unacceptably toxic to an object to which the formulation would be administered.

[00395] A pharmaceutically acceptable vehicle refers to a Petition 870260042208, dated 05 / 05 / 2026, page 100 / 1364 84 / 448 ingredient in a pharmaceutical formulation, other than an active ingredient, that is not toxic to an object. A pharmaceutically acceptable vehicle includes, but is not limited to, a buffer, excipient, stabilizer, or preservative.

[00396] An effective amount of an agent, for example, a pharmaceutical formulation, refers to an effective quantity, in dosages and for periods of time necessary, to achieve the desired therapeutic or prophylactic result.

[00397] An individual or object is a mammal. Mammals include, but are not limited to, domesticated animals (e.g., cows, sheep, cats, dogs, and horses), primates (e.g., humans and non-human primates such as monkeys), rabbits, and rodents (e.g., mice and rats). In certain embodiments, the individual or object is a human being.

[00398] The term CD137, as used herein, refers to any CD137 native to any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise indicated. The term encompasses unprocessed full-length CD137 as well as any form of CD137 resulting from processing in the cell. The term also encompasses naturally occurring variants of CD137, for example, spliced ​​variants or allelic variants.

[00399] An amino acid sequence of a full-length human CD137 specimen is shown in SEQ ID NO: 1 (NCBI Reference Sequence: NP_001552.2) and an amino acid sequence of an extracellular region of a human CD137 specimen is shown in SEQ ID NO: 2. An amino acid sequence of a full-length mouse CD137 specimen is shown in SEQ ID NO: 3 (NCBI Reference Sequence: NP_035742.1) and an amino acid sequence of an extracellular region of a CD137 specimen Petition 870260042208, dated 05 / 05 / 2026, page 101 / 1364 An amino acid sequence from a full-length CD137 of a monkey specimen is shown in SEQ ID NO: 4. An amino acid sequence from a full-length CD137 of a monkey specimen is shown in SEQ ID NO: 5 (NCBI Reference Sequence: ABY47575.1), and an amino acid sequence from an extracellular region of a monkey CD137 is shown in SEQ ID NO: 6.

[00400] CD137 is a member of the tumor necrosis factor (TNF) receptor family. Its alternative names are tumor necrosis factor receptor superfamily member 9 (TNFRSF9), 4-1BB, and ILA. In addition to its expression on activated CD4+ and CD8+ T cells, CD137 is expressed on B cells, dendritic cells, natural killer (NK) cells and NK-T cells, macrophages, monocytes, neutrophils, CD4-CD25+ regulatory T cells, and vascular endothelial cells. Expression on cancer cells is also reported (Labiano, et al. Oncoimmunology, vol. 24: e1062967 (2015)). The natural ligand of CD137, CD137L, is presented by antigen-presenting cells, such as B cells, monocytes / macrophages, and dendritic cells (Watts, et al., Annu. Rev. Immunol., Vol. 23: p. 23-68 (2005)).Through its interaction with the ligand, CD137 causes increased TCR-induced T cell proliferation, cytokine production, functional maturation, apoptosis suppression, and long-term survival of CD8+ T cells (Nam, et al., Curr. Cancer Drug Targets, vol. 5: p. 357-363 (2005); Watts, et al., Annu. Rev. Immunol., vol. 23: p. 23-68 (2005)).

[00401] The terms carcinoma, cancer, and cancerous refer to or describe the physiological condition in mammals that is typically characterized by unregulated cell growth / proliferation.

[00402] The term tumor refers to any growth and proliferation of neoplastic cells, whether malignant or benign, and all precancerous and cancerous cells and tissues. The terms carcinoma, cancer, cancerous, proliferative cell disorder, disorder Petition 870260042208, dated 05 / 05 / 2026, page 102 / 1364 86 / 448 proliferative and tumor are not mutually exclusive, as referred to in this document.

[00403] The terms cell proliferative disorder and proliferative disorder refer to disorders that are associated with some degree of abnormal cell proliferation. In one embodiment, the cell proliferative disorder is cancer.

[00404] As used herein, treatment (and its grammatical variations, such as treat or treating) refers to clinical intervention in an attempt to alter the natural course of the individual being treated and may be performed for prophylaxis or during the course of clinical pathology. Desirable effects of treatment include, but are not limited to, prevention of the occurrence or recurrence of the disease, relief of symptoms, reduction of any direct or indirect pathological consequences of the disease, prevention of metastasis, reduction of the rate of disease progression, improvement or palliation of the disease state, and remission or improved prognosis. In some embodiments, the antibodies of this disclosure are used to delay the development of a disease or to slow the progression of a disease. II. Compositions and Methods (anti-CD137 agonist antigen-binding molecules)

[00405] In one aspect, the present disclosure is based, in part, on anti-CD137 agonist antigen-binding molecules and their uses. In certain embodiments, antibodies that bind to CD137 are provided. The antibodies in the present disclosure may exhibit activation action on immune cells, cytotoxicity, or antitumor activity and are therefore useful, for example, in the diagnosis or treatment of cancer. A. Antigen-binding molecules anti-CD137 Samples or Antibodies

[00406] In one aspect, the present disclosure provides molecules of Petition 870260042208, dated 05 / 05 / 2026, page 103 / 1364 87 / 448 Isolated antigen-binding molecules or antibodies that bind to CD137. In certain embodiments, anti-CD137 antigen-binding molecules or antibodies - They possess CD137-binding activity dependent on a small molecule compound; - They bind to the extracellular region of CD137; - They form a ternary complex together with a low molecular weight compound and CD137; - They bind to human-derived CD137 and monkey-derived CD137; - They are agonistic for CD137 activity; - They show agonistic activity for CD137 in the presence of a low molecular weight compound; - They have low agonist activity for CD137 in the absence of the low molecular weight compound; and / or - They substantially show no agonistic activity for CD137 in the absence of the low molecular weight compound. Binding activity of antigen- or antibody-binding molecules

[00407] In certain embodiments, the binding activity of the antigen- or antibody-binding molecules provided herein is, in the presence of a low molecular weight compound, with a dissociation constant (KD) of 1 micromolar or less, 100 nM or less, 10 nM or less, 1 nM or less, 0.1 nM or less, 0.01 nM or less, or 0.001 nM or less (e.g., 10-6M or less, 10-7M or less, 10-8M or less, 10-9M or less, 10-10M or less, e.g. 106M to 10-10M, 10-7M to 10-9M, e.g. 10-7M to 10-8M).

[00408] In one embodiment, the binding activity of an antigen- or antibody-binding molecule is measured by a radiolabeled antigen binding assay (RIA) and represented by KD. In one embodiment, an RIA is performed with the Fab version of an antibody of interest and its antigen. For example, the solution binding affinity of Fabs for the antigen is measured by equilibrizing Fab with a minimum concentration of antigen labeled with (125I) in the presence of Petition 870260042208, dated 05 / 05 / 2026, page 104 / 1364 88 / 448 a series of titrations of unlabeled antigen, followed by capturing the bound antigen with an anti-Fab antibody-coated plate (see, for example, Chen et al., J. Mol. Biol. 293: 865-881 (1999)). To establish assay conditions, MICROTITER multi-well plates (trademark) (Thermo Scientific) are coated overnight with 5 micrograms / ml of a capture anti-Fab antibody (Cappel Labs) in 50 mM sodium carbonate (pH 9.6), and subsequently blocked with 2% (w / v) bovine serum albumin in PBS for two to five hours at room temperature (approximately 23 degrees C). On a non-adsorbent plate (Nunc # 269620), 100 pM or 26 pM [125I]-antigen are mixed with serial dilutions of a Fab of interest (e.g., consistent with the evaluation of the anti-VEGF antibody, Fab-12, in Presta et al., Cancer Res. 57: 4593-4599 (1997)).The Fab of interest is then incubated overnight; however, incubation may continue for a longer period (e.g., approximately 65 hours) to ensure equilibrium is reached. After this, the mixtures are transferred to a capture plate for incubation at room temperature (e.g., for one hour). The solution is then removed and the plate washed eight times with 0.1% polysorbate 20 (TWEEN-20 (trademark)) in PBS. Once the plates are dry, 150 microliters / well of scintillating agent (MICROSCINT-20™; Packard) are added and the plates are counted on a TOPCOUNT™ gamma counter (Packard) for ten minutes. Concentrations of each Fab that give less than or equal to 20% of the maximum binding are selected for use in competitive binding assays.

[00409] In one embodiment, to measure the binding activity of an antibody, ligand capture methods, for example, use BIACORE (registered trademark) T200 or BIACORE (registered trademark) 4000 (GE Healthcare, Uppsala, Sweden), which rely on methods of Petition 870260042208, dated 05 / 05 / 2026, page 105 / 1364 89 / 448 surface plasmon resonance analysis is used as the measurement principle. The BIACORE control software (registered trademark) is used for device operation. In one embodiment, the amine coupling kit (GE Healthcare, Uppsala, Sweden) is used according to the manufacturer's instructions to allow a ligand-capturing molecule, e.g., an anti-label antibody, an anti-IgG antibody, protein A, etc., to be fixed onto a sensor chip (GE Healthcare, Uppsala, Sweden) coated with carboxymethyldextran. The ligand-capturing molecule is diluted with 10 mM of a sodium acetate solution to an appropriate pH and injected at an appropriate flow rate and for an appropriate injection time.Binding activity measurements are performed using a buffer containing 0.05% polysorbate 20 (also known as Tween (trademark)-20) as the measuring buffer, at a flow rate of 10–30 microliters / minute and at a measurement temperature preferably of 25°C or 37°C. For measurements performed with an antibody captured by the ligand-capturing molecule as a ligand, an antibody is injected to allow a target quantity of the antibody to be captured, and then a serial dilution of an antigen and / or an Fc receptor (analyte) prepared using measuring buffer is injected. For measurements performed with an antigen and / or an Fc receptor captured by the ligand-capturing molecule as a ligand, an antigen and / or an Fc receptor is injected to allow a target quantity of the same to be captured, and then a serial dilution of the antibody (analyte) prepared using measuring buffer is injected.

[00410] In one embodiment, the measurement results are analyzed using the BIACORE evaluation software (registered trademark). The kinetic parameter calculation is performed by fitting association and dissociation sensograms simultaneously using Petition 870260042208, dated 05 / 05 / 2026, page 106 / 1364 90 / 448 A 1:1 binding model and an association rate (kon or ka), a dissociation rate (koff or kd), and an equilibrium dissociation constant (KD) can be calculated. For the case of weak binding activity, in particular, for cases where dissociation is rapid and kinetic parameters are difficult to calculate, the steady-state model can be used to calculate the equilibrium dissociation constant (KD). As additional parameters relating to binding activity, the amount of analyte binding per unit amount of ligand can be calculated by dividing an amount of analyte binding (resonance unit: RU) at a specific concentration by an amount of ligand captured. [Small Molecule Compound-Dependent Binding Activity]

[00411] In one aspect, the anti-CD137 antigen-binding molecule or antibody has small molecule compound-dependent CD137 binding activity. In a non-limiting embodiment, the anti-CD137 antigen-binding molecule or antibody has a higher CD137 binding activity in the presence of a small molecule compound compared to the CD137 binding activity in the absence of the small molecule compound. In a different embodiment, the anti-CD137 antigen-binding molecule or antibody has a higher CD137 binding activity in the presence of a high concentration of a small molecule compound compared to the CD137 binding activity in the presence of a low concentration of the small molecule compound.In a preferred embodiment, the binding activity of the antigen-binding molecule to anti-CD137 or antibody to CD137 in the presence of a small molecule compound is 2 times or more, 3 times or more, 5 times or more, 10 times or more, 15 times or more, 20 times or more, 25 times or more, 30 times or more, 50 times or more, 100 times or more, 200 times or more, 300 times or more, 500. Petition 870260042208, dated 05 / 05 / 2026, page 107 / 1364 91 / 448 times or more, 1 x 103 times or more, 2 x 103 times or more, 3 x 10 times or more, 5 x 103 times or more, 1 x 104 times or more, 2 x 10 times or more, 3 x 104 times or more, 5 x 104 times or more, or 1 x 10 times or more, compared to the binding activity in the absence of the small molecule compound.In a different preferred embodiment, the binding activity of the anti-CD137 antigen-binding molecule or antibody to CD137 in the presence of a small molecule compound is greater than 2 times, greater than 3 times, greater than 5 times, greater than 10 times, greater than 15 times, greater than 20 times, greater than 25 times, greater than 30 times, greater than 50 times, greater than 100 times, greater than 200 times, greater than 300 times, greater than 500 times, greater than 1 x 10³ times, greater than 2 x 10³ times, greater than 3 x 10³ times, greater than 5 x 10³ times, greater than 1 x 10⁴ times, greater than 2 x 10⁴ times, greater than 3 x 10⁴ times, greater than 5 x 10⁴ times, or greater than 1 x 105 times, compared to the binding activity in the absence of the small molecule compound.

[00412] The concentration of the small molecule compound can be any arbitrary concentration, as long as the difference in binding activity of the antigen-binding molecule to anti-CD137 or antibody is detected. In one embodiment, the concentration of the small molecule compound in the presence of a small molecule compound and / or in the presence of a high concentration of a small molecule compound is, for example, 100 nM or more, 500 nM or more, 1 μM or more, 3 μM or more, 5 μM or more, 10 μM or more, 50 μM or more, 100 μM or more, 150 μM or more, 200 μM or more, 250 μM or more, 300 μM or more, 400 μM or more, 500 μM or more, or 1 mM or more. Alternatively, the concentration may be defined as an amount that is sufficient for the antigen-binding molecule to show maximum binding activity to the antiCD137 antigen or antibody. Furthermore, Petition 870260042208, dated 05 / 05 / 2026, p. 108 / 1364 92 / 448 In one embodiment, the concentration of the small molecule compound in the presence of a low concentration of a small molecule compound may be, for example, 500 μM or less, 250 μM or less, 200 pM or less, 150 pM or less, 100 pM or less, 50 pM or less, 10 pM or less, 1 pM or less, 500 nM or less, 100 nM or less, 50 nM or less, or 10 nM or 1 nM or less. The case where the concentration of the small molecule compound is zero, or the substantial concentration is zero, may also be selected as a low concentration embodiment.

[00413] Here, the term substantial concentration is zero means, for example, a concentration that is so minute that it cannot be detected by current technology, even though the small molecule compound is present.

[00414] In one embodiment, the binding activity with respect to CD137 in the presence of a small molecule compound at a concentration of 10 pM, 50 pM, 100 pM, 150 pM, 200 pM or 250 pM is 2 times or more, 5 times or more, 10 times or more, 15 times or more, 16 times or more, 17 times or more, 18 times or more, 19 times or more, or 20 times or more, compared with the binding activity for CD137 in the absence of the small molecule compound. In one embodiment, the binding activity of the antigen-binding molecule to anti-CD137 or an antibody against CD137 in the presence of a small molecule compound at 10 pM or more is 2 times or more, 5 times or more, 10 times or more, 15 times or more, 16 times or more, 17 times or more, 18 times or more, 19 times or more, or 20 times or more, compared to the binding activity to CD137 in the absence of the small molecule compound.In one embodiment, the binding activity of the antigen-binding molecule against anti-CD137 or an antibody against CD137 in the presence of a small molecule compound at 100 pM or more is 2 times or more, 5 times or more, 10. Petition 870260042208, dated 05 / 05 / 2026, page 109 / 1364 93 / 448 times or more, 15 times or more, 16 times or more, 17 times or more, 18 times or more, 19 times or more, or 20 times or more, compared to the binding activity for CD137 in the absence of the small molecule compound.

[00415] In one embodiment, the binding activity (KD) of the antigen-binding molecule to anti-CD137 or antibody to CD137 in the presence of a small molecule compound of 10 μM or more is, a dissociation constant (KD) of 9 x 10-7M or less, 8 x 10-7M or less, 7 x 10-7M or less, 6 x 10-7M or less, 5 x 10-7M or less, or 4 x 10-7M or less, or preferably, a dissociation constant (KD) of 5 x 10-7M or less. In another embodiment, the binding activity (KD) of the antigen-binding molecule to anti-CD137 or antibody to CD137 in the absence of a small molecule compound is too large to be calculated by Biacore (weak binding activity), or is a dissociation constant (KD) of 1 x 10-7M or more, 5 x 10-7M or more, 7 x 10-7M or more, 8 x 10-7M or more, 9 x 10-7M or more, 1 x 10-6M or more, 2 x 10-6M or more, 3 x 10-6M or more, or 4 x 10-6M or more, or preferably, is a dissociation constant (KD) of 1 x 10-6M or more.In another embodiment, the binding activity (KD) of the antigen-binding molecule to anti-CD137 or an antibody to CD137 in the presence of a small molecule compound at 100 μM or more is, 9 x 10-7M or less, 8 x 10-7M or less, 7 x 10-7M or less, 6 x 10-7M or less, 5 x 10-7M or less, 4 x 10-7M or less, 3 x 10-7M or less, 2 x 10-7M or less, or 1 x 10-7M or less, or preferably, the dissociation constant (KD) is 2 x 10-7M or less. In another embodiment, the binding activity (KD) of the antigen-binding molecule to anti-CD137 or an antibody to CD137 in the absence of the small molecule compound is too large to be calculated by Biacore (weak binding activity), or is a dissociation constant (KD) of 1 x 10-7M or more, 5 x 10-7M or more. Petition 870260042208, dated 05 / 05 / 2026, p. 110 / 1364 94 / 448 x 10-7M or more, 8 x 10-7M or more, 9 x 10-7M or more, 1 x 10-6M or more, 2 x 10-6M or more, 3 x 10-6M or more, or 4 x 10-6M or more, or preferably, a dissociation constant (KD) of 1 x 10-6M or more.

[00416] In one embodiment, the binding activity (KD) of the anti-CD137 antigen-binding molecule or antibody to CD137 in the presence of a small molecule compound at 10 μM or more is a dissociation constant (KD) of 8 x 10⁻⁸ M or less, and the binding activity (KD) to CD137 in the absence of the compound is too large to be calculated by Biacore (weak binding activity). In another embodiment, the binding activity (KD) of the anti-CD137 antigen-binding molecule or antibody to CD137 in the presence of a small molecule compound at 100 pM is a dissociation constant (KD) of 2 x 10⁻⁸ M or less, and the binding activity to CD137 in the absence of the small molecule compound is too large to be calculated by Biacore (weak binding activity).

[00417] In one aspect, the present disclosure provides an anti-CD137 antigen-binding molecule or antibody in which the value of, [binding activity (amount of binding) to CD137 in the presence of a low molecular weight compound at 10 pM or more] / [binding activity (amount of binding) to CD137 in the absence of the small molecule compound)] is equal to or greater than the value of a reference anti-CD137 antigen-binding molecule. In a different aspect, the present disclosure provides an anti-CD137 antigen-binding molecule or antibody in which the value of [binding activity (amount of binding) to CD137 in the presence of a low molecular weight compound at 100 pM or more] / [binding activity (amount of binding) to CD137 in the absence of the small molecule compound] is equal to or greater than the value of an anti-CD137 antigen-binding molecule reference. Petition 870260042208, dated 05 / 05 / 2026, page 111 / 1364 95 / 448 CD137. In any of the above aspects, the reference anti-CD137 antigen-binding molecule can be selected from anti-CD137 antibodies containing HVR-H1, HVR-H2, HVR-H3, HVR-L1, HVR-L2, and HVR-L3 having the same amino acid sequences as HVR-H1, HVR-H2, HVR-H3, HVR-L1, HVR-L2, and HVR-L3 contained in A375 / B167, A372 / B040, A356 / B040, A486 / B167, A487 / B167. A488 / B226, A489 / B223, A548 / B376, A551 / B256, A551 / B379, A555 / B379, A548 / B256 or A549 / B167 described in Table 17.

[00418] In one embodiment, the reference antiCD137 antigen-binding molecule is an antibody comprising the amino acid sequence A375 / B167, A372 / B040, A356 / B040, A486 / B167, A487 / B167, A488 / B226, A489 / B223, A548 / B376, A551 / B256, A551 / B379, A555 / B379, A548 / B256, or A549 / B167 described in Table 17 as a combination of variable heavy chain region / variable light chain region. In a preferred embodiment, the reference antigen-binding molecule is an anti-CD137 antibody comprising HVR-H1, HVR-H2, HVR-H3, HVR-L1, HVR-L2, and HVR-L3 having the same amino acid sequences as HVR-H1, HVR-H2, HVR-H3, HVR-L1, HVR-L2, and HVR-L3 comprised in A375 / B167. In another embodiment, the reference anti-CD137 antigen-binding molecule is an anti-CD137 antibody comprising A375 / B167 as a combination of variable heavy chain region / variable light chain region. In a different preferred embodiment, the reference antigen-binding molecule is the anti-CD137 antibody comprising HVR-H1, HVR-H2, HVR-H3, HVR-L1, HVR-L2 and HVR-L3 with the same amino acid sequences as HVR-H1, HVR-H2, HVR-H3, HVR-L1, HVR-L2 and HVR-L3 included in A551 / B379.In another embodiment, the reference anti-CD137 antigen-binding molecule is an anti-CD137 antibody comprising A551 / B379 as a combination of. Petition 870260042208, dated 05 / 05 / 2026, page 112 / 1364 96 / 448 variable heavy chain region / variable light chain region. In a preferred embodiment, the reference antigen-binding molecule comprises constant heavy and light chain regions of human origin (e.g., G1T3 (SEQ ID NO: 138) as the constant heavy chain region, Lamlib of human λ chain (SEQ ID NO: 63) as the constant light chain region).

[00419] In one aspect, the present disclosure provides an anti-CD137 antigen-binding molecule or antibody wherein the binding activity (amount of binding) to CD137 in the absence of a small molecule compound is equal to or less than that of a reference anti-CD137 antigen-binding molecule, and also, the binding activity (amount of binding) to CD137 in the presence of the small molecule compound at 10 μM or more is equal to or greater than that of the reference anti-CD137 antigen-binding molecule for CD137 under the same conditions.In a different aspect, the present disclosure provides an anti-CD137 antigen-binding molecule or antibody wherein the binding activity for CD137 in the absence of a small molecule compound is equal to or less than that of a reference anti-CD137 antigen-binding molecule, and also, the binding activity (amount of binding) for CD137 in the presence of the small molecule compound at 10 pM or more is equal to or greater than the binding activity (amount of binding) of the reference anti-CD137 antigen-binding molecule for CD137 under the same conditions. In any of the above aspects, the reference anti-CD137 antigen-binding molecule can be selected from anti-CD137 antibodies containing HVR-H1, HVR-H2, HVR-H3, HVR-L1, HVR-L2, and HVR-L3 having the same amino acid sequences as HVR-H1, HVR-H2, HVR-H3, HVR-L1, HVR-L2, and HVR-L3 contained in A375 / B167, A372 / B040, A356 / B040, A486 / B167, A487 / B167, A488 / B226, A489 / B223, A548 / B376. Petition 870260042208, dated 05 / 05 / 2026, page 113 / 1364 97 / 448 A551 / B256, A551 / B379, A555 / B379, A548 / B256 or A549 / B167 described in Table 17.

[00420] In one embodiment, the reference anti-CD137 antigen-binding molecule is an anti-CD137 antibody comprising the amino acid sequence A375 / B167, A372 / B040, A356 / B040, A486 / B167, A487 / B167, A488 / B226, A489 / B223, A548 / B376, A551 / B256, A551 / B379, A555 / B379, A548 / B256, or A549 / B167 described in Table 17 as a combination of variable heavy chain region / variable light chain region. In a different preferred embodiment, the reference antigen-binding molecule is an anti-CD137 antibody comprising HVR-H1, HVR-H2, HVR-H3, HVR-L1, HVR-L2, and HVR-L3 having the same amino acid sequences as HVR-H1, HVR-H2, HVR-H3, HVR-L1, HVR-L2, and HVR-L3 comprised in A375 / B167. In another embodiment, the reference anti-CD137 antigen-binding molecule is an anti-CD137 antibody comprising A375 / B167 as a combination of variable heavy chain region / variable light chain region. In a preferred embodiment, the reference antigen-binding molecule is an anti-CD137 antibody comprising HVR-H1, HVR-H2, HVR-H3, HVR-L1, HVR-L2 and HVR-L3 having the same amino acid sequences as HVR-H1, HVR-H2, HVR-H3, HVR-L1, HVR-L2 and HVR-L3 included in A551 / B379.In another embodiment, the reference anti-CD137 antigen-binding molecule is an anti-CD137 antibody comprising A551 / B379 as a combination of heavy chain variable region / light chain variable region. In a preferred embodiment, the reference antigen-binding molecule comprises human-derived heavy and light chain constant regions (e.g., G1T3 (SEQ ID NO: 138) as the heavy chain constant region, human λ-chain Lamlib (SEQ ID NO: 63) as the light chain constant region). Petition 870260042208, dated 05 / 05 / 2026, page 114 / 1364 98 / 448

[00421] In one aspect, the present disclosure provides an anti-CD137 antigen-binding molecule or antibody in which the value of [binding activity (KD) for CD137 in the presence of a low molecular weight compound at 1 μM] / [binding activity (KD) for CD137 in the presence of the compound at 10 μM or more] is equal to or greater than the value of a reference antigen-binding molecule. In a different aspect, the present disclosure provides an anti-CD137 antigen-binding molecule or antibody in which the value of [binding activity (KD) for CD137 in the presence of a low molecular weight compound at 1 μM] / [binding activity (KD) for CD137 in the presence of the low molecular weight compound at 100 μM or more] is equal to or greater than the value of a reference antigen-binding molecule.In any of the above aspects, the reference antigen-binding molecule can be selected from anti-CD137 antibodies containing HVR-H1, HVR-H2, HVR-H3, HVR-L1, HVR-L2, and HVR-L3 possessing the same amino acid sequences as HVR-H1, HVR-H2, HVR-H3, HVR-L1, HVR-L2, and HVR-L3 contained in A375 / B167, A372 / B040, A356 / B040, A486 / B167, A487 / B167, and A488 / B226. A489 / B223, A548 / B376, A551 / B256, A551 / B379, A555 / B379, A548 / B256 or A549 / B167 described in Table 17.

[00422] In one embodiment, the reference antigen-binding molecule is an antibody comprising the amino acid sequence A375 / B167, A372 / B040, A356 / B040, A486 / B167, A487 / B167, A488 / B226, A489 / B223, A548 / B376, A551 / B256, A551 / B379, A555 / B379, A548 / B256 or A549 / B167 described in Table 17 as a combination of variable heavy chain region / variable light chain region. In a preferred embodiment, the reference antigen-binding molecule is an antibody comprising HVR-H1, HVR-H2, HVR-H3, HVR-L1, HVR-L2 and HVR-L3 having the same amino acid sequences as HVR-H1, HVR-H2, HVR-H3, HVR-L1, Petition 870260042208, dated 05 / 05 / 2026, p. 115 / 1364 99 / 448 In one embodiment, the reference antigen-binding molecule is an antibody comprising A375 / B167 as a combination of variable heavy chain region / variable light chain region. In another embodiment, the reference antigen-binding molecule is an antibody comprising HVR-H1, HVR-H2, HVR-H3, HVR-L1, HVR-L2, and HVR-L3 having the same amino acid sequences as HVR-H1, HVR-H2, HVR-H3, HVR-L1, HVR-L2, and HVR-L3 comprised in A551 / B379. In yet another embodiment, the reference antigen-binding molecule is an antibody comprising A551 / B379 as a combination of variable heavy chain region / variable light chain region.In a preferred embodiment, the reference antigen-binding molecule comprises constant regions of human-derived heavy and light chains (e.g., G1T3 (SEQ ID NO: 138) as the heavy chain constant region, human λ-chain Lamlib (SEQ ID NO: 63) as the light chain constant region.

[00423] In one embodiment, the binding activity of anti-CD137 antibody to CD137, with the presence, absence, high concentration and / or low concentration of a small molecule compound, is measured, for example, by a ligand capture method using BIACORE (registered trademark) T200 with surface plasmon resonance spectroscopy as the measurement principle.

[00424] Details of an exemplary method for measuring the binding activity of anti-CD137 antibody to CD137 are described below. In one embodiment, the binding activity of anti-CD137 antibody to CD137 is assessed by BIACORE (registered trademark) T200. In a preferred embodiment, this assay uses 20 mM ACES (pH 7.4), 150 mM NaCl, 2 mM MgCl2, and 0.05% Tween 20 as running buffer and is performed at 37 °C. In one embodiment, this measurement is Petition 870260042208, dated 05 / 05 / 2026, page 116 / 1364 100 / 448 performed after capturing an antibody as a ligand on the ligand capture molecule. Specifically, an appropriate amount (e.g., approximately 100 RU, 200 RU, 300 RU, 400 RU, or 500 RU) of antibody is captured, interacting with an antibody solution prepared using the running buffer with a chip prepared first by immobilizing Sure A Protein (GE Healthcare) on a CM3 Series S Chip sensor (GE Healthcare).

[00425] In a preferred embodiment, approximately 100 to 500 RU, preferably approximately 250 to 400 RU of antibody are captured. Then, the binding activity for CD137 in the presence and absence of a small molecule compound is evaluated by the interaction of a CD137 solution prepared using a running buffer supplemented with a small molecule compound to a target concentration (e.g., 1 μM, 10 μM, 50 μM, or 100 μM), or a CD137 solution prepared using a running buffer that does not contain the small molecule compound. Although the concentration of CD137 in the CD137 solution can be appropriately determined, for example, when hCD137-HisBAP (see Example 1-1) is used as the antigen, the measurement is performed using an antigen concentration of 0 nM, 15.625 nM, 62.5 nM, 250 nM, and 1000 nM, respectively. In one embodiment, the dissociation constant (KD) of the anti-CD137 antibody to human CD137 is calculated using the Biacore T200 Evaluation 2.0 software.Specifically, the bonding rate constant ka (L / mol / s) and the dissociation rate constant kd (L / s) are calculated by global fitting of a sensorgram obtained by measurement using the 1:1 Langmuir bonding model. The dissociation constant KD (mol / L) is calculated from the values.

[00426] Other exemplary assay methods for measuring the binding activity of anti-CD137 antibody to CD137 will be described in detail below. Binding of anti-CD137 antibodies to CD137 Petition 870260042208, dated 05 / 05 / 2026, page 117 / 1364 Human CD137 binding was assessed using Biacore T200. Binding to human CD137 was measured using 20 mM ACES (pH 7.4), 150 mM NaCl, 2 mM MgCl2, and 0.05% Tween 20 as a running buffer, and this was performed at 37 °C. First, an antibody of approximately 250 to 400 RU is captured by the interaction of an antibody solution prepared using the running buffer with a chip where the Sure A protein (GE Healthcare) is immobilized on a CM3 Series S Sensor Chip (GE Healthcare). Next, a solution of human CD137 prepared using an ATP-supplemented running buffer at a target concentration (e.g., 1 pM, 10 pM, 50 pM, or 100 pM), or a solution of human CD137 prepared using an ATP-free running buffer, is interacted to assess CD137 binding activity in the presence and absence of ATP.hCD137-HisBAP prepared by the method of Example (1-1) is used as the human CD137 antigen, and the measurement is performed at antigen concentrations of 0 nM, 15.625 nM, 62.5 nM, 250 nM, and 1000 nM, respectively. The chip is regenerated using 25 mM NaOH and 10 mM Glycine-HCl (pH 1.5), and the measurement is conducted by repeatedly capturing the antibodies. The dissociation constant of each antibody for human CD137 is calculated using Biacore T200 Evaluation 2.0 software. Specifically, the binding rate constant ka (L / mol / s) and the dissociation rate constant kd (L / s) are calculated by global fitting of the sensorgram obtained by measurement using the 1:1 Langmuir binding model. The dissociation constant KD (mol / L) is calculated from the values.

[00427] In one embodiment, the binding activity of anti-CD137 antibody to CD137 (preferably human CD137) can also be reformulated as the amount of CD137 binding per antibody unit quantity. Specifically, using the sensorgrams obtained by the assay method above using BIACORE Petition 870260042208, dated 05 / 05 / 2026, page 118 / 1364 102 / 448 (registered trademark) T200, the amount of CD137 binding to the antibody (RU) is divided by the amount of antibody captured to calculate the amount of CD137 binding per antibody unit quantity. In one embodiment, the binding activity of anti-CD137 antibody to CD137 (preferably human CD137) can also be measured by the method described in Example 5-3 or 6-2.

[00428] The terms small molecule and small molecule compound refer to a naturally occurring chemical substance other than biopolymers present in the living body or a non-naturally occurring chemical substance. Preferably, it is a target tissue-specific compound or a non-naturally occurring compound, but is not limited to the same. In one embodiment, the small molecule compound in the present disclosure is a cancerous tissue-specific compound or a cancerous tissue-specific metabolite. The term in the present disclosure a cancerous tissue-specific compound (cancerous tissue-specific compound) refers to a compound that exists differentially in tumor tissue compared to non-tumor tissue. As used in this document, the term cancer is generally used to describe a malignant neoplasm and may be metastatic or non-metastatic.The term metabolism refers to chemical changes that occur within an organism's tissues, including assimilation and catabolism. Assimilation refers to the biosynthesis or accumulation of molecules, and catabolism refers to the breakdown of molecules. A metabolite is an intermediate or product resulting from the metabolism of a substance.

[00429] The term target tissue means any tissue in the living body to which the antigen-binding molecule of the present invention is intended to be delivered. The target tissue may be a histologically distinguishable tissue, such as various organs, or a pathologically distinguishable tissue, such as normal tissues and diseased tissues. Petition 870260042208, dated 05 / 05 / 2026, p. 119 / 1364 103 / 448 In certain modalities, the target tissue is tumor tissue. In contrast, non-target tissue(s) means tissues of the living body that are not the target tissue.

[00430] The term tumor tissue means a tissue comprising at least one tumor cell. Generally, a tumor tissue is made up of a population of tumor cells that constitute the main body of the tumor (parenchyma) and connective tissues and blood vessels existing between the tumor cells and supporting the tumor (stroma). In some cases, they are clearly distinguishable, but there are cases where they are mixed. In some cases, there are cells, such as immune system cells, that have infiltrated the tumor tissue. In contrast, non-tumor tissue means a tissue in the living body different from tumor tissue(s). Healthy non-diseased tissues / normal tissues are representatives of such non-tumor tissues.

[00431] As a non-limiting embodiment of a cancer tissue-specific compound, or a cancer tissue-specific metabolite, as used in this disclosure, at least one compound selected from the compounds detailed below may be suitably exemplified. The meaning of at least one compound includes, in addition to the case where the binding activity against the antigen by the same antigen-binding domains described below depends on one type of cancer tissue-specific compound or cancer tissue-specific metabolite, the case where the binding activity depends on several types of cancer tissue-specific compounds or cancer tissue-specific metabolites.

[00432] As used herein, the term target tissue-specific compound refers to a compound that is differentially present in the target tissue compared to non-target tissue. In various embodiments, the target tissue-specific compound may be Petition 870260042208, dated 05 / 05 / 2026, p. 120 / 1364 104 / 448 a compound defined by a qualitative specificity of the target tissue, such as being present in the target tissue but not in non-target tissues, or being present in non-target tissue but not in the target tissue. In a different embodiment, the target tissue-specific compound may be a compound defined by a quantitative specificity of the target tissue, such as being present in the target tissue at a concentration that is different (e.g., a higher or lower concentration) compared to the target tissue. In a specific embodiment, the target tissue-specific compound is present in the target tissue at a concentration that is, for example, 1.05 times or more, 1.1 times or more, 1.15 times or more, 1.2 times or more, 1.25 times or more, 1.3 times or more, 1.35 times or more, 1.4 times or more, 1.45 times or more, 1.5 times or more, 1.55 times or more, 1.6 times or more, 1.65 times or more, 1.7 times or more, 1.75 times or more, 1.8 times or more, 1.85 times or more, 1.9 times or more, 1.95 times or more, 2 times or more, 2.1 times or more, 2.2 times or more, 2.3 times or more, 2.4 times or more, 2.5 times or more, 3 times or more, 5 times or more, 10 times or more, 50 times or more, 100 times or more, 103 times or more, 104 times or more, 105 times or more, 106 times or more compared to non-target tissue.In another embodiment, the specific target tissue compound is present in the target tissue at a concentration that is, for example, 1.05 times or more, 1.1 times or more, 1.15 times or more, 1.2 times or more, 1.25 times or more, 1.3 times or more, 1.35 times or more, 1.4 times or more, 1.45 times or more, 1.5 times or more, 1.55 times or more, 1.6 times or more, 1.65 times or more, 1.7 times or more, 1.75 times or more, 1.8 times or more, 1.85 times or more, 1.9 times or more, 1.95 times or more, 2 times or more, 2.1 times or more, 2.2 times or more, 2.3 times or more, 2.4 times or more, 2.5 times or more, 3 times or more, 5 times or more, 10 times or more, 50 times or more, 100 times or more, 103 times or more, 104 times or more, 105 times or more, 106 times or more, or more in comparison. Petition 870260042208, dated 05 / 05 / 2026, page 121 / 1364 105 / 448 with non-target tissue. In a specific embodiment, a target tissue-specific compound, compared to non-target tissue, is present in a target tissue at a concentration that is statistically significantly higher or lower (i.e., as determined using one of Welch's t-tests or Wilcoxon rank-sum test, the p-value is less than 0.05 and / or the q-value is less than 0.10). In a specific embodiment, the target tissue-specific compound is a tumor tissue-specific compound.

[00433] In a specific embodiment, a tumor tissue-specific compound is a metabolite produced by a specific metabolism of a tumor cell. The metabolite may be a product generated by metabolism essential for vital activities (primary metabolite) or a product generated by metabolism not necessarily necessary for vital activities (secondary metabolite). Examples of primary metabolites may include sugars, proteins, lipids, nucleic acids, and the like. Examples of secondary metabolites include antibiotics and dyes. The metabolite may be a biopolymer or a small molecule. In a specific embodiment, the biopolymer is a molecule with a molecular weight of about 5000 or more that consists of one or more types of repeating units, including, for example, polysaccharides, polypeptides, and polynucleotides.In one specific embodiment, small molecules are molecules with a molecular weight of about 500 or less and are chemical substances present in the living body. In another embodiment, the tumor tissue-specific compound is a small molecule metabolite produced specifically in tumor cells (Eva Gottfried, Katrin Peter and Marina P. Kreutz, From Molecular and Modular Tumor Therapy (2010) 3 (2), 111-132). In yet another embodiment, the tumor tissue-specific compound is a metabolite that is produced specifically by a cell that... Petition 870260042208, dated 05 / 05 / 2026, p. 122 / 1364 106 / 448 infiltrates tumor tissue (e.g., an immune cell) or a stromal cell (e.g., a cancer-associated fibroblast (CAF)) present in tumor tissue. Examples of immune cells that infiltrate tumor tissue are dendritic cells, suppressor dendritic cells, regulatory T cells, depleted T cells, myeloma-derived suppressor cells (MDSCs), and the like. In another embodiment, a metabolite that is produced by cells present in tumor tissue (e.g., tumor cells, immune cells, stromal cells, etc.), which is released to the exterior of the cell when the cells die by apoptosis or necrosis, or the like, may also be included in the tumor tissue-specific compounds of the present disclosure.

[00434] To identify a specific tumor tissue compound, transcriptome-level analysis (e.g., Dhanasekaran et al. (Nature (2001) 41 2, 822-826), Lapointe et al. (Proc. Natl. Acad. Sci. USA (2004)) 101, 811-816), or Perou et al. (Nature (2000) 406, 747-752)), proteome-level analysis (e.g., Ahram et al. (Mol. Carcinog. (2002) 33, 9-15), and Hood et al. (Mol. Cell. Proteomics (2005) 4, 1741-1753)), and metabolomics-focused metabology analysis (metabolomics) can be appropriately used. That is, in order to identify a metabolite in a test sample, high-performance liquid chromatography (HPLC), nuclear magnetic resonance (NMR) (Brindle et al. (J. Mol. Recognit. (1997) 10, 182-187)), mass spectrometry (GC / MS and LC / MS) (Gates and Sweeley (Clin. Chem. (1978) 24, 1663-1673)) and metabolic profiling using ELISA, and such, can be appropriately used alone and / or in combination.

[00435] In a specific embodiment, the specific tumor tissue compound is at least one compound selected from the group consisting of: nucleosides possessing a ring structure of Petition 870260042208, dated 05 / 05 / 2026, page 123 / 1364 107 / 448 purine, amino acids and their metabolites, lipids and their metabolites, primary metabolites of carbohydrate metabolism, also as nicotinamide and its metabolites. In another embodiment, the tumor tissue-specific compound is at least one compound selected from (1) to (6) below: (1) nucleosides possessing a purine structure, such as adenosine (ADO), adenosine triphosphate (ATP), adenosine diphosphate (ADP), adenosine monophosphate (AMP) and inosine; (2) amino acids, such as alanine, glutamic acid and aspartic acid; (3) amino acid metabolites, such as kynurenine, anthranilic acid, 3-hydroxykynurenine and kynurenic acid; (4) arachidonic acid metabolites, such as prostaglandin E2; (5) primary metabolites of the glycolytic pathway or Krebs cycle, such as lactic acid, succinic acid and citric acid; and, (6) nicotinamide metabolites, such as 1-methyl nicotinamide. (1) Nucleosides possessing purine structure, such as adenosine (ADO), adenosine triphosphate (ATP), adenosine diphosphate (ADP), adenosine monophosphate (AMP) and inosine

[00436] It is known that when tumor cells die, a large amount of intracellular ATP leaks out. Therefore, the concentration of ATP in tumor tissue is significantly higher than in normal tissue (PLoS One. (2008) 3, e2599). AMP is metabolized by enzymes on the cell surface, such as extracellular 5'-nucleotidase (eco5'-nucleotidase) (CD73) (Resta and Thompson (Immunol. Rev. (1998) 161, 95-109) and Sadej et al. (Melanoma Res. (2006) 16, 213-222)). Adenosine is a purine nucleoside that is constitutively present in the extracellular environment at low concentrations, but a Petition 870260042208, dated 05 / 05 / 2026, pp. 124 / 1364 108 / 448 A marked increase in extracellular adenosine concentration has been reported in hypoxic tissues found in solid tumors (Blay and Hoskin (Cancer Res. (1997) 57, 260 2-2605)). CD73 is expressed on the surface of tumors and immune cells (Kobie et al. (J. Immunol. (2006) 17 7, 6780-6786)) and elevated activity has been found in breast cancer (Canbolat et al. (Breast Cancer Res. Treat. (1996) 37, 189-193)), stomach cancer (Durak et al. (Cancer Lett. (1994) 84, 199-202)), pancreatic cancer (Flocke and Mannherz (Biochim. Biophys. Acta (1991) 1076, 273-281)) and glioblastoma (Bardot et al. (Br. J. Cancer (1994) 70, 212-218)). It has been proposed that the accumulation of adenosine in tumor tissue is the result of increased AMP dephosphorylation by cytoplasmic 5'nucleotidase (Headrick and Willis (Biochem. J. (1989) 261, 541-550)). Furthermore, regulatory T cells that infiltrate tumor tissue also express ATPase and produce adenosine (Proc. Natl. Acad. Sci.).USA (2006) 103 (35), 13132-13137; Curr. Med. Chem. (2011) 18: 5217-5223). It is believed that the adenosine produced maintains tumor tissues in an immunosuppressive environment through adenosine receptors, such as the A2A receptor (Curr. Med. Chem. (2011) 18, 5217-5223). Thus, ATP, ADP, AMP, adenosine, and others, which are considered to accumulate in high concentrations in tumor tissue through purine nucleotide metabolism, are examples of tumor tissue-specific compounds used in this disclosure. Furthermore, since adenosine is degraded to inosine by adenosine deaminase, inosine accumulates in high concentrations.

[00437] In a specific embodiment, nucleosides possessing a purine ring structure include compounds containing adenosine. In specific embodiments, compounds containing adenosine include, for example, adenosine (ADO), adenosine triphosphate (ATP), adenosine diphosphate (ADP), adenosine monophosphate Petition 870260042208, dated 05 / 05 / 2026, page 125 / 1364 109 / 448 adenosine (AMP), cyclic adenosine monophosphate (cAMP), deoxyadenosine (dADO), deoxyadenosine triphosphate (dATP), deoxyadenosine diphosphate (dADP), deoxyadenosine monophosphate (dAMP), [Y-thio]adenosine triphosphate (ATPyS), and so on. In another embodiment, nucleosides possessing a purine ring structure include inosine, which is a metabolite of adenosine.

[00438] In addition, in a specific embodiment, nucleosides possessing a purine ring structure include commercially available nucleosides possessing a purine ring structure, such as ADPbetaS (Sigma Inc.) and others like them. (2) Amino acids, such as alanine, glutamic acid, and aspartic acid

[00439] The rate of glutamine intake, which acts as a nitrogen carrier in the living body, is increased in tumor cells, and such incorporation of glutamine and the resulting conversion into glutamic acid and lactic acid (glutamine degradation (glutaminolysis)) are considered characteristic of tumor cells (Mazurek and Eigenbrodt (Anticancer Res. (2003) 23, 1149-1154, and Mazurek et al. (J. cell. Physiol. (1999) 181, 136-146). Plasma glutamine levels are decreased in cancer patients, while glutamic acid concentration is increased (Droge et al. (Immunobiology (1987) 174, 473-479)), and studies of glucose metabolism labeled Studies with 13C in lung cancer tissue showed a correlation between the concentrations of 13C-labeled succinic acid, 13C-labeled alanine, 13C-labeled glutamic acid, and 13C-labeled citrate.For these reasons, alanine, glutamic acid, aspartic acid, and others similar, which are thought to accumulate in high concentrations in tumor tissue due to, for example, glutamine degradation, are examples of tumor tissue-specific compounds used in this disclosure. (3) Amino acid metabolites, such as kynurenine, acid Petition 870260042208, dated 05 / 05 / 2026, pp. 126 / 1364 110 / 448 anthranilic acid, 3-hydroxykynurenine and kynurenic acid

[00440] Indoleamine 2,3-dioxygenase (IDO) is an enzyme that metabolizes tryptophan and is highly expressed in many types of cancer, such as melanoma, colon cancer, kidney cancer, and others (Uyttenhove et al. (Nat. Med. (2003) 9, 1269-1274)). IDO catalyzes the conversion of tryptophan to kynurenine. In gliomas that do not express IDO, kynurenine is produced from tryptophan by liver tryptophan 2,3-dioxygenase (TDO) (Opitz et al. (Nature (2011) 478 (7368), 197-203)). IDO is also expressed in dendritic cells that infiltrate tumor tissue, and dendritic cells also produce kynurenine (J. Immunol. (2008) 181, 5396-5404). Furthermore, IDO is also expressed in myeloid-derived suppressor cells (MDSCs) of tumor tissues, and MDSCs also produce kynurenine (Yu et al. (J. Immunol. (2013) 190, 3783-3797)). Kynurenine is converted to anthranilic acid by kynureninase and to 3-hydroxykynurenine by kynurenine 3-hydroxylase.Both anthranilic acid and 3-hydroxykynurenine are converted to 3-hydroxyanthranilic acid, a precursor of NAD. Kynurenine is converted to kynurenic acid by kynurenine aminotransferase. For these reasons, kynurenine and its metabolites, i.e., anthranilic acid, 3-hydroxykynurenine, kynurenic acid, and the like, are examples of tumor tissue-specific compounds used in this disclosure, in particular, tumor cell-specific metabolites. (4) Arachidonic acid metabolites, such as prostaglandin E2

[00441] Prostaglandin E2 (PGE2) promotes the growth of colon cancer cells and suppresses their apoptosis (Sheng et al. (Cancer Res. (1998) 58, 362-366)). Of the PGE2 synthases, it has been found that COX-1 is constitutively expressed in almost all tissues, while COX-2 is induced by certain inflammatory cytokines and oncogenes in tumors (Warner and Mitchell (FASEB J. (2004) 18, 790-804)). Overexpression of COX-2 has been reported to be Petition 870260042208, dated 05 / 05 / 2026, p. 127 / 1364 111 / 448 is associated with a poor prognosis in breast cancer (Denkert et al. (Clin. Breast Cancer (2004) 4, 428-433)) and rapid disease progression in ovarian cancer (Denker et al. (Mod. Pathol. (2006) 19, 1261-1269)). Furthermore, regulatory T cells that infiltrate tumor tissue also produce PGE2 (Curr. Med. Chem. (2011) 18, 5217-5223). Due to these reasons, arachidonic acid metabolites, such as PGE2, are examples of tumor tissue-specific compounds, in particular tumor cell-specific metabolites or immune cell-specific metabolites that infiltrate tumor tissues. In addition to PGE2, thromboxane A2 (TXA2) production is increased in tumor tissues, such as those in colon cancer (J. Lab. Clin. Med. (1993) 122, 518-523). (5) Primary metabolites of the glycolytic pathway or Krebs cycle, such as lactic acid, succinic acid and citric acid.

[00442] The glycolytic phenotype characterized by the upregulation of glycolytic enzymes (via Embden-Meyerhof), such as pyruvate kinase, hexokinase, and lactate dehydrogenase (LDH), has been conventionally known as the Warburg effect, a characteristic of solid tumors. Lactic acid, which is the end product of glycolysis, and succinic acid and citric acid produced by the Krebs cycle are known to accumulate in tumor tissues (Teresa et al. (Mol. Cancer (2009) 8, 41-59)). Due to these reasons, lactic acid, succinic acid, citric acid, and others, which are primary metabolites produced by glycolysis, are examples of tumor tissue-specific compounds, in particular tumor cell-specific metabolites, used in this disclosure. Furthermore, it is known that due to cell death, succinate, which is present in high concentration in cells, leaks out of the cells (Nature Immunology, (2008) 9, 1261-1269).This is believed to be the reason for the increased concentration of succinic acid in tumor tissues. Petition 870260042208, dated 05 / 05 / 2026, pp. 128 / 1364 112 / 448 where cell death occurs frequently. (6) Nicotinamide metabolites, such as 1-methyl nicotinamide

[00443] Nicotinamide N-methyltransferase is known to be highly expressed in a plurality of human tumor tissues. It is also known that 1-methyl-nicotinamide, which is a stable metabolite of nicotinamide produced by this enzyme, is secreted to the exterior of tumor cells (Yamada et al. (J. Nutr. Sci. Vitaminol. (2010) 56, 83-86)). Due to this reason, 1-methyl-nicotinamide and similar compounds, which are thought to accumulate in tumor tissue at a high concentration as a result of nicotinamide metabolism, are examples of tumor tissue-specific compounds used in this disclosure.

[00444] An antigen-binding molecule of the present disclosure comprises an antigen-binding domain. As the antigen-binding domain, a domain of any structure may be used, provided it binds to the target antigen.In one embodiment, the antigen-binding domains of this disclosure include, for example, variable regions of heavy chains and / or light chains of antibodies, Avimers containing a module (domain A) of approximately 35 amino acids contained in various cell membrane proteins in the living body (International Publications WO2004 / 044011 and WO2005 / 040229), Adnectins containing the 10Fn3 domain of fibronectin, which is a glycoprotein expressed in the cell membrane (International Publication WO2002 / 032925), Afibodies using as a base a 58-amino acid IgG-binding domain of Protein A (WO1995 / 001937), DARPins (ankyrin engineered repeat proteins) using an ankyrin repeat (AR), which is a 33-amino acid repeat sequence, as a base (International Publication WO2002 / 020565), Anti-kalins containing a lipocalin, such as neutrophil gelatinase-associated lipocalin (NGAL) as a base. Petition 870260042208, dated 05 / 05 / 2026, pp. 129 / 1364 113 / 448 (International Publication WO2003 / 029462), variable lymphocyte receptors (VLRs) which are proteins that function in adaptive immune systems of jawless vertebrates, such as Lampetra japonica and Eptatretus, and contain a leucine-rich repeat module (LRR) (International Publication WO2008 / 016854), and others similar. In one specific embodiment, the antigen-binding domain of this disclosure comprises variable heavy and light chain regions of an antibody. In another embodiment, the antigen-binding domain of this disclosure includes, for example, scFv (single-chain Fv), single-chain antibodies, Fv, scFv2 (single-chain Fv2), Fab or F(ab')2. HVR and variable region

[00445] In one aspect, the present disclosure provides an anti-CD137 antigen-binding molecule or antibody comprising at least one, at least two, or all three VH HVR sequences selected from: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7; (b) HVRH2 comprising any of the amino acid sequences selected from SEQ ID NOs: 8, 9, 10, 11, 12, 13, 14, 15, and 16; and (c) HVR-H3 comprising any of the amino acid sequences selected from SEQ ID NO: 17, 18, 19, or 20. In one embodiment, the anti-CD137 antigen-binding molecule or antibody comprises: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7; (b) HVR-H2 comprising any of the amino acid sequences selected from SEQ ID NOs: 8, 9, 10, 11, 12, 13, 14, 15 and 16; and (c) HVR-H3 comprising any of the amino acid sequences selected from SEQ ID NO: 17, 18, 19 or 20.

[00446] In one embodiment, the antiCD137 antigen-binding molecule is an antibody comprising the amino acid sequence of Petition 870260042208, dated 05 / 05 / 2026, pages 130 / 1364 114 / 448 A375 / B167, A372 / B040, A356 / B040, A486 / B167, A487 / B167, A488 / B226, A489 / B223, A548 / B376, A551 / B256, A551 / B379, A555 / B379, A548 / B256, or A549 / B167 described in Table 17 as a combination of variable heavy chain region / variable light chain region. In a preferred embodiment, the antigen-binding molecule is an anti-CD137 antibody comprising HVR-H1, HVR-H2, HVR-H3, HVR-L1, HVR-L2, and HVR-L3 having the same amino acid sequences as HVR-H1, HVR-H2, HVR-H3, HVR-L1, HVR-L2, and HVR-L3 comprised in A375 / B167. In another embodiment, the anti-CD137 antigen-binding molecule is an anti-CD137 antibody comprising A375 / B167 as a combination of variable heavy chain region / variable light chain region. In a different preferred embodiment, the antigen-binding molecule is an anti-CD137 antibody comprising HVRH1, HVR-H2, HVR-H3, HVR-L1, HVR-L2 and HVR-L3 having the same amino acid sequences as HVR-H1, HVR-H2, HVR-H3, HVR-L1, HVR-L2 and HVR-L3 included in A551 / B379.

[00447] In one embodiment, the antiCD137 antigen-binding molecule or antibody comprises (a) HVR-H1 comprising the amino acid sequence SEQ ID NO: 7; (b) HVR-H2 comprising the amino acid sequence SEQ ID NO: 8; and (c) HVRH3 comprising the amino acid sequence SEQ ID NO: 17.

[00448] In one embodiment, the antiCD137 antigen-binding molecule or antibody comprises (a) HVR-H1 comprising the amino acid sequence SEQ ID NO: 7; (b) HVR-H2 comprising the amino acid sequence SEQ ID NO: 9; and (c) HVRH3 comprising the amino acid sequence SEQ ID NO: 17.

[00449] In one embodiment, the antiCD137 antigen-binding molecule or antibody comprises (a) HVR-H1 comprising the amino acid sequence SEQ ID NO: 7; (b) HVR-H2 that Petition 870260042208, dated 05 / 05 / 2026, pp. 131 / 1364 115 / 448 comprises the amino acid sequence of SEQ ID NO: 10; and (c) HVRH3 comprising the amino acid sequence of SEQ ID NO: 17.

[00450] In one embodiment, the antiCD137 antigen-binding molecule or antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 11; and (c) HVRH3 comprising the amino acid sequence of SEQ ID NO: 18.

[00451] In one embodiment, the antiCD137 antigen-binding molecule or antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 8; and (c) HVRH3 comprising the amino acid sequence of SEQ ID NO: 18.

[00452] In one embodiment, the antiCD137 antigen-binding molecule or antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 12; and (c) HVRH3 comprising the amino acid sequence of SEQ ID NO: 18.

[00453] In one embodiment, the antiCD137 antigen-binding molecule or antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 13; and (c) HVRH3 comprising the amino acid sequence of SEQ ID NO: 18.

[00454] In one embodiment, the antiCD137 antigen-binding molecule or antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 14; and (c) HVRH3 comprising the amino acid sequence of SEQ ID NO: 19.

[00455] In one embodiment, the antiCD137 antigen-binding molecule or antibody comprises (a) HVR-H1 comprising the amino acid sequence SEQ ID NO: 7; (b) HVR-H2 which. Petition 870260042208, dated 05 / 05 / 2026, pp. 132 / 1364 116 / 448 comprises the amino acid sequence with SEQ ID NO: 15; and (c) HVRH3 comprising the amino acid sequence with SEQ ID NO: 20.

[00456] In one embodiment, the antiCD137 antigen-binding molecule or antibody comprises (a) HVR-H1 comprising the amino acid sequence SEQ ID NO: 7; (b) HVR-H2 comprising the amino acid sequence SEQ ID NO: 16; and (c) HVRH3 comprising the amino acid sequence SEQ ID NO: 20.

[00457] In one embodiment, the antiCD137 antigen-binding molecule or antibody comprises (a) HVR-H1 comprising the amino acid sequence SEQ ID NO: 7; (b) HVR-H2 comprising the amino acid sequence SEQ ID NO: 14; and (c) HVRH3 comprising the amino acid sequence SEQ ID NO: 17.

[00458] In a different aspect, the present disclosure provides an anti-CD137 antigen-binding molecule or antibody comprising at least one, at least two, or all three HVR VL sequences selected from: (a) HVR-L1 comprising any of the amino acid sequences selected from SEQ ID NOs: 21, 22, 23, 24, and 25; (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 26; and (c) HVR-L3 comprising any of the amino acid sequences selected from SEQ ID NOs: 27, 28, and 29. In one embodiment, the anti-CD137 antigen-binding molecule or antibody comprises (a) HVR-L1 comprising any of the amino acid sequences selected from SEQ ID NOs: 21, 22, 23, 24, and 25; (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 26; and (c) HVR-L3 comprising any of the selected amino acid sequences of SEQ ID NOs: 27, 28 and 29.

[00459] In one embodiment, the antiCD137 antigen-binding molecule or antibody comprises (a) HVR-L1 comprising Petition 870260042208, dated 05 / 05 / 2026, pp. 133 / 1364 117 / 448 amino acid sequence of SEQ ID NO: 21; (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 26; and (c) HVRL3 comprising the amino acid sequence of SEQ ID NO: 27.

[00460] In one embodiment, the antiCD137 antigen-binding molecule or antibody comprises (a) HVR-L1 comprising the amino acid sequence SEQ ID NO: 22; (b) HVR-L2 comprising the amino acid sequence SEQ ID NO: 26; and (c) HVRL3 comprising the amino acid sequence SEQ ID NO: 27.

[00461] In one embodiment, the antiCD137 antigen-binding molecule or antibody comprises (a) HVR-L1 comprising the amino acid sequence SEQ ID NO: 21; (b) HVR-L2 comprising the amino acid sequence SEQ ID NO: 26; and (c) HVRL3 comprising the amino acid sequence SEQ ID NO: 28.

[00462] In one embodiment, the antiCD137 antigen-binding molecule or antibody comprises (a) HVR-L1 comprising the amino acid sequence SEQ ID NO: 21; (b) HVR-L2 comprising the amino acid sequence SEQ ID NO: 26; and (c) HVRL3 comprising the amino acid sequence SEQ ID NO: 29.

[00463] In one embodiment, the antiCD137 antigen-binding molecule or antibody comprises (a) HVR-L1 comprising the amino acid sequence SEQ ID NO: 23; (b) HVR-L2 comprising the amino acid sequence SEQ ID NO: 26; and (c) HVRL3 comprising the amino acid sequence SEQ ID NO: 27.

[00464] In one embodiment, the antiCD137 antigen-binding molecule or antibody comprises (a) HVR-L1 comprising the amino acid sequence SEQ ID NO: 24; (b) HVR-L2 comprising the amino acid sequence SEQ ID NO: 26; and (c) HVRL3 comprising the amino acid sequence SEQ ID NO: 27.

[00465] In one embodiment, the antiCD137 antigen-binding molecule or antibody comprises (a) HVR-L1 comprising Petition 870260042208, dated 05 / 05 / 2026, pp. 134 / 1364 118 / 448 amino acid sequence of SEQ ID NO: 25; (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 26; and (c) HVRL3 comprising the amino acid sequence of SEQ ID NO: 27.

[00466] In another aspect, the antiCD137 antigen-binding molecule or antibody of this disclosure comprises (a) a VH domain comprising at least one, at least two, or all three HVR VH sequences selected from (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7; (ii) HVRH2 comprising any of the amino acid sequences selected from SEQ ID NOs: 8, 9, 10, 11, 12, 13, 14, 15, and 16; and (iii) HVR-H3 comprising any of the amino acid sequences selected from SEQ ID NOs: 17, 18, 19, or 20; and (b) a VL domain comprising at least one, at least two, or all three selected HVR VL sequences from (i) HVRL1 comprising any of the selected amino acid sequences from SEQ ID NOs: 21, 22, 23, 24, and 25; (ii) HVR-L2 comprising the amino acid sequence from SEQ ID NO: 26;and (iii) HVR-L3 comprising any of the selected amino acid sequences from SEQ ID NOS: 27, 28 and 29.;

[00467] In another aspect, the present disclosure provides an anti-CD137 antigen-binding molecule or antibody comprising (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 8; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 17; (d) HVRL1 comprising the amino acid sequence of SEQ ID NO: 21; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 26; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 27.

[00468] In another aspect, the present disclosure provides a Petition 870260042208, dated 05 / 05 / 2026, pp. 135 / 1364 119 / 448 Anti-CD137 antigen-binding molecule or antibody comprising (a) HVR-H1 comprising amino acid sequence SEQ ID NO: 7; (b) HVR-H2 comprising amino acid sequence SEQ ID NO: 9; (c) HVR-H3 comprising amino acid sequence SEQ ID NO: 17; (d) HVRL1 comprising amino acid sequence SEQ ID NO: 22; (e) HVR-L2 comprising amino acid sequence SEQ ID NO: 26; and (f) HVR-L3 comprising amino acid sequence SEQ ID NO: 27.

[00469] In another aspect, the present disclosure provides an anti-CD137 antigen-binding molecule or antibody comprising (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 10; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 17; (d) HVRL1 comprising the amino acid sequence of SEQ ID NO: 22; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 26; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 27.

[00470] In another aspect, the present disclosure provides an anti-CD137 antigen-binding molecule or antibody comprising (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 11; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 18; (d) HVRL1 comprising the amino acid sequence of SEQ ID NO: 21; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 26; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 27.

[00471] In another aspect, the present disclosure provides a Petition 870260042208, dated 05 / 05 / 2026, pp. 136 / 1364 120 / 448 Anti-CD137 antigen-binding molecule or antibody comprising (a) HVR-H1 comprising amino acid sequence SEQ ID NO: 7; (b) HVR-H2 comprising amino acid sequence SEQ ID NO: 8; (c) HVR-H3 comprising amino acid sequence SEQ ID NO: 18; (d) HVRL1 comprising amino acid sequence SEQ ID NO: 21; (e) HVR-L2 comprising amino acid sequence SEQ ID NO: 26; and (f) HVR-L3 comprising amino acid sequence SEQ ID NO: 27.

[00472] In another aspect, the present disclosure provides an anti-CD137 antigen-binding molecule or antibody comprising (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 12; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 18; (d) HVRL1 comprising the amino acid sequence of SEQ ID NO: 21; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 26; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 28.

[00473] In another aspect, the present disclosure provides an anti-CD137 antigen-binding molecule or antibody comprising (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 13; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 18; (d) HVRL1 comprising the amino acid sequence of SEQ ID NO: 21; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 26; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 29.

[00474] In another aspect, the present disclosure provides a Petition 870260042208, dated 05 / 05 / 2026, pp. 137 / 1364 121 / 448 Anti-CD137 antigen-binding molecule or antibody comprising (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 14; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 19; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 23; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 26; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 27.

[00475] In another aspect, the present disclosure provides an anti-CD137 antigen-binding molecule or antibody comprising (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 15; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 20; (d) HVRL1 comprising the amino acid sequence of SEQ ID NO: 24; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 26; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 27.

[00476] In another aspect, the present disclosure provides an anti-CD137 antigen-binding molecule or antibody comprising (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 15; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 20; (d) HVRL1 comprising the amino acid sequence of SEQ ID NO: 25; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 26; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 27.

[00477] In another aspect, the present disclosure provides a Petition 870260042208, dated 05 / 05 / 2026, pp. 138 / 1364 122 / 448 Anti-CD137 antigen-binding molecule or antibody comprising (a) HVR-H1 comprising amino acid sequence SEQ ID NO: 7; (b) HVR-H2 comprising amino acid sequence SEQ ID NO: 16; (c) HVR-H3 comprising amino acid sequence SEQ ID NO: 20; (d) HVRL1 comprising amino acid sequence SEQ ID NO: 25; (e) HVR-L2 comprising amino acid sequence SEQ ID NO: 26; and (f) HVR-L3 comprising amino acid sequence SEQ ID NO: 27.

[00478] In another aspect, the present disclosure provides an anti-CD137 antigen-binding molecule or antibody comprising (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 14; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 19; (d) HVRL1 comprising the amino acid sequence of SEQ ID NO: 24; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 26; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 27.

[00479] In another aspect, the present disclosure provides an anti-CD137 antigen-binding molecule or antibody comprising (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 14; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 17; (d) HVRL1 comprising the amino acid sequence of SEQ ID NO: 21; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 26; and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 27.

[00480] In a specific modality, any one or more Petition 870260042208, dated 05 / 05 / 2026, pp. 139 / 1364 123 / 448 amino acids of the anti-CD137 antibody described above are substituted at the following HVR positions:

[00481] In HVR-H2 (SEQ ID NO: 30): positions 5, 6, 7, 10, 13, 14 and / or 17

[00482] In HVR-H3 (SEQ ID NO: 31): position 3 and / or 6

[00483] In HVR-L1 (SEQ ID NO: 32): positions 4, 5, 9 and / or 11

[00484] In HVR-L3 (SEQ ID NO: 33): positions 6, 7 and / or 8.

[00485] In a specific embodiment, the substitutions provided by this specification are conservative substitutions. In a specific embodiment, any one or more of the following substitutions may be made in any combination:

[00486] In HVR-H2 (SEQ ID NO: 8): K5H or S; S6G; T7S; E10Y; D13E; S14Q; V17G or L

[00487] In HVR-H3 (SEQ ID NO: 17): A3P, K or I; F6E

[00488] In HVR-L1 (SEQ ID NO: 21): R4S; Y5T; Y9F; E11N

[00489] In HVR-L3 (SEQ ID NO: 27): E6P; H7A; Q8I

[00490] All possible combinations of the substitutions mentioned above are encompassed in the consensus sequences of SEQ ID NOs: 30, 31, 32 and 33 for HVR-H2, HVR-H3, HVR-L1 and HVR-L3, respectively.

[00491] In any of the above embodiments, an anti-CD137 antigen-binding molecule or antibody is humanized. In one embodiment, an anti-CD137 antigen-binding molecule or antibody comprises HVRs as in any of the previous embodiments and additionally comprises a human acceptor structure, for example, a human immunoglobulin structure or a human consensus structure. In another embodiment, an anti-CD137 antigen-binding molecule or antibody comprises HVRs as in any of the above embodiments and additionally comprises a variable heavy chain (VH) region. Petition 870260042208, dated 05 / 05 / 2026, page 140 / 1364 124 / 448 or a variable light chain (VL) region comprising a structure sequence (FR). In one embodiment, FR1 in the variable heavy chain region comprises the amino acid sequence of SEQ ID NO: 35, FR2 in the variable heavy chain region comprises the amino acid sequence of SEQ ID NO: 36, FR3 in the variable heavy chain region comprises the amino acid sequence of SEQ ID NO: 37, and FR4 in the variable heavy chain region comprises the amino acid sequence of SEQ ID NO: 38. In another embodiment, FR1 in the variable light chain region comprises the amino acid sequence of SEQ ID NO: 39, FR2 in the variable light chain region comprises the amino acid sequence of SEQ ID NO: 40, FR3 in the variable light chain region comprises the amino acid sequence of SEQ ID NO: 41, and FR4 in the variable light chain region comprises the amino acid sequence of SEQ ID NO: 42.

[00492] In another aspect, an antiCD137 antigen-binding molecule or antibody comprises a variable heavy chain (VH) domain sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with the amino acid sequence of SEQ ID NO: 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, or 53. In certain embodiments, the VH sequence with at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but a Anti-CD137 antigen-binding molecule or antibody comprising this sequence retains the ability to bind to CD137. In certain embodiments, a total of 1 to 10 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO: 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, or 53. In certain embodiments, substitutions, insertions, or deletions occur in regions outside the HVRs (i.e., in the FRs).Optionally, the anti-CD137 antibody comprises the sequence. Petition 870260042208, dated 05 / 05 / 2026, pp. 141 / 1364 125 / 448 VH in SEQ ID NO: 43, 44, 45, 46, 47, 48, 49, 50, 51, 52 or 53, including post-translational modifications of that sequence. In a particular embodiment, the VH comprises one, two or three HVRs selected from: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 7; (b) HVR-H2 comprising any amino acid sequence selected from SEQ ID NOs: 8, 9, 10, 11, 12, 13, 14, 15 and 16; and (c) HVR-H3 comprising any amino acid sequence selected from SEQ ID NO: 17, 18, 19 or 20. Post-translational modifications include, but are not limited to, a modification of glutamine or glutamate at the N-terminal heavy chain or light chain to pyroglutamic acid by pyroglutamylation.

[00493] In another aspect, an anti-CD137 antigen-binding molecule or antibody is provided, wherein the antibody comprises a variable light chain (VL) domain having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity with the amino acid sequence of SEQ ID NO: 54, 55, 56, 57, 58, 59 or 60. In certain embodiments, a VL sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity contains substitutions (e.g., conservative substitutions), insertions or deletions relative to the reference sequence, but an anti-CD137 antigen-binding molecule or antibody comprising this sequence retains the ability to bind to CD137. In certain embodiments, a total of 1 to 10 amino acids have been substituted, inserted and / or deleted in SEQ ID NO: 54, 55, 56, 57, 58, 59 or 60. In certain embodiments, the substitutions, insertions, or deletions occur in regions outside the HVRs (i.e., in the FRs).Optionally, the anti-CD137 antigen-binding molecule or antibody comprises the VL sequence in SEQ ID NO: 54, 55, 56, 57, 58, 59, or 60, including post-translational modifications of that sequence. In one particular embodiment, the VL comprises one, two, or three HVRs. Petition 870260042208, dated 05 / 05 / 2026, p. 142 / 1364 126 / 448 selected from (a) HVR-L1 comprising any amino acid sequence selected from SEQ ID NOs: 21, 22, 23, 24 and 25; (b) HVR-L2 comprising the amino acid sequence from SEQ ID NO: 26; and (c) HVR-L3 comprising any amino acid sequence selected from SEQ ID NO: 27, 28 and 29. Post-translational modifications include, but are not limited to, a modification of glutamine or glutamate at the N-terminal heavy chain or light chain to pyroglutamic acid by pyroglutamylation.

[00494] In another aspect, an antiCD137 antigen-binding molecule or antibody is provided, wherein the antigen-binding molecule or antibody comprises a VH as in any of the embodiments provided above, and a VL as in any of the embodiments provided above.

[00495] - In one embodiment, the antiCD137 antigen-binding molecule or antibody comprises the VH and VL sequences at SEQ ID NO: 43 and SEQ ID NO: 54, respectively, including post-translational modifications of these sequences.

[00496] - In one embodiment, the antiCD137 antigen-binding molecule or antibody comprises the VH and VL sequences at SEQ ID NO: 44 and SEQ ID NO: 55, respectively, including post-translational modifications of these sequences.

[00497] - In one embodiment, the antiCD137 antigen-binding molecule or antibody comprises the VH and VL sequences at SEQ ID NO: 45 and SEQ ID NO: 55, respectively, including post-translational modifications of these sequences.

[00498] - In one embodiment, the antiCD137 antigen-binding molecule or antibody comprises the VH and VL sequences at SEQ ID NO: 46 and SEQ ID NO: 54, respectively, including post-translational modifications of these sequences.

[00499] - In one embodiment, the antigen-binding molecule anti Petition 870260042208, dated 05 / 05 / 2026, pp. 143 / 1364 127 / 448 CD137 or antibody comprises the VH and VL sequences at SEQ ID NO: 47 and SEQ ID NO: 54, respectively, including post-translational modifications of these sequences.

[00500] - In one embodiment, the antiCD137 antigen-binding molecule or antibody comprises the VH and VL sequences in SEQ ID NO: 48 and SEQ ID NO: 56, respectively, including post-translational modifications of these sequences.

[00501] - In one embodiment, the antiCD137 antigen-binding molecule or antibody comprises the VH and VL sequences in SEQ ID NO: 49 and SEQ ID NO: 57, respectively, including post-translational modifications of these sequences.

[00502] - In one embodiment, the antiCD137 antigen-binding molecule or antibody comprises the VH and VL sequences in SEQ ID NO: 50 and SEQ ID NO: 58, respectively, including post-translational modifications of these sequences.

[00503] - In one embodiment, the antiCD137 antigen-binding molecule or antibody comprises the VH and VL sequences in SEQ ID NO: 51 and SEQ ID NO: 59, respectively, including post-translational modifications of these sequences.

[00504] - In one embodiment, the antiCD137 antigen-binding molecule or antibody comprises the VH and VL sequences in SEQ ID NO: 51 and SEQ ID NO: 60, respectively, including post-translational modifications of these sequences.

[00505] - In one embodiment, the antiCD137 antigen-binding molecule or antibody comprises the VH and VL sequences in SEQ ID NO: 52 and SEQ ID NO: 60, respectively, including post-translational modifications of these sequences.

[00506] - In one embodiment, the antiCD137 antigen-binding molecule or antibody comprises the VH and VL sequences in SEQ ID NO: 50 and SEQ ID NO: 59, respectively, including post-Petition 870260042208 modifications, dated 05 / 05 / 2026, page 144 / 1364 128 / 448 translation of these sequences.

[00507] - In one embodiment, the antiCD137 antigen-binding molecule or antibody comprises the VH and VL sequences in SEQ ID NO: 53 and SEQ ID NO: 54, respectively, including post-translational modifications of these sequences.

[00508] The aforementioned post-translational modifications include, but are not limited to, a modification of glutamine or glutamate at the N-terminal heavy chain or light chain into pyroglutamic acid by pyroglutamylation.

[00509] The SEQ ID numbers corresponding to the amino acid sequences of the preferred heavy chain variable region and light chain variable region and their HVR1, 2 and 3 for each anti-CD137 antigen-binding molecule or antibody of the present disclosure are shown in the table below. Table 1 Variable Regions of Heavy / Light Chain SEQ ID No. of Variable Region SEQ ID No. of Hypervariable Region (HVR) Heavy Chain Light Chain H1 H2 H3 L1 L2 L3 A375 / B167 43 54 7 8 17 21 26 27 A372 / B040 44 55 7 9 17 22 26 27 A356 / B040 45 55 7 10 17 22 26 27 A486 / B167 46 54 7 11 18 21 26 27 A487 / B167 47 54 7 8 18 21 26 27 A488 / B226 48 56 7 12 18 21 26 28 A489 / B223 49 57 7 13 18 21 26 29 A548 / B376 50 58 7 14 19 23 26 27 A551 / B256 51 59 7 15 20 24 26 27 A551 / B379 51 60 7 15 20 25 26 27 A555 / B379 52 60 7 16 20 25 26 27 A548 / B256 50 59 7 14 19 24 26 27 A549 / B167 53 54 7 14 17 21 26 27

[00510] When an anti-CD137 antigen-binding molecule or antibody provided here has glutamine as its N-terminal amino acid Petition 870260042208, dated 05 / 05 / 2026, pp. 145 / 1364 129 / 448 of the heavy or light chain, this amino acid can be replaced by glutamic acid. When an anti-CD137 antibody provided herein has glutamic acid as the N-terminal amino acid of the heavy or light chain, this amino acid can be replaced by glutamine.

[00511] In a preferred embodiment, the anti-CD137 antigen-binding molecules or antibodies comprising the above-described HVRs, variable heavy chain regions and / or variable light chain regions, all possess low molecular weight compound-dependent binding activity for CD137 as described above. Constant Regions

[00512] In another aspect, anti-CD137 antigen-binding molecules or antibodies comprise a constant region. The constant region may be a heavy chain constant region (including an Fc region), a light chain constant region, or both. In another aspect, anti-CD137 antigen-binding molecules or antibodies comprise an Fc region. In some embodiments, the constant region is that with the native sequence. Examples of heavy chain constant regions derived from native antibodies include, for example, a human IgG1 heavy chain constant region (SEQ ID NOs: 61, 62), human IgG2, human IgG3, human IgG4, and the like. Examples of light chain constant regions derived from native antibodies include, for example, human kappa chain, human lambda chain (e.g., SEQ ID NO: 63), and the like.

[00513] A parental constant region or parental Fc region used herein refers to a constant region or an Fc region before introducing amino acid change(s) described herein. A parental antigen-binding molecule refers to an antigen-binding molecule comprising the parental constant region or parental Fc region. In some embodiments, the parental Fc region is an Fc region possessing a native sequence (or an Fc region of an antibody). Petition 870260042208, dated 05 / 05 / 2026, pages 146 / 1364 130 / 448 native). Antibodies include, for example, IgA (IgA1, IgA2), IgD, IgE, IgG (IgG1, IgG2, IgG3, IgG4), IgM, etc. Antibodies may be derived from humans or monkeys (e.g., cynomolgus, rhesus monkey, marmoset, chimpanzee, or baboon). Native antibodies may also include naturally occurring mutations. A plurality of IgG allotype sequences due to genetic polymorphism are described in Sequences of protein of immunological interest, NIH Publication No. 91-3242, and any of them may be used in the present disclosure. In one embodiment, the parental Fc region is an Fc region derived from a human IgG1 heavy chain constant region, shown in SEQ ID NO: 61, 62, or 182.

[00514] In one aspect, anti-CD137 antigen-binding molecules or antibodies possess an increased isoelectric point (pI) compared to anti-CD137 antigen-binding molecules or antibodies comprising a native sequence Fc region or a parental Fc region. In some embodiments, the variant Fc regions include at least one amino acid alteration. In other embodiments, the amino acid alteration leads to an elevation of the isoelectric point (pI) of the variant Fc region compared to the parental Fc region. Without being constrained by any particular theory, the pH of biological fluids (e.g., plasma) is believed to be in a neutral pH range.In biological fluids, the net positive charge of an antigen-binding molecule or antibody with an increased pI is increased due to the increased pI, and as a result, the antigen-binding molecule or antibody is more strongly attracted by the Coulomb physicochemical interaction to the endothelial cell surface, which has a net negative charge, compared to an antigen-binding molecule or antibody that does not have an increased pI. Therefore, antigen-binding molecules (or antibodies) are called agonistic molecules, or bound antigen-binding molecules. Petition 870260042208, dated 05 / 05 / 2026, page 147 / 1364 131 / 448 antigen-binding molecules (or antibodies) can approach the surface of cells expressing the Fc-gamma receptor, resulting in increased binding of antigen-binding molecules or antibodies to cells expressing the Fc-gamma receptor. For those anti-CD137 agonist antigen-binding molecules or antibodies that show CD137 agonist activity based on the contribution of Fc-gamma receptor binding activity, anti-CD137 agonist antigen-binding molecules or antibodies possessing increased binding to Fc-gamma receptor-expressing cells due to pI-enhancing amino acid changes may exhibit stronger CD137 agonist activity compared to anti-CD137 agonist antigen-binding molecules or antibodies that do not possess pI-enhancing amino acid changes.

[00515] In the present disclosure, pI may be a theoretical pI or experimentally determined. The pI value may be determined, for example, by isoelectric focusing known to those skilled in the art. The value of a theoretical pI may be calculated, for example, using gene and amino acid sequence analysis software (Genetyx, etc.). In the calculation, the properties of an antibody may be reflected in a calculation formula. For example, (i) generally, conserved Cys within an antibody forms a disulfide bond and does not carry an electrical charge from the side chain; therefore, such Cys may be excluded from the calculation and only the free-form Cys that does not form a disulfide bond may be included in the calculation.Alternatively, (ii) the charge state or isoelectric point of antibodies may change due to post-translational modifications; therefore, a calculation formula may be modified as follows, taking into account such post-translational modifications: (a) when the N-terminal heavy chain is Q (glutamine), the N-terminal amino group is excluded from the calculation, assuming that pyroglutamylation occurs, (b) when the C-terminal chain. Petition 870260042208, dated 05 / 05 / 2026, pages 148 / 1364 132 / 448 heavy is K (lysine), K (only one residue) is excluded from the calculation, assuming truncation occurs; and (c) the side chains of all C (cysteine) present in the generally conserved positions are excluded from the calculation, assuming that all these C form disulfide bonds within the molecule. In a preferred embodiment, both of the above described (i) and (ii) can be reflected in the calculation formula.

[00516] In one embodiment, the value of pI may be increased, for example, by at least 0.01, 0.03, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5 or more, by at least 0.6, 0.7, 0.8, 0.9 or more, by at least 1.0, 1.1, 1.2, 1.3, 1.4, 1.5 or more, or by at least 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 3.0 or more, compared with before modification.

[00517] In one embodiment, amino acid changes related to increased pI and methods for increasing the pI of an antigen-binding molecule or antibody are described herein in detail in III. Compositions and Methods (Agonistic Antigen-binding Molecules Comprising a Variant Fc Region with Increased Isoelectric Point (pI)). Those skilled in the art would understand that any amino acid changes and methods for increasing pI described in III. Compositions and Methods (Agonistic Antigen-binding Molecules Comprising a Variant Fc Region with Increased Isoelectric Point (pI)) can be applied to anti-CD137 antigen-binding molecules or antibodies.

[00518] In one embodiment, the anti-CD137 antigen-binding molecules or antibodies possess a variant Fc region with increased pI, and the variant Fc region comprises at least one amino acid change at at least one position selected from the group consisting of positions 285, 311, 312, 315, 318, 333, 335, 337, 341, 342, 343, 384, 385, 388, 390, 399, 400, 401, 402, 413, 420, 422, and 431, according to EU numbering. In other embodiments, the variant Fc regions with increased pI comprise Petition 870260042208, dated 05 / 05 / 2026, page 149 / 1364 133 / 448 Arg or Lys in each selected position.

[00519] In other embodiments, the anti-CD137 antigen-binding molecules or antibodies have a variant Fc region with increased pI, and the variant Fc region comprises at least one amino acid change at at least one position selected from the group consisting of positions 311, 343, and 413 according to EU numbering. In other embodiments, the variant Fc regions with increased pI comprise an amino acid change at position 311, 343, or 413 according to EU numbering. In another embodiment, the variant Fc regions with increased pI comprise Arg or Lys at each selected position.

[00520] In another aspect, the present disclosure provides anti-CD137 antigen-binding molecules or antibodies comprising a pI-variant Fc region comprising amino acid changes at any of the following (1) to (3): (1) at positions 311 and 343; (2) at positions 311 and 413; and (3) at positions 343 and 413, according to EU numbering. In other embodiments, the pI-variant Fc regions comprise Arg or Lys at each selected position.

[00521] In one embodiment, the anti-CD137 antigen-binding molecules or antibodies of the present disclosure comprise a variant Fc region comprising amino acid change(s) identified in Table 2 below.

[00522] Amino acid changes to increase the pI of an Fc region Petition 870260042208, dated 05 / 05 / 2026, pp. 150 / 1364 134 / 448 Table 2 Number of Amino Acid Substitutions (EU Numbering) 1 P343R / D413K 2 Q311R / P343R 3 P343R 4 D413K 5 Q311R 6 Q311R / D413K

[00523] In one embodiment, the anti-CD137 antigen-binding molecules or antibodies comprise a variant Fc region prepared by making amino acid alteration(s) to an Fc region possessing a native sequence. In one embodiment, the variant Fc regions have increased binding activity for at least one Fc-gamma receptor selected from the group consisting of Fc-gamma RIa, Fc-gamma RIIa, Fc-gamma RIIb, Fc-gamma RIIIa, and Fc-gamma RIIIb, compared to an Fc region possessing a native sequence or a parental Fc region. Preferably, the variant Fc regions have increased binding activity for Fc-gamma RIIb, compared to an Fc region possessing a native sequence or a parental Fc region.It is reported that an anti-CD137 antibody comprising a variant Fc region with increased binding activity for Fc-gamma RIIb possesses increased agonist activity compared to the anti-CD137 antibody comprising an Fc region with a native sequence. In one embodiment, such as amino acid changes to increase binding activity for Fc-gamma RIIb, the amino acid changes taught in WO2012 / 115241, WO2014 / 030728, WO2014 / 163101 and / or WO2017 / 104783 can be used, for example. In a preferred embodiment, the changes to increase binding activity for Fc-gamma RIIb are amino acid change(s) of at least one selected position from the group consisting of the positions. Petition 870260042208, dated 05 / 05 / 2026, pp. 151 / 1364 135 / 448 234, 235, 236, 237, 238, 264, 268, 295, 326 and 330, according to the EU numbering system.

[00524] Fc gamma receptors (hereinafter referred to as Fc gamma receptors, Fc gamma R or FcgR) refer to receptors that can bind to the Fc region of IgG1, IgG2, IgG3 and IgG4 monoclonal antibodies, and practically means any member of the family of proteins encoded by the Fc gamma receptor genes. In humans, this family includes Fc gamma RI (CD64) including Fc gamma RIa, Fc gamma RIb and Fc gamma RIc isoforms; Fc gamma RII (CD32) including Fc gamma RIIa isoforms (including H131 (type H) and R131 (type R) allotypes), Fc gamma RIIb (including Fc gamma RIIb-1 and Fc gamma RIIb-2) and Fc gamma RIIc; and Fc gamma RIII (CD16) including Fc gamma RIIIa isoforms (including V158 and F158 allotypes), and Fc gamma RIIIb isoforms (including Fc gamma RIIIb-NA1 and Fc gamma RIIIb-NA2 allotypes) and any human Fc gamma Rs, Fc gamma R isoforms or allotypes yet to be discovered, but are not limited to them. Fc gamma RIIb1 and Fc gamma RIIb2 have been reported as splicing variants of human Fc gamma RIIb.Furthermore, a splicing variant called Fc gamma RIIb3 has been reported (J Exp Med, 1989, 170: 1369-1385). In addition to these splicing variants, human Fc gamma RIIb includes NCBI-registered AAI46679.1 and all NCBI-registered splicing variants, which are NP_001002273.1, NP_001002274.1, NP_001002275.1, NP_001177757.1 and... NP_003992.3. Furthermore, human Fc gamma RIIb includes all previously reported genetic polymorphisms, as well as Fc gamma RIIb (Arthritis Rheum. 48: 3242-3252 (2003); Kono et al., Hum. Mol. Genet. 14: 2881-2892 (2005); and Kyogoju et al., Arthritis Rheum. 46: 1242-1254 (2002)), and every genetic polymorphism that will be reported in the future.

[00525] In Fc gamma RIIa, there are two allotypes, one where the amino acid at position 131 of Fc gamma RIIa is histidine (type H) and the other Petition 870260042208, dated 05 / 05 / 2026, p. 152 / 1364 136 / 448 where the amino acid at position 131 is replaced by arginine (R type) (Warrmerdam, J Exp. Med. 172: 19-25 (1990)).

[00526] Fc gamma R includes Fc gamma Rs derived from humans, mice, rats, rabbits, and monkeys, but is not limited to them and can be derived from any organism. Mouse Fc gamma Rs include Fc gamma RI (CD64), Fc gamma RII (CD32), Fc gamma RIII (CD16), and Fc gamma RIII-2 (CD16-2), and any mouse Fc gamma Rs, or isoforms of Fc gamma R, but are not limited to them.

[00527] In another aspect, the present disclosure provides anti-CD137 antigen-binding molecules or antibodies comprising a variant Fc region with increased binding activity for Fc-gamma RIIb, comprising amino acid changes of any of the following (1) to (8): (1) at positions 234, 238, 264 and 330; (2) at positions 234, 238 and 330; (3) at positions 234, 237, 238 and 330; (4) at positions 236, 268 and 330; (5) at positions 235, 236, 268, 295, 326 and 330; in accordance with EU numbering.

[00528] In one embodiment, the anti-CD137 antigen-binding molecules or antibodies of the present disclosure comprise a variant Fc region comprising amino acid changes identified in Table 3 below. In another embodiment, the anti-CD137 antigen-binding molecules or antibodies of the present disclosure comprise a variant Fc region that additionally comprises, in addition to the amino acid change(s) described in Table 2 (amino acid changes involving increased pI of an Fc region), any combination of amino acid changes identified in Table 3 below.

[00529] Amino acid modifications to increase the Fc-gamma RIIb binding activity of an Fc region Petition 870260042208, dated 05 / 05 / 2026, page 153 / 1364 137 / 448 Table 3 Amino acid substitution number (EU numbering) 1 L234Y / P238D / V264I / A330K 2 L234Y / P238D / A330K 3 L234Y / G237D / P238D / A330K 4 G236N / H268D / A330K 5 L235W / G236N / H268D / Q295L / K326T / A330K

[00530] In one embodiment, the present disclosure provides variant Fc regions, including those variant Fc regions that have at least one amino acid change and have RIIb Fc-gamma binding activity equivalent to or greater than that of a reference Fc region. In one embodiment, the reference Fc region is an Fc region comprising any combination of amino acid changes identified in Table 3 above. In a preferred embodiment, the reference Fc region is an Fc region comprised within a TT14 (SEQ ID NO: 149), TT16 (SEQ ID NO: 150), MY201 (SEQ ID NO: 153), or MY518 (SEQ ID NO: 154) heavy chain constant region. In a preferred embodiment, the reference Fc region is an Fc region comprised within a MY201 (SEQ ID NO: 153) or MY518 (SEQ ID NO: 154) heavy chain constant region.

[00531] In another aspect, the present disclosure provides isolated agonist-binding molecules or antibodies comprising a variant Fc region with increased binding activity for the Fc-gamma receptor (preferably, Fc-gamma RIIb) and increased pI. In a particular embodiment, the variant Fc regions described herein comprise at least two amino acid changes in the parental Fc regions. As described above, an antigen-binding molecule or antibody with an increased pI is more strongly attracted by the Coulomb physicochemical interaction to the endothelial cell surface that has a net negative charge compared to an antigen-binding molecule or antibody. Petition 870260042208, dated 05 / 05 / 2026, page 154 / 1364 138 / 448 which does not have an increased pI. Therefore, for those agonistic antigen-binding molecules or antibodies that show agonistic activity based on the contribution of binding activity to the Fc-gamma receptor (preferably Fc-gamma RIIb), the agonistic activity of the antigen-binding molecules or antibodies can be increased by the combination of amino acid alteration(s) to increase the Fc-gamma receptor (preferably Fc-gamma RIIb) and amino acid alteration(s) to increase pI.

[00532] In one embodiment, anti-CD137 antigen-binding molecules or antibodies comprise a variant Fc region comprising both amino acid alteration(s) to increase binding activity for the Fc-gamma receptor (e.g., Fc-gamma RIIb) and amino acid alteration(s) to increase the isoelectric point (pI), as described above. As described above, an antigen-binding molecule or antibody with an increased pI is more strongly attracted by the Coulomb physicochemical interaction to the endothelial cell surface that has a net negative charge compared to an antigen-binding molecule or antibody that does not have an increased pI.Therefore, for those anti-CD137 agonist antigen-binding molecules or antibodies that show CD137 agonist activity based on the contribution of binding activity to the Fc-gamma receptor (preferably Fc-gamma RIIb), the agonist activity of the anti-CD137 antigen-binding molecules or antibodies can be increased by combining amino acid alteration(s) to increase the Fc-gamma receptor (preferably Fc-gamma RIIb) and amino acid alteration(s) to increase pI.

[00533] In one aspect, the present disclosure provides polypeptides comprising a variant Fc region with increased binding activity for Fc-gamma RIIb and with an increased pI, comprising at least three amino acid changes including (a) Petition 870260042208, dated 05 / 05 / 2026, pp. 155 / 1364 139 / 448 at least one amino acid change of at least one position selected from the group consisting of positions 234, 235, 236, 237, 238, 264, 268, 295, 326 and 330, according to EU numbering, and (b) at least two amino acid changes of at least two positions selected from the group consisting of positions 311, 343 and 413, according to EU numbering.

[00534] In another aspect, the present disclosure provides polypeptides comprising a variant Fc region with an increased binding activity for Fc-gamma RIIb and an increased pI, comprising amino acid changes of any of the following (1) to (26): (1) positions 235, 236, 268, 295, 326, 330, 343 and 413; (2) positions 214, 235, 236, 268, 295, 326, 330, 343 and 413; (3) positions 234, 238, 250, 264, 307, 330, 343 and 413; (4) positions 234, 238, 264, 330, 343 and 413; (5) positions 234, 237, 238, 250, 307, 330, 343 and 413; (6) positions 234, 237, 238, 330, 343 and 413; (7) positions 235, 236, 268, 295, 326, 330, 311 and 343; (8) positions 234, 238, 250, 264, 307, 330, 311 and 343; (9) positions 234, 238, 264, 330, 311 and 343; (10) positions 234, 237, 238, 250, 307, 330, 311 and 343; (11) positions 234, 237, 238, 330, 311 and 343; (12) positions 235, 236, 268, 295, 326, 330 and 343; (13) positions 214, 235, 236, 268, 295, 326, 330 and 343; (14) positions 235, 236, 268, 295, 326, 330 and 413; (15) positions 214, 236, 268, 330 and 343; (16) positions 214, 235, 236, 268, 330 and 343; (17) positions 214, 236, 268, 330 and 413; (18) positions 214, 236, 268, 330, 343 and 413; (19) positions 214, 235, 236, 268, 330, 343 and 413; Petition 870260042208, dated 05 / 05 / 2026, pages 156 / 1364 140 / 448 (20) positions 214, 236, 268, 330 and 311; (21) positions 214, 235, 236, 268, 330 and 311; (22) positions 214, 236, 268, 330, 311 and 343; (23) positions 214, 235, 236, 268, 330, 311 and 343; (24) positions 214, 236, 268, 330, 311 and 413; (25) positions 214, 235, 236, 268, 330, 311 and 413; (26) positions 214, 235, 236, 268, 295, 326, 330 and 311, according to EU numbering.

[00535] In one embodiment, the variant Fc regions of the present disclosure comprise any combination of amino acid changes identified in Table 4 below. Table 4 Number of Amino Acid Substitutions (EU Numbering) 1 L235W / G236N / H268D / Q295L / K326T / A330K / P343R / D413K 2 K214R / L235W / G236N / H268D / Q295L / K326T / A330K / P343R / D413K 3 L234Y / P238D / T250V / V264I / T307P / A330K / P343R / D413K 4 L234Y / P238D / V264I / A330K / P343R / D413K 5 L234Y / G237D / P238D / T250V / T307P / A330K / P343R / D413K 6 L234Y / G237D / P238D / A330K / P343R / D413K 7 L235W / G236N / H268D / Q295L / K326T / A330K / Q311R / P343R 8 L234Y / P238D / T250V / V264I / T307P / A330K / Q311R / P343R 9 L234Y / P238D / V264I / A330K / Q311R / P343R 10 L234Y / G237D / P238D / T250V / T307P / A330K / Q311R / P343R 11 L234Y / G237D / P238D / A330K / Q311R / P343R 12 L235W / G236N / H268D / Q295L / K326T / A330K / P343R 13 K214R / L235W / G236N / H268D / Q295L / K326T / A330K / P343R 14 L235W / G236N / H268D / Q295L / K326T / A330K / D413K 15 K214R / G236N / H268D / A330K / P343R 16 K214R / G236N / H268D / A330K / P343R / D413K 19 K214R / L235W / G236N / H268D / A330K / P343R / D413K Petition 870260042208, dated 05 / 05 / 2026, page 157 / 1364 141 / 448 Number of Amino Acid Substitutions (EU Numbering) 20 K214R / G236N / H268D / A330K / Q311R 21 K214R / L235W / G236N / H268D / A330K / Q311R 22 K214R / G236N / H268D / A330K / Q311R / P343R 23 K214R / L235W / G236N / H268D / A330K / Q311R / P343R 24 K214R / G236N / H268D / A330K / Q311R / D413K 25 K214R / L235W / G236N / H268D / A330K / Q311R / D413K 26 K214R / L235W / G236N / H268D / Q295L / K326T / A330K / Q311R

[00536] In one embodiment, the variant Fc regions comprising any combination of amino acid changes described in Table 4 above do not have the amino acid at position 447 according to EU numbering. In a preferred embodiment, the variant Fc regions comprising any combination of amino acid changes described in Table 4 above do not have the amino acids at positions 446 and 447 according to EU numbering.

[00537] Those skilled in the art would understand that at least one amino acid change to increase the binding activity for Fcgamma R (including Fc-gamma RIIb) compared to the parental Fc region as described or suggested, for example, in WO2013 / 047752, WO2013 / 125667, WO2014 / 030728, WO2014 / 163101 or WO2017104783, and at least one amino acid change to increase pI compared to the parental Fc region as described or suggested, for example, in WO2017 / 104783, WO2017 / 046994, and any combination of these amino acid changes may be used, in addition to the changes provided in the illustrations above.

[00538] Furthermore, amino acid changes made for other purpose(s) can be combined in a variant Fc region described herein. For example, amino acid substitutions that increase FcRn binding activity (Hinton et al., J. Immunol. 176 (1): 346-356 (2006); Dall'Acqua et al., J. Biol. Chem. 281 (33): 23514 Petition 870260042208, dated 05 / 05 / 2026, pp. 158 / 1364 142 / 448 23524 (2006); Petkova et al., Intl. Immunol. 18 (12): 1759-1769 (2006); Zalevsky et al., Nat. Biotechnol. 28 (2): 157-159 (2010); WO 2006 / 019447; WO 2006 / 053301; and WO 2009 / 086320) and amino acid substitutions may be added to improve antibody heterogeneity or stability (WO 2009 / 041613). Alternatively, polypeptides with the property of promoting antigen removal, which are described in WO 2011 / 122011, WO 2012 / 132067, WO 2013 / 046704 or WO 2013 / 180201, polypeptides with the property of specific binding to a target tissue, which are described in WO 2013 / 180200, polypeptides with the property of repeated binding to a plurality of antigen molecules, which are described in WO 2009 / 125825, WO 2012 / 073992 or WO 2013 / 047752, can be combined with a variant Fc region described herein.Alternatively, in order to confer binding activity to other antigens, the amino acid changes disclosed in EP1752471 and EP1772465 can be combined in CH3 of a variant Fc region described herein.

[00539] In one embodiment, the anti-CD137 antigen-binding molecules or antibodies of the present disclosure comprise a heavy chain constant region comprising any amino acid sequence selected from SEQ ID NOs: 64-85. Preferably, the anti-CD137 antigen-binding molecules or antibodies of the present disclosure comprise a heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 75 or 82.

[00540] In a preferred embodiment, anti-CD137 antigen-binding molecules or antibodies comprising the variant Fc region described above possess CD137-binding activity described above that is dependent on a small molecule compound.

[00541] In one embodiment, antigen-binding molecules Petition 870260042208, dated 05 / 05 / 2026, page 159 / 1364 143 / 448 anti-CD137 or antibodies of the present disclosure comprise the following variable region and constant region: a variable region comprising the HVR described above, a variable heavy chain region and / or a variable light chain region; and the Fc variant region described above. In a preferred embodiment, the antigen-binding molecules of the anti-CD137 or antibodies of the present disclosure may be any anti-CD137 antibody selected from the antibodies described in Table 52.

[00542] In another aspect, the present disclosure provides antigen-binding molecules or antibodies that bind to the same epitope on CD137 as the anti-CD137 antigen-binding molecules or antibodies provided herein, in the presence of a low molecular weight compound (for example, in the presence of 10 micromolar or more, 50 micromolar or more, 100 micromolar or more, 150 micromolar or more, 200 micromolar or more, or 250 micromolar or more of the low molecular weight compound). For example, in a particular embodiment, these antigen-binding molecules or antibodies are provided that bind to the same epitope as the anti-CD137 antigen-binding molecules or antibodies comprising A375 / B167, A372 / B040, A356 / B040, A486 / B167, A487 / B167, A488 / B226, A489 / B223, A548 / B376, A551 / B256, A551 / B379, A555 / B379, A548 / B256 and / or A549 / B167 described in Table 17, as a combination of variable heavy chain region / variable light chain region. In one embodiment, the anti-CD137 antigen-binding molecules or antibodies of the present disclosure possessing CD137-binding activity that is dependent on small molecule compound-dependent antigen-binding activity recognize an epitope formed by a complex formed from the antigen (e.g., CD137) and the low molecular weight compound (e.g., ATP). Petition 870260042208, dated 05 / 05 / 2026, page 160 / 1364 144 / 448

[00543] In a further aspect, the present disclosure provides antigen-binding molecules or antibodies that compete for binding to CD137 with the anti-CD137 antigen-binding molecules or antibodies provided herein, in the presence of a low molecular weight compound (e.g., in the presence of 10 micromolar or more, 50 micromolar or more, 100 micromolar or more, 150 micromolar or more, 200 micromolar or more, or 250 micromolar or more of the low molecular weight compound). For example, in one particular embodiment, these compete for the CD137 binding site with the anti-CD137 antigen-binding molecules or antibodies comprising A375 / B167, A372 / B040, A356 / B040, A486 / B167, A487 / B167, A488 / B226, A489 / B223, A548 / B376, A551 / B256, A551 / B379, A555 / B379, A548 / B256 and / or A549 / B167 described in Table 17, as a combination of variable heavy chain region / variable light chain region.

[00544] In a further aspect of this disclosure, an anti-CD137 antigen-binding molecule or antibody according to any of the above embodiments is a monoclonal antibody, including a chimeric, humanized or human antibody. In one embodiment, an anti-CD137 antibody is an antibody fragment, for example, an Fv, Fab, Fab', scFv, diabody or F(ab')2 fragment. In another embodiment, the antibody is a full-length antibody, for example, an intact IgG1 antibody or other antibody class or isotype as defined herein.

[00545] In another aspect, an anti-CD137 antigen-binding molecule or antibody according to any of the above embodiments may incorporate any of the features, individually or in combination, as described in Sections 1-7 below: 1. Agonistic activity of anti-CD137 antigen-binding molecule Petition 870260042208, dated 05 / 05 / 2026, pp. 161 / 1364 145 / 448 or antibody

[00546] In a specific embodiment, the anti-CD137 antigen-binding molecule or antibody in the present disclosure possesses CD137 agonist activity. CD137 signaling not only stimulates IFN-γ secretion and NK cell proliferation (Buechele et al., 2012; Lin et al., 2008; Melero et al., 1998), but also increases DC survival and activation indicated by the upregulation of costimulatory molecules and cytokine secretion (Choi et al., 2009; Futagawa et al., 2002; Wilcox et al., 2002). However, CD137 is best characterized as a costimulatory molecule that regulates TCR-induced activation in both CD4+ and CD8+ T cell subsets. In combination with TCR activation, anti-CD137 agonist antibodies increase T cell proliferation, stimulate lymphokine secretion, and reduce the sensitivity of T lymphocytes to activation-induced cell death (reviewed in Snel et al., 2011).Of these phenomena, the physiological phenomena observed after CD137 signaling in T cells are mediated by downstream signals activated by CD137 signaling, such as TRAF2, TRAF1, in particular NF-kappaB, JNK, Erk, Akt, survivin, Bcl-XL and / or Bcl-2 (Ward-Kavanagh et al., Immunity, 44: 1005 (2016)).

[00547] In one embodiment, the anti-CD137 agonist antigen-binding molecule or anti-CD137 agonist antibody is an antigen-binding molecule or antibody that, by binding to CD137, transduces the CD137 signal and induces or significantly increases IFN-gamma secretion, proliferation and increased survival of NK cells; DC activation indicated by upregulation of cytokine and co-stimulatory molecule secretion; TCR induction; T cell proliferation; and / or lymphokine secretion. In a different embodiment, the anti-CD137 agonist antigen-binding molecule or anti-CD137 agonist antibody is a molecule of Petition 870260042208, dated 05 / 05 / 2026, page 162 / 1364 146 / 448 antigen- or antibody-binding that transduces the CD137 signal by binding to CD137 on T cells and significantly induces NFkappaB activation of T cells. Furthermore, the antigen- or antibody-binding molecule showing CD137 agonist activity means that any of the aforementioned physiological phenomena are observed when the antigen- or antibody-binding molecule binds to CD137. The method for measuring CD137 agonist activity is described in detail in section C. Assays below.

[00548] In a specific embodiment, the anti-CD137 antigen-binding molecule or antibody in the present disclosure possesses small molecule compound-dependent CD137 agonist activity. In a non-limiting embodiment, the CD137 agonist activity of the anti-CD137 antigen-binding molecule or antibody against CD137 in the presence of a small molecule compound is greater than the CD137 agonist activity in the absence of the small molecule compound. In a different embodiment, the CD137 agonist activity of the anti-CD137 antigen-binding molecule or antibody in the presence of a high concentration of a small molecule compound is greater compared to the CD137 agonist activity in the presence of a low concentration of the small molecule compound.In another embodiment, the CD137 agonist activity of the anti-CD137 antigen-binding molecule or antibody in the presence of a small molecule compound is 2 times or more, 3 times or more, 5 times or more, 10 times or more, 20 times or more, 30 times or more, 50 times or more, 100 times or more, 200 times or more, 300 times or more, 500 times or more, 1 x 103 times or more, 2 x 103 times or more, 3 x 103 times or more, 5 x 103 times or more, 1 x 104 times or more, 2 x 104 times or more, 3 x 104 times or more, 5 x 104 times or more, or 1 x 105 times or more, compared with the CD137 agonist activity in the absence of the molecule compound. Petition 870260042208, dated 05 / 05 / 2026, pp. 163 / 1364 147 / 448 small.

[00549] Any suitable concentration can be selected as the concentration of the small molecule compound, provided that a difference in the binding activity of the anti-CD137 antigen-binding molecule or antibody is detected. In one embodiment, the anti-CD137 antigen-binding molecule or antibody transduces the CD137 signal by binding to CD137 on the cell surface. Therefore, one skilled in the art would understand that the anti-CD137 antigen-binding molecule or antibody that has small molecule compound-dependent CD137 binding activity has small molecule compound-dependent CD137 agonist activity.However, on the other hand, since the methods for measuring binding activity and agonist activity are different, one skilled in the art would understand that the concentration of a small molecule compound for which a difference in binding activity is detected may be different from the concentration of the small molecule compound for which a difference in agonist activity is detected (e.g., for an anti-CD137 antigen-binding molecule or antibody whose CD137 binding activity in the presence of a small molecule compound at 10 μM is 2 times or more compared to the CD137 binding activity in the absence of the small molecule compound, the CD137 agonist activity (assay value) in the presence of the small molecule compound at 10 μM may be less than 2 times compared to the CD137 agonist activity (assay value) in the absence of the small molecule compound).Furthermore, it will be understood by those skilled in the art that the determination of agonist activity may vary depending on the CD137 agonist activity assay (see C. Assays).

[00550] In one embodiment, the antiCD137 antigen-binding molecule or antibody (i) shows agonistic activity for CD137 in Petition 870260042208, dated 05 / 05 / 2026, pages 164 / 1364 148 / 448 presence of a small molecule compound at 10 μM, 50 μM, 100 μM, 150 μM, 200 μM or 250 μM, and (ii) shows substantially no agonist activity towards CD137 in the absence of the small molecule compound, or has low agonist activity towards CD137 in the absence of the small molecule compound (compared to the presence of the small molecule compound).

[00551] In one embodiment, when the agonistic activity of the anti-CD137 antigen-binding molecule or antibody is evaluated by a) Agonistic Activity Assay (PBMC) explained in detail in C. Assays, the anti-CD137 antigen-binding molecule or antibody (i) exhibits agonistic activity for CD137 in the presence of a small molecule compound at 250 μM, and (ii) has low agonistic activity for CD137 in the absence of the small molecule compound (compared to the presence of the small molecule compound).In another embodiment, the anti-CD137 antigen-binding molecule or antibody (i) shows agonistic activity for CD137 in the presence of a small molecule compound at 250μM, and (ii) shows substantially no agonistic activity for CD137 in the absence of the small molecule compound.

[00552] In one embodiment, when the agonistic activity of the anti-CD137 antigen-binding molecule or antibody is evaluated by b) Agonistic activity assay (reporter gene assay) explained in detail in C. Assays, the anti-CD137 antigen-binding molecule or antibody (i) exhibits agonistic activity for CD137 in the presence of 10 μM, 50 μM, 100 μM, 150 μM, 200 μM, or 250 μM of a small molecule compound, and (ii) has substantially no agonistic activity toward CD137 or has less agonistic activity in the absence of the small molecule compound (compared to that in the presence of the small molecule compound). Antibody concentrations in the gene assay Petition 870260042208, dated 05 / 05 / 2026, pages 165 / 1364 149 / 448 reporters can be arbitrarily selected, for example, the final antibody concentration is 0, 0.001, 0.01, 0.1, 1 or 10 μg / mL. In a preferred embodiment, the final antibody concentration is 0.1 μg / mL or 1 μg / mL.

[00553] In one embodiment, when the final antibody concentration is 0.1 μg / mL in b) Agonistic activity assay (reporter gene assay) explained in detail in C. Assays, (i) the CD137 agonist activity (relative light unit) of the anti-CD137 antigen-binding molecule or antibody in the presence of 10 μM of a small molecule compound is 2 times or more, 3 times or more, 5 times or more, 10 times or more, 20 times or more, 30 times or more, 50 times or more, 60 times or more, 70 times or more, 80 times or more, or 90 times or more, compared with (ii) the CD137 agonist activity (relative light unit) in the absence of the small molecule compound. In one embodiment, when the final antibody concentration is 0.1 μg / mL in b) Agonistic activity assay (reporter gene assay) explained in detail in C.In assays, (i) the CD137 agonist activity (relative light unit) of the anti-CD137 antigen-binding molecule or antibody in the presence of 100 μM of a small molecule compound is 2 times or more, 3 times or more, 5 times or more, 10 times or more, 20 times or more, 30 times or more, 50 times or more, 60 times or more, 70 times or more, 80 times or more, or 90 times or more, compared with (ii) the CD137 agonist activity (relative light unit) in the absence of the small molecule compound. In one embodiment, when the final antibody concentration is 0.1 μg / mL in b) Agonistic activity assay (reporter gene assay) explained in detail in C. Assays, (i) the agonistic activity of CD137 (relative light unit) of the antigen-binding molecule anti-CD137 or antibody in the presence of 250 μM of a small molecule compound is 2 times or more, 3 times or more. Petition 870260042208, dated 05 / 05 / 2026, pages 166 / 1364 150 / 448 times or more, 10 times or more, 20 times or more, 30 times or more, 50 times or more, 60 times or more, 70 times or more, 80 times or more, or 90 times or more, compared with (ii) the agonistic activity of CD137 (relative light unit) in the absence of the small molecule compound. In either of the above embodiments, however, 0.1 μg / mL of an anti-CD137 antigen-binding molecule or antibody exhibits substantially no CD137 agonistic activity in the absence of the small molecule compound.

[00554] In one embodiment, when the final antibody concentration is 1 pg / mL in b) Agonistic activity assay (reporter gene assay) explained in detail in C. Assays, (i) the CD137 agonist activity (relative light unit) of the anti-CD137 antigen-binding molecule or antibody in the presence of 10 μM of a small molecule compound is 2 times or more, 3 times or more, 5 times or more, 10 times or more, 20 times or more, 30 times or more, 50 times or more, 60 times or more, 70 times or more, 80 times or more, or 90 times or more, compared with (ii) the CD137 agonist activity (relative light unit) in the absence of the small molecule compound.In one embodiment, when the final antibody concentration is 0.1 pg / mL in b) Agonistic activity assay (reporter gene assay) explained in detail in Assays (i) the CD137 agonist activity (relative light unit) of the anti-CD137 antigen-binding molecule or antibody in the presence of 100 μM of a small molecule compound is 2 times or more, 3 times or more, 5 times or more, 10 times or more, 20 times or more, 30 times or more, 50 times or more, 60 times or more, 70 times or more, 80 times or more or 90 times or more higher, compared with (ii) the CD137 agonist activity (relative light unit) in the absence of the small molecule compound. In one embodiment, when the final antibody concentration is 0.1 pg / mL in b) Agonistic activity assay. Petition 870260042208, dated 05 / 05 / 2026, page 167 / 1364 151 / 448 (reporter gene assay) explained in detail in C. Assays, (i) the CD137 agonist activity (relative light unit) of the anti-CD137 antigen-binding molecule or antibody in the presence of 250 μM of a small molecule compound is 2 times or more, 3 times or more, 5 times or more, 10 times or more, 20 times or more, 30 times or more, 50 times or more, 60 times or more, 70 times or more, 80 times or more, or 90 times or more, compared with (ii) the CD137 agonist activity (relative light unit) in the absence of the small molecule compound. In either of the foregoing embodiments, moreover, 1 μg / mL of an anti-CD137 antigen-binding molecule or antibody exhibits substantially no CD137 agonist activity in the absence of the small molecule compound. 2. Antibody Fragments

[00555] In certain embodiments, an antibody provided here is an antibody fragment. Antibody fragments include, but are not limited to, Fab, Fab', Fab'-SH, F(ab')2, Fv, and scFv fragments and other fragments described below. For a review of certain antibody fragments, see Hudson et al. Nat. Med. 9: 129-134 (2003). For a review of scFv fragments, see, for example, Pluckthun, in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds., (Springer-Verlag, New York), pp. 269-315 (1994); see also WO 93 / 16185; and U.S. Patents Nos. 5,571,894 and 5,587,458. For a discussion of the Fab and F(ab')2 fragments comprising salvage receptor-binding epitope residues and possessing an increased in vivo half-life, see U.S. Patent No. 5,869,046.

[00556] Diabodies are antibody fragments with two antigen-binding sites that can be bivalent or bispecific. See, for example, EP 404097; WO 1993 / 01161; Hudson et al., Nat. Med. 9: 129134 (2003); and Hollinger et al., Proc. Natl. Acad. Sci. USA 90: 6444-6448 Petition 870260042208, dated 05 / 05 / 2026, pages 168 / 1364 152 / 448 (1993). Triabodies and tetrabodies are also described in Hudson et al., Nat. Med. 9: 129-134 (2003).

[00557] Single-domain antibodies are antibody fragments comprising all or part of the variable heavy-chain domain or all or part of the variable light-chain domain of an antibody. In certain embodiments, a single-domain antibody is a human single-domain antibody (Domantis, Inc., Waltham, MA; see, for example, U.S. Patent No. 6,248,516 B1).

[00558] Antibody fragments can be made by various techniques, including, but not limited to, proteolytic digestion of an intact antibody, as well as production by recombinant host cells (e.g., E. coli or phage), as described herein. 3. Chimeric and Humanized Antibodies

[00559] In certain embodiments, an antibody provided herein is a chimeric antibody. Certain chimeric antibodies are described, for example, in U.S. Patent No. 4,816,567; and Morrison et al., Proc. Natl. Acad. Sci. USA, 81: 6851-6855 (1984)). In one example, a chimeric antibody comprises a non-human variable region (e.g., a variable region derived from a mouse, rat, hamster, rabbit, or non-human primate such as a monkey) and a human constant region. In another example, a chimeric antibody is a class-switched antibody in which the class or subclass has been altered from that of the parental antibody. Chimeric antibodies include antigen-binding fragments thereof.

[00560] In certain embodiments, a chimeric antibody is a humanized antibody. Typically, a non-human antibody is humanized to reduce immunogenicity to humans while retaining the specificity and affinity of the parent non-human antibody. Generally, a humanized antibody comprises one or more Petition 870260042208, dated 05 / 05 / 2026, page 169 / 1364 153 / 448 variable domains in which HVRs, for example, CDRs, (or portions thereof) are derived from a non-human antibody, and FRs (or portions thereof) are derived from human antibody sequences. A humanized antibody will optionally also comprise at least a portion of a human constant region. In some embodiments, some FR residues in a humanized antibody are replaced by corresponding residues from a non-human antibody (e.g., the antibody from which the HVR residues are derived), for example, to restore or improve the antibody's specificity or affinity.

[00561] Humanized antibodies and methods for producing them are reviewed, for example, in Almagro and Fransson, Front. Biosci. 13: 1619-1633 (2008), and are further described, for example, in Riechmann et al., Nature 332: 323-329 (1988); Queen et al., Proc. Nat'l Acad. Sci. USA 86: 10029-10033 (1989); US Patents Nos. 5,821,337, 7,527,791, 6,982,321 and 7,087,409; Kashmiri et al., Methods 36: 25-34 (2005) (which describes specificity-determining region (SDR) grafting); Padlan, Mol. Immunol. 28: 489-498 (1991) (describing regeneration); Dall'Acqua et al., Methods 36: 43-60 (2005) (describing FR shuffling); and Osbourn et al., Methods 36: 61-68 (2005) and Klimka et al., Br. J. Cancer, 83: 252-260 (2000) (describing the guided selection approach for FR shuffling).

[00562] Human structural regions that can be used for humanization include, but are not limited to: structural regions selected using the best-fit method (see, for example, Sims et al. J. Immunol. 151: 2296 (1993)); structural regions derived from the consensus sequence of human antibodies from a particular subgroup of variable light or heavy chain regions (see, for example, Carter et al. Proc. Natl. Acad. Sci. USA, 89: 4285 (1992); Petition 870260042208, dated 05 / 05 / 2026, page 170 / 1364 154 / 448 and Presta et al. J. Immunol., 151: 2623 (1993)); mature human structural regions (somatically mutated) or human germline structural regions (see, for example, Almagro and Fransson, Front. Biosci. 13: 1619-1633 (2008)); and structural regions derived from screening RF libraries (see, for example, Baca et al., J. Biol. Chem. 272: 10678-10684 (1997) and Rosok et al., J. Biol. Chem. 271: 22611-22618 (1996)). 4. Human Antibodies

[00563] In certain embodiments, an antibody provided here is a human antibody. Human antibodies can be produced using various techniques known in the art. Human antibodies are generally described in van Dijk and van de Winkel, Curr. Opin. Pharmacol. 5: 368-74 (2001) and Lonberg, Curr. Opin. Immunol. 20: 450-459 (2008).

[00564] Human antibodies can be prepared by administering an immunogen to a transgenic animal that has been modified to produce intact human antibodies or intact antibodies with variable human regions in response to antigenic challenge. These animals typically contain all or a portion of the human immunoglobulin loci, which replace endogenous immunoglobulin loci, or which are present extrachromosomally or randomly integrated into the animal's chromosomes. In such transgenic mice, the endogenous immunoglobulin loci have generally been inactivated. For a review of methods for obtaining human antibodies from transgenic animals, see Lonberg, Nat. Biotech. 23: 1117-1125 (2005). See also, for example, U.S. Patents Nos. 6,075,181 and 6,150,584 describing the XENOMOUSE™ technology; U.S. Patent No. 5,770,429 describing the HuMab technology (trademark); US Patent No. 7,041,870 describing KM MOUSE technology. Petition 870260042208, dated 05 / 05 / 2026, p. 171 / 1364 155 / 448 (registered trademark), and Publication of US Patent Application No. US 2007 / 0061900, describing the VelociMouse technology (registered trademark). The human variable regions of intact antibodies generated by such animals can be further modified, for example, by combining them with a different human constant region.

[00565] Human antibodies can also be produced by hybridoma-based methods. Human myeloma and human mouse heteromyeloma cell lines have been described for the production of human monoclonal antibodies. (see, for example, Kozbor J. Immunol., 133: 3001 (1984); Brodeur et al., Monoclonal Techniques and Applications of Antibody Production, pp. 51-63 (Marcel Dekker, Inc., New York, 1987); and Boerner et al., J. Immunol., 147: 86 (1991).) Human antibodies generated using human B-cell hybridoma technology are also described in Li et al., Proc. Natl. Acad. Sci. USA, 103: 3557-3562 (2006). Additional methods include those described, for example, in US Patent No. 7,189,826 (describing the production of human monoclonal IgM antibodies from hybridoma cell lines) and Ni, Xiandai Mianyixue, 26 (4): 265-268 (2006) (describing human-human hybridomas). Human hybridoma technology (triome technology) is also described in Vollmers and Brandlein, Histology and Histopathology, 20 (3): 927-937 (2005) and Vollmers and Brandlein, Methods and Findings in Experimental and Clinical Pharmacology, 27 (3): 185-91 (2005).

[00566] Human antibodies can also be generated by isolating variable domain sequences from selected Fv clones from human-derived phage display libraries. These variable domain sequences can then be combined with a desired human constant domain. Techniques for selecting human antibodies from antibody libraries are described below. 5. Library-Derived Antibodies Petition 870260042208, dated 05 / 05 / 2026, page 172 / 1364 156 / 448

[00567] The antibodies of the present disclosure can be isolated by combinatorial library screening for antibodies with the desired activity or activities. For example, a variety of methods are known in the art for generating phage display libraries and screening such libraries for antibodies possessing the desired binding characteristics. Such methods are reviewed, for example, in Hoogenboom et al. in Methods in Molecular Biology 178: 1-37 (O'Brien et al., ed., Human Press, Totowa, NJ, 2001) and further described, for example, in McCafferty et al., Nature 348: 552-554; Clackson et al., Nature 352: 624-628 (1991); Marks et al., J. Mol. Biol. 222: 581-597 (1992); Marks and Bradbury, in Methods in Molecular Biology 248: 161-175 (Lo, ed., Human Press, Totowa, NJ, 2003); Sidhu et al., J. Mol. Biol. 338 (2): 299-310 (2004); Lee et al., J. Mol. Biol. 340 (5): 1073-1093 (2004); Fellouse, Proc. Natl. Academic. Sci. USA 101 (34): 12467-12472 (2004); and Lee et al., J.Immunol. Methods 284 (1-2): 119-132 (2004).

[00568] In certain phage display methods, VH and VL gene repertoires are cloned separately by polymerase chain reaction (PCR) and randomly recombined into phage libraries, which can then be screened for antigen-binding phage as described in Winter et al., Ann. Rev. Immunol., 12: 433-455 (1994). The phage typically displays antibody fragments, such as single-chain Fv fragments (scFv) or Fab fragments. Libraries from immunized sources provide high-affinity antibodies for the immunogen without the need to construct hybridomas. Alternatively, the virgin repertoire can be cloned (e.g., from human) to provide a single source of antibodies for a wide range of non-self and also self antigens without any immunization, as described by Griffiths et al., EMBO J, 12: 725-734 (1993). Finally, virgin libraries can also be Petition 870260042208, dated 05 / 05 / 2026, page 173 / 1364 157 / 448 synthetically made by cloning non-rearranged V gene segments from stem cells, and using PCR primers containing random sequences to encode the highly variable CDR3 regions and to perform in vitro rearrangement, as described by Hoogenboom and Winter, J. Mol. Biol., 227: 381-388 (1992). Patent publications describing human antibody phage libraries include, for example: US Patent No. 5,750,373, and US Patent Publication Nos. 2005 / 0079574, 2005 / 0119455, 2005 / 0266000, 2007 / 0117126, 2007 / 0160598, 2007 / 0237764, 2007 / 0292936 and 2009 / 0002360.

[00569] Antibodies or antibody fragments isolated from human antibody libraries are considered human antibodies or human antibody fragments herein.

[00570] Antigen-binding molecules or antibodies with antigen-binding activity dependent on a low molecular weight compound of the present disclosure can be selected by screening over a library of antigen-binding molecules. As such, the combinatorial libraries described above can be used. The library of antigen-binding molecules can be unbiased (naive library) or biased. Examples of the latter type of library include a library of antigen-binding molecules to which binding activity for a specified compound is conferred in advance.In a particular embodiment, an antigen-binding molecule library is a library of antigen-binding molecules to which amino acid alteration(s) to confer binding activity for a specified compound are introduced in advance. Examples of such a library include libraries described, for example, in International Publication WO 2015 / 083764. Petition 870260042208, dated 05 / 05 / 2026, page 174 / 1364 158 / 448 6. Multispecific Antibodies

[00571] In certain embodiments, an antibody provided here is a multispecific antibody, for example, a bispecific antibody. Multispecific antibodies are monoclonal antibodies that have binding specificities for at least two different sites. In certain embodiments, one of the binding specificities is for CD137 and the other is for any other antigen. In certain embodiments, bispecific antibodies can bind to two different epitopes of CD137. Bispecific antibodies can also be used to target cytotoxic agents in cells expressing CD137. Bispecific antibodies can be prepared as full-length antibodies or antibody fragments.

[00572] In one embodiment, the anti-CD137 antigen-binding molecules or antibodies of the present disclosure are bispecific antibodies, one arm of which has CD137-binding activity dependent on a small molecule compound and the other arm of which binds to an antigen other than CD137. The antigen other than CD137 is not particularly limited in structure. In other words, the antigen can be inorganic or organic substances. Exemplary antigens are disclosed in the present descriptive report (e.g., IV. Compositions and methods (antigen-binding molecules whose antigen-binding activity changes depending on the concentration of a small molecule compound), B. Antigen). In one embodiment, the antigens are preferably antigens expressed on cancer cells, immune cells, stromal cells, or such in cancerous tissues or inflammatory tissues.

[00573] Techniques for the production of multispecific antibodies include, but are not limited to, recombinant co-expression of two immunoglobulin heavy-chain-light chain pairs with different specificities (see Milstein and Cuello, Nature 305: 537). Petition 870260042208, dated 05 / 05 / 2026, page 175 / 1364 159 / 448 (1983)), WO 93 / 08829, and Traunecker et al., EMBO J. 10: 3655 (1991)), and knob-in-hole engineering (see, for example, US Patent No. 5,731,168). Multispecific antibodies can also be produced by engineering electrostatic targeting effects for the production of heterodimeric Fc antibody molecules (WO 2009 / 089004A1); crosslinking of two or more antibodies or fragments (see, for example, US Patent No. 4,676,980, and Brennan et al., Science, 229: 81 (1985)); using leucine zippers to produce bispecific antibodies (see, for example, Kostelny et al., J. Immunol., 148 (5): 1547-1553 (1992)); using diacorpo technology to make bispecific antibody fragments (see, for example, Hollinger et al., Proc. Natl. Acad. Sci. USA, 90: 6444-6448 (1993)); and using single-chain Fv dimers (scFv) (see, for example, Gruber et al., J. Immunol., 152: 5368 (1994)); and the preparation of trispecific antibodies as described, for example, in Tutt et al. J. Immunol. 147: 60 (1991).

[00574] Genetically constructed antibodies with three or more functional antigen-binding sites, including octopus antibodies, are also included here (see, for example, US 2006 / 0025576A1).

[00575] The antibody or fragment here also includes a dual-action Fab or DAF comprising an antigen-binding site that binds to CD137 as well as another different antigen (see, for example, US 2008 / 0069820). 7. Antibody Variants

[00576] In certain embodiments, variants of the amino acid sequence of the antibodies provided herein are contemplated. For example, it may be desirable to improve the binding affinity and / or other biological properties of the antibody. Variants of the amino acid sequence of an antibody can be prepared by introducing appropriate modifications to the nucleotide sequence. Petition 870260042208, dated 05 / 05 / 2026, page 176 / 1364 160 / 448 which encodes the antibody or by peptide synthesis. These modifications include, for example, deletions and / or insertions and / or substitutions of residues in the antibody amino acid sequences. Any combination of deletion, insertion, and substitution can be made to arrive at the final construct, provided that the final construct possesses the desired characteristics, for example, antigen binding. a) Replacement, insertion, and deletion variants

[00577] In certain embodiments, antibody variants with one or more amino acid substitutions are provided. Sites of interest for substitutional mutagenesis include HVRs and FRs. Conservative substitutions are shown in Table 1 under the heading of preferred substitutions. More substantial changes are provided in Table 1 under the heading of exemplary substitutions and as further described below with reference to amino acid side chain classes. Amino acid substitutions can be introduced into an antibody of interest and the products selected for a desired activity, e.g., retained / enhanced antigen binding, decreased immunogenicity, or enhanced ADCC or CDC. Table 5 Original Residue Exemplary Substitutions Preferred Substitutions Ward (A) Val; Read; Ile Val Arg (R) Lys; Gln; Asn Lys Asn(N)Gln; His; Asp, Lys; Arg Gln Asp (D) Glu; Asn Glu Cys (C) Ser; Ala Ser Gln (Q) Asn; Glu Asn Glu (E) Asp; Gln Asp Gly (G) Ala Ala His (H) Asn; Gln; Lys; Arg Arg Petition 870260042208, dated 05 / 05 / 2026, page 177 / 1364 161 / 448 Original Residue Exemplary Substitutions Preferred Substitutions Ile (I) Leu; Val; Met; Allah; Phe; Norleucine Leu Leu (L) Norleucine; Ile; Val; Met; Allah; Phe Ile Lys (K) Arg; Gln; Asn Arg Met (M) Leu; Phe; Ile Leu Phe (F) Trp; Read; Val; Ile; Allah; Tyr Tyr Pro (P) Ala Ala Ser(S) Thr Thr Thr (T) Val; Ser Ser Trp (W) Tyr; Phe Tyr Tyr (Y) Trp; Phe; Thr; Ser Phe Val (V) Ile; Read; Met; Phe; Allah; Norleucine Leu

[00578] Amino acids can be grouped according to common side chain properties: (1) hydrophobic: Norleucine, Met, Ala, Val, Leu, Ile; (2) neutral hydrophilic: Cys, Ser, Thr, Asn, Gln; (3) acid: Asp, Glu; (4) basic: His, Lys, Arg; (5) residues that influence chain orientation: Gly, Pro; (6) aromatic: Trp, Tyr, Phe.

[00579] Non-conservative substitutions will involve replacing a member of one of these classes with another class.

[00580] One type of substitution variant involves the replacement of one or more residues in the hypervariable region of a parental antibody (e.g., a humanized or human antibody). Generally, the resulting variants selected for further study will have modifications (e.g., enhancements) in certain biological properties (e.g., increased affinity, immunogenicity). Petition 870260042208, dated 05 / 05 / 2026, page 178 / 1364 162 / 448 reduced) relative to the parental antibody and / or will have retained substantially certain biological properties of the parental antibody. An exemplary replacement variant is an affinity-matured antibody, which can be conveniently generated, for example, using phage display-based affinity maturation techniques, such as those described herein. Briefly, one or more HVR residues are mutated and the variant antibodies are displayed on the phage and screened for a particular biological activity (e.g., binding affinity).

[00581] Alterations (e.g., substitutions) can be made to HVRs, for example, to improve antibody affinity. Such alterations can be made to HVR hotspots, i.e., residues encoded by codons that mutate at high frequency during the somatic maturation process (see, for example, Chowdhury, Methods Mol. Biol. 207: 179-196 (2008)), and / or residues that contact the antigen, with the resulting VH or VL variant being tested for binding affinity. Affinity maturation by constructing and reselecting secondary libraries has been described, for example, in Hoogenboom et al. in Methods in Molecular Biology 178: 1-37 (O'Brien et al., ed., Human Press, Totowa, NJ, (2001).) In some affinity maturation modalities, diversity is introduced into the variable genes chosen for maturation by any one of a variety of methods (e.g., error-object PCR, strand shuffling, or oligonucleotide-directed mutagenesis).A secondary library is then created. The library is then screened to identify any antibody variants with the desired affinity. Another method for introducing diversity involves HVR-targeted approaches, in which multiple HVR residues (e.g., 4-6 residues at a time) are randomized. The HVR residues involved in antigen binding can be specifically identified by... Petition 870260042208, dated 05 / 05 / 2026, page 179 / 1364 163 / 448 example, using mutagenesis or alanine screening modeling. CDR-H3 and CDR-L3 in particular are frequently targeted.

[00582] In certain embodiments, substitutions, insertions, or deletions may occur within one or more HVRs, provided that such alterations do not substantially reduce the antibody's ability to bind to the antigen. For example, conservative alterations (e.g., conservative substitutions as provided herein) that do not substantially reduce binding affinity may be made in HVRs. Such alterations may, for example, be outside the antigen-contact residues in HVRs. In certain embodiments of the VH and VL variant sequences provided above, each HVR is unchanged or contains no more than one, two, or three amino acid substitutions.

[00583] A useful method for identifying antibody residues or regions that can be targeted for mutagenesis is termed alanine screening mutagenesis, as described by Cunningham and Wells (1989) Science, 244: 1081-1085. In this method, a target residue or group of residues (e.g., charged residues such as arg, asp, his, lys, and glu) are identified and replaced with a neutral or negatively charged amino acid (e.g., alanine or polyalanine) to determine if the antibody interaction with the antigen is affected. Additional substitutions can be introduced at amino acid sites demonstrating functional sensitivity to the initial substitutions. Alternatively or additionally, a crystal structure of an antigen-antibody complex can be analyzed to identify contact points between the antibody and the antigen. These contact residues and neighboring residues can be targeted or eliminated as candidates for substitution.The variants can be tracked to determine if they contain the desired properties. Petition 870260042208, dated 05 / 05 / 2026, page 180 / 1364 164 / 448

[00584] Amino acid sequence insertions include amino- and / or carboxyl-terminal fusions ranging in length from one residue to polypeptides containing one hundred or more residues, as well as intra-sequence insertions of single or multiple amino acid residues. Examples of terminal insertions include an antibody with an N-terminal methionyl residue. Other variants of antibody molecule insertion include the fusion of an enzyme (e.g., for ADEPT) or a polypeptide that increases the plasma half-life of the antibody to the N- or C-terminal of the antibody. b) Glycosylation Variants

[00585] In certain embodiments, an antibody provided here is altered to increase or decrease the extent to which the antibody is glycosylated. The addition or deletion of glycosylation sites to an antibody can be conveniently accomplished by altering the amino acid sequence so that one or more glycosylation sites are created or removed.

[00586] When the antibody comprises an Fc region, the carbohydrate attached to it can be altered. Native antibodies produced by mammalian cells typically comprise a branched biantennary oligosaccharide that is usually linked by an N-linkage to the Asn297 of the CH2 domain of the Fc region. See, for example, Wright et al. TIBTECH 15: 26-32 (1997). The oligosaccharide may include various carbohydrates, for example, mannose, N-acetylglucosamine (GlcNAc), galactose, and sialic acid, as well as a fucose linked to a GlcNAc in the stem of the biantennary oligosaccharide structure. In some embodiments, modifications of the oligosaccharide in an antibody of the present disclosure can be made in order to create antibody variants with certain enhanced properties.

[00587] In one embodiment, the antibody variants are provided with a carbohydrate structure that lacks fucose. Petition 870260042208, dated 05 / 05 / 2026, page 181 / 1364 165 / 448 linked (directly or indirectly) to an Fc region. For example, the amount of fucose in such an antibody may be 1% to 80%, 1% to 65%, 5% to 65%, or 20% to 40%. The amount of fucose is determined by calculating the average amount of fucose within the sugar chain in Asn297, relative to the sum of all glycostructures attached to Asn297 (e.g., complex, hybrid, and high-mannose structures) as measured by MALDI-TOF mass spectrometry, as described in WO 2008 / 077546, for example. Asn297 refers to the asparagine residue located near position 297 in the Fc region (EU numbering of Fc region residues); However, Asn297 can also be located approximately + / - 3 amino acids upstream or downstream of position 297, that is, between positions 294 and 300, due to small sequence variations in antibodies. These fucosylation variants may have improved ADCC function. See, for example, US Patent Publication Nos.US 2003 / 0157108 (Presta, L.); US 2004 / 0093621 (Kyowa Hakko Kogyo Co., Ltd). Examples of publications related to defucosylated or fucose-deficient antibody variants include: US 2003 / 0157108; WO 2000 / 61739; WO 2001 / 29246; US 2003 / 0115614; US 2002 / 0164328; US 2004 / 0093621; US ​​2004 / 0132140; US 2004 / 0110704; US. 2004 / 0110282; US 2004 / 0109865; WO 2003 / 085119; WO 2003 / 084570; WO 2005 / 035586; WO 2005 / 035778; WO2005 / 053742; WO2002 / 031140; Okazaki et al. J. Mol. Biol. 336: 1239-1249 (2004); Yamane-Ohnuki et al. Biotech. Bioeng. 87:614 (2004). Examples of cell lines capable of producing defucosylated antibodies include Lec13 CHO cells deficient in protein fucosylation (Ripka et al. Arch. Biochem. Biophys. 249: 533-545 (1986); US Patent Application 2003 / 0157108 A1, Presta, L; and WO 2004 / 056312 A1, Adams et al., especially in Example 11), and knockout cell lines, such as the alpha-1,6-fucosyltransferase gene, FUT8, CHO knockout cells. Petition 870260042208, of 05 / 05 / 2026, p. 182 / 1364 166 / 448 (vide, for example, Yamane-Ohnuki et al. Biotech. Bioeng. 87: 614 (2004); Kanda, Y. et al., Biotechnol. Bioeng., 94 (4): 680-688 (2006); and WO2003 / 08517).

[00588] Antibody variants are further provided with bisected oligosaccharides, for example, where a biantennary oligosaccharide linked to the Fc region of the antibody is bisected by GlcNAc. Such antibody variants may have reduced fucosylation and / or improved ADCC function. Examples of such antibody variants are described, for example, in WO 2003 / 011878 (Jean-Mairet et al.); US Patent No. 6,602,684 (Umana et al.); and US 2005 / 0123546 (Umana et al.). Antibody variants with at least one galactose residue in the oligosaccharide linked to the Fc region are also provided. Such antibody variants may have improved CDC function. Such antibody variants are described, for example, in WO 1997 / 30087 (Patel et al.); WO 1998 / 58964 (Raju, S.); and WO 1999 / 22764 (Raju, S.). c) Variants of the Fc region

[00589] In certain embodiments, one or more amino acid modifications may be introduced into the Fc region of an antibody provided in this document, thus generating an Fc region variant (which may also be called an altered Fc region). The Fc region variant may comprise a human Fc region sequence (e.g., an Fc region of human IgG1, IgG2, IgG3, or IgG4) comprising an amino acid modification (e.g., a substitution) at one or more amino acid positions.

[00590] In certain embodiments, the present disclosure contemplates an antibody variant that possesses some, but not all, effector functions, making it a desirable candidate for applications where the in vivo antibody half-life is important, but certain effector functions (such as complement and ADCC) are unnecessary or deleterious. In vitro and / or in vivo cytotoxicity assays may be Petition 870260042208, dated 05 / 05 / 2026, pp. 183 / 1364 167 / 448 conducted to confirm the reduction / depletion of CDC and / or ADCC activities. For example, Fc receptor (FcR) binding assays can be conducted to ensure that the antibody does not bind to Fc gamma R (therefore, it likely lacks ADCC activity) but retains FcRn binding activity. The primary cells for mediating ADCC, NK cells, express only Fc gamma RIII, while monocytes express Fc gamma RI, Fc gamma RII, and Fc gamma RIII. FcR expression in hematopoietic cells is summarized in Table 3 on page 464 of Ravetch and Kinet, Annu. Rev. Immunol. 9: 457-492 (1991). Non-limiting examples of in vitro assays to evaluate the ADCC activity of a molecule of interest are described in US Patent No. 5,500,362 (see, for example, Hellstrom, I. et al. Proc. Nat'l Acad. Sci. USA 83: 7059-7063 (1986)) and Hellstrom, I et al., Proc. Nat'l Acad. Sci. USA 82: 1499-1502 (1985); 5,821,337 (see Bruggemann, M. et al., J. Exp. Med. 166: 1351-1361 (1987)).Alternatively, non-radioactive assay methods may be employed (see, for example, ACT1™ non-radioactive cytotoxicity assay for flow cytometry (CellTechnology, Inc., Mountain View, CA); and CytoTox 96 (registered trademark) non-radioactive cytotoxicity assay (Promega, Madison, WI)). Useful effector cells for such assays include peripheral blood mononuclear cells (PBMCs) and Natural Killer (NK) cells. Alternatively or additionally, the ADCC activity of the molecule of interest may be evaluated in vivo, for example, in an animal model, such as that disclosed in Clynes et al. Proc. Nat'l Acad. Sci. USA 95: 652-656 (1998). 2006 / 029879 and WO 2005 / 100402.To assess complement activation, a CDC assay can be performed (see, for example, Gazzano-Santoro et al., J. Immunol. Petition 870260042208, dated 05 / 05 / 2026, p. 184 / 1364 168 / 448 Methods 202: 163 (1996); Cragg, MS et al., Blood 101: 1045-1052 (2003); and Cragg, MS and MJ Glennie, Blood 103: 2738-2743 (2004)). FcRn binding and in vivo elimination / half-life determinations can also be performed using methods known in the art (see, for example, Petkova, SB et al., Int'l. Immunol. 18 (12): 1759-1769 (2006)).

[00591] Antibodies with reduced effector function include those with substitution of one or more of the residues in the Fc region 238, 265, 269, 270, 297, 327 and 329 (US Patent No. 6,737,056). Such Fc mutants include Fc mutants with substitutions at two or more amino acid positions 265, 269, 270, 297 and 327, including the so-called Fc DANA mutant with substitution of residues 265 and 297 for alanine (US Patent No. 7,332,581).

[00592] Certain antibody variants with increased or decreased binding to FcRs are described. (see, for example, US Patent No. 6,737,056; WO 2004 / 056312 and Shields et al., J. Biol. Chem. 9 (2): 65916604 (2001)).

[00593] In certain embodiments, an antibody variant comprises an Fc region with one or more amino acid substitutions that enhance ADCC, for example, substitutions at positions 298, 333, and / or 334 of the Fc region (EU residue numbering).

[00594] In some embodiments, alterations are made to the Fc region that result in altered (i.e., increased or decreased) C1q binding and / or Complement-Dependent Cytotoxicity (CDC), for example, as described in U.S. Patent No. 6,194,551, WO 99 / 51642 and Idusogie et al. J. Immunol. 164: 4178-4184 (2000).

[00595] Antibodies with increased half-life and increased binding to the neonatal Fc receptor (FcRn), which is responsible for the transfer of maternal IgGs to the fetus (Guyer et al., J. Immunol. 117: 587 (1976) and Kim et al., J. Immunol. 24: 249 (1994)), are described in Petition 870260042208, dated 05 / 05 / 2026, p. 185 / 1364 169 / 448 US2005 / 0014934A1 (Hinton et al.). These antibodies comprise an Fc region with one or more substitutions that enhance the binding of the Fc region to FcRn. Such Fc variants include those with substitutions at one or more of the residues in the Fc region: 238, 256, 265, 272, 286, 303, 305, 307, 311, 311, 312, 317, 340, 356, 360, 362, 376, 378, 380, 382, ​​413, 424 or 434, for example, substitution of the Fc region residue 434 (US Patent No. 7,371,826).

[00596] See also Duncan & Winter, Nature 322: 738-40 (1988); US Patent No. 5,648,260; US Patent No. 5,624,821; and WO 94 / 29351 relating to other examples of variants of the Fc region.

[00597] In one embodiment, the binding activity with respect to each human Fc-gamma receptor (Fc-gamma R) of an antibody Fc region (including a variant Fc region (the same applies hereafter)) can be measured by a ligand capture method using, for example, BIACORE (registered trademark) T200, which is based on surface plasmon resonance analysis methods as the measurement principle.

[00598] Details of an exemplary method for measuring the binding activity of the antibody Fc region to various human Fc gamma receptors (Fc-gamma Rs) are described below. In one embodiment, the binding activity of an antibody Fc region to Fc-gamma R is assessed using BIACORE (trademark) T200. In a preferred embodiment, this measurement is performed at 25 degrees C, using 50 mM of a phosphate measuring buffer, 150 mM NaCl, 0.05 w / v%-P20, pH 7.4. Specifically, approximately 1000 RU of an antibody comprising a variant Fc region is first captured on a sensor chip, with CaptureSelect (trademark) Fab-lambda Kinetics Human Biotin Conjugate (ThermoFisher scientific) immobilized as a ligand capture molecule. Human Fc-gamma Rs are diluted with the measuring buffer to 8 nM for Fc-gamma. Petition 870260042208, dated 05 / 05 / 2026, page 186 / 1364 170 / 448 RIa and 1000 nM for other Fc-gamma Rs, and can bind to the captured antibody. The binding activity of each antibody with respect to each Fc-gamma R is evaluated by calculating the amount of Fc-gamma R bound per amount of antibody unit (RU) using Biacore T200 Evaluation 2.0 software. In one embodiment, the binding activity of an antibody Fc region to various human Fc-gamma receptors (Fc-gamma Rs) can be measured by the method described in Example 7-4.

[00599] In a preferred embodiment, the Fc-gamma Rs used for the measurement method described above can be an extracellular domain of an Fc-gamma R prepared by the method described below. First, the synthesis of a gene from an extracellular domain of an Fc-gamma R is performed by a method known to those skilled in the art. For this synthesis, sequences of each Fc-gamma R are prepared based on information registered in the NCBI.More specifically, the sequence for Fc-gamma RI is prepared based on NCBI accession sequence # NM_000566.3, the sequence for Fc-gamma RIIa is prepared based on NCBI accession sequence # NM_001136219.1, the sequence for Fc-gamma RIIb is prepared based on NCBI accession sequence # NM_004001.3, and the sequence for Fc-gamma RIIIa is prepared based on NCBI accession sequence # NM_001127593.1, with the His tag appended to the C-terminal. Polymorphic sites for Fc-gamma RIIa are prepared with reference to J. Exp. Med., 1990, 172, 19-25, and polymorphic sites for Fc-gamma RIIIa are prepared with reference to J. Clin. Invest., 1997, 100, 1059-1070. Gene fragments obtained are inserted into an expression vector for animal cells to prepare expression vectors. The prepared expression vectors are transiently introduced into FreeStyle293 cells (Invitrogen) derived from human embryonic renal cancer cells, and a protein of interest can be expressed.The culture supernatant is collected and filtered through a filter. Petition 870260042208, dated 05 / 05 / 2026, page 187 / 1364 171 / 448 The protein is 0.22 micrometers in size and then purified through four steps described below. The first step is cation-exchange column chromatography (SP Sefarose FF); the second step is affinity column chromatography for His markers (HisTrap HP); the third step is gel filtration column chromatography (Superdex200); and the fourth step is aseptic filtration. Note that for Fc-gamma RI, anion-exchange column chromatography using Q sefarose FF is performed as the first step. The concentration of the purified protein is calculated based on the absorbance coefficient, obtained by measuring the absorption at 280 nm using a spectrophotometer and using PACE or another method for the measured values ​​(Protein Science, 1995, 4, 2411-2423).

[00600] In one embodiment, the binding activity of an antibody Fc region to human FcRn can be measured by a ligand capture method using, for example, BIACORE (registered trademark) T200, which is based on surface plasmonic resonance analysis methods as the measurement principle.

[00601] Details of an exemplary method for measuring the binding activity of the antibody Fc region to human FcRn are described below. In one embodiment, the binding activity of an antibody Fc region to human FcRn is assessed using BIACORE (trademark) T200. In a preferred embodiment, this measurement is performed at 25 degrees C, using 50 mM of a phosphate measuring buffer, 150 mM NaCl, 0.05 w / v%-P20, pH 6.0. Specifically, approximately 400 RU of an antibody comprising an Fc region is first captured on a sensor chip, on which CaptureSelect (trademark) Biotin Kinetics Fab Conjugate (ThermoFisher Scientific) is immobilized as a ligand capture molecule, and then diluted human FcRn using the measuring buffer is allowed to bind to it. The binding activity of each Petition 870260042208, dated 05 / 05 / 2026, page 188 / 1364 The 172 / 448 antibody to FcRn is evaluated by calculating KD(M) using the steady-state model in Biacore T200 Evaluation 2.0 software. In a preferred embodiment, the human FcRn protein used in this measurement is prepared according to the method described in Reference Example 2 of WO2010107110. In another embodiment, the binding activity of an antibody Fc region relative to several human FcRn can be measured by the method described in Example 7-5. d) Cysteine-engineered antibody variants

[00602] In certain embodiments, it may be desirable to create cysteine-engineered antibodies, for example, tioMAbs, in which one or more residues of an antibody are replaced by cysteine ​​residues. In particular embodiments, the replaced residues occur at accessible sites of the antibody. By replacing these residues with cysteine, reactive thiol groups are positioned at accessible sites of the antibody and can be used to conjugate the antibody with other moieties, such as drug moieties or drug-linkant moieties, to create an immunoconjugate, as described further in this document. In certain embodiments, any one or more of the following residues may be replaced by cysteine: V205 (Kabat numbering) of the light chain; A118 (EU numbering) of the heavy chain; and S400 (EU numbering) of the Fc region of the heavy chain.Cysteine-based engineered antibodies can be generated as described, for example, in U.S. Patent No. 7,521,541. e) Antibody Derivatives

[00603] In certain embodiments, an antibody provided in this document may be further modified to contain additional non-protein moieties that are known in the art and readily available. Suitable moieties for antibody derivatization include, but are not limited to, water-soluble polymers. Non-limiting examples of water-soluble polymers include, but Petition 870260042208, dated 05 / 05 / 2026, page 189 / 1364 173 / 448 are not limited to, polyethylene glycol (PEG), ethylene glycol / propylene glycol copolymers, carboxymethylcellulose, dextran, polyvinyl alcohol, polyvinylpyrrolidone, poly-1,3-dioxolane, poly-1,3,6-trioxane, ethylene / maleic anhydride copolymer, polyamino acids (homopolymers or random copolymers) and dextran or poly(n-vinylpyrrolidone)polyethylene glycol, polypropylene glycol homopolymers, polypropylene oxide / ethylene oxide copolymers, polyoxyethylated polyols (e.g., glycerol), polyvinyl alcohol and mixtures thereof. Polyethylene glycol propionaldehyde may offer manufacturing advantages due to its stability in water. The polymer may have any molecular weight and may be branched or unbranched. The number of polymers attached to the antibody can vary, and if more than one polymer is attached, they may be the same or different molecules.In general, the number and / or type of polymers used for derivatization can be determined based on considerations including, but not limited to, the particular properties or functions of the antibody to be improved, whether the antibody derivative will be used in a therapy under defined conditions, etc.

[00604] In another embodiment, conjugates of an antibody and a non-protein moiety are provided that can be selectively heated by exposure to radiation. In one embodiment, the non-protein moiety is a carbon nanotube (Kam et al., Proc. Natl. Acad. Sci. USA 102: 11600-11605 (2005)). The radiation can be of any wavelength and includes, but is not limited to, wavelengths that do not harm ordinary cells but heat the non-protein moiety to a temperature at which cells proximal to the non-protein moiety of the antibody are killed. B. Recombinant methods and compositions

[00605] Antibodies can be produced using recombinant methods and compositions, for example, as described in Petition 870260042208, dated 05 / 05 / 2026, page 190 / 1364 174 / 448 US Patent No. 4,816,567. In one embodiment, isolated ...

Claims

1. Antibody, characterized in that it comprises a combination of VH, CH, VL, and CL selected from (a) to (l) below: (a) a VH comprising the amino acid sequence of SEQ ID NOs: 51, a constant region of the heavy chain (CH) comprising the amino acid sequence of SEQ ID NO: 72, a VL comprising the amino acid sequence of SEQ ID NO: 60, and a constant region of the light chain (CL) comprising the amino acid sequence of SEQ ID NO: 63; (b) a VH comprising the amino acid sequence of SEQ ID NO: 51, a CH comprising the amino acid sequence of SEQ ID NO: 74, a VL comprising the amino acid sequence of SEQ ID NO: 60, and a CL comprising the amino acid sequence of SEQ ID NO: 63;(c) a VH comprising the amino acid sequence of SEQ ID NO: 51, a CH comprising the amino acid sequence of SEQ ID NO: 75, a VL comprising the amino acid sequence of SEQ ID NO: 60 and a CL comprising the amino acid sequence of SEQ ID NO: 63; (d) a VH comprising the amino acid sequence of SEQ ID NO: 51, a CH comprising the amino acid sequence of SEQ ID NO: 77, a VL comprising the amino acid sequence of SEQ ID NO: 60 and a CL comprising the amino acid sequence of SEQ ID NO: 63; (e) a VH comprising the amino acid sequence of SEQ ID NO: 51, a CH comprising the amino acid sequence of SEQ ID NO: 78, a VL comprising the amino acid sequence of SEQ ID NO: 60 and a CL comprising the amino acid sequence of SEQ ID NO: 63;(f) a VH comprising the amino acid sequence of Petition 870260042208, dated 05 / 05 / 2026, p. 466 / 1364 2 / 3 SEQ ID NO: 51, a CH comprising the amino acid sequence of SEQ ID NO: 79, a VL comprising the amino acid sequence of SEQ ID NO: 60 and a CL comprising the amino acid sequence of SEQ ID NO: 63; (g) a VH comprising the amino acid sequence of SEQ ID NO: 51, a CH comprising the amino acid sequence of SEQ ID NO: 80, a VL comprising the amino acid sequence of SEQ ID NO: 60 and a CL comprising the amino acid sequence of SEQ ID NO: 63; (h) a VH comprising the amino acid sequence of SEQ ID NO: 51, a CH comprising the amino acid sequence of SEQ ID NO: 81, a VL comprising the amino acid sequence of SEQ ID NO: 60 and a CL comprising the amino acid sequence of SEQ ID NO: 63;(i) a VH comprising the amino acid sequence of SEQ ID NO: 51, a CH comprising the amino acid sequence of SEQ ID NO: 82, a VL comprising the amino acid sequence of SEQ ID NO: 60 and a CL comprising the amino acid sequence of SEQ ID NO: 63; (j) a VH comprising the amino acid sequence of SEQ ID NO: 51, a CH comprising the amino acid sequence of SEQ ID NO: 83, a VL comprising the amino acid sequence of SEQ ID NO: 60 and a CL comprising the amino acid sequence of SEQ ID NO: 63; (k) a VH comprising the amino acid sequence of SEQ ID NO: 51, a CH comprising the amino acid sequence of SEQ ID NO: 84, a VL comprising the amino acid sequence of SEQ ID NO: 60 and a CL comprising the amino acid sequence of SEQ ID NO: 63;and (l) a VH comprising the amino acid sequence of Petition 870260042208, dated 05 / 05 / 2026, page 467 / 1364 3 / 3 SEQ ID NO: 51, a CH comprising the amino acid sequence of SEQ ID NO: 85, a VL comprising the amino acid sequence of SEQ ID NO: 60 and a CL comprising the amino acid sequence of SEQ ID NO: 63.; 2. Isolated nucleic acid, characterized in that it encodes the antibody, as defined in claim 1.

3. Vector, characterized in that the nucleic acid, as defined in claim 2.

4. Host cell, characterized in that it comprises nucleic acid, as defined in claim 2.

5. A method for producing an antibody, characterized in that it comprises culturing a host cell comprising the nucleic acid encoding the antibody, as defined in claim 1, such that the antibody is produced.

6. Immunoconjugate, characterized in that it comprises the antibody, as defined in claim 1, and a cytotoxic agent.

7. Pharmaceutical formulation, characterized in that it comprises the antibody, as defined in claim 1, and a pharmaceutically acceptable carrier.

8. Use of the antibody, as defined in claim 1, characterized in that it is in the manufacture or preparation of a medicament for the treatment of a solid tumor.

9. Use of the antibody, as defined in claim 1, characterized in that it is in the manufacture or preparation of a medicament to activate the immune cells of an individual, wherein the immune cells are B cells, dendritic cells, natural killer cells, macrophages and / or T cells.

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