Formulations of antibodies that bind to human CD137 and their uses

Abstract

The present disclosure relates to a stable formulation comprising an antibody or an antigen-binding fragment thereof that binds to human CD137, and to the use of said formulation in methods of treating or ameliorating various diseases and disorders (including cancer) suitable for treatment with a CD137 antibody, among other things.

Description

[0001] Cross - reference to related applications

[0002] This application claims the benefit of U.S. Provisional Patent Application Serial No. 62 / 793,342, filed on January 16, 2019, and U.S. Provisional Patent Application Serial No. 62 / 960,501, filed on January 13, 2020, the entire contents of each of which are incorporated herein by reference in their entirety. Background of the Invention

[0003] In recent years, increasing evidence has shown that the immune system functions as an important barrier to tumor formation and development. The recognition of the existence of naturally occurring T cells with anti - tumor potential or activity in cancer patients reasonably explains the development of immunotherapy in oncology. Immune cells, such as T cells, macrophages, and natural killer cells, can exhibit anti - tumor activity and effectively control the occurrence and growth of malignant tumors. Tumor - specific antigens or tumor - associated antigens can induce immune cells to recognize and eliminate malignant tumors (Chen & Mellman, (2013) Immunity 39(1):1 - 10). Despite the existence of tumor - specific immune responses, malignant tumors often evade or avoid immune attacks through multiple immune regulatory mechanisms, leading to the inability to control tumor occurrence and development (Motz & Coukos, (2013) Immunity 39(1):61 - 730). In fact, an emerging direction in cancer is to develop these immune regulatory mechanisms and disable the anti - tumor immune responses that lead to tumor escape and immune evasion (Hanahan and Weinberg (2011) Cell 144(5):646 - 674).

[0004] New approaches to cancer immunotherapy include counteracting these immune escape and evasion mechanisms and inducing the innate immune system to fight tumors. CD137 (also known as "tumor necrosis factor receptor superfamily member 9" (TNFRSF9), 4 - 1BB, and "induced by lymphocyte activation" (ILA)) is a transmembrane co - stimulatory receptor protein belonging to the tumor necrosis factor superfamily. CD137 is a T - cell co - stimulatory receptor induced upon TCR activation (Nam et al., (2005) Curr Cancer Drug Targets 5:357 - 363; Watts et al., (2005) Annu Rev Immunol 23:23 - 68). In addition to being expressed on activated CD4+ and CD8+ T cells, CD137 is also expressed on CD4+CD25+ regulatory T cells, activated natural killer (NK) and NK - T cells, monocytes, neutrophils, and dendritic cells.

[0005] Under physiological conditions, CD137 ligates to CD137 ligand (CD137L), an agonist membrane molecule present on antigen-presenting cells including B cells, monocytes, macrophages, and dendritic cells (Watts et al., (2005) Annu Rev Immunol 23:23-68). When interacting with its ligand, CD137 results in increased TCR-induced T cell proliferation, cytokine production, functional maturation, and extended CD8+ T cell survival. The potential of CD137 co-stimulation using various agonists such as agonistic antibodies, recombinant CD137L protein, and CD137-specific aptamers to enable the immune system to attack tumors has been documented in many models (Dharmadhikari et al., (2016) Oncoimmunology 5(4):e1113367 and references therein). A recent report on the clinical evaluation of an agonistic CD137 antibody (urelumab, BMS-663513; Bristol-Myers Squibb) documented treatment-related adverse events observed in human subjects, including severe liver toxicity symptoms (transaminitis) related to antibody dose (Segal et al., (2016) Clin Cancer Res 23(8):1929-1936). In contrast, a different agonistic CD137 antibody (utomilumab, PF-05082566; Pfizer) tested in combination with an anti-PD-1 antibody (pembrolizumab) showed comparable results to anti-PD-1 antibody therapy alone, although no dose-limiting toxicities were observed (Tolcher, A. et al., (2017) Clin Cancer Res 23(18):5349-5357).

[0006] One of the challenges in developing therapeutic antibodies is protein stability. Antibodies have a three-dimensional structure, called the tertiary structure, which is sensitive to the balance of intramolecular and intermolecular interactions between amino acid functional groups and the external environment. Non-covalent interactions are crucial for maintaining the native folded structure of antibodies. Additionally, the folded antibody structure is in a dynamic equilibrium state, and any factor that alters the interaction balance can cause a structural change. This results in an unstable macromolecule. For example, when the native antibody structure unfolds to an intermediate or denatured state, protein variants tend to aggregate (Awwad et al., Pharmaceutics, 2018, 10, 83; doi:10.3390 / pharmaceutics10030083).

[0007] Many factors can cause protein aggregation, including temperature, mechanical, physical, and freeze / thaw stress. Thus, one of the obstacles in antibody development is the development of stable antibody formulations. Excipients have been used to increase the stability of antibody formulations by reducing protein motility and movement, increasing the conformational stability of the antibody, and inhibiting aggregation. However, due to differences in antibody sequences, excipients used in one antibody formulation may not be suitable for another antibody (Awwad et al.). Thus, there remains an unmet need for the development of stable antibody formulations, including those comprising novel agonist antibodies that bind to human CD137. SUMMARY OF THE INVENTION

[0008] The present disclosure is at least in part based on the discovery of novel agonist anti-CD137 antibodies and their formulations that exhibit protective anti-tumor immunity in animals. Notably, the antibodies described herein are effective against multiple tumor types and over a wide dose range. In addition, as illustrated in the working examples, the antibodies described herein have a therapeutic effect on very large tumors. For example, treating tumor-bearing mice with the agonist anti-CD137 antibody described herein resulted in complete regression of tumors as large as 1,800 mm 3 . As Figure 15 shown, treatment of such mice also generated protective immunity. Consistent with the observed efficacy was a positive immunophenotypic change in the tumor microenvironment, such as increased immune cell infiltration and a concomitant decrease in regulatory T cell and exhausted T cell populations (see, e.g., Figures 22A - 22D ).

[0009] As noted above, agonism of CD137 has been associated with certain adverse events, including deaths related to hepatotoxicity in humans (see, e.g., Segal et al. (2017) Clin Cancer Res 23(8) :1929-1935). Similar toxicity has also been observed in animal models following treatment with agonist anti-CD137 antibodies (e.g., the 3H3 antibody) (see, e.g., Bartkowiak et al. (2018) Clin Cancer Res 24(5) :1138-1151). However, the agonist anti-CD137 antibodies described herein have little effect on the liver, as determined by, for example, plasma levels of liver enzymes (e.g., alanine transaminase (ALT)) and immune cell infiltration. For example, there is no evidence of increased immune cell infiltration in the liver or spleen of mice treated with the antibody. Thus, the antibodies described herein are not only highly effective but also avoid certain toxicities associated with CD137 agonism.

[0010] Although the present disclosure is not bound by any particular theory or mechanism of action, it is believed that the excellent therapeutic properties and moderately toxic properties of the antibodies described herein stem in part from one or both of their affinity and the novel epitopes to which they bind. That is, the antibodies described herein share a common novel epitope that is different from the epitopes of other agonist anti-CD137 antibodies. And, as illustrated in the working examples, binding of the antibodies described herein to this epitope results in differentiated in vitro activities, such as effects on regulatory T cell proliferation, cytokines produced by CD8 + T cells and macrophages, and intracellular signaling, compared to agonist antibodies that bind to different epitopes of CD137. In addition, it has been demonstrated that the affinity range ("sweet spot") of the antibodies is optimal for anti-tumor activity. For example, medium-affinity antibodies have been shown to be more effective against large tumors compared to antibodies with higher or lower affinity.

[0011] The present disclosure relates at least in part to stable anti-CD137 antibody formulations. Notably, the anti-CD137 antibody formulations of the present disclosure maintain the stability of the antibody or its antigen-binding fragment, minimize the formation of antibody aggregates (high molecular weight species) and particulates, reduce the percentage of charge variants, and maintain the structural integrity of the antibody.

[0012] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises the heavy chain CDR3 of SEQ ID NO: 126, wherein X is any amino acid, and a buffer comprising histidine.

[0013] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises the heavy chain CDR3 of SEQ ID NO: 126, wherein X is any amino acid, a buffer comprising histidine, and a disaccharide.

[0014] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises the heavy chain CDR3 of SEQ ID NO: 126, wherein X is any amino acid, and a buffer comprising histidine, wherein the formulation has a pH of about 5.0 - 7.0.

[0015] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises the heavy chain CDR3 of SEQ ID NO: 126, wherein X is any amino acid, a buffer comprising histidine, a disaccharide, a nonionic surfactant, and a salt, wherein the pH of the formulation is about 5.0 to about 7.0.

[0016] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises the heavy chain CDR3 of SEQ ID NO: 126, wherein X is any amino acid, a buffer comprising histidine, about 5% to about 15% (weight / volume) of a disaccharide, about 0.01% to about 0.1% (weight / volume (w / v)) of a nonionic surfactant, and about 50 mM to 200 mM of a salt, wherein the pH of the formulation is about 5.0 to about 7.0.

[0017] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises the heavy chain CDR3 of SEQ ID NO: 126, wherein X is any amino acid, a buffer comprising about 10 mM to about 100 mM of histidine, about 5% to about 15% (weight / volume) of sucrose, about 0.01% to about 0.1% (weight / volume (w / v)) of polysorbate-80, and about 50 mM to 200 mM of NaCl, wherein the pH of the formulation is about 5.0 to about 7.0.

[0018] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises the heavy chain CDR3 of SEQ ID NO: 126, wherein X is any amino acid, a buffer comprising histidine, a disaccharide, a nonionic surfactant, and a salt, wherein the pH of the formulation is about 5.0 to about 7.4.

[0019] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises the heavy chain CDR3 of SEQ ID NO: 126, wherein X is any amino acid, a buffer comprising histidine, about 5% to about 15% (weight / volume) of a disaccharide, about 0.01% to about 0.1% (weight / volume (w / v)) of a nonionic surfactant, and about 50 mM to 200 mM of a salt, wherein the pH of the formulation is about 5.0 to about 7.4.

[0020] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises the heavy chain CDR3 of SEQ ID NO:126, wherein X is any amino acid, a buffer comprising about 10 mM to about 100 mM histidine, about 5% to about 15% (weight / volume) sucrose, about 0.01% to about 0.1% (weight / volume (w / v)) polysorbate-80, and about 50 mM to 200 mM NaCl, wherein the pH of the formulation is about 5.0 to about 7.4.

[0021] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises the heavy chain CDR3 of SEQ ID NO:126, wherein X is any amino acid, a buffer comprising about 20 mM histidine, about 10% (weight / volume (w / v)) sucrose, about 0.03% (weight / volume (w / v)) polysorbate-80, and about 100 mM NaCl, wherein the pH of the formulation is about 6.0.

[0022] In any of the foregoing or related aspects of the present disclosure, the anti-CD137 antibody comprises the heavy chain CDR3 of DXPFXLDXXYYYYYX (SEQ ID NO:127), wherein X is any amino acid. In any of the foregoing or related aspects of the present disclosure, the anti-CD137 antibody comprises DX1X2X3X4LX5X6X7X8YX9YYX 10 (SEQ ID NO:128) of the heavy chain CDR3, wherein X1 is any amino acid, wherein X2 is a nonpolar amino acid, wherein X3 is a nonpolar amino acid, wherein X4 is any amino acid, wherein X5 is a polar amino acid, wherein X6 is any amino acid, wherein X7 is any amino acid, wherein X8 is a polar amino acid, wherein X9 is a polar amino acid, and wherein X 10 is any amino acid. In any of the foregoing or related aspects of the present disclosure, X2 is proline, X3 is phenylalanine or tryptophan, X5 is aspartic acid or glutamic acid, X8 is tyrosine, and X9 is tyrosine.

[0023] In any of the foregoing or related aspects of the present disclosure, the anti-CD137 antibody of the formulation of the present disclosure comprises the heavy chain CDR3 of SEQ ID NO:68.

[0024] In any of the foregoing or related aspects of the present disclosure, the anti-CD137 antibody comprises a heavy chain CDR1 of SEQ ID NO:48, a heavy chain CDR2 of SEQ ID NO:56, and a heavy chain CDR3 of SEQ ID NO:68, and a light chain CDR1 of SEQ ID NO:69, a light chain CDR2 of SEQ ID NO:78, and a light chain CDR3 of SEQ ID NO:89.

[0025] In any of the foregoing or related aspects of the present disclosure, the anti-CD137 antibody comprises a heavy chain CDR1 of SEQ ID NO:51, a heavy chain CDR2 of SEQ ID NO:108, and a heavy chain CDR3 of SEQ ID NO:68, and a light chain CDR1 of SEQ ID NO:69, a light chain CDR2 of SEQ ID NO:78, and a light chain CDR3 of SEQ ID NO:89.

[0026] In any of the foregoing or related aspects of the present disclosure, the anti-CD137 antibody comprises heavy and light chain sequences that each contain an amino acid sequence having at least 90% identity to SEQ ID NOs: 4 and 6, respectively. In some aspects, the anti-CD137 antibody comprises heavy and light chain sequences having the amino acid sequences as set forth in SEQ ID NOs: 4 and 6, respectively.

[0027] In any of the foregoing or related aspects of the present disclosure, the anti-CD137 antibody comprises heavy and light chain sequences that each contain an amino acid sequence having at least 90% identity to SEQ ID NOs: 101 and 6, respectively. In some aspects, the anti-CD137 antibody comprises heavy and light chain sequences having the amino acid sequences as set forth in SEQ ID NOs: 101 and 6, respectively.

[0028] In any of the foregoing or related aspects of the present disclosure, the antibody comprises an IgG1 heavy chain constant region. In some aspects, the IgG1 heavy chain constant region is a wild-type human IgG1 heavy chain constant region. In some aspects, the IgG1 heavy chain constant region comprises an amino acid substitution relative to the wild-type human IgG1 heavy chain constant region.

[0029] In any of the foregoing or related aspects of the present disclosure, the antibody comprises an IgG4 heavy chain constant region. In some aspects, the IgG4 heavy chain constant region is a wild-type human IgG4 heavy chain constant region. In some aspects, the IgG4 heavy chain constant region comprises an amino acid substitution relative to the wild-type human IgG4 heavy chain constant region.

[0030] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NOs: 48, 56, and 68, respectively, and light chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NOs: 69, 78, and 89, respectively, and a buffer comprising histidine.

[0031] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NOs: 135, 139, and 143, respectively, and light chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NOs: 144, 147, and 150, respectively, and a buffer comprising histidine.

[0032] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NOs: 48, 154, and 159, respectively, and light chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NOs: 144, 147, and 150, respectively, and a buffer comprising histidine.

[0033] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NOs: 48, 56, and 68, respectively, and light chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NOs: 69, 78, and 89, respectively, a buffer comprising histidine, and a disaccharide.

[0034] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NOs: 135, 139, and 143, respectively, and light chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NOs: 144, 147, and 150, respectively, a buffer comprising histidine, and a disaccharide.

[0035] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NO: 48, 154, and 159, respectively, and light chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NO: 144, 147, and 150, respectively, a buffer comprising histidine, and a disaccharide.

[0036] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NO: 48, 56, and 68, respectively, and light chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NO: 69, 78, and 89, respectively, and a buffer comprising histidine, wherein the formulation has a pH of about 5.0 - 7.0.

[0037] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NO: 135, 139, and 143, respectively, and light chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NO: 144, 147, and 150, respectively, and a buffer comprising histidine, wherein the formulation has a pH of about 5.0 - 7.0.

[0038] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NO: 48, 154, and 159, respectively, and light chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NO: 144, 147, and 150, respectively, and a buffer comprising histidine, wherein the formulation has a pH of about 5.0 - 7.0.

[0039] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NO: 48, 56, and 68, respectively, and light chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NO: 69, 78, and 89, respectively, a buffer comprising histidine, a disaccharide, a nonionic surfactant, and a salt, wherein the pH of the formulation is from about 5.0 to about 7.0.

[0040] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NO: 135, 139, and 143, respectively, and light chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NO: 144, 147, and 150, respectively, a buffer comprising histidine, a disaccharide, a nonionic surfactant, and a salt, wherein the pH of the formulation is from about 5.0 to about 7.0.

[0041] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NO: 48, 154, and 159, respectively, and light chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NO: 144, 147, and 150, respectively, a buffer comprising histidine, a disaccharide, a nonionic surfactant, and a salt, wherein the pH of the formulation is from about 5.0 to about 7.0.

[0042] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NO: 48, 56, and 68, respectively, and light chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NO: 69, 78, and 89, respectively, a buffer comprising histidine, about 5% to about 15% (weight / volume) of a disaccharide, about 0.01% to about 0.1% (weight / volume (w / v)) of a nonionic surfactant, and about 50 mM to 200 mM of a salt, wherein the pH of the formulation is from about 5.0 to about 7.0.

[0043] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy chain CDR1, CDR2, and CDR3 sequences as shown in SEQ ID NO: 135, 139, and 143, respectively, and light chain CDR1, CDR2, and CDR3 sequences as shown in SEQ ID NO: 144, 147, and 150, respectively, a buffer comprising histidine, about 5% to about 15% (weight / volume) of a disaccharide, about 0.01% to about 0.1% (weight / volume (w / v)) of a non-ionic surfactant, and about 50 mM to 200 mM of a salt, wherein the pH of the formulation is about 5.0 to about 7.0.

[0044] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy chain CDR1, CDR2, and CDR3 sequences as shown in SEQ ID NO: 48, 154, and 159, respectively, and light chain CDR1, CDR2, and CDR3 sequences as shown in SEQ ID NO: 144, 147, and 150, respectively, a buffer comprising histidine, about 5% to about 15% (weight / volume) of a disaccharide, about 0.01% to about 0.1% (weight / volume (w / v)) of a non-ionic surfactant, and about 50 mM to 200 mM of a salt, wherein the pH of the formulation is about 5.0 to about 7.0.

[0045] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy chain CDR1, CDR2, and CDR3 sequences as shown in SEQ ID NO: 48, 56, and 68, respectively, and light chain CDR1, CDR2, and CDR3 sequences as shown in SEQ ID NO: 69, 78, and 89, respectively, a buffer comprising about 10 mM to about 100 mM of histidine, about 5% to about 15% (weight / volume) of sucrose, about 0.01% to about 0.1% (weight / volume (w / v)) of polysorbate-80, and about 50 mM to 200 mM of NaCl, wherein the pH of the formulation is about 5.0 to about 7.0.

[0046] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NO: 135, 139, and 143, respectively, and light chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NO: 144, 147, and 150, respectively, a buffer comprising about 10 mM to about 100 mM histidine, about 5% to about 15% (weight / volume) sucrose, about 0.01% to about 0.1% (weight / volume (w / v)) polysorbate-80, and about 50 mM to 200 mM NaCl, wherein the pH of the formulation is about 5.0 to about 7.0.

[0047] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NO: 48, 154, and 159, respectively, and light chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NO: 144, 147, and 150, respectively, a buffer comprising about 10 mM to about 100 mM histidine, about 5% to about 15% (weight / volume) sucrose, about 0.01% to about 0.1% (weight / volume (w / v)) polysorbate-80, and about 50 mM to 200 mM NaCl, wherein the pH of the formulation is about 5.0 to about 7.0.

[0048] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NO: 48, 56, and 68, respectively, and light chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NO: 69, 78, and 89, respectively, and a buffer comprising histidine, wherein the formulation has a pH of about 5.0 - 7.4.

[0049] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NO: 135, 139, and 143, respectively, and light chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NO: 144, 147, and 150, respectively, and a buffer comprising histidine, wherein the formulation has a pH of about 5.0 - 7.4.

[0050] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy chain CDR1, CDR2, and CDR3 sequences as shown in SEQ ID NO: 48, 154, and 159, respectively, and light chain CDR1, CDR2, and CDR3 sequences as shown in SEQ ID NO: 144, 147, and 150, respectively, and a buffer comprising histidine, wherein the formulation has a pH of about 5.0 - 7.4.

[0051] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy chain CDR1, CDR2, and CDR3 sequences as shown in SEQ ID NO: 48, 56, and 68, respectively, and light chain CDR1, CDR2, and CDR3 sequences as shown in SEQ ID NO: 69, 78, and 89, respectively, a buffer comprising histidine, a disaccharide, a nonionic surfactant, and a salt, wherein the pH of the formulation is about 5.0 to about 7.4.

[0052] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy chain CDR1, CDR2, and CDR3 sequences as shown in SEQ ID NO: 135, 139, and 143, respectively, and light chain CDR1, CDR2, and CDR3 sequences as shown in SEQ ID NO: 144, 147, and 150, respectively, a buffer comprising histidine, a disaccharide, a nonionic surfactant, and a salt, wherein the pH of the formulation is about 5.0 to about 7.4.

[0053] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy chain CDR1, CDR2, and CDR3 sequences as shown in SEQ ID NO: 48, 154, and 159, respectively, and light chain CDR1, CDR2, and CDR3 sequences as shown in SEQ ID NO: 144, 147, and 150, respectively, a buffer comprising histidine, a disaccharide, a nonionic surfactant, and a salt, wherein the pH of the formulation is about 5.0 to about 7.4.

[0054] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy chain CDR1, CDR2, and CDR3 sequences as shown in SEQ ID NO: 48, 56, and 68, respectively, and light chain CDR1, CDR2, and CDR3 sequences as shown in SEQ ID NO: 69, 78, and 89, respectively, a buffer comprising histidine, about 5% to about 15% (weight / volume) of a disaccharide, about 0.01% to about 0.1% (weight / volume (w / v)) of a nonionic surfactant, and about 50 mM to 200 mM of a salt, wherein the pH of the formulation is about 5.0 to about 7.4.

[0055] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy chain CDR1, CDR2, and CDR3 sequences as shown in SEQ ID NO: 135, 139, and 143, respectively, and light chain CDR1, CDR2, and CDR3 sequences as shown in SEQ ID NO: 144, 147, and 150, respectively, a buffer comprising histidine, about 5% to about 15% (weight / volume) of a disaccharide, about 0.01% to about 0.1% (weight / volume (w / v)) of a nonionic surfactant, and about 50 mM to 200 mM of a salt, wherein the pH of the formulation is about 5.0 to about 7.4.

[0056] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy chain CDR1, CDR2, and CDR3 sequences as shown in SEQ ID NO: 48, 154, and 159, respectively, and light chain CDR1, CDR2, and CDR3 sequences as shown in SEQ ID NO: 144, 147, and 150, respectively, a buffer comprising histidine, about 5% to about 15% (weight / volume) of a disaccharide, about 0.01% to about 0.1% (weight / volume (w / v)) of a nonionic surfactant, and about 50 mM to 200 mM of a salt, wherein the pH of the formulation is about 5.0 to about 7.4.

[0057] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NO: 48, 56, and 68, respectively, and light chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NO: 69, 78, and 89, respectively, a buffer comprising about 10 mM to about 100 mM histidine, about 5% to about 15% (weight / volume) sucrose, about 0.01% to about 0.1% (weight / volume (w / v)) polysorbate-80, and about 50 mM to 200 mM NaCl, wherein the pH of the formulation is about 5.0 to about 7.4.

[0058] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NO: 135, 139, and 143, respectively, and light chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NO: 144, 147, and 150, respectively, a buffer comprising about 10 mM to about 100 mM histidine, about 5% to about 15% (weight / volume) sucrose, about 0.01% to about 0.1% (weight / volume (w / v)) polysorbate-80, and about 50 mM to 200 mM NaCl, wherein the pH of the formulation is about 5.0 to about 7.4.

[0059] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NO: 48, 154, and 159, respectively, and light chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NO: 144, 147, and 150, respectively, a buffer comprising about 10 mM to about 100 mM histidine, about 5% to about 15% (weight / volume) sucrose, about 0.01% to about 0.1% (weight / volume (w / v)) polysorbate-80, and about 50 mM to 200 mM NaCl, wherein the pH of the formulation is about 5.0 to about 7.4.

[0060] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NOs: 48, 56, and 68, respectively, and light chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NOs: 69, 78, and 89, respectively, a buffer comprising about 20 mM histidine, about 10% (weight / volume (w / v)) sucrose, about 0.03% (weight / volume (w / v)) polysorbate-80, and about 100 mM NaCl, wherein the pH of the formulation is about 6.0.

[0061] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NOs: 135, 139, and 143, respectively, and light chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NOs: 144, 147, and 150, respectively, a buffer comprising about 20 mM histidine, about 10% (weight / volume (w / v)) sucrose, about 0.03% (weight / volume (w / v)) polysorbate-80, and about 100 mM NaCl, wherein the pH of the formulation is about 6.0.

[0062] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NOs: 48, 154, and 159, respectively, and light chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NOs: 144, 147, and 150, respectively, a buffer comprising about 20 mM histidine, about 10% (weight / volume (w / v)) sucrose, about 0.03% (weight / volume (w / v)) polysorbate-80, and about 100 mM NaCl, wherein the pH of the formulation is about 6.0.

[0063] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy chain and light chain variable sequences as set forth in SEQ ID NOs: 4 and 6, respectively, and a buffer comprising histidine.

[0064] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy and light chain variable sequences as shown in SEQ ID NO: 4 and 6 respectively, a buffer comprising histidine, and a disaccharide.

[0065] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy and light chain variable sequences as shown in SEQ ID NO: 4 and 6 respectively, and a buffer comprising histidine, wherein the formulation has a pH of about 5.0 - 7.0.

[0066] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy and light chain variable sequences as shown in SEQ ID NO: 4 and 6 respectively, a buffer comprising histidine, a disaccharide, a non-ionic surfactant, and a salt, wherein the pH of the formulation is about 5.0 to about 7.0.

[0067] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy and light chain variable sequences as shown in SEQ ID NO: 4 and 6 respectively, a buffer comprising histidine, about 5% to about 15% (weight / volume) of a disaccharide, about 0.01% to about 0.1% (weight / volume (w / v)) of a non-ionic surfactant, and about 50 mM to 200 mM of a salt, wherein the pH of the formulation is about 5.0 to about 7.0.

[0068] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy and light chain variable sequences as shown in SEQ ID NO: 4 and 6 respectively, a buffer comprising about 10 mM to about 100 mM of histidine, about 5% to about 15% (weight / volume) of sucrose, about 0.01% to about 0.1% (weight / volume (w / v)) of polysorbate-80, and about 50 mM to 200 mM of NaCl, wherein the pH of the formulation is about 5.0 to about 7.0.

[0069] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy and light chain variable sequences as shown in SEQ ID NO: 4 and 6 respectively, and a buffer comprising histidine, wherein the formulation has a pH of about 5.0 - 7.4.

[0070] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy and light chain variable sequences as shown in SEQ ID NOs: 4 and 6, respectively, a buffer comprising histidine, a disaccharide, a nonionic surfactant, and a salt, wherein the pH of the formulation is from about 5.0 to about 7.4.

[0071] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy and light chain variable sequences as shown in SEQ ID NOs: 4 and 6, respectively, a buffer comprising histidine, about 5% to about 15% (weight / volume) of a disaccharide, about 0.01% to about 0.1% (weight / volume (w / v)) of a nonionic surfactant, and about 50 mM to 200 mM of a salt, wherein the pH of the formulation is from about 5.0 to about 7.4.

[0072] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy and light chain variable sequences as shown in SEQ ID NOs: 4 and 6, respectively, a buffer comprising about 10 mM to about 100 mM of histidine, about 5% to about 15% (weight / volume) of sucrose, about 0.01% to about 0.1% (weight / volume (w / v)) of polysorbate-80, and about 50 mM to 200 mM of NaCl, wherein the pH of the formulation is from about 5.0 to about 7.4. In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy and light chain amino acid sequences as shown in SEQ ID NOs: 129 and 133, respectively, and a buffer comprising histidine.

[0073] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises heavy and light chain amino acid sequences as shown in SEQ ID NOs: 129 and 133, respectively, and a buffer comprising histidine.

[0074] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises the heavy and light chain amino acid sequences shown in SEQ ID NOs: 129 and 133, a buffer comprising histidine, and a disaccharide.

[0075] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises the heavy and light chain amino acid sequences shown in SEQ ID NOs: 129 and 133, and a buffer comprising histidine, wherein the formulation has a pH of about 5.0 - 7.0.

[0076] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises the heavy and light chain amino acid sequences shown in SEQ ID NOs: 129 and 133, a buffer comprising histidine, a disaccharide, a nonionic surfactant, and a salt, wherein the pH of the formulation is about 5.0 to about 7.0.

[0077] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises the heavy and light chain amino acid sequences shown in SEQ ID NOs: 129 and 133, a buffer comprising histidine, about 5% to about 15% (weight / volume) of a disaccharide, about 0.01% to about 0.1% (weight / volume (w / v)) of a nonionic surfactant, and about 50 mM to 200 mM of a salt, wherein the pH of the formulation is about 5.0 to about 7.0.

[0078] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises the heavy and light chain amino acid sequences shown in SEQ ID NOs: 129 and 133, a buffer comprising about 10 mM to about 100 mM of histidine, about 5% to about 15% (weight / volume) of sucrose, about 0.01% to about 0.1% (weight / volume (w / v)) of polysorbate-80, and about 50 mM to 200 mM of NaCl, wherein the pH of the formulation is about 5.0 to about 7.0.

[0079] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises the heavy and light chain amino acid sequences as set forth in SEQ ID NOs: 129 and 133, and a buffer comprising histidine, wherein the formulation has a pH of about 5.0 - 7.4.

[0080] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises the heavy and light chain amino acid sequences as set forth in SEQ ID NOs: 129 and 133, a buffer comprising histidine, a disaccharide, a non-ionic surfactant, and a salt, wherein the pH of the formulation is about 5.0 to about 7.4.

[0081] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises the heavy and light chain amino acid sequences as set forth in SEQ ID NOs: 129 and 133, a buffer comprising histidine, about 5% to about 15% (weight / volume) of a disaccharide, about 0.01% to about 0.1% (weight / volume (w / v)) of a non-ionic surfactant, and about 50 mM to 200 mM of a salt, wherein the pH of the formulation is about 5.0 to about 7.4.

[0082] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises the heavy and light chain amino acid sequences as set forth in SEQ ID NOs: 129 and 133, a buffer comprising about 10 mM to about 100 mM of histidine, about 5% to about 15% (weight / volume) of sucrose, about 0.01% to about 0.1% (weight / volume (w / v)) of polysorbate-80, and about 50 mM to 200 mM of NaCl, wherein the pH of the formulation is about 5.0 to about 7.4.

[0083] In some aspects, the present disclosure provides a formulation comprising: an anti-CD137 antibody at a concentration of about 1 mg / ml to about 100 mg / ml, wherein the anti-CD137 antibody comprises the heavy and light chain amino acid sequences as set forth in SEQ ID NOs: 129 and 133, a buffer comprising about 20 mM of histidine, about 10% (weight / volume (w / v)) of sucrose, about 0.03% (weight / volume (w / v)) of polysorbate-80, and about 100 mM of NaCl, wherein the pH of the formulation is about 6.0.

[0084] In any of the foregoing or related aspects of the present disclosure, the formulations of the present disclosure contain histidine. In some aspects, the formulations of the present disclosure contain from about 10 mM histidine to about 100 mM histidine. In some aspects, the formulation contains about 20 mM histidine.

[0085] In any of the foregoing or related aspects of the present disclosure, the formulations of the present disclosure contain a disaccharide. In some embodiments, the disaccharide is selected from sucrose, lactose, maltose, and trehalose. In some embodiments, the disaccharide is sucrose. In some embodiments, the formulations of the present disclosure contain from about 5% to about 15% (weight / volume) of the disaccharide. In some embodiments, the formulations of the present disclosure contain about 10% (weight / volume) of the disaccharide.

[0086] In any of the foregoing or related aspects of the present disclosure, the formulations of the present disclosure contain a salt. In some aspects, the salt is NaCl. In some aspects, the formulations of the present disclosure contain the salt at a concentration of about 50 mM to 200 mM. In some aspects, the formulations of the present disclosure contain the salt at a concentration of about 100 mM.

[0087] In any of the foregoing or related aspects of the present disclosure, the formulations of the present disclosure have a pH of about 5.0 - 7.0. In some embodiments, the formulations of the present disclosure have a pH of about 6.0.

[0088] In any of the foregoing or related aspects of the present disclosure, the formulations of the present disclosure have a pH of about 5.0 - 8.0. In some embodiments, the formulations of the present disclosure have a pH of about 5.0 - 7.4.

[0089] In any of the foregoing or related aspects of the present disclosure, the formulations of the present disclosure contain a nonionic surfactant. In some aspects, the nonionic surfactant is polysorbate. In some aspects, the polysorbate is polysorbate - 80. In some aspects, the formulations of the present disclosure contain from about 0.01% to about 0.1% (weight / volume (w / v)) of the nonionic surfactant. In some aspects, the formulations of the present disclosure contain about 0.03% (weight / volume (w / v)) of the nonionic surfactant.

[0090] In any of the foregoing or related aspects of the present disclosure, the formulations of the present disclosure comprise an anti-CD137 antibody at a concentration of from about 1 mg / ml to about 100 mg / ml. In some aspects, the formulations of the present disclosure comprise an anti-CD137 antibody at a concentration of from about 5 mg / ml to about 15 mg / ml. In some aspects, the formulations of the present disclosure comprise an anti-CD137 antibody at a concentration of from about 15 mg / ml to about 30 mg / ml. In some aspects, the formulations of the present disclosure comprise an anti-CD137 antibody at a concentration of from about 30 mg / ml to about 45 mg / ml. In some aspects, the formulations of the present disclosure comprise an anti-CD137 antibody at a concentration of from about 45 mg / ml to about 60 mg / ml. In some aspects, the formulations of the present disclosure comprise an anti-CD137 antibody at a concentration of from about 60 mg / ml to about 75 mg / ml. In some aspects, the formulations of the present disclosure comprise an anti-CD137 antibody at a concentration of from about 75 mg / ml to about 90 mg / ml. In some aspects, the formulations of the present disclosure comprise an anti-CD137 antibody at a concentration of from about 85 mg / ml to about 100 mg / ml. In some aspects, the formulations of the present disclosure comprise an anti-CD137 antibody at a concentration of about 5 mg / ml. In some aspects, the formulations of the present disclosure comprise an anti-CD137 antibody at a concentration of about 10 mg / ml. In some aspects, the formulations of the present disclosure comprise an anti-CD137 antibody at a concentration of about 15 mg / ml. In some aspects, the formulations of the present disclosure comprise an anti-CD137 antibody at a concentration of about 20 mg / ml.

[0091] In some aspects, the present disclosure provides a method of inducing or enhancing the dimerization of human CD137 trimers in a subject, comprising administering to a subject in need thereof an effective amount of a formulation of the present disclosure.

[0092] In some aspects, the present disclosure provides a method of inducing or enhancing the multimerization of human CD137 trimers in a subject, comprising administering to a subject in need thereof an effective amount of a formulation of the present disclosure.

[0093] In some aspects, the present disclosure provides a method of inducing or enhancing T cell activation in a subject, comprising administering to a subject in need thereof an effective amount of a formulation of the present disclosure. In some aspects, T cell activation occurs in the tumor microenvironment.

[0094] In some aspects, the present disclosure provides a method of inducing or enhancing a cytotoxic T cell response in a subject, comprising administering to a subject in need thereof an effective amount of a formulation of the present disclosure. In some aspects, the cytotoxic T cell response occurs in the tumor microenvironment.

[0095] In some aspects, the present disclosure provides a method of inducing or enhancing cytokine production in immune cells in a subject, comprising administering to a subject in need thereof an effective amount of a preparation of the present disclosure. In some aspects, the cytokines produced are IL-2, TNFα, IL-13, IFN-γ, or a combination thereof. In some aspects, cytokine production occurs in the tumor microenvironment.

[0096] In some aspects, the present disclosure provides a method of inducing or enhancing T cell proliferation in a subject, comprising administering to a subject in need thereof an effective amount of a preparation of the present disclosure. In some aspects, T cell proliferation occurs in the tumor microenvironment.

[0097] In some aspects, the present disclosure provides a method of reducing or inhibiting tumor growth, comprising administering to a subject in need thereof an effective amount of a preparation of the present disclosure.

[0098] In some aspects, the present disclosure provides a method of treating a disorder mediated by human CD137 in a subject, comprising administering to a subject in need thereof an effective amount of a preparation of the present disclosure.

[0099] In some aspects, the present disclosure provides a method of treating cancer in a subject, comprising administering to a subject in need thereof an effective amount of a preparation of the present disclosure.

[0100] In any of the foregoing aspects, after administration of the preparation, infiltration of immune cells into the tumor microenvironment is increased. In some aspects, the immune cells express CD45.

[0101] In any of the foregoing aspects, after administration of the preparation, the number of regulatory T (Treg) cells in the tumor microenvironment is reduced. In some aspects, the Treg cells express CD4, FOXP-3, and CD25.

[0102] In any of the foregoing aspects, after administration of the isolated monoclonal antibody or antigen-binding portion, the number of macrophages in the tumor microenvironment is reduced. In some aspects, the macrophages express CD45 and CD11b.

[0103] In any of the foregoing aspects, after administration of the isolated monoclonal antibody or antigen-binding portion, T cell exhaustion in the tumor microenvironment is reduced. In some aspects, the reduction of T cell exhaustion includes a reduction in the expression of TIGIT, PD-1, LAG-3, or a combination thereof.

[0104] In any of the foregoing aspects, the cancer is selected from the group consisting of melanoma, glioma, renal cancer, breast cancer, blood cancer, and head and neck cancer. In some aspects, the blood cancer is B cell lymphoma.

[0105] In some aspects, the present disclosure provides a method of inducing an anti-tumor memory immune response, comprising administering to a subject in need thereof an effective amount of a preparation of the present disclosure.

[0106] In any of the foregoing aspects, the anti-CD137 antibody binds to the Fcγ receptor.

[0107] In any of the foregoing aspects, depletion of CD4+ T cells, CD8+ T cells, natural killer cells, or a combination thereof reduces the efficacy of the preparation.

[0108] In some aspects, the present disclosure provides a kit comprising a container containing a preparation of the present disclosure and a package insert containing instructions for administering the preparation for treating or delaying the development of cancer in a subject in need thereof, or for reducing or inhibiting tumor growth in a subject in need thereof.

[0109] In some aspects, the present disclosure provides a kit comprising a container containing a preparation of the present disclosure and a package insert containing instructions for administering the preparation alone or in combination with another agent for treating or delaying the development of cancer in a subject in need thereof, or for reducing or inhibiting tumor growth in a subject in need thereof.

[0110] In some aspects, the present disclosure provides the use of a preparation of the present disclosure in the manufacture of a medicament for treating or delaying the development of cancer in a subject in need thereof, or for reducing or inhibiting tumor growth in a subject in need thereof.

[0111] In some aspects, the present disclosure provides a preparation of the present disclosure for use in the manufacture of a medicament for treating or delaying the development of cancer in a subject in need thereof, or for reducing or inhibiting tumor growth in a subject in need thereof.

[0112] In some aspects, the present disclosure provides a preparation of the present disclosure for use as a medicament.

[0113] In view of the foregoing, in some aspects, the preparation described herein comprises an isolated monoclonal antibody or an antigen-binding portion thereof that specifically binds to human CD137, wherein the antibody or antigen-binding portion binds to human CD137 with an affinity (K D ) of about 40 nM to about 100 nM. In some aspects, the present disclosure provides an isolated monoclonal antibody or an antigen-binding portion thereof that specifically binds to human CD137, wherein the antibody or antigen-binding portion binds to human CD137 with an affinity (K D)Binds to human CD137. In some aspects, the anti-CD137 antibody has at least twice the affinity for human CD137 as mAb10 has for murine CD137 (e.g., at least three-fold, four-fold, five-fold, six-fold, seven-fold, eight-fold, nine-fold, or ten-fold). In some aspects, the affinity of the anti-CD137 antibody is not higher than 500 nM, 450 nM, 400 nM, 350 nM, 300 nM, 250 nM, 200 nM, 250 nM, 200 nM, 175 nM, 150 nM, 125 nM, 110 nM, or 100 nM. In some aspects, the anti-CD137 antibody has at least twice the affinity for human CD137 as mAb10 has for murine CD137 (e.g., at least three-fold, four-fold, five-fold, six-fold, seven-fold, eight-fold, nine-fold, or ten-fold), but is not higher than 500 nM, 450 nM, 400 nM, 350 nM, 300 nM, 250 nM, 200 nM, 250 nM, 200 nM, 175 nM, 150 nM, 125 nM, 110 nM, or 100 nM.

[0114] In some aspects, the formulations described herein comprise an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, wherein the antibody or antigen-binding portion binds to an epitope on human CD137, and the epitope comprises one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or all 25) of amino acids 111-132 of SEQ ID NO:3. In some aspects, the present disclosure provides an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, wherein the antibody or antigen-binding portion binds to an epitope within amino acids 111-132 of SEQ ID NO:3. In some aspects, the present disclosure provides an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, wherein the antibody or antigen-binding portion binds to all or part of amino acids 111-132 of SEQ ID NO:3. In some aspects, the epitope comprises K114 of SEQ ID NO:3. In some aspects, the epitope comprises residues E111, T113, and K114 of SEQ ID NO:3. In some aspects, the epitope comprises residues E111, T113, K114, N126, and I132 of SEQ ID NO:3. In some aspects, the epitope comprises residues E111, T113, K114, and P135 of SEQ ID NO:3. In some aspects, the epitope comprises residues E111, T113, K114, N126, I132, and P135 of SEQ ID NO:3. In some aspects, the antibody or antigen-binding portion thereof binds to human CD137 with an affinity between about 30 nM and about 100 nM (e.g., between about 30 nM and about 110 nM).

[0115] In some aspects, the formulations described herein comprise an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, wherein the antibody or antigen-binding portion binds to human CD137 with an affinity (K D ) between about 40 - 100 nM (e.g., between about 40 nM and about 100 nM) and binds to an epitope on human CD137, and the epitope comprises K114 of SEQ ID NO:3. In some aspects, the present disclosure provides an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, wherein the antibody or antigen-binding portion binds to human CD137 with an affinity (K D) binds to human CD137 and binds to an epitope on human CD137, said epitope comprising K114 of SEQ ID NO:3. In some aspects, the epitope comprises residues E111, T113, and K114 of SEQ ID NO:3. In some aspects, the epitope comprises residues E111, T113, K114, N126, and I132 of SEQ ID NO:3. In some aspects, the epitope comprises residues E111, T113, K114, and P135 of SEQ ID NO:3. In some aspects, the epitope comprises residues E111, T113, K114, N126, I132, and P135 of SEQ ID NO:3.

[0116] In some aspects, the formulations described herein comprise an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, wherein the antibody or antigen-binding portion has an affinity (K D ) binds to human CD137 and binds to an epitope on human CD137, said epitope comprising a sequence of one or more amino acid residues corresponding to amino acid positions 111 to 135 of SEQ ID NO:3. In some aspects, the epitope comprises 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 amino acids corresponding to amino acid positions 111 to 135 of SEQ ID NO:3.

[0117] In some aspects, the formulations described herein comprise an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, wherein the antibody or antigen-binding portion has an affinity (K D ) binds to human CD137 and binds to an epitope on human CD137, said epitope being within amino acid residues 111 - 135 of SEQ ID NO:3. In some aspects, the epitope is at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 amino acids. In some aspects, the epitope is less than 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, or 5 amino acids.

[0118] In some aspects, the formulations described herein comprise an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, wherein the antibody or antigen-binding portion binds to human CD137 with an affinity (K D ) of about 30 - 100 nM (e.g., between about 30 nM and about 100 nM), and binds to an epitope on human CD137 that comprises ELTK (corresponding to amino acid residues 111 - 114 of SEQ ID NO:3). In some aspects, the epitope further comprises one or more of residues N126, I132, and P135 of SEQ ID NO:3.

[0119] In any of the foregoing aspects, the epitope is a non-linear epitope. In any of the foregoing aspects, mutation of residue K114 of SEQ ID NO:3 abolishes the binding of the antibody or its antigen-binding portion to human CD137.

[0120] In any of the foregoing aspects, the antibody or antigen-binding portion described herein binds to human CD137 with an affinity (K D ) of about 30 - 100 nM, 30 - 95 nM, 45 - 95 nM, 50 - 90 nM, 55 - 85 nM, 60 - 80 nM, 65 - 75 nM, 55 - 75 nM, 40 - 70 nM, 50 - 80 nM, or 60 - 90 nM. In some aspects, the antibody or antigen-binding portion binds to the non-ligand-binding region of the extracellular domain of human CD137. In some aspects, the antibody or antigen-binding portion does not inhibit the interaction between CD137 and CD137L. In some aspects, the non-ligand-binding region spans cysteine rich domain (CRD) III and CRDIV. In any of the foregoing aspects, the antibody or antigen-binding portion does not inhibit the formation of trimers of CD137:CD137L monomers.

[0121] In some aspects, the formulations described herein comprise an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, wherein the antibody or antigen-binding portion:

[0122] (i) binds to human CD137 with an affinity (K D ) of about 30 - 100 nM (e.g., between about 30 nM and about 100 nM);

[0123] (ii) binds to the non-ligand-binding region of the extracellular domain of human CD137; and

[0124] (iii) binds to an epitope on human CD137 that comprises K114 of SEQ ID NO:3.

[0125] In some aspects, the formulations described herein comprise an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, wherein the antibody or antigen-binding portion:

[0126] (i) binds to human CD137 with an affinity (K D ) of about 30 - 100 nM (e.g., between about 30 nM and about 100 nM);

[0127] (ii) does not inhibit the interaction between human CD137 and the human CD137 ligand; and

[0128] (iii) binds to an epitope on human CD137 that comprises K114 of SEQ ID NO:3.

[0129] In some aspects, the formulations described herein comprise an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, wherein the antibody or antigen-binding portion: (i) binds to human CD137 with an affinity (K D ) of about 30 - 100 nM, and (ii) does not inhibit the formation of the trimer of the CD137:CD137L monomer (i.e., the CD137:CD137L trimer:trimer complex). In some aspects, the present disclosure features the following: an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, wherein the antibody or antigen-binding portion: (i) binds to human CD137 with an affinity (K D ) of about 30 - 100 nM, and (ii) binds to a non-ligand-binding region of the extracellular domain of human CD137. In some aspects, the present disclosure features an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, wherein the antibody or antigen-binding portion: (i) binds to human CD137 with an affinity (K D ) of about 30 - 100 nM, and (ii) does not inhibit the interaction between human CD137 and the CD137 ligand.

[0130] In any of the foregoing aspects, the antibody or antigen-binding portion comprises a heavy chain CDR3 containing the amino acid sequence DXXXXLXXXXYXYYX (SEQ ID NO: 126), where X is any amino acid. In some aspects, the antibody or antigen-binding portion comprises a heavy chain CDR3 containing the amino acid sequence DXPFXLDXXYYYYYX (SEQ ID NO: 127), where X is any amino acid. In any of the foregoing aspects, mutation of the residues D95, L100, Y100E, Y100G, Y100H or combinations thereof of the heavy chain CDR3 to alanine results in loss of binding to human CD137. In any of the foregoing aspects, mutation of the residues P97, F98, D100A, Y100D, Y100F or combinations thereof to alanine results in reduced binding to human CD137. In any of the foregoing aspects, the antibody or antigen-binding portion comprises heavy and light chain CDRs, wherein the heavy chain CDR3 comprises the amino acid sequence as set forth in SEQ ID NO: 68.

[0131] In other aspects, the formulations described herein comprise an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, wherein

[0132] (i) the antibody or antigen-binding portion binds to human CD137 with an affinity (K D ) of about 30 - 100 nM (e.g., between about 30 nM and about 100 nM); and

[0133] (ii) the antibody or antigen-binding portion comprises a heavy chain CDR3 containing the amino acid sequence DXXXXLXXXXYXYYX (SEQ ID NO: 126), where X is any amino acid. In some embodiments, X is any amino acid other than alanine.

[0134] In another aspect, the formulations described herein comprise an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, wherein

[0135] (i) the antibody or antigen-binding portion binds to human CD137 with an affinity (K D ) of about 30 - 100 nM (e.g., between about 30 nM and about 100 nM); and

[0136] (ii) the antibody or antigen-binding portion comprises a heavy chain CDR3 containing the amino acid sequence DX1X2X3X4LX5X6X7X8YX9YYX 10The heavy chain CDR3 of (SEQ ID NO:128), wherein X1 is any amino acid, wherein X2 is a non-polar amino acid, wherein X3 is a non-polar amino acid, wherein X4 is any amino acid, wherein X5 is a polar amino acid, wherein X6 is any amino acid, wherein X7 is any amino acid, wherein X8 is a polar amino acid, wherein X9 is a polar amino acid, and wherein X 10 is any amino acid. In some aspects, X2 is proline, X3 is phenylalanine or tryptophan, X5 is aspartic acid or glutamic acid, X8 is tyrosine, and X9 is tyrosine.

[0137] In other aspects, the formulations described herein comprise an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, wherein

[0138] (i) the antibody or antigen-binding portion binds to human CD137 with an affinity (K D ) of about 30 - 100 nM (e.g., between about 30 nM and about 100 nM); and

[0139] (ii) the antibody or its antigen-binding portion specifically binds to an epitope on human CD137 that comprises one or more of the residues E111, T113, K114, N126, I132, and P135 of SEQ ID NO:3.

[0140] In other aspects, the formulations described herein comprise an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, wherein

[0141] (i) the antibody or antigen-binding portion binds to human CD137 with an affinity (K D ) of about 30 - 100 nM (e.g., between about 30 nM and about 100 nM);

[0142] (ii) the antibody or its antigen-binding portion specifically binds to an epitope on human CD137 that comprises one or more of the residues E111, T113, K114, N126, I132, and P135 of SEQ ID NO:3;

[0143] (iii) the antibody or antigen-binding portion comprises a heavy chain CDR3 containing the amino acid sequence DXXXXLXXXXYXYYX (SEQ ID NO:126), wherein X is any amino acid; or

[0144] (iv) a combination of the above. In some aspects, X is any amino acid other than alanine.

[0145] In other aspects, the formulations described herein comprise an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, wherein

[0146] (i) The antibody or antigen-binding portion binds to human CD137 with an affinity (K D ) of about 30 - 100 nM (e.g., between about 30 nM and about 100 nM);

[0147] (ii) The antibody or its antigen-binding portion specifically binds to an epitope on human CD137, said epitope comprising one or more of the residues E111, T113, K114, N126, I132, and P135 of SEQ ID NO:3;

[0148] (iii) The antibody or antigen-binding portion comprises a heavy chain CDR3 containing the amino acid sequence DX1X2X3X4LX5X6X7X8YX9YYX 10 (SEQ ID NO:128), wherein X1 is any amino acid, wherein X2 is a nonpolar amino acid, wherein X3 is a nonpolar amino acid, wherein X4 is any amino acid, wherein X5 is a polar amino acid, wherein X6 is any amino acid, wherein X7 is any amino acid, wherein X8 is a polar amino acid, wherein X9 is a polar amino acid, and wherein X 10 is any amino acid; or

[0149] (iv) A combination of the above. In some aspects, X2 is proline, X3 is phenylalanine or tryptophan, X5 is aspartic acid or glutamic acid, X8 is tyrosine, and X9 is tyrosine.

[0150] In other aspects, the formulations described herein comprise an isolated monoclonal antibody or its antigen-binding portion that specifically binds to human CD137, wherein

[0151] (i) The antibody or antigen-binding portion binds to human CD137 with an affinity (K D ) of about 30 - 100 nM (e.g., between about 30 nM and about 100 nM);

[0152] (ii) The antibody or its antigen-binding portion specifically binds to an epitope on human CD137, said epitope comprising one or more of the residues E111, T113, K114, N126, I132, and P135 of SEQ ID NO:3; and

[0153] (iii) The antibody or antigen-binding portion comprises a heavy chain CDR3 containing the amino acid sequence DXXXXLXXXXYXYYX (SEQ ID NO:126), wherein X is any amino acid. In some aspects, X is any amino acid other than alanine.

[0154] In other aspects, the formulations described herein comprise an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, wherein

[0155] (i) the antibody or antigen-binding portion binds to human CD137 with an affinity (K D ) of about 30 - 100 nM (e.g., between about 30 nM and about 100 nM);

[0156] (ii) the antibody or its antigen-binding portion specifically binds to an epitope on human CD137, said epitope comprising one or more of the residues E111, T113, K114, N126, I132, and P135 of SEQ ID NO:3; and

[0157] (iii) the antibody or antigen-binding portion comprises a heavy chain CDR3 containing the amino acid sequence DX1X2X3X4LX5X6X7X8YX9YYX 10 (SEQ ID NO:128), wherein X1 is any amino acid, wherein X2 is a nonpolar amino acid, wherein X3 is a nonpolar amino acid, wherein X4 is any amino acid, wherein X5 is a polar amino acid, wherein X6 is any amino acid, wherein X7 is any amino acid, wherein X8 is a polar amino acid, wherein X9 is a polar amino acid, and wherein X 10 is any amino acid. In some aspects, X2 is proline, X3 is phenylalanine or tryptophan, X5 is aspartic acid or glutamic acid, X8 is tyrosine, and X9 is tyrosine.

[0158] In any of the foregoing aspects, the epitope comprises K114. In any of the foregoing aspects, the epitope comprises E111, T113, and K114. In any of the foregoing aspects, the epitope comprises E11, T113, K114, N126, and I132. In any of the foregoing aspects, the epitope comprises the residues E111, T113, K114, N126, I132, and P135 of SEQ ID NO:3.

[0159] In other aspects, the formulations described herein comprise an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, wherein

[0160] (i) the antibody or antigen-binding portion binds to human CD137 with an affinity (K D ) of about 30 - 100 nM (e.g., between about 30 nM and about 100 nM); and

[0161] (ii) the antibody or its antigen-binding portion specifically binds to an epitope that comprises a sequence of one or more amino acid residues corresponding to amino acid positions 111 to 135 of SEQ ID NO:3.

[0162] In other aspects, the formulations described herein comprise an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, wherein

[0163] (i) the antibody or antigen-binding portion binds to human CD137 with an affinity (K D ) of about 30 - 100 nM (e.g., between about 30 nM and about 100 nM);

[0164] (ii) the antibody or its antigen-binding portion specifically binds to an epitope comprising a sequence of one or more amino acid residues corresponding to amino acid positions 111 to 135 of SEQ ID NO:3;

[0165] (iii) the antibody or antigen-binding portion comprises a heavy chain CDR3 containing the amino acid sequence DXXXXLXXXXYXYYX (SEQ ID NO:126), wherein X is any amino acid; or

[0166] (iv) a combination of the above. In some aspects, X is any amino acid other than alanine.

[0167] In other aspects, the formulations described herein comprise an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, wherein

[0168] (i) the antibody or antigen-binding portion binds to human CD137 with an affinity (K D ) of about 30 - 100 nM (e.g., between about 30 nM and about 100 nM);

[0169] (ii) the antibody or its antigen-binding portion specifically binds to an epitope comprising a sequence of one or more amino acid residues corresponding to amino acid positions 111 to 135 of SEQ ID NO:3;

[0170] (iii) the antibody or antigen-binding portion comprises a heavy chain CDR3 containing the amino acid sequence DX1X2X3X4LX5X6X7X8YX9YYX 10 (SEQ ID NO:128), wherein X1 is any amino acid, wherein X2 is a non-polar amino acid, wherein X3 is a non-polar amino acid, wherein X4 is any amino acid, wherein X5 is a polar amino acid, wherein X6 is any amino acid, wherein X7 is any amino acid, wherein X8 is a polar amino acid, wherein X9 is a polar amino acid, and wherein X 10 is any amino acid; or

[0171] (iv) a combination of the above. In some aspects, X2 is proline, X3 is phenylalanine or tryptophan, X5 is aspartic acid or glutamic acid, X8 is tyrosine, and X9 is tyrosine.

[0172] In another aspect, the formulations described herein comprise an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, wherein

[0173] (i) the antibody or antigen-binding portion binds to human CD137 with an affinity (K D ) of about 30 - 100 nM (e.g., between about 30 nM and about 100 nM);

[0174] (ii) the antibody or antigen-binding portion thereof specifically binds to an epitope comprising a sequence of one or more amino acid residues corresponding to amino acid positions 111 to 135 of SEQ ID NO:3; and

[0175] (iii) the antibody or antigen-binding portion comprises a heavy chain CDR3 containing the amino acid sequence DXXXXLXXXXYXYYX (SEQ ID NO:126), wherein X is any amino acid. In some aspects, X is any amino acid other than alanine.

[0176] In another aspect, the formulations described herein comprise an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, wherein

[0177] (i) the antibody or antigen-binding portion binds to human CD137 with an affinity (K D ) of about 30 - 100 nM (e.g., between about 30 nM and about 100 nM);

[0178] (ii) the antibody or antigen-binding portion thereof specifically binds to an epitope comprising a sequence of one or more amino acid residues corresponding to amino acid positions 111 to 135 of SEQ ID NO:3; and

[0179] (iii) the antibody or antigen-binding portion comprises a heavy chain CDR3 containing the amino acid sequence DX1X2X3X4LX5X6X7X8YX9YYX 10 (SEQ ID NO:128), wherein X1 is any amino acid, wherein X2 is a nonpolar amino acid, wherein X3 is a nonpolar amino acid, wherein X4 is any amino acid, wherein X5 is a polar amino acid, wherein X6 is any amino acid, wherein X7 is any amino acid, wherein X8 is a polar amino acid, wherein X9 is a polar amino acid, and wherein X 10 is any amino acid. In some aspects, X2 is proline, X3 is phenylalanine or tryptophan, X5 is aspartic acid or glutamic acid, X8 is tyrosine, and X9 is tyrosine.

[0180] In any of the foregoing aspects, the epitope comprises 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 amino acid residues corresponding to amino acid positions 111 to 135 of SEQ ID NO:3.

[0181] In some aspects, the formulations described herein comprise an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, wherein

[0182] (i) the antibody or antigen-binding portion binds to human CD137 with an affinity of about 30 - 100 nM (e.g., between about 30 nM and about 100 nM); and

[0183] (ii) the antibody or its antigen-binding portion specifically binds to an epitope that comprises ELTK (corresponding to amino acid residues 111 - 114 of SEQ ID NO:3).

[0184] In some aspects, the formulations described herein comprise an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, wherein

[0185] (i) the antibody or antigen-binding portion binds to human CD137 with an affinity (K D ) of about 30 - 100 nM (e.g., between about 30 nM and about 100 nM);

[0186] (ii) the antibody or its antigen-binding portion specifically binds to an epitope that comprises ELTK (corresponding to amino acid residues 111 - 114 of SEQ ID NO:3);

[0187] (iii) the antibody or antigen-binding portion comprises a heavy chain CDR3 containing the amino acid sequence DXXXXLXXXXYXYYX (SEQ ID NO:126), where X is any amino acid; or

[0188] (iv) a combination of the above. In some aspects, X is any amino acid other than alanine.

[0189] In some aspects, the formulations described herein comprise an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, wherein

[0190] (i) the antibody or antigen-binding portion binds to human CD137 with an affinity (K D ) of about 30 - 100 nM (e.g., between about 30 nM and about 100 nM);

[0191] (ii) The antibody or its antigen-binding portion specifically binds to an epitope comprising ELTK (amino acid residues 111 - 114 corresponding to SEQ ID NO:3);

[0192] (iii) The antibody or antigen-binding portion comprises a heavy chain CDR3 containing the amino acid sequence DX1X2X3X4LX5X6X7X8YX9YYX 10 (SEQ ID NO:128), wherein X1 is any amino acid, wherein X2 is a nonpolar amino acid, wherein X3 is a nonpolar amino acid, wherein X4 is any amino acid, wherein X5 is a polar amino acid, wherein X6 is any amino acid, wherein X7 is any amino acid, wherein X8 is a polar amino acid, wherein X9 is a polar amino acid, and wherein X 10 is any amino acid; or

[0193] (iv) A combination of the above. In some aspects, X2 is proline, X3 is phenylalanine or tryptophan, X5 is aspartic acid or glutamic acid, X8 is tyrosine, and X9 is tyrosine.

[0194] In some aspects, the present disclosure provides an isolated monoclonal antibody or its antigen-binding portion that specifically binds to human CD137, wherein

[0195] (i) The antibody or antigen-binding portion binds to human CD137 with an affinity (K D ) of about 30 - 100 nM (e.g., between about 30 nM and about 100 nM);

[0196] (ii) The antibody or its antigen-binding portion specifically binds to an epitope comprising ELTK (amino acid residues 111 - 114 corresponding to SEQ ID NO:3); and

[0197] (iii) The antibody or antigen-binding portion comprises a heavy chain CDR3 containing the amino acid sequence DXXXXLXXXXYXYYX (SEQ ID NO:126), wherein X is any amino acid. In some aspects, X is any amino acid other than alanine.

[0198] In some aspects, the formulations described herein comprise an isolated monoclonal antibody or its antigen-binding portion that specifically binds to human CD137, wherein

[0199] (i) The antibody or antigen-binding portion binds to human CD137 with an affinity (K D ) of about 30 - 100 nM (e.g., between about 30 nM and about 100 nM);

[0200] (ii) the antibody or its antigen-binding portion specifically binds to an epitope comprising ELTK (amino acid residues 111-114 corresponding to SEQ ID NO: 3); and

[0201] (iii) the antibody or antigen-binding portion comprises a heavy chain CDR3 containing the amino acid sequence DX1X2X3X4LX5X6X7X8YX9YYX 10 (SEQ ID NO: 128), wherein X1 is any amino acid, wherein X2 is a non-polar amino acid, wherein X3 is a non-polar amino acid, wherein X4 is any amino acid, wherein X5 is a polar amino acid, wherein X6 is any amino acid, wherein X7 is any amino acid, wherein X8 is a polar amino acid, wherein X9 is a polar amino acid, and wherein X 10 is any amino acid. In some aspects, X2 is proline, X3 is phenylalanine or tryptophan, X5 is aspartic acid or glutamic acid, X8 is tyrosine, and X9 is tyrosine.

[0202] In any of the foregoing aspects, the epitope comprises the residues ELTK of SEQ ID NO: 3 (amino acid residues 111-114 corresponding to SEQ ID NO: 3). In some aspects, the epitope comprises ELTK of SEQ ID NO: 3 (amino acid residues 111-114 corresponding to SEQ ID NO: 3) and the residues N126, I132, and P135 of SEQ ID NO: 3.

[0203] In any of the foregoing aspects, the epitope is a non-linear epitope. In some aspects, mutation of the residue K114 of human CD137 (SEQ ID NO: 3) abolishes the binding of the antibody or its antigen-binding portion to human CD137.

[0204] In any of the foregoing aspects, the antibody or its antigen-binding portion comprises a heavy chain CDR3 containing the amino acid sequence DXPFXLDXXYYYYYX (SEQ ID NO: 128), wherein X is any amino acid. In some aspects, mutation of the residues D95, L100, Y100E, Y100G, Y100H, or a combination thereof of the heavy chain CDR3 of the antibody or antigen-binding portion described herein results in loss of binding to human CD137. In some aspects, mutation of the residues P97, F98, D100A, Y100D, Y100F, or a combination thereof of the heavy chain CDR3 of the antibody or antigen-binding portion described herein to alanine results in reduced binding to human CD137. In other aspects, mutation of the residues P97, F98, D100A, Y100D, Y100F, or a combination thereof of the heavy chain CDR3 of the antibody or antigen-binding portion described herein to any residue other than alanine results in increased binding to human CD137.

[0205] In any of the foregoing aspects, the antibody or antigen-binding portion thereof binds to human CD137 with a K of about 45-95 nM, 50-90 nM, 55-85 nM, 60-80 nM, 65-75 nM, 55-75 nM, 40-70 nM, 50-80 nM or 60-90 nM. D In any of the foregoing aspects, the antibody or antigen-binding portion thereof binds to human CD137 with a K of about 45 nM to about 95 nM, about 50 to about 90 nM, about 55 to about 85 nM, about 60 to about 80 nM, about 65 to about 75 nM, about 55 to about 75 nM, about 40 to about 70 nM, about 50 to about 80 nM or about 60 to about 90 nM. D In any of the foregoing aspects, the antibody or antigen-binding portion thereof binds to human CD137.

[0206] In any of the foregoing aspects, the antibody or antigen-binding portion thereof comprises heavy and light chain CDRs, wherein the heavy chain CDR3 comprises the amino acid sequence shown in SEQ ID NO: 68.

[0207] In any of the foregoing aspects, the antibody or antigen-binding portion thereof comprises heavy and light chain CDRs selected from the group consisting of:

[0208] (a) heavy chain CDR1, CDR2 and CDR3 sequences as shown in SEQ ID NOs: 48, 56 and 68 respectively, and light chain CDR1, CDR2 and CDR3 sequences as shown in SEQ ID NOs: 69, 78 and 89 respectively; and

[0209] (b) heavy chain CDR1, CDR2 and CDR3 sequences as shown in SEQ ID NOs: 51, 108 and 68 respectively, and light chain CDR1, CDR2 and CDR3 sequences as shown in SEQ ID NOs: 69, 78 and 89 respectively.

[0210] In some embodiments, the agonistic isolated monoclonal antibody or antigen-binding portion thereof comprises heavy and light chain CDRs selected from the group consisting of:

[0211] (a) heavy chain CDR1, CDR2 and CDR3 sequences as shown in SEQ ID NOs: 135, 139 and 143 respectively, and light chain CDR1, CDR2 and CDR3 sequences as shown in SEQ ID NOs: 144, 147 and 150 respectively; and

[0212] (b) heavy chain CDR1, CDR2 and CDR3 sequences as shown in SEQ ID NOs: 137, 141 and 143 respectively, and light chain CDR1, CDR2 and CDR3 sequences as shown in SEQ ID NOs: 144, 147 and 150 respectively.

[0213] In some embodiments, the agonistic isolated monoclonal antibody or antigen-binding portion thereof comprises heavy and light chain CDRs selected from the group consisting of:

[0214] (a) heavy chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NO:48, 154, and 159, respectively, and light chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NO:144, 147, and 150, respectively; and

[0215] (b) heavy chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NO:51, 156, and 159, respectively, and light chain CDR1, CDR2, and CDR3 sequences as set forth in SEQ ID NO:144, 147, and 150, respectively.

[0216] In any of the foregoing aspects, the antibody or antigen-binding portion thereof comprises heavy and light chain variable regions, wherein the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:4 and 101; and wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO:6.

[0217] In any of the foregoing aspects, the antibody or antigen-binding portion thereof comprises heavy and light chain variable regions comprising amino acid sequences selected from the group consisting of:

[0218] (a) SEQ ID NO:4 and 6, respectively; and

[0219] (b) SEQ ID NO:101 and 6, respectively.

[0220] In any of the foregoing aspects, the antibody or antigen-binding portion thereof comprises heavy and light chain variable regions, wherein the heavy chain variable region comprises an amino acid sequence that is at least 90% identical to an amino acid sequence selected from the group consisting of SEQ ID NO:4 and 101; and wherein the light chain variable region comprises an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:6.

[0221] In any of the foregoing aspects, the antibody or antigen-binding portion thereof comprises heavy and light chain variable regions comprising amino acid sequences that are at least 90% identical to amino acid sequences selected from the group consisting of:

[0222] (a) SEQ ID NO:4 and 6, respectively; and

[0223] (b) SEQ ID NO:101 and 6, respectively.

[0224] In any of the foregoing aspects, the antibody or antigen-binding portion thereof comprises a heavy chain and a light chain, and the heavy chain and the light chain comprise amino acid sequences selected from the group consisting of:

[0225] (a) SEQ ID NO: 129 and 133, respectively; and

[0226] (b) SEQ ID NO: 131 and 133, respectively.

[0227] In any of the foregoing aspects, the isolated monoclonal antibody or antigen-binding portion thereof described herein is an agonist of human CD137 activity.

[0228] In any of the foregoing aspects, the isolated monoclonal antibody or antigen-binding portion thereof described herein competes with mAb1 or an antigen-binding fragment of mAb1 for binding to an epitope of human CD137.

[0229] In some aspects, the formulation described herein comprises an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to CD137, wherein the antibody or antigen-binding portion thereof comprises heavy and light chain CDRs selected from the group consisting of:

[0230] (a) Heavy chain CDR1, CDR2, and CDR3 sequences as shown in SEQ ID NOs: 48, 56, and 68, respectively, and light chain CDR1, CDR2, and CDR3 sequences as shown in SEQ ID NOs: 69, 78, and 89, respectively;

[0231] (b) Heavy chain CDR1, CDR2, and CDR3 sequences as shown in SEQ ID NOs: 48, 56, and 68, respectively, and light chain CDR1, CDR2, and CDR3 sequences as shown in SEQ ID NOs: 70, 79, and 90, respectively;

[0232] (c) Heavy chain CDR1, CDR2, and CDR3 sequences as shown in SEQ ID NOs: 48, 56, and 68, respectively, and light chain CDR1, CDR2, and CDR3 sequences as shown in SEQ ID NOs: 71, 80, and 91, respectively;

[0233] (d) Heavy chain CDR1, CDR2, and CDR3 sequences as shown in SEQ ID NOs: 48, 56, and 68, respectively, and light chain CDR1, CDR2, and CDR3 sequences as shown in SEQ ID NOs: 72, 81, and 92, respectively;

[0234] (e) Heavy chain CDR1, CDR2, and CDR3 sequences as shown in SEQ ID NOs: 48, 56, and 68, respectively, and light chain CDR1, CDR2, and CDR3 sequences as shown in SEQ ID NOs: 73, 82, and 91, respectively;

[0235] (f) The heavy chain CDR1, CDR2 and CDR3 sequences as shown in SEQ ID NO:48, 56 and 68 respectively, and the light chain CDR1, CDR2 and CDR3 sequences as shown in SEQ ID NO:74, 83 and 93 respectively;

[0236] (g) The heavy chain CDR1, CDR2 and CDR3 sequences as shown in SEQ ID NO:48, 56 and 68 respectively, and the light chain CDR1, CDR2 and CDR3 sequences as shown in SEQ ID NO:75, 84 and 91 respectively;

[0237] (h) The heavy chain CDR1, CDR2 and CDR3 sequences as shown in SEQ ID NO:48, 56 and 68 respectively, and the light chain CDR1, CDR2 and CDR3 sequences as shown in SEQ ID NO:74, 85 and 94 respectively;

[0238] (i) The heavy chain CDR1, CDR2 and CDR3 sequences as shown in SEQ ID NO:48, 56 and 68 respectively, and the light chain CDR1, CDR2 and CDR3 sequences as shown in SEQ ID NO:76, 86 and 95 respectively;

[0239] (j) The heavy chain CDR1, CDR2 and CDR3 sequences as shown in SEQ ID NO:48, 56 and 68 respectively, and the light chain CDR1, CDR2 and CDR3 sequences as shown in SEQ ID NO:77, 87 and 93 respectively;

[0240] (k) The heavy chain CDR1, CDR2 and CDR3 sequences as shown in SEQ ID NO:48, 56 and 68 respectively, and the light chain CDR1, CDR2 and CDR3 sequences as shown in SEQ ID NO:69, 88 and 90 respectively;

[0241] (l) The heavy chain CDR1, CDR2 and CDR3 sequences as shown in SEQ ID NO:49, 57 and 68 respectively, and the light chain CDR1, CDR2 and CDR3 sequences as shown in SEQ ID NO:69, 78 and 89 respectively;

[0242] (m) The heavy chain CDR1, CDR2 and CDR3 sequences as shown in SEQ ID NO:49, 58 and 68 respectively, and the light chain CDR1, CDR2 and CDR3 sequences as shown in SEQ ID NO:69, 78 and 89 respectively;

[0243] (n) The heavy chain CDR1, CDR2 and CDR3 sequences as shown in SEQ ID NO:49, 59 and 68 respectively, and the light chain CDR1, CDR2 and CDR3 sequences as shown in SEQ ID NO:69, 78 and 89 respectively;

[0244] (o) The heavy chain CDR1, CDR2 and CDR3 sequences as shown in SEQ ID NO:49, 60 and 68 respectively, and the light chain CDR1, CDR2 and CDR3 sequences as shown in SEQ ID NO:69, 78 and 89 respectively;

[0245] (p) The heavy chain CDR1, CDR2 and CDR3 sequences as shown in SEQ ID NO:50, 61 and 68 respectively, and the light chain CDR1, CDR2 and CDR3 sequences as shown in SEQ ID NO:69, 78 and 89 respectively;

[0246] (q) The heavy chain CDR1, CDR2 and CDR3 sequences as shown in SEQ ID NO:50, 58 and 68 respectively, and the light chain CDR1, CDR2 and CDR3 sequences as shown in SEQ ID NO:69, 78 and 89 respectively;

[0247] (r) The heavy chain CDR1, CDR2 and CDR3 sequences as shown in SEQ ID NO:51, 62 and 68 respectively, and the light chain CDR1, CDR2 and CDR3 sequences as shown in SEQ ID NO:69, 78 and 89 respectively;

[0248] (s) The heavy chain CDR1, CDR2 and CDR3 sequences as shown in SEQ ID NO:52, 63 and 68 respectively, and the light chain CDR1, CDR2 and CDR3 sequences as shown in SEQ ID NO:69, 78 and 89 respectively;

[0249] (t) The heavy chain CDR1, CDR2 and CDR3 sequences as shown in SEQ ID NO:50, 64 and 68 respectively, and the light chain CDR1, CDR2 and CDR3 sequences as shown in SEQ ID NO:69, 78 and 89 respectively;

[0250] (u) The heavy chain CDR1, CDR2 and CDR3 sequences as shown in SEQ ID NO:50, 65 and 68 respectively, and the light chain CDR1, CDR2 and CDR3 sequences as shown in SEQ ID NO:69, 78 and 89 respectively;

[0251] (v) The heavy chain CDR1, CDR2, and CDR3 sequences as shown in SEQ ID NO:51, 108, and 68 respectively, and the light chain CDR1, CDR2, and CDR3 sequences as shown in SEQ ID NO:69, 78, and 89 respectively;

[0252] (w) The heavy chain CDR1, CDR2, and CDR3 sequences as shown in SEQ ID NO:107, 56, and 68 respectively, and the light chain CDR1, CDR2, and CDR3 sequences as shown in SEQ ID NO:69, 78, and 89 respectively; and

[0253] (x) The heavy chain CDR1, CDR2, and CDR3 sequences as shown in SEQ ID NO:48, 56, and 68 respectively, and the light chain CDR1, CDR2, and CDR3 sequences as shown in SEQ ID NO:109, 110, and 92 respectively.

[0254] In other aspects, the formulations described herein comprise an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, wherein the antibody or antigen-binding portion thereof comprises heavy and light chain variable regions, wherein the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:4, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 101, and 103; and wherein the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:6, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, and 105.

[0255] In other aspects, the formulations described herein comprise an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, wherein the antibody or antigen-binding portion comprises heavy and light chain variable regions encoded by nucleotide sequences selected from the group consisting of:

[0256] (a) SEQ ID NO:5 and 7 respectively; and

[0257] (b) SEQ ID NO:102 and 7 respectively.

[0258] In other aspects, the formulations described herein comprise an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, wherein the antibody or antigen-binding portion comprises heavy and light chain variable regions encoded by nucleotide sequences selected from the group consisting of:

[0259] (a) SEQ ID NO:5 and 7 respectively;

[0260] (b) SEQ ID NO:5 and 29 respectively;

[0261] (c) are SEQ ID NO:5 and 31 respectively;

[0262] (d) are SEQ ID NO:5 and 33 respectively;

[0263] (e) are SEQ ID NO:5 and 35 respectively;

[0264] (f) are SEQ ID NO:5 and 37 respectively;

[0265] (g) are SEQ ID NO:5 and 39 respectively;

[0266] (h) are SEQ ID NO:5 and 41 respectively;

[0267] (i) are SEQ ID NO:5 and 43 respectively;

[0268] (j) are SEQ ID NO:5 and 45 respectively;

[0269] (k) are SEQ ID NO:5 and 47 respectively;

[0270] (l) are SEQ ID NO:9 and 7 respectively;

[0271] (m) are SEQ ID NO:11 and 7 respectively;

[0272] (n) are SEQ ID NO:13 and 7 respectively;

[0273] (o) are SEQ ID NO:15 and 7 respectively;

[0274] (p) are SEQ ID NO:17 and 7 respectively;

[0275] (q) are SEQ ID NO:19 and 7 respectively;

[0276] (r) are SEQ ID NO:21 and 7 respectively;

[0277] (s) are SEQ ID NO:23 and 7 respectively;

[0278] (t) are SEQ ID NO:25 and 7 respectively;

[0279] (u) are SEQ ID NO:27 and 7 respectively;

[0280] (v) are SEQ ID NO:102 and 7 respectively;

[0281] (w) are SEQ ID NO:104 and 7 respectively; and

[0282] (x) are SEQ ID NO:5 and 106, respectively.

[0283] In other aspects, the formulations described herein comprise an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, wherein the antibody or antigen-binding portion thereof comprises heavy and light chain CDRs, and wherein the heavy chain CDR3 comprises the amino acid sequence shown in SEQ ID NO:68.

[0284] In another aspect, the formulations described herein comprise an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, wherein the antibody or antigen-binding portion thereof comprises heavy and light chain CDRs, and wherein the heavy chain CDR3 comprises the amino acid sequence DXXXXLXXXXYXYYX (SEQ ID NO:126), where X is any amino acid. In some aspects, X is any amino acid other than alanine.

[0285] In some aspects, the formulations described herein comprise an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, wherein the antibody or antigen-binding portion thereof comprises heavy and light chain CDRs, and wherein the heavy chain CDR3 comprises the amino acid sequence DXPFXLDXXYYYYYX (SEQ ID NO:127), where X is any amino acid. In some aspects, X is any amino acid other than alanine.

[0286] In other aspects, the formulations described herein comprise an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, wherein the antibody or antigen-binding portion thereof comprises heavy and light chain CDRs, and wherein the heavy chain CDR3 comprises the amino acid sequence DXXXXLXXXXYXYYX (SEQ ID NO:126), where X is any amino acid, and wherein mutation of the residues D95, L100, Y100E, Y100G, Y100H or combinations thereof results in loss of binding to human CD137.

[0287] In other aspects, the formulations described herein comprise an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, wherein the antibody or antigen-binding portion thereof comprises heavy and light chain CDRs, and wherein the heavy chain CDR3 comprises the amino acid sequence DXPFXLDXXYYYYYX (SEQ ID NO:127), where X is any amino acid, and wherein mutation of the residues P97, F98, D100A, Y100D, Y100F or combinations thereof to alanine results in reduced binding to human CD137.

[0288] In some aspects, the formulations described herein comprise an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, wherein the antibody or antigen-binding portion thereof comprises heavy and light chain CDRs, wherein the heavy chain CDR3 comprises the amino acid sequence DXPFXLDXXYYYYYX (SEQ ID NO:127), where X is any amino acid, and wherein mutation of residue P97, F98, D100A, Y100D, Y100F, or combinations thereof to any residue other than alanine results in increased binding to human CD137.

[0289] In other aspects, the formulations described herein comprise an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, wherein the antibody or antigen-binding portion thereof comprises heavy and light chain CDRs, wherein the heavy chain CDR3 comprises the amino acid sequence DX1X2X3X4LX5X6X7X8YX9YYX10 (SEQ ID NO:128), where X1 is any amino acid, where X2 is a nonpolar amino acid, where X3 is a nonpolar amino acid, where X4 is any amino acid, where X5 is a polar amino acid, where X6 is any amino acid, where X7 is any amino acid, where X8 is a polar amino acid, where X9 is a polar amino acid, and where X10 is any amino acid. In some aspects, X2 is proline, X3 is phenylalanine or tryptophan, X5 is aspartic acid or glutamic acid, X8 is tyrosine, and X9 is tyrosine.

[0290] In some aspects, the formulations described herein comprise an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, wherein the antibody or antigen-binding portion thereof comprises heavy and light chain variable regions, and the heavy and light chain variable regions comprise amino acid sequences selected from the group consisting of:

[0291] (a) SEQ ID NO:4 and 6, respectively;

[0292] (b) SEQ ID NO:4 and 28, respectively;

[0293] (c) SEQ ID NO:4 and 30, respectively;

[0294] (d) SEQ ID NO:4 and 32, respectively;

[0295] (e) SEQ ID NO:4 and 34, respectively;

[0296] (f) SEQ ID NO:4 and 36, respectively;

[0297] (g) SEQ ID NO:4 and 38, respectively;

[0298] (h) are SEQ ID NO:4 and 40 respectively;

[0299] (i) are SEQ ID NO:4 and 42 respectively;

[0300] (j) are SEQ ID NO:4 and 44 respectively;

[0301] (k) are SEQ ID NO:4 and 46 respectively;

[0302] (l) are SEQ ID NO:8 and 6 respectively;

[0303] (m) are SEQ ID NO:10 and 6 respectively;

[0304] (n) are SEQ ID NO:12 and 6 respectively;

[0305] (o) are SEQ ID NO:14 and 6 respectively;

[0306] (p) are SEQ ID NO:16 and 6 respectively;

[0307] (q) are SEQ ID NO:18 and 6 respectively;

[0308] (r) are SEQ ID NO:20 and 6 respectively;

[0309] (s) are SEQ ID NO:22 and 6 respectively;

[0310] (t) are SEQ ID NO:24 and 6 respectively;

[0311] (u) are SEQ ID NO:26 and 6 respectively;

[0312] (v) are SEQ ID NO:101 and 6 respectively;

[0313] (w) are SEQ ID NO:103 and 6 respectively; and

[0314] (x) are SEQ ID NO:4 and 105 respectively.

[0315] In other aspects, the formulations described herein comprise an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, wherein the antibody or antigen-binding portion thereof comprises heavy and light chain variable regions, wherein the heavy chain variable region comprises an amino acid sequence that is at least 90% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 4, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 101, and 103; and wherein the light chain variable region comprises an amino acid sequence that is at least 90% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 6, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, and 105.

[0316] In some aspects, the formulations described herein comprise an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, wherein the antibody or antigen-binding portion thereof comprises heavy and light chain variable regions that comprise an amino acid sequence that is at least 90% identical to an amino acid sequence selected from the group consisting of:

[0317] (a) SEQ ID NO: 4 and 6, respectively;

[0318] (b) SEQ ID NO: 4 and 28, respectively;

[0319] (c) SEQ ID NO: 4 and 30, respectively;

[0320] (d) SEQ ID NO: 4 and 32, respectively;

[0321] (e) SEQ ID NO: 4 and 34, respectively;

[0322] (f) SEQ ID NO: 4 and 36, respectively;

[0323] (g) SEQ ID NO: 4 and 38, respectively;

[0324] (h) SEQ ID NO: 4 and 40, respectively;

[0325] (i) SEQ ID NO: 4 and 42, respectively;

[0326] (j) SEQ ID NO: 4 and 44, respectively;

[0327] (k) SEQ ID NO: 4 and 46, respectively;

[0328] (l) SEQ ID NO: 8 and 6, respectively;

[0329] (m) SEQ ID NO: 10 and 6, respectively;

[0330] (n) are SEQ ID NO:12 and 6, respectively;

[0331] (o) are SEQ ID NO:14 and 6, respectively;

[0332] (p) are SEQ ID NO:16 and 6, respectively;

[0333] (q) are SEQ ID NO:18 and 6, respectively;

[0334] (r) are SEQ ID NO:20 and 6, respectively;

[0335] (s) are SEQ ID NO:22 and 6, respectively;

[0336] (t) are SEQ ID NO:24 and 6, respectively;

[0337] (u) are SEQ ID NO:26 and 6, respectively;

[0338] (v) are SEQ ID NO:101 and 6, respectively;

[0339] (w) are SEQ ID NO:103 and 6, respectively; and

[0340] (x) are SEQ ID NO:4 and 105, respectively.

[0341] In some aspects, the formulations described herein comprise an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, wherein the antibody or antigen-binding portion thereof comprises heavy and light chain sequences, and the heavy and light chain sequences comprise amino acid sequences selected from the group consisting of:

[0342] (a) are SEQ ID NO:129 and 133, respectively; and

[0343] (b) are SEQ ID NO:131 and 133, respectively.

[0344] In some aspects, the formulations described herein comprise an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, wherein the antibody or antigen-binding portion thereof comprises heavy and light chain sequences, and the heavy and light chain sequences have the amino acid sequences shown in SEQ ID NO:129 and 133, respectively.

[0345] In some aspects, the formulations described herein comprise an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, wherein the antibody or antigen-binding portion thereof comprises heavy and light chain sequences, and the heavy and light chain sequences have the amino acid sequences shown in SEQ ID NO:131 and 133, respectively.

[0346] In any of the foregoing aspects, the antibody or antigen-binding portion specifically binds to and agonizes human CD137.

[0347] In any of the foregoing aspects, the isolated monoclonal antibody or its antigen-binding portion exhibits at least one or more of the properties selected from the group consisting of:

[0348] (a) inducing or enhancing dimerization of CD137 trimers;

[0349] (b) inducing or enhancing multimerization of CD137 trimers;

[0350] (c) inducing or enhancing T cell activation;

[0351] (d) inducing or enhancing cytotoxic T cell responses;

[0352] (e) inducing or enhancing T cell proliferation;

[0353] (f) inducing or enhancing cytokine production; and

[0354] (g) any combination of properties (a)-(f).

[0355] In any of the foregoing aspects, the isolated monoclonal antibody or its antigen-binding portion, relative to a reference antibody that binds to human CD137, exhibits at least one or more of the properties selected from the group consisting of:

[0356] (a) not inducing or enhancing intrahepatic T cell activation;

[0357] (b) not inducing or enhancing intrahepatic T cell proliferation;

[0358] (c) not inducing or enhancing splenic T cell activation;

[0359] (d) not inducing or enhancing splenic T cell proliferation;

[0360] (e) not inducing or enhancing macrophage activation;

[0361] (f) not inducing or enhancing macrophage differentiation;

[0362] (g) not inducing or enhancing alanine transaminase (ALT) activity; and

[0363] (h) any combination of properties (a)-(g). In some aspects, the reference antibody is urelumab.

[0364] In any of the foregoing aspects, the isolated monoclonal antibody or antigen-binding portion thereof induces or enhances human CD137-mediated T cell activation in the tumor microenvironment, but does not significantly induce or enhance human CD137-mediated T cell activation in the spleen and / or liver.

[0365] In any of the foregoing aspects, the isolated monoclonal antibody or antigen-binding portion thereof induces or enhances T cell activation in the tumor microenvironment, but does not significantly induce or enhance T cell activation in the spleen and / or liver.

[0366] In any of the foregoing aspects, the isolated monoclonal antibody or antigen-binding portion thereof induces or enhances human CD137-mediated cytotoxic T cell responses in the tumor microenvironment, but does not significantly induce or enhance human CD137-mediated cytotoxic T cell responses in the spleen and / or liver.

[0367] In any of the foregoing aspects, the isolated monoclonal antibody or antigen-binding portion thereof induces or enhances cytotoxic T cell responses in the tumor microenvironment, but does not significantly induce or enhance T cell responses in the spleen and / or liver.

[0368] In any of the foregoing aspects, the isolated monoclonal antibody or antigen-binding portion thereof induces human CD137-mediated T cell proliferation in the tumor microenvironment, but does not significantly induce human CD137-mediated T cell proliferation in the spleen and / or liver.

[0369] In any of the foregoing aspects, the isolated monoclonal antibody or antigen-binding portion thereof induces T cell proliferation in the tumor microenvironment, but does not significantly induce T cell proliferation in the spleen and / or liver.

[0370] In any of the foregoing aspects, the isolated monoclonal antibody or antigen-binding portion thereof induces human CD137-mediated T cell infiltration in the tumor microenvironment, but does not significantly induce human CD137-mediated T cell infiltration in the spleen and / or liver.

[0371] In any of the foregoing aspects, the isolated monoclonal antibody or antigen-binding portion thereof induces T cell infiltration in the tumor microenvironment, but does not significantly induce T cell infiltration in the spleen and / or liver.

[0372] In any of the foregoing aspects, the isolated monoclonal antibody or antigen-binding fragment thereof induces or enhances human CD137-mediated cytokine production in the tumor microenvironment, but does not significantly induce or enhance human CD137-mediated cytokine production in the spleen and / or liver.

[0373] In any of the foregoing aspects, the properties of the antibodies or antigen-binding portions described herein are not Fcγ receptor binding-dependent. In some aspects, the properties of the antibodies or antigen-binding portions described herein are enhanced by Fcγ receptor binding.

[0374] In any of the foregoing aspects, the isolated monoclonal antibody or antigen-binding portion thereof cross-competes with mAb1 (i.e., an antibody comprising heavy and light chain variable sequences of SEQ ID NOs: 4 and 6, respectively). In some aspects, the isolated monoclonal antibody or antigen-binding portion thereof cross-competes with mAb1 (i.e., an antibody comprising heavy and light chain variable sequences of SEQ ID NOs: 4 and 6, respectively), mab8 (i.e., an antibody comprising heavy and light chain variable sequences of SEQ ID NOs: 101 and 6, respectively), or mAb10 (i.e., an antibody comprising heavy and light chain variable sequences of SEQ ID NOs: 26 and 6, respectively). In some aspects, the isolated monoclonal antibody or antigen-binding portion thereof cross-competes with mab8 (i.e., an antibody comprising heavy and light chain variable sequences of SEQ ID NOs: 101 and 6, respectively). In some aspects, the isolated monoclonal antibody or antigen-binding portion thereof cross-competes with mAb10 (i.e., an antibody comprising heavy and light chain variable sequences of SEQ ID NOs: 26 and 6, respectively).

[0375] In any of the foregoing aspects, the isolated monoclonal antibody or antigen-binding portion thereof comprises at least the functional properties of mAb1 (i.e., an antibody comprising heavy and light chain variable sequences of SEQ ID NOs: 4 and 6, respectively). In some aspects, the isolated monoclonal antibody or antigen-binding portion thereof comprises at least the functional properties of mAb1 (i.e., an antibody comprising heavy and light chain variable sequences of SEQ ID NOs: 4 and 6, respectively), mab8 (i.e., an antibody comprising heavy and light chain variable sequences of SEQ ID NOs: 101 and 6, respectively), or mAb10 (i.e., an antibody comprising heavy and light chain variable sequences of SEQ ID NOs: 26 and 6, respectively). In some aspects, the isolated monoclonal antibody or antigen-binding portion thereof comprises at least the functional properties of mab8 (i.e., an antibody comprising heavy and light chain variable sequences of SEQ ID NOs: 101 and 6, respectively). In some aspects, the isolated monoclonal antibody or antigen-binding portion thereof comprises at least the functional properties of mAb10 (i.e., an antibody comprising heavy and light chain variable sequences of SEQ ID NOs: 26 and 6, respectively).

[0376] In any of the foregoing aspects, the isolated monoclonal antibody or antigen-binding portion thereof has at least a K equivalent to that of mAb1 (i.e., an antibody comprising heavy and light chain variable sequences of SEQ ID NOs: 4 and 6, respectively) DValue. In some aspects, the isolated monoclonal antibody or antigen-binding portion thereof has a K that is at least equivalent to that of mAb1 (i.e., an antibody comprising heavy and light chain variable sequences of SEQ ID NOs: 4 and 6, respectively), mab8 (i.e., an antibody comprising heavy and light chain variable sequences of SEQ ID NOs: 101 and 6, respectively), or mAb10 (i.e., an antibody comprising heavy and light chain variable sequences of SEQ ID NOs: 26 and 6, respectively). D Value. In some aspects, the isolated monoclonal antibody or antigen-binding portion thereof has a K that is at least equivalent to that of mab8 (i.e., an antibody comprising heavy and light chain variable sequences of SEQ ID NOs: 101 and 6, respectively). D Value. In some aspects, the isolated monoclonal antibody or antigen-binding portion thereof has a K that is at least equivalent to that of mAb10 (i.e., an antibody comprising heavy and light chain variable sequences of SEQ ID NOs: 26 and 6, respectively). D Value.

[0377] In any of the foregoing aspects, the isolated monoclonal antibody or antigen-binding portion thereof cross-reacts with cynomolgus CD137 and / or murine CD137.

[0378] In any of the foregoing aspects, the isolated monoclonal antibody or antigen-binding portion thereof is selected from the group consisting of: IgGl, IgG2, and IgG3, IgG4, and IgM, and IgAl, and IgA2, and IgD, and IgE antibodies. In some aspects, the isolated monoclonal antibody or antigen-binding portion thereof is an IgG1 antibody or an IgG4 antibody.

[0379] In any of the foregoing aspects, the isolated monoclonal antibody comprises a wild-type IgGl or wild-type IgG4 heavy chain constant region. In some aspects, the isolated monoclonal antibody comprises a mutated IgGl heavy chain constant region. In some aspects, the isolated monoclonal antibody comprises a mutated IgG4 heavy chain constant region. In some aspects, the mutated IgG4 heavy chain constant region comprises a substitution at Ser228. In some aspects, the mutated IgG4 heavy chain constant region comprises the substitution S228P.

[0380] In any of the foregoing aspects, the isolated monoclonal antibody or antigen-binding portion thereof binds to an epitope of CD137, wherein the amino acid residues of the epitope bound by the antibody are within 4 angstroms of the amino acid residues of the paratope of the mAb1 antibody, as described herein.

[0381] In any of the foregoing aspects, the isolated monoclonal antibody or antigen-binding portion thereof binds to an epitope of CD137, wherein a mutation of the epitope bound by the antibody inhibits, reduces, or blocks binding to both the antibody and the antibody mAb1.

[0382] In any of the foregoing aspects, the isolated antibody or antigen-binding portion thereof is fully human or humanized (i.e., a fully human or humanized antibody or antigen-binding portion thereof).

[0383] In some aspects, the present disclosure provides a pharmaceutical composition comprising the isolated monoclonal antibody or antigen-binding portion thereof described herein, and a pharmaceutically acceptable carrier.

[0384] In other aspects, the present disclosure provides a nucleic acid comprising a nucleotide sequence encoding the light chain, heavy chain, or both the light and heavy chains of the isolated monoclonal antibody or antigen-binding portion thereof described herein. In some aspects, the nucleic acid comprises SEQ ID NO: 5 and 7. In some aspects, the nucleic acid comprises SEQ ID NO: 102 and 7. In some aspects, the present disclosure provides an expression vector comprising the nucleic acid described herein. In other aspects, the present disclosure provides a cell transformed with the expression vector described herein.

[0385] In another aspect, the present disclosure provides a method of generating an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137, the method comprising maintaining the cell described herein under conditions that permit expression of the monoclonal antibody or antigen-binding portion thereof. In some aspects, the method of generating an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to human CD137 further comprises obtaining the monoclonal antibody or antigen-binding portion thereof.

[0386] In another aspect, the present disclosure provides a method of inducing or enhancing dimerization of human CD137 trimers in a subject, comprising administering to a subject in need thereof an effective amount of the isolated monoclonal antibody or antigen-binding portion thereof described herein, or the pharmaceutical composition described herein.

[0387] In another aspect, the present disclosure provides a method of inducing or enhancing multimerization of human CD137 trimers in a subject, comprising administering to a subject in need thereof an effective amount of the isolated monoclonal antibody or antigen-binding portion thereof described herein, or the pharmaceutical composition described herein.

[0388] In other aspects, the present disclosure provides a method of inducing or enhancing T cell activation mediated by human CD137 in a subject, comprising administering to a subject in need thereof an effective amount of the isolated monoclonal antibody or antigen-binding portion thereof described herein, or the pharmaceutical composition described herein. In some aspects, the T cell activation occurs in the tumor microenvironment. In other aspects, the T cell activation does not significantly occur in the spleen and / or liver of the subject.

[0389] In another aspect, the present disclosure provides a method of inducing or enhancing a human CD137-mediated cytotoxic T cell response in a subject, comprising administering to a subject in need thereof an effective amount of the isolated monoclonal antibody or antigen-binding portion thereof described herein, or a pharmaceutical composition described herein. In some aspects, the cytotoxic T cell response occurs in the tumor microenvironment. In other aspects, the cytotoxic T cell response does not significantly occur in the spleen and / or liver of the subject.

[0390] In some aspects, the present disclosure provides a method of inducing or enhancing human CD137-mediated cytokine production in a subject, comprising administering to a subject in need thereof an effective amount of the isolated monoclonal antibody or antigen-binding portion thereof described herein, or a pharmaceutical composition described herein. In some embodiments, the cytokines produced are IL-2, TNFα, IL-13, IFN-γ, or a combination thereof. In some embodiments, the cytokine produced is IL-2. In some embodiments, the cytokine produced is TNFα. In some embodiments, the cytokine produced is IL-13. In some embodiments, the cytokine produced is IFN-γ. In some embodiments, the cytokines produced are IL-2 and TNFα. In some embodiments, the cytokines produced are IL-2 and IL-13. In some embodiments, the cytokines produced are IL-2 and IFN-γ. In some embodiments, the cytokines produced are TNFα and IL-13. In some embodiments, the cytokines produced are TNFα and IFN-γ. In some embodiments, the cytokines produced are IL-13 and IFN-γ. In some embodiments, the cytokines produced are IL-2, TNFα, and IL-13. In some embodiments, the cytokines produced are IL-2, TNFα, and IFN-γ. In some embodiments, the cytokines produced are IFN-γ, TNFα, and IL-13. In other embodiments, the cytokine production occurs in the tumor microenvironment. In other embodiments, the cytokine production does not significantly occur in the spleen and / or liver of the subject.

[0391] In another aspect, the present disclosure provides a method of inducing or enhancing human CD137-mediated T cell proliferation in a subject, comprising administering to a subject in need thereof an effective amount of the isolated monoclonal antibody or antigen-binding portion thereof described herein, or a pharmaceutical composition described herein. In some embodiments, the T cell proliferation occurs in the tumor microenvironment. In other embodiments, the T cell proliferation does not significantly occur in the spleen and / or liver of the subject.

[0392] In another aspect, the present disclosure provides a method of reducing or inhibiting tumor growth, comprising administering to a subject in need thereof an effective amount of the isolated monoclonal antibody or antigen-binding portion thereof described herein, or the pharmaceutical composition described herein.

[0393] In another aspect, the present disclosure provides a method of treating a condition mediated by human CD137 in a subject, comprising administering to a subject in need thereof an effective amount of the isolated monoclonal antibody or antigen-binding portion thereof described herein, or the pharmaceutical composition described herein.

[0394] In some aspects, the present disclosure provides a method of treating cancer in a subject, comprising administering to a subject in need thereof an effective amount of the isolated monoclonal antibody or antigen-binding portion thereof described herein, or the pharmaceutical composition described herein. In some embodiments, the cancer is selected from the group consisting of melanoma, glioma, renal cancer, breast cancer, blood cancer, and head and neck cancer. In some embodiments, the blood cancer is B cell lymphoma.

[0395] In some aspects, the present disclosure provides a method of inducing an anti-tumor memory immune response, comprising administering to a subject in need thereof an effective amount of the isolated monoclonal antibody or antigen-binding portion thereof described herein, or the pharmaceutical composition described herein.

[0396] In any of the foregoing aspects, after administration of the antibody or antigen-binding portion, infiltration of immune cells into the tumor microenvironment is increased. In some aspects, the immune cells express CD45.

[0397] In any of the foregoing aspects, after administration of the antibody or antigen-binding portion, the number of regulatory T (Treg) cells in the tumor microenvironment is reduced. In some aspects, the Treg cells express CD4, FOXP-3, and CD24.

[0398] In any of the foregoing aspects, after administration of the monoclonal antibody or antigen-binding portion, the number of macrophages in the tumor microenvironment is reduced. In some aspects, the macrophages express CD45 and CD11b.

[0399] In any of the foregoing aspects, after administration of the antibody or antigen-binding portion, T cell exhaustion is reduced. In some aspects, the reduction of T cell exhaustion includes a reduction in the expression of TIGIT, PD-1, LAG-3, or a combination thereof. In some aspects, the reduction of T cell exhaustion includes a reduction in the expression of TIGIT and PD-1.

[0400] In any of the foregoing aspects, depletion of CD4+ T cells, CD8+ T cells, natural killer cells, or a combination thereof will reduce the efficacy of the antibody or its antigen-binding portion.

[0401] In another aspect, the present invention provides a method for detecting the presence or absence of human CD137 in a biological sample, comprising:

[0402] (a) contacting the biological sample with an antibody or antigen-binding portion described herein, wherein the antibody or antigen-binding portion is labeled with a detectable substance; and

[0403] (b) detecting the antibody or antigen-binding portion that binds to human CD137, thereby detecting the presence or absence of human CD137 in the biological sample.

[0404] In another aspect, the present disclosure provides a kit comprising a container containing an antibody or antigen-binding portion described herein and optionally a pharmaceutically acceptable carrier or a pharmaceutical composition described herein, and a package insert containing instructions for administering the antibody or pharmaceutical composition for treating or delaying the development of cancer in a subject in need thereof, or for reducing or inhibiting tumor growth in a subject in need thereof.

[0405] In another aspect, the present disclosure provides a kit comprising a container containing an antibody or antigen-binding portion described herein and optionally a pharmaceutically acceptable carrier or a pharmaceutical composition described herein, and a package insert containing instructions for administering the antibody or pharmaceutical composition alone or in combination with another agent for treating or delaying the development of cancer in a subject in need thereof, or for reducing or inhibiting tumor growth in a subject in need thereof.

[0406] In another aspect, the present disclosure provides the use of an isolated monoclonal antibody or antigen-binding portion thereof described herein for inducing or enhancing T cell activation mediated by human CD137 in a subject. In other aspects, the present disclosure provides the use of an isolated monoclonal antibody or antigen-binding portion thereof described herein for inducing or enhancing the multimerization of human CD137 trimers in a subject. In another aspect, the present disclosure provides the use of an isolated monoclonal antibody or antigen-binding portion thereof described herein for inducing or enhancing cytotoxic T cell responses mediated by human CD137 in a subject. In other aspects, the present disclosure provides the use of an isolated monoclonal antibody or antigen-binding portion thereof described herein for inducing or enhancing cytokine production mediated by human CD137 in a subject. In another aspect, the present disclosure provides the use of an isolated monoclonal antibody or antigen-binding portion thereof described herein for inducing or enhancing T cell proliferation mediated by human CD137 in a subject.

[0407] In another aspect, the present disclosure provides the use of the isolated monoclonal antibody or antigen-binding portion thereof described herein for reducing or inhibiting tumor growth in a subject in need thereof. In other aspects, the present disclosure provides the use of the isolated monoclonal antibody or antigen-binding portion thereof described herein for treating a disorder mediated by human CD137 in a subject in need thereof. In another aspect, the present disclosure provides the use of the isolated monoclonal antibody or antigen-binding portion thereof described herein for treating cancer in a subject in need thereof.

[0408] In another aspect, the present disclosure provides the use of the isolated monoclonal antibody or antigen-binding portion thereof described herein in the manufacture of a medicament for treating or delaying the development of cancer or reducing or inhibiting tumor growth in a subject in need thereof. In other aspects, the present disclosure provides the isolated monoclonal antibody or antigen-binding portion thereof described herein for the manufacture of a medicament for treating or delaying the development of cancer or reducing or inhibiting tumor growth in a subject in need thereof. In another aspect, the present disclosure provides the isolated monoclonal antibody or antigen-binding portion thereof described herein for use as a medicament.

[0409] BRIEF DESCRIPTION OF THE DRAWINGS

[0410] This patent or application file contains at least one color drawing. Copies of this patent or patent application publication with color drawing(s) will be provided by the Patent Office upon request and payment of the necessary fee.

[0411] Figure 1 Depicts the distribution of the binding affinities of affinity matured clones of the parental anti-CD137 antibody mAb1.

[0412] Figure 2 Schematic diagram of shows the results of mAb1 CDRH3 alanine scanning, measured by the binding affinity (K D ) to human or mouse CD137.

[0413] Figure 3A Shows the amino acid sequence of human CD137, with the residues comprising the epitopes bound by mAb1, mAb4 or mAb5 shown in bold.

[0414] Figure 3B Depicts the kinetic binding data of mAb1 to the extracellular domain of mouse and rat CD137, determined by surface plasmon resonance.

[0415] Figure 3C Provides an X-ray crystallography image of human CD137 bound to CD137L (shown in grey), and the residues E111, T113, K114 and P135 are shown as spheres.

[0416] Figure 3D An X-ray crystallography image of the trimeric form of human CD137 bound to CD137L (shown in gray) is provided, and residues E111, T113, K114, and P135 are shown as spheres.

[0417] Figure 4A A scatter plot of flow cytometry data is provided, which depicts an increase in the expression of TIGIT (top) or PD-1 (bottom) on CD44+ T cells in response to an anti-CD137 antibody.

[0418] Figure 4B Quantification of CD8+CD44+ T cells expressing TIGIT (top) or PD-1 (bottom) in the spleens of mice after treatment with an anti-CD137 antibody is depicted.

[0419] Figure 4C Quantification of CD8+ T cells in the spleens of mice after treatment with an anti-CD137 antibody is depicted, expressed as a percentage of CD45+ cells (left) or the number of cells per spleen (right).

[0420] Figure 5A Individual CT26 tumor volumes in mice after treatment with the indicated doses of an anti-CD137 antibody are shown.

[0421] Figure 5B Shown is Figure 5A the mean tumor volume provided in

[0422] Figure 5C The Kaplan-Meier plot of shows the overall survival rate of mice with tumors after treatment with an anti-CD137 antibody.

[0423] Figure 5D Tumor volumes in mice re-challenged with oncogenic CT26 cells are shown.

[0424] Figure 6A Individual CT26 tumor volumes in mice after treatment with parental and affinity-matured anti-CD137 antibodies are shown.

[0425] Figure 6B Provided is Figure 6A the mean tumor volume provided in

[0426] Figure 7 Percentages of CD8+ or CD4+ T cells from splenic T cells (top) and tumor-infiltrating leukocytes (bottom) after treatment with the indicated doses of an anti-CD137 antibody are depicted.

[0427] Figure 8Shows individual tumor volumes in mice treated with mAb1 with or without lymphodepleting antibodies. CD4+ T cells were depleted with GK1.5 (middle panel), CD8+ T cells were depleted with YTS169.4 (second right panel), and NK cells were depleted with anti-asialo-GM1 antibody (far right panel).

[0428] Figure 9 Shows individual tumor volumes in mice bearing CT26 tumors (colon cancer), EMT-6 tumors (breast cancer), A20 tumors (B cell lymphoma), or MC38 tumors (colon cancer) and treated with mAb8 or isotype control antibody.

[0429] Figures 10A - 10C Shows the in vivo anti-tumor efficacy of anti-CD137 antibody administered at 150 μg / mouse. Individual tumor volumes are shown in 10A, mean tumor volumes are shown in 10B, and percent survival is shown in 10C.

[0430] Figures 11A - 11C Shows the in vivo anti-tumor efficacy of anti-CD137 antibody administered at 20 μg / mouse. Individual tumor volumes are shown in 11A, mean tumor volumes are shown in 11B, and percent survival is shown in 11C.

[0431] Figure 12 Shows individual tumor volumes in mice bearing CT26 tumors and treated with different doses of mAb1 (i.e., 12.5, 25, 50, 100, or 200 μg) or isotype control.

[0432] Figure 13A and 13B Shows the role of Fc binding in the anti-tumor efficacy of mAb1. Figure 13A Shows mAb1 as an IgG4 isotype or IgG4 aglycosylated isotype. Mean tumor volumes are shown above and individual tumor volumes are shown below. Figure 13B Shows mAb1 as an IgG4 isotype or IgG1 aglycosylated isotype. Mean tumor volumes are shown above and individual tumor volumes are shown below.

[0433] Figures 14A - 14D Shows the in vivo anti-tumor efficacy of anti-CD137 antibody in mice with large established tumors (i.e., 500 mm 3 ). Individual tumor volumes are shown in 14A and 14D, mean tumor volumes are shown in 14B, and percent survival is shown in 14C.

[0434] Figure 15The Kaplan-Meier survival plots show protective anti-tumor immunity in mice rechallenged with CT26 cells on the contralateral side that had been previously treated with mAb1, mAb8, or isotype control from Figures 14A - 14C and were considered cured.

[0435] Figure 16A Scatter plots providing flow cytometry data show the expansion of CD45+ intrahepatic T cells after treatment with the indicated doses of anti-CD137 antibody.

[0436] Figure 16B Depicts the quantification of intrahepatic CD8+ T cells (left) and CD4+ T cells (right) after treatment with the indicated doses of anti-CD137 antibody.

[0437] Figure 17A Depicts the percentages of CD3+, CD4+, or CD8+ T cells from splenic T cells after treatment of mice with affinity matured anti-CD137 antibody.

[0438] Figure 17B Depicts the percentages of CD3+, CD4+, or CD8+ T cells from hepatic T cells after treatment of mice with affinity matured anti-CD137 antibody.

[0439] Figure 18A Depicts the percentages of splenic CD8+CD44+ T cells expressing TIGIT, PD-1, or LAG3 after treatment of mice with affinity matured anti-CD137 antibody.

[0440] Figure 18B Depicts the percentages of hepatic CD8+CD44+ T cells expressing TIGIT, PD-1, or LAG3 after treatment of mice with affinity matured anti-CD137 antibody.

[0441] Figure 19A Depicts the percentages of splenic CD4+CD44+ T cells expressing TIGIT, PD-1, or LAG3 after treatment of mice with affinity matured anti-CD137 antibody.

[0442] Figure 19B Depicts the percentages of hepatic CD4+CD44+ T cells expressing TIGIT, PD-1, or LAG3 after treatment of mice with affinity matured anti-CD137 antibody.

[0443] Figures 20A - 20C Provides in vivo toxicity metrics resulting from multiple administrations of different doses of anti-CD137 antibodies mAb1, mAb8, or 3H3. Figure 20A Shows the percentage of CD8+ T cells in the liver after administration of anti-CD137 antibody. Figure 20BShows the alanine transaminase (ALT) activity in the plasma of mice administered with anti-CD137 antibody. Figure 20C Shows the TNFα level in the plasma of mice administered with anti-CD137 antibody.

[0444] Figure 21 Provides representative pictures of liver sections from mice treated with Figures 20A - 20C mAb1, mAb8, 3H3 or isotype control described in

[0445] Figures 22A - 22D hematoxylin and eosin (H&E) stained. Arrows indicate infiltration of immune cells. Figure 22A Shows the overall immune cell infiltration based on CD45 expression. Figure 22B Shows the reduction of Treg cells, measured by FOXP-3 and CD25 expression. Figure 22C Shows the reduction of T cell exhaustion, measured by PD-1 and TIGIT expression. Figure 22D Shows the reduction of tumor-associated macrophages, measured by F4 / 80 and CD11b expression.

[0446] Figure 23 Shows the immunophenotypic analysis of spleens from mice with CT26 tumors treated with anti-CD137 antibodies mAb1 and 3H3 or isotype control.

[0447] Figure 24 Shows the concentration (pg / ml) of IL-2 produced by murine T cells in the OVA stimulation assay when stimulated with the indicated anti-CD137 antibodies. Along with atezolizumab (anti-PD-L1 antibody), murine anti-PD-1 (RMP1-14) was used as a comparator.

[0448] Figure 25A and 25B Shows the percentage of murine CD8+ T cells expressing CD25 (25A) or TIGIT (25B) when stimulated with the indicated anti-CD137 antibodies in the OVA stimulation assay. Along with atezolizumab (anti-PD-L1 antibody), murine anti-PD-1 (RMP1-14) and murine anti-CD137 (3H3) were used as comparators.

[0449] Figure 26The bar graph depicts the quantification of cytokines (IL-2, TNFα, IL-13, and IFN-γ) produced by CD3+ T cells after incubation with plate-bound anti-CD137 antibody. Cytokine levels are shown as fold increase relative to baseline activation by anti-CD3 antibody.

[0450] Figures 27A - 27C Depicts the dose response of IFN-γ production in the mixed lymphocyte reaction after treatment with anti-CD137 antibody. Anti-PD1 antibody (Keytruda; Merck) was used as a control.

[0451] Figure 28 Shows IFN-γ production by human T cells co-cultured with CHO cells engineered to express CD32 (CHO-CD32 cells) in the presence of anti-CD137 antibodies mAb1, mAb8, mAb4, or mAb5, or isotype control.

[0452] Figure 29 Shows the proliferation of Treg cells when co-cultured with CHO cells engineered to express CD32 (CHO-CD32 cells) in the presence or absence of anti-CD137 antibodies mAb1, mAb8, mAb4, or mAb5, and isotype control.

[0453] Figure 30 Shows NFκβ and SRF signaling in CCL-119 cells transduced with luciferase reporters for NFκβ or SRF in the presence of different concentrations of mAb1, mAB8, mAb4, or mAb5.

[0454] Figure 31 Shows the induction of IL-6, TNFα, or IL-27 by bone marrow-derived murine macrophages stimulated with TLR9 agonist CpG in the presence of anti-CD137 antibodies mAb1, 3H3, or LOB12.3, or isotype control.

[0455] Figure 32 Shows the induction of TNFα by human monocyte-derived macrophages stimulated with LPS in the presence of anti-CD137 antibodies mAb1, mAb4, or mAb5, or isotype control.

[0456] Figure 33 Shows the effect of anti-CD137 antibody on macrophage differentiation, as determined by CD64 expression in THP1 monocytes cultured with PMA in the presence of anti-CD137 antibodies mAb1, mAb4, or mAb5, or isotype control.

[0457] Figures 34A - 34CShows the percentages of hCD45+, hCD8+ or hCD4+ in immunocompetent mice of recipient PBMCs and anti-CD137 antibodies mAb1, mAb4 or mAb5 or isotype control.

[0458] Figure 35 Illustrates cIEF charge variant analysis of high concentration (100 mg / mL) mAb1 in three buffers at different pH values and stored at 4 °C or 25 °C for up to 4 weeks.

[0459] Figure 36 Illustrates cIEF charge variant analysis of 5 mg / mL mAb1 in three buffers at different pH values at 40 °C for up to 4 weeks.

[0460] Figure 37 Depicts the size exclusion chromatogram of mAb1 diluted 10-fold in 5%, 0.5% or 0.1% dextran solution, 0.9% saline or pure water and incubated at room temperature for three days. The term dextran is used interchangeably with the term glucose.

[0461] Figure 38 Illustrates the dynamic light scattering (DLS) results of mAb1 diluted 10-fold in 5%, 0.5% or 0.1% dextran solution, 0.9% saline or pure water and incubated at room temperature for three days.

[0462] Figure 39 Illustrates microfluidic imaging analysis of subvisible particles from 2 - 80 mm. The analysis was performed using the disclosed formulation containing 10 mg / mL mAb1 that underwent three cycles of freeze-thaw at -30 °C / room temperature and three passes through a needle with a built-in 5 μm filter or a vented needle with a built-in 5 μm filter. Detailed Description

[0463] The present disclosure provides various formulations of anti-CD137 antibodies or antigen-binding fragments thereof that specifically bind to and agonize human CD137. In some embodiments, the formulations of the present disclosure comprise (i) an anti-CD137 antibody or antigen-binding fragment thereof, (ii) a buffer (e.g., histidine), (iii) a disaccharide (e.g., sucrose); (iv) a nonionic surfactant (e.g., polysorbate 80); (v) a salt (e.g., NaCl). In some embodiments, the formulations of the present disclosure have a pH of from about 5.0 to about 7.0. In some embodiments, the formulations of the present disclosure have a pH of from about 5.0 to about 7.4. The present disclosure also provides methods of treating cancer in a patient or reducing or inhibiting tumor growth in a patient, comprising administering to the patient a formulation of the present disclosure. The present disclosure also provides methods of inducing or enhancing T cell activation in a patient, comprising administering to the patient a formulation of the present disclosure.

[0464] The present disclosure is at least in part based on the discovery that the formulations of the present disclosure are stable and minimize the formation of antibody aggregates and particulates. In particular, it is shown that the formulations of the present disclosure provide the anti-CD137 agonist antibody mAb1, which has excellent stability, no significant loss of monomeric antibody, and no significant amount of degradation. In particular, it has surprisingly been found that the anti-CD137 antibody is stable at high concentrations and under forced degradation conditions (e.g., elevated temperature) when formulated in a histidine buffer at pH 5.8. Similarly, it has been found that when the anti-CD137 antibody is formulated in buffers at pH 6.0 and pH 6.5, the acidic / basic species are reduced. It has also been found that the anti-CD137 formulations of the present disclosure have excellent stability with no significant loss of monomeric antibody. In addition, it has been found that the addition of sucrose to the anti-CD137 formulations of the present disclosure results in improved antibody stability upon elevated temperature.

[0465] Accordingly, in some embodiments, the present disclosure provides stable anti-CD137 antibody formulations. In some embodiments, the present disclosure provides stable anti-CD137 antibody formulations comprising (i) a buffer containing from about 10 mM to about 100 mM histidine, (ii) from about 5% to about 15% (weight / volume) sucrose, (iii) from about 0.01% to about 0.1% (weight / volume (w / v)) polysorbate-80, and (iv) from about 50 mM - 200 mM NaCl, wherein the pH of the formulation is from about 5.0 to about 7.0. In some embodiments, the pH of the formulation is from about 5.0 to about 7.4. In some embodiments, the pH of the formulation is from about 5.0 to about 8.0.

[0466] It has been shown that cancer treatment with an agonist anti-CD137 antibody induces immune-mediated tumor rejection in mice, and similar agents of this type are currently being tested in cancer patients. Previous reports have shown that administration of an anti-CD137 antibody can induce a significant accumulation of polyclonal infiltration of T lymphocytes in the liver (Dubrot et al., (2010) Cancer Immunology, Immunotherapy 59(8):1223-1233), suggesting the possibility of liver inflammation and drug-induced hepatotoxicity. A recent clinical evaluation report on the agonistic anti-CD137 antibody (Urelumab, BMS-663513; Bristol-Myers Squibb) documented treatment-related adverse events observed in human subjects, including signs of severe hepatotoxicity (transaminitis) related to the antibody dose (Segal et al., (2016) Clin Cancer Res 23(8):1929-1936).

[0467] The present disclosure provides an isolated monoclonal antibody or antigen-binding portion thereof that specifically binds to an epitope of human CD137 and agonizes human CD137, and formulations thereof. In some embodiments, the antibody or antigen-binding portion thereof competes with mAb1 for binding to the epitope of human CD137. In some aspects, compared to the anti-mouse CD137 3H3 antibody (Melero et al. (1997) Nature Medicine 3(6):682-685; Uno et al. (2006) Nature Medicine 12(6):693-696) and at least two anti-human CD137 antibodies in clinical development (BMS-663513 / Urelumab, Bristol-Meyers Squibb, and PF-05082566 / Utomilumab, Pfizer), the anti-CD137 agonist antibody of the present disclosure induces cytokine production and CD8+ T cell expansion in the tumor microenvironment, as well as protective anti-tumor immunity in vivo, with a reduced likelihood of toxicity-related events.

[0468] Definitions

[0469] Unless otherwise indicated, the terms used in the claims and the specification are defined as follows.

[0470] It must be noted that the singular forms "a", "an", and "the" used in the specification and the appended claims include plural referents unless the context clearly dictates otherwise. Further, unless the context requires otherwise, singular terms shall include plurals and plural terms shall include singulars.

[0471] As used herein, "about" will be understood by one of ordinary skill in the art and will vary to some extent depending on the context in which it is used. If the use of the term by one of ordinary skill in the art is not clear based on the context in which the term is used, "about" will mean up to plus or minus 10% of a particular value.

[0472] As used herein, the term "agonist" refers to any molecule that partially or fully promotes, induces, increases, and / or activates the biological activity of a native polypeptide (e.g., CD137) disclosed herein. Suitable agonist molecules specifically include agonist antibodies or antibody fragments, fragments or amino acid sequence variants of native polypeptides, peptides, antisense oligonucleotides, small organic molecules, etc. In some embodiments, activation in the presence of an agonist is dose-dependent. In some embodiments, the measured signal (e.g., biological activity) is at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 100% higher than the signal measured in a negative control under comparable conditions. Methods for identifying agonists suitable for use in the methods of the present disclosure are also disclosed herein. For example, these methods include, but are not limited to, binding assays such as enzyme-linked immunosorbent assay (ELISA), systems and radioimmunoassay (RIA). These assays measure the ability of an agonist to bind to a target polypeptide (e.g., a receptor or ligand, e.g., CD137) and thus show the ability of the agonist to promote, increase, or activate polypeptide activity. The efficacy of an agonist can also be determined using functional assays, such as the ability of an agonist to activate or promote polypeptide function. For example, a functional assay can include contacting a polypeptide with a candidate agonist molecule and measuring a detectable change in one or more biological activities normally associated with the polypeptide. The potency of an agonist is generally defined by its EC 50 value (the concentration required to activate 50% of the agonist response). The lower the EC 50 value, the greater the potency of the agonist and the lower the concentration required to activate the maximum biological response.

[0473] As used herein, the term "alanine scanning" refers to a technique for determining the contribution of a particular wild-type residue to the stability or function (e.g., binding affinity) of a given protein or polypeptide. The technique involves replacing the wild-type residue in the polypeptide with an alanine residue and then assessing the stability or function (e.g., binding affinity) of the alanine-substituted derivative or mutant polypeptide and comparing it to the wild-type polypeptide. Techniques for replacing wild-type residues in polypeptides with alanine are known in the art.

[0474] The term "ameliorate" refers to any therapeutically beneficial result in the treatment of a disease state such as cancer, including its prevention, reduction in severity or development, remission, or cure.

[0475] As used herein, the term "amino acid" refers to naturally occurring and synthetic amino acids, as well as amino acid analogs and amino acid mimetics that act in a manner similar to naturally occurring amino acids. Naturally occurring amino acids are those encoded by the genetic code, as well as those amino acids that are subsequently modified, such as hydroxyproline, γ-carboxyglutamate, and O-phosphoserine. Amino acid analogs refer to compounds that have the same basic chemical structure as a naturally occurring amino acid, i.e., a carbon bonded to hydrogen, a carboxyl group, an amino group, and an R group, such as homoserine, norleucine, methionine sulfoxide, methionine methylsulfonium. Such analogs have modified R groups (e.g., norleucine) or modified peptide backbones, but retain the same basic chemical structure as the naturally occurring amino acids. Amino acid mimetics refer to compounds that have a structure different from the general chemical structure of amino acids, but act in a manner similar to naturally occurring amino acids.

[0476] Amino acids can be referred to herein by their well-known three-letter symbols or by the one-letter symbols recommended by the IUPAC-IUB Commission on Biochemical Nomenclature. Similarly, nucleotides can be represented by their commonly accepted single-letter codes. As used herein, "polar amino acids" refer to amino acids that contain side chains that are more prone to be in an aqueous environment. In some embodiments, polar amino acids are selected from the group consisting of arginine, asparagine, aspartic acid, glutamic acid, glutamine, histidine, lysine, serine, threonine, and tyrosine. Polar amino acids can be positively charged, negatively charged, or neutrally charged. As used herein, "nonpolar amino acids" refer to amino acids selected from the group consisting of alanine, cysteine, glycine, isoleucine, leucine, methionine, phenylalanine, proline, tryptophan, and valine.

[0477] As used herein, "amino acid substitution" refers to the replacement of at least one existing amino acid residue in a predetermined amino acid sequence (the amino acid sequence of the starting polypeptide) with another different "substituting" amino acid residue. "Amino acid insertion" refers to the incorporation of at least one additional amino acid into a predetermined amino acid sequence. While an insertion typically consists of the insertion of one or two amino acid residues, larger "peptide insertions" can also be created, for example, insertions of about three to about five, or even up to about ten, fifteen, or twenty amino acid residues. As disclosed above, the inserted residues can be naturally occurring or non-naturally occurring. "Amino acid deletion" refers to the removal of at least one amino acid residue from a predetermined amino acid sequence.

[0478] As used herein, the terms "amount" or "level" refer to the detectable amount, level, or abundance of a substance (e.g., a protein). When referring to a polypeptide, such as those described herein, the terms "expression level" or "level of expression" are generally used interchangeably and typically refer to the detectable amount of the polypeptide in a biological sample (e.g., on the cell surface).

[0479] As used herein, the term "anti-CD137 agonist antibody" (used interchangeably with the term "anti-CD137 antibody") refers to an antibody that specifically binds to CD137 and partially or fully promotes, induces, increases, and / or activates the biological activity, response, and / or downstream pathways mediated by CD137 signaling or other CD137-mediated functions. In some embodiments, the anti-CD137 agonist antibody binds to CD137 and permits the binding of CD137L. In some embodiments, the anti-CD137 agonist antibody binds to CD137 and induces the multimerization of CD137. In some embodiments, the anti-CD137 agonist antibody binds to CD137 and induces the dimerization of CD137 trimers. In some embodiments, the anti-CD137 agonist antibody binds to CD137 and induces the multimerization of CD137 trimers. Examples of anti-CD137 agonist antibodies are provided herein. Methods for detecting trimers: Methods for detecting trimer complex formation are known to those of skill in the art. For example, it has been shown that electron microscopy can detect such complexes, see, e.g., Won, E. The Journal of Biological Chemistry, Vol. 285(12):9202 - 9210 (2010).

[0480] As used herein, the term "anti-CD137 mAb1" (used interchangeably with "mAb1") refers to an exemplary anti-CD137 agonist antibody that comprises the following variable heavy chain (V H ) amino acid sequence:

[0481] EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDSPFLLDDYYYYYYMDVWGKGTTVTVSS(SEQ ID NO:4),

[0482] and the variable light chain (V L ) amino acid sequence:

[0483] DIQMTQSPSSVSASVGDRVTITCRASQGISSWLAWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQGHLFPITFGGGTKVEIK(SEQ ID NO:6).

[0484] As used herein, the term "anti-CD137 mAb8" (used interchangeably with "mAb8") refers to an exemplary anti-CD137 agonist antibody that comprises a variable heavy chain (V H ) amino acid sequence:

[0485] EVQLLESGGGLVQPGGSLRLSCAASGFTFRNYAMSWVRQAPGKGLEWVSAISGSGDTTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDSPFLLDDYYYYYYMDVWGKGTTVTVSS(SEQ ID NO:101);

[0486] and the variable light chain (V L ) amino acid sequence:

[0487] DIQMTQSPSSVSASVGDRVTITCRASQGISSWLAWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQGHLFPITFGGGTKVEIK(SEQ ID NO:6).

[0488] As used herein, the term "anti-CD137 mAb10" (used interchangeably with "mAb10") refers to an exemplary anti-CD137 agonist antibody that comprises the following variable heavy chain (V H ) amino acid sequence:

[0489] EVQLLESGGGLVQPGGSLRLSCAASGFTFYGYAMSWVRQAPGKGLEWVAAISGSGDSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDSPFLLDDYYYYYYMDVWGKGTTVTVSS(SEQ ID NO:26);

[0490] and the following variable light chain (V L ) amino acid sequence:

[0491] DIQMTQSPSSVSASVGDRVTITCRASQGISSWLAWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQGHLFPITFGGGTKVEIK(SEQ ID NO:6).

[0492] As used herein, the term "antibody" refers to a complete antibody comprising two light chain polypeptides and two heavy chain polypeptides. Complete antibodies include different antibody isotypes, including IgM, IgG, IgA, IgD, and IgE antibodies. The term "antibody" includes polyclonal antibodies, monoclonal antibodies, chimerized or chimeric antibodies, humanized antibodies, primatized antibodies, deimmunized antibodies, and fully human antibodies. Antibodies can be prepared or derived from any of a variety of species, such as mammals, such as humans, non-human primates (such as orangutans, baboons, or chimpanzees), horses, cows, pigs, sheep, goats, dogs, cats, rabbits, guinea pigs, gerbils, hamsters, rats, and mice. Antibodies can be purified antibodies or recombinant antibodies.

[0493] As used herein, the terms "antibody fragment", "antigen-binding fragment", "antigen-binding portion", or like terms refer to an antibody fragment that retains the ability to bind to a target antigen (e.g., CD137) and inhibit the activity of the target antigen. Such fragments include, for example, single-chain antibodies, single-chain Fv fragments (scFv), Fd fragments, Fab fragments, Fab' fragments, or F(ab')2 fragments. An scFv fragment is a single polypeptide chain comprising the variable regions of the heavy and light chains of the antibody from which the scFv is derived. In addition, intrabodies, minibodies, triabodies, and diabodies are also included within the definition of an antibody and are applicable to the methods described herein. See, e.g., Todorovska et al., (2001) J. Immunol. Methods 248(1):47-66; Hudson and Kortt, (1999) J. Immunol. Methods 231(1):177-189; Poljak, (1994) Structure 2(12):1121-1123; Rondon and Marasco, (1997) Annu. Rev. Microbiol. 51:257-283, the disclosures of each of which are hereby incorporated by reference in their entireties.

[0494] As used herein, the term "antibody fragment" also includes, for example, single-domain antibodies, such as camelized single domain antibodies. See, e.g., Muyldermans et al., (2001) Trends Biochem. Sci. 26:230-235; Nuttall et al., (2000) Curr. Pharm. Biotech. 1:253-263; Reichmann et al., (1999) J. Immunol. Meth. 231:25-38; PCT application publication nos. WO 94 / 04678 and WO 94 / 25591; and U.S. Patent No. 6,005,079, all of which are hereby incorporated by reference in their entireties. In some embodiments, the present disclosure provides single-domain antibodies comprising two VH domains with modifications that allow for the formation of a single-domain antibody.

[0495] In some embodiments, the antigen-binding fragment comprises the variable region of a heavy chain polypeptide and the variable region of a light chain polypeptide. In some embodiments, the antigen-binding fragment described herein comprises the CDRs of the light and heavy chain polypeptides of an antibody.

[0496] The term "antigen-presenting cell" or "APC" is a cell that presents exogenous antigen complexed with MHC on its surface. T cells use the T cell receptor (TCR) to recognize this complex. Examples of APCs include, but are not limited to, dendritic cells (DCs), peripheral blood mononuclear cells (PBMCs), monocytes (e.g., THP-1), B lymphoblastoid cells (e.g., C1R.A2, 1518B-LCL), and monocyte-derived dendritic cells (DCs). Some APCs internalize antigens by phagocytosis or receptor-mediated endocytosis.

[0497] The term "antigen presentation" refers to the process by which APCs capture antigens and enable them to be recognized by T cells, e.g., as components of MHC-I and / or MHC-II conjugates.

[0498] As used herein, the term "apoptosis" refers to the process of programmed cell death that occurs in multicellular organisms (e.g., humans). The highly regulated biochemical and molecular events that lead to apoptosis can result in observable and characteristic morphological changes in the cell, including membrane blebbing, cell volume shrinkage, chromosomal DNA condensation and fragmentation, and mRNA decay. A common method for identifying cells (including T cells) that are undergoing apoptosis is to expose the cells to a fluorophore-conjugated protein (Annexin V). Annexin V is typically used to detect apoptotic cells by its ability to bind to phosphatidylserine on the outer leaflet of the plasma membrane, which is an early indicator that the cell is undergoing the apoptotic process.

[0499] As used herein, the term "binding to immobilized CD137" refers to the ability of the human antibodies of the present disclosure to bind to CD137 (e.g., CD137 expressed on the cell surface or CD137 bound to a solid support).

[0500] As used herein, the term "bispecific" or "bifunctional antibody" refers to an artificial hybrid antibody that has two different heavy / light chain pairs and two different binding sites. Bispecific antibodies can be produced by a variety of methods, including the fusion of hybridomas or the ligation of Fab' fragments. See, e.g., Songsivilai & Lachmann, (1990) Clin. Exp. Immunol. 79:315-321; Kostelny et al., (1992) J. Immunol. 148:1547-1553.

[0501] Traditionally, the recombinant production of bispecific antibodies has been based on the co-expression of two immunoglobulin heavy chain / light chain pairs, where the two heavy chain / light chain pairs have different specificities (Milstein and Cuello, (1983) Nature 305:537-539). Antibody variable domains with the desired binding specificities (antibody-antigen binding sites) can be fused to immunoglobulin constant domain sequences. The fusion of the heavy chain variable region is preferably to an immunoglobulin heavy chain constant region, including at least part of the hinge region, CH2 region, and CH3 region. For further details of exemplary currently known methods of generating bispecific antibodies, see, for example, Suresh et al., (1986) Methods Enzymol. 121:210; PCT Publication No. WO96 / 27011; Brennan et al., (1985) Science 229:81; Shalaby et al., J. Exp. Med. (1992) 175:217-225; Kostelny et al., (1992) J. Immunol. 148(5):1547-1553; Hollinger et al., (1993) Proc. Natl. Acad. Sci. USA 90:6444-6448; Gruber et al., (1994) J. Immunol. 152:5368; and Tutt et al., (1991) J. Immunol. 147:60. Bispecific antibodies also include cross-linked or heteroconjugate antibodies. Heteroconjugate antibodies can be prepared using any convenient cross-linking method. Suitable cross-linking agents are well known in the art and are disclosed, along with some cross-linking techniques, in U.S. Patent No. 4,676,980.

[0502] Also described are various techniques for preparing and isolating bispecific antibody fragments directly from recombinant cell cultures. For example, leucine zippers have been used to produce bispecific antibodies. See, e.g., Kostelny et al. (1992) J Immunol 148(5):1547-1553. Leucine zipper peptides from the Fos and Jun proteins can be genetically fused to the Fab' portions of two different antibodies. Antibody homodimers can be reduced at the hinge region to form monomers, which are then re-oxidized to form antibody heterodimers. This method can also be used to produce antibody homodimers. The "diabody" technology described by Hollinger et al. (1993) Proc Natl Acad Sci USA 90:6444-6448 provides an alternative mechanism for preparing bispecific antibody fragments. The fragments contain a heavy chain variable domain (VH) linked to a light chain variable domain (VL) by a linker that is too short to allow pairing between the two domains on the same chain. Thus, the VH and VL domains of one fragment are forced to pair with the complementary VL and VH domains of another fragment, thereby forming two antigen-binding sites. Another strategy for preparing bispecific antibody fragments by using single-chain Fv (scFv) dimers has also been reported. See, e.g., Gruber et al. (1994) J Immunol 152:5368. Alternatively, the antibody can be a "linear antibody" such as that described by Zapata et al. (1995) Protein Eng. 8(10):1057-1062. Briefly, these antibodies contain a pair of tandem Fd fragments (VH-CH1-VH-CH1) that form a pair of antigen-binding regions. Linear antibodies can be bispecific or monospecific.

[0503] Antibodies having more than two valencies (e.g., trispecific antibodies) are involved and described, e.g., in Tutt et al. (1991) J Immunol 147:60.

[0504] The present disclosure also encompasses variant forms of multispecific antibodies, such as the bispecific variable domain immunoglobulin (DVD-Ig) molecules described in Wu et al. (2007) Nat Biotechnol 25(11):1290-1297. The DVD-Ig molecules are designed such that two different light chain variable domains (VL) from two different parental antibodies are tandemly linked directly or via a short linker by recombinant DNA techniques, followed by a light chain constant domain. Similarly, the heavy chain comprises two different heavy chain variable domains (VH) tandemly linked, followed by the constant domains CH1 and the Fc region. Methods for preparing DVD-Ig molecules from two parental antibodies are further described, for example, in PCT Publication Nos. WO 08 / 024188 and WO 07 / 024715. In some embodiments, the bispecific antibody is a Fab-in-Tandem immunoglobulin, in which the light chain variable region with a second specificity is fused to the heavy chain variable region of a complete antibody. Such antibodies are described, for example, in International Patent Application Publication No. WO 2015 / 103072.

[0505] As used herein, "cancer antigen" or "tumor antigen" refers to (i) a tumor-specific antigen, (ii) a tumor-associated antigen, (iii) a cell expressing a tumor-specific antigen, (iv) a cell expressing a tumor-associated antigen, (v) an embryonic antigen on a tumor, (vi) an autologous tumor cell, (vii) a tumor-specific membrane antigen, (viii) a tumor-associated membrane antigen, (ix) a growth factor receptor, (x) a growth factor ligand, (xi) any other type of antigen or antigen-presenting cell or substance associated with cancer.

[0506] The term "cancer" is well recognized in the art and refers to malignancies of epithelial or endocrine tissues, including carcinomas of the respiratory system, gastrointestinal system, urogenital system, testicular cancer, breast cancer, prostate cancer, endocrine system cancer, and melanoma. The anti-CD137 antibodies described herein can be used to treat patients suffering from, suspected of having, or at high risk of developing any type of cancer, including renal cancer or melanoma. Exemplary cancers include those formed from cervical, lung, prostate, breast, head and neck, colon, and ovarian tissues. The term also includes carcinosarcoma, which includes a malignant tumor composed of carcinomatous and sarcomatous tissues. "Adenocarcinoma" refers to a carcinoma derived from glandular tissue or a carcinoma in which the tumor cells form recognizable glandular structures.

[0507] As used herein, the term "competes," when used in the context of antigen-binding proteins (e.g., immunoglobulins, antibodies, or antigen-binding fragments thereof) that compete to bind the same epitope, refers to the interaction between the antigen-binding proteins, as determined by an assay (e.g., a competitive binding assay; a cross-blocking assay), wherein a test antigen-binding protein (e.g., a test antibody) inhibits (e.g., reduces or blocks) the specific binding of a reference antigen-binding protein (e.g., a reference antibody, such as mAb1) to a common antigen (e.g., CD137 or a fragment thereof). In some embodiments, the antibodies described herein cross-compete with mAb1 (i.e., the antibody comprising the heavy and light chain variable sequences of SEQ ID NOs: 4 and 6, respectively), mab8 (i.e., the antibody comprising the heavy and light chain variable sequences of SEQ ID NOs: 101 and 6, respectively), or mAb10 (i.e., the antibody comprising the heavy and light chain variable sequences of SEQ ID NOs: 26 and 6, respectively).

[0508] A polypeptide or amino acid sequence "derived from" a designated polypeptide or protein refers to the source of the polypeptide. Preferably, a polypeptide or amino acid sequence derived from a particular sequence has an amino acid sequence that is substantially the same as this sequence or a portion thereof, where the portion consists of at least 10 to 20 amino acids, preferably at least 20 to 30 amino acids, more preferably at least 30 to 50 amino acids, or it can be recognized by one of ordinary skill in the art as having its origin in the sequence. A polypeptide derived from another peptide may have one or more mutations relative to the starting polypeptide, e.g., one or more amino acid residues have been replaced by another amino acid residue, or it has one or more amino acid residues inserted or deleted.

[0509] A polypeptide may comprise a non-naturally occurring amino acid sequence. Such variants will necessarily have less than 100% sequence identity or similarity to the starting molecule. In certain embodiments, the variant will have an amino acid sequence that has about 75% to less than 100%, more preferably about 80% to less than 100%, more preferably about 85% to less than 100%, more preferably about 90% to less than 100% (e.g., 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%), and most preferably about 95% to less than 100% amino acid sequence identity or similarity to the amino acid sequence of the starting polypeptide, e.g., over the length of the variant molecule.

[0510] In certain embodiments, there is a difference of one amino acid between the starting polypeptide sequence and the sequence derived therefrom. The identity or similarity of a sequence is defined herein as the percentage of amino acid residues in a candidate sequence that are identical (i.e., the same residue) to the starting amino acid residues after aligning the sequences and introducing gaps (if necessary) to achieve the maximum percentage of sequence identity. In certain embodiments, the polypeptide consists of, consists essentially of, or comprises an amino acid sequence selected from the sequences shown in any of Tables 22-27. In certain embodiments, the polypeptide comprises an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to an amino acid sequence selected from the sequences shown in any of Tables 22-27. In certain embodiments, the polypeptide comprises a contiguous amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a contiguous amino acid sequence selected from the sequences shown in any of Tables 22-27. In certain embodiments, the polypeptide comprises an amino acid sequence having at least 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 200, 300, 400, or 500 (or any integer within these numbers) contiguous amino acids of an amino acid sequence selected from the sequences shown in any of Tables 22-27.

[0511] In certain embodiments, the antibodies of the present disclosure are encoded by nucleotide sequences. The nucleotide sequences of the invention can be used in a variety of applications, including: cloning, gene therapy, protein expression and purification, mutagenesis introduction, DNA vaccination of hosts in need thereof, antibody production for, e.g., passive immunization, PCR, primer and probe generation, and the like. In certain embodiments, the nucleotide sequences of the invention comprise or consist of or consist essentially of a nucleotide sequence selected from the sequences shown in any one of Tables 22-27. In certain embodiments, the nucleotide sequence comprises a nucleotide sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleotide sequence selected from the sequences shown in any one of Tables 22-27. In certain embodiments, the nucleotide sequence comprises a continuous nucleotide sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a continuous nucleotide sequence selected from the sequences shown in any one of Tables 22-27. In certain embodiments, the nucleotide sequence comprises an amino acid sequence having at least 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 200, 300, 400 or 500 (or any integer within these numbers) consecutive nucleotides of a nucleotide sequence selected from the sequences shown in any one of Tables 22-27.

[0512] One of ordinary skill in the art will also understand that antibodies suitable for use in the methods disclosed herein can be altered such that they are different in sequence from the naturally occurring or native sequences from which they are derived, while still retaining the activity of the desired native sequence. For example, nucleotide or amino acid substitutions can be made that result in conservative substitutions or alterations of "non-essential" amino acid residues. Mutations can be introduced by standard techniques such as site-directed mutagenesis and PCR-mediated mutagenesis.

[0513] Antibodies suitable for the methods disclosed herein may contain conservative amino acid substitutions at one or more amino acid residues, e.g., at essential or non-essential amino acid residues. A "conservative amino acid substitution" is an amino acid substitution in which the amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art and include basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), β-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Accordingly, non-essential amino acid residues in the binding polypeptide are preferably replaced with another amino acid residue from the same side chain family. In certain embodiments, an amino acid segment (string) may be replaced with a segment that is structurally similar but different in the order and / or composition of side chain family members. Alternatively, in certain embodiments, mutations may be introduced randomly along all or part of the coding sequence, e.g., by saturation mutagenesis, and the resulting mutants may be incorporated into the binding polypeptides of the invention and screened for their ability to bind to the target.

[0514] As used herein, the term antigen "cross-presentation" refers to the presentation of exogenous protein antigens to T cells by MHC class I and II molecules on APCs.

[0515] As used herein, the term "cross-reactivity" refers to the ability of the antibodies of the present disclosure to bind to CD137 from different species. For example, an antibody that binds to human CD137 of the present disclosure may also bind to CD137 of another species. As used herein, cross-reactivity is determined by detecting specific reactivity with a purified antigen in a binding assay (e.g., SPR, ELISA) or binding to cells physiologically expressing CD137 or functionally interacting. Methods for determining cross-reactivity include the standard binding assays described herein, e.g., by performing BIACORE TM surface plasmon resonance (SPR) analysis, or flow cytometry using a BIACORE TM 2000 SPR instrument (Biacore AB, Uppsala, Sweden).

[0516] As used herein, the term "cytotoxic T lymphocyte (CTL) response" refers to an immune response induced by cytotoxic T cells. The CTL response is mainly mediated by CD8 + T cells.

[0517] As used herein, the term "dimerization" refers to the formation of a macromolecular complex from two macromolecules, such as proteins or protein multimers, that are typically non-covalently associated. Homodimerization refers to the process of dimerization when the macromolecules (such as proteins) are identical in nature. Heterodimerization refers to the process of dimerization when the macromolecules (such as proteins) are different in nature. Methods for determining dimerization are known to those of skill in the art. For example, such methods include, but are not limited to, yeast two-hybrid assays, fluorescence resonance energy transfer (FRET), bioluminescence resonance energy transfer (BRET), protein mass spectrometry, evanescent wave methods, size exclusion chromatography, analytical ultracentrifugation, scattering techniques, NMR spectroscopy, isothermal titration calorimetry, fluorescence anisotropy, fluorescence correlation spectroscopy (FCS), fluorescence recovery after photobleaching (FRAP), proximity imaging (PRIM), and bimolecular fluorescence complementation (BiFC) (see, e.g., Gell D.A., Grant R.P., Mackay J.P. (2012) The Detection and Quantitation of Protein Oligomerization. In: Matthews J.M. (eds) Protein Dimerization and Oligomerization in Biology. Advances in Experimental Medicine and Biology, vol 747. Springer, New York, NY; and Xie, Q. et al. Methods Mol Biol, 2011; 680:3-28).

[0518] As used herein, the term "dimerization of CD137" refers to the dimerization of two CD137 trimers. In some embodiments, the anti-CD137 agonist antibodies described herein induce or enhance the dimerization of CD137. In some embodiments, the anti-CD137 agonist antibodies described herein induce or enhance the dimerization of CD137 relative to the amount of dimerization in the absence of the anti-CD137 agonist antibody. In some embodiments, the anti-CD137 agonist antibodies described herein induce or enhance the dimerization of CD137 relative to the amount of dimerization in the presence of a reference anti-CD137 agonist antibody. In some embodiments, the dimerization is increased by at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100%.

[0519] As used herein, the term "EC" 50 refers to the concentration of an antibody or an antigen-binding portion thereof when the response induced in an in vitro or in vivo assay is 50% of the maximal response, i.e., the response that is halfway between the maximal response and the baseline.

[0520] As used herein, the term "effective dose" or "effective dosage" is defined as an amount sufficient to achieve or at least partially achieve the desired effect. The term "therapeutically effective dose" is defined as an amount sufficient to cure or at least partially arrest a disease and its complications in a patient already suffering from the disease. The effective amount for this purpose will depend on the severity of the condition being treated and the general state of the patient's own immune system.

[0521] As used herein, the term "epitope" or "antigenic determinant" refers to a determinant or site on an antigen (e.g., CD137) that is specifically bound by an antigen-binding protein (e.g., an immunoglobulin, an antibody, or an antigen-binding fragment). Epitopes of protein antigens can be classified as "linear epitopes" and "conformational epitopes". As used herein, the term "linear epitope" refers to an epitope formed by a continuous linear sequence of contiguous amino acids. Linear epitopes of protein antigens are generally retained upon exposure to chemical denaturants (e.g., acids, bases, solvents, cross-linking agents, chaotropic agents, disulfide bond reducing agents) or physical denaturants (e.g., heat, radioactivity, or mechanical shear or stress). In some embodiments, the epitope is non-linear, also referred to as an interrupted epitope. As used herein, the term "conformational epitope" or "non-linear epitope" refers to an epitope formed by non-contiguous amino acids juxtaposed by the tertiary folding of a polypeptide. Conformational epitopes are generally lost upon treatment with a denaturant. An epitope typically comprises at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 amino acids in a unique spatial conformation. In some embodiments, the epitope comprises fewer than 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, or 5 amino acids in a unique spatial conformation. Generally, an antibody or an antigen-binding fragment thereof specific for a particular target molecule will preferentially recognize and bind a specific epitope on the target molecule within a complex mixture of proteins and / or macromolecules. In some embodiments, the epitope does not comprise all of the amino acids of the extracellular domain of human CD137.

[0522] The present disclosure also encompasses antibodies that bind to an epitope on CD137, said epitope comprising all or part of the epitope recognized by a specific antibody described herein (e.g., the same or overlapping regions, or regions between or spanning regions).

[0523] As used herein, the term "epitope mapping" refers to the process or method of identifying the binding site or epitope of an antibody or its antigen-binding fragment on its target protein antigen. Epitope mapping methods and techniques are provided herein.

[0524] As used herein, the term "CD137" refers to a specific member of the tumor necrosis factor receptor (TNFR) family of transmembrane proteins. Alternative names and acronyms for CD137 in the art include "tumor necrosis factor receptor superfamily member 9" (TNFRSF9), 4-1BB, and "induced by lymphocyte activation" (ILA) (Alderson et al., (1994) Eur J Immunol 24(9):2219-2227; Schwarz et al., (1993) Gene 134(2):295-298). The exemplary amino acid sequence of full-length human CD137 (comprising the leader, transmembrane, and cytoplasmic domains) is shown in Table 23 (SEQ ID NO:3) and herein:

[0525] MGNSCYNIVATLLLVLNFERTRSLQDPCSNCPAGTFCDNNRNQICSPCPPNSFSSAGGQRTCDICRQCKGVFRTRKECSSTSNAECDCTPGFHCLGAGCSMCEQDCKQGQELTKKGCKDCCFGTFNDQKRGICRPWTNCSLDGKSVLVNGTKERDVVCGPSPADLSPGASSVTPPAPAREPGHSPQIISFFLALTSTALLFLLFFLTLRFSVVKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCEL

[0526] As used herein, the term "CD137L" or "CD137 ligand" refers to a member of the tumor necrosis factor (TNF) family of transmembrane proteins. Alternative names and acronyms for CD137L in the art include "tumor necrosis factor superfamily member 9" (TNFSF9) and 4-1BB ligand (4-1BBL) (Alderson et al., (1994) Eur J Immunol 24(9):2219-2227). The exemplary amino acid sequence of full-length CD137L is shown in Table 23 (SEQ ID NO:97).

[0527] As used herein, the term "Fc-mediated effector function" or "Fc effector function" refers to the biological activity of an antibody that is apart from the primary function and purpose of the antibody. For example, the effector function of a therapeutic agonist antibody is a biological activity apart from activation of the target protein or pathway. Examples of antibody effector functions include C1q binding and complement-dependent cytotoxicity; Fc receptor binding; antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis; downregulation of cell surface receptors (such as the B cell receptor); lack of activation of platelets expressing Fc receptors; and B cell activation. Many effector functions begin with the binding of Fc to an Fcγ receptor.

[0528] As used herein, the term "Fc receptor" refers to a polypeptide found on the surface of an immune effector cell that is bound by the Fc region of an antibody. In some embodiments, the Fc receptor is an Fcγ receptor. Fcγ receptors are divided into three subclasses, FcγRI (CD64), FcγRII (CD32), and FγcRIII (CD16). All four IgG isotypes (IgG1, IgG2, IgG3, and IgG4) bind to and activate the Fc receptors FcγRI, FcγRIIA, and FcγRIIIA. FcγRIIB is an inhibitory receptor, and thus an antibody that binds to this receptor does not activate complement and cellular responses. FcγRI is a high-affinity receptor that binds IgG in monomeric form, while FcγRIIA and FcγRIIA are low-affinity receptors that bind IgG only in polymeric form and with a slightly lower affinity. The binding of an antibody to an Fc receptor and / or Clq is controlled by specific residues or domains within the Fc region. Binding also depends on residues located within the hinge region and within the CH2 portion of the antibody. In some embodiments, the agonistic and / or therapeutic activity of an antibody described herein depends on the binding of the Fc region to an Fc receptor (e.g., FcγR). In some embodiments, the agonistic and / or therapeutic activity of an antibody described herein is enhanced by the binding of the Fc region to an Fc receptor (e.g., FcγR).

[0529] As used herein, the term "glycosylation profile" is defined as the profile of carbohydrate units covalently linked to a protein, more specifically to an immunoglobulin. When a person of ordinary skill in the art recognizes the glycosylation profile of a heterologous antibody as being more similar to the glycosylation pattern in a non-human transgenic animal species than to the species from which the CH gene of the transgenic is derived, the glycosylation map of the heterologous antibody can be characterized as being substantially similar to the glycosylation profile naturally present on antibodies produced by a non-human transgenic animal species.

[0530] As used herein, the term "blood cancer" includes lymphoma, leukemia, myeloma, or lymphoid malignancies, as well as cancers of the spleen and lymph nodes. Exemplary lymphomas include B-cell lymphoma (B-cell blood cancer) and T-cell lymphoma. B-cell lymphomas include Hodgkin lymphoma and most non-Hodgkin lymphomas. Non-limiting examples of B-cell lymphomas include diffuse large B-cell lymphoma, follicular lymphoma, mucosa-associated lymphoid tissue lymphoma, small cell lymphocytic lymphoma (which overlaps with chronic lymphocytic leukemia), mantle cell lymphoma (MCL), Burkitt's lymphoma, mediastinal large B cell lymphoma, Waldenstrom macroglobulinemia, nodal marginal zone B cell lymphoma, splenic marginal zone lymphoma, intravascular large B cell lymphoma, primary effusion lymphoma, and lymphomatoid granulomatosis. Non-limiting examples of T-cell lymphomas include extranodal T-cell lymphoma, cutaneous T-cell lymphoma, anaplastic large cell lymphoma, and angioimmunoblastic T cell lymphoma. Hematological malignancies also include leukemias such as, but not limited to, secondary leukemia, chronic lymphocytic leukemia, acute myeloid leukemia, chronic myeloid leukemia, and acute lymphocytic leukemia. Hematological malignancies further include myelomas such as, but not limited to, multiple myeloma and smoldering multiple myeloma. Other blood cancers and / or B-cell or T-cell related cancers are encompassed by the term hematological malignancies.

[0531] As used herein, the term "human antibody" includes antibodies having variable and constant regions (if present) with human germline immunoglobulin sequences. The human antibodies of the disclosure may include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by in vitro random or site-specific mutagenesis or by in vivo somatic mutation) (see, e.g., Lonberg et al., (1994) Nature 368(6474):856-859); Lonberg, (1994) Handbook of Experimental Pharmacology 113:49-101; Lonberg & Huszar, (1995) Intern. Rev. Immunol. 13:65-93 and Harding & Lonberg, (1995) Ann. N.Y. Acad. Sci. 764:536-546). However, the term "human antibody" does not include antibodies in which CDR sequences derived from the germline of another mammalian species (e.g., mouse) have been grafted onto human framework sequences (i.e., humanized antibodies).

[0532] As used herein, the term "xenogeneic antibody" is defined in terms of the transgenic non-human organism that produces such antibodies. This term refers to an antibody having an amino acid sequence or an encoding nucleic acid sequence corresponding to an amino acid sequence or an encoding nucleic acid sequence found in an organism that is not composed of transgenic non-human animals and is typically from a species of non-transgenic non-human animal.

[0533] The terms "induce an immune response" and "enhance an immune response" are used interchangeably and refer to stimulating an immune response (i.e., passive or adaptive) to a particular antigen. The term "induce" as used for inducing CDC or ADCC refers to stimulating a particular direct cell killing mechanism.

[0534] As used herein, a subject "in need of prophylaxis", "in need of treatment", or "in need thereof" refers to a subject who would reasonably benefit from a given treatment (e.g., treatment with a composition comprising an anti-CD137 antibody) according to a suitable healthcare worker (e.g., a physician, nurse, or nurse practitioner for humans; a veterinarian for non-human mammals).

[0535] The term "in vivo" refers to a process that occurs in a living organism.

[0536] As used herein, the term "isolated antibody" is intended to refer to an antibody that is substantially free of other antibodies having different antigen specificities (e.g., an isolated antibody that specifically binds to human CD137 is substantially free of antibodies that specifically bind to antigens other than CD137). However, an isolated antibody that specifically binds to an epitope may be cross-reactive with other CD137 proteins from different species. However, in the specific binding assays described herein, the antibody continues to exhibit specific binding to human CD137. In addition, an isolated antibody is generally substantially free of other cellular materials and / or chemicals. In some embodiments, combinations of "isolated" antibodies having different CD137 specificities are combined in a well-defined composition.

[0537] As used herein, the term "isolated nucleic acid molecule" refers to a nucleic acid encoding an antibody or antibody portion that binds to CD137 (e.g., V H 、V L 、CDR3), and is intended to refer to such a nucleic acid molecule in which the nucleotide sequence encoding the antibody or antibody portion does not contain other nucleotide sequences encoding antibodies or antibody portions that bind to antigens other than CD137, and in the human genomic DNA, such other sequences may be naturally flanked by the nucleic acid. For example, a sequence selected from any of the sequences shown in Tables 22-27 corresponds to the nucleotide sequences of the heavy chain (V H ) and light chain (V L ) variable regions of the anti-CD137 antibody monoclonal antibodies described herein.

[0538] As used herein, "isotype" refers to the class of antibody encoded by the heavy chain constant region gene (e.g., IgM or IgGl). In some embodiments, the human monoclonal antibodies of the present disclosure are of the IgGl isotype. In some embodiments, the human monoclonal antibodies of the present disclosure are of the IgG1 isotype and contain mutations. In some embodiments, the human monoclonal antibodies of the present disclosure are of the IgG2 isotype. In some embodiments, the human monoclonal antibodies of the present disclosure are of the IgG3 isotype. In some embodiments, the human monoclonal antibodies of the present disclosure are of the IgG4 isotype. In some embodiments, the human monoclonal antibodies of the present disclosure are of the IgG4 isotype and contain mutations. In some embodiments, the mutation is a substitution at Ser228. In some embodiments, the substitution at Ser228 is S228P.

[0539] As used herein, the term "isotype switching" refers to the phenomenon in which the class or isotype of an antibody changes from one Ig class to another different Ig class.

[0540] As used herein, the term "KD" or "K D" refers to the equilibrium dissociation constant of the binding reaction between an antibody and an antigen. K D The value is a numerical representation of the ratio of the antibody dissociation rate constant (kd) to the antibody association rate constant (ka). K D The value is inversely correlated with the binding affinity of the antibody for the antigen. K D The smaller the value, the greater the affinity of the antibody for its antigen. Affinity is the strength of binding of an individual molecule to its ligand and is typically measured and reported by the equilibrium dissociation constant (K D ), which is used to evaluate and rank the strength of bimolecular interactions.

[0541] As used herein, the terms "kd" or "k d " (or "koff" or "k off ") are intended to refer to the dissociation rate constant at which an antibody dissociates from an antibody / antigen complex. The value of kd is a numerical representation of the fraction of the complex that decays or dissociates per second and is expressed in the unit sec -1 .

[0542] As used herein, the terms "ka" or "k a " (or "kon" or "k on ") are intended to refer to the association rate constant at which an antibody binds to an antigen. The value of ka is a numerical representation of the number of antibody / antigen complexes formed per second in a 1 molar (1M) solution of antibody and antigen and is expressed in the units M -1 sec -1 .

[0543] As used herein, the terms "linked," "fused," or "fusion" are used interchangeably. These terms refer to the joining together of two or more elements or components or domains by any means, including chemical coupling or recombinant means. Methods of chemical coupling (e.g., using heterobifunctional crosslinkers) are known in the art.

[0544] As used herein, "local administration" or "local delivery" refers to delivery that does not rely on the transport of a composition or agent via the vascular system to its target tissue or site. For example, a composition can be delivered by injection or implantation of the composition or agent or by injection or implantation of a device containing the composition or agent. After local administration near the target tissue or site, the composition or agent or one or more of its components can diffuse to the target tissue or site.

[0545] As used herein, the term "MHC molecule" refers to two types of molecules, MHC class I and MHC class II. MHC class I molecules present antigens to specific CD8+ T cells, and MHC class II molecules present antigens to specific CD4+ T cells. Antigens delivered exogenously to an APC are processed primarily for binding to MHC class II. In contrast, antigens delivered endogenously to an APC are processed primarily for binding to MHC class I.

[0546] As used herein, the term "monoclonal antibody" refers to an antibody that exhibits a single binding specificity and affinity for a particular epitope. Thus, the term "human monoclonal antibody" refers to an antibody that exhibits a single binding specificity and has variable and optionally constant regions derived from human germline immunoglobulin sequences. In some embodiments, a human monoclonal antibody is produced by a hybridoma that includes B cells obtained from a transgenic non-human animal, such as a transgenic mouse, having a genome that includes a human heavy chain transgene and a light chain transgene, which are fused with an immortalized cell.

[0547] As used herein, the term "oligomerization" refers to the formation of a macromolecular complex that includes more than two macromolecules (e.g., proteins) that typically bind through non-covalent interactions. Methods for assaying oligomerization are known to those of skill in the art and are described above for dimerization. In some embodiments, the anti-CD137 agonist antibodies described herein induce or enhance the oligomerization of CD137. In some embodiments, the anti-CD137 agonist antibodies described herein induce or enhance the oligomerization of CD137 relative to the amount of oligomerization in the absence of the anti-CD137 agonist antibody. In some embodiments, the anti-CD137 agonist antibodies described herein induce or enhance the oligomerization of CD137 relative to the amount of oligomerization in the presence of a reference anti-CD137 agonist antibody. In some embodiments, the oligomerization is increased by at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100%.

[0548] As used herein, the term "naturally occurring" when applied to an object refers to the fact that the object can be found in nature. For example, a polypeptide or polynucleotide sequence that exists in a living organism (including a virus) that can be isolated from a natural source and has not been intentionally modified by humans in the laboratory is naturally occurring.

[0549] As used herein, the term "nonswitched isotype" refers to the heavy chain isotype class that is produced when isotype switching has not occurred; the CH gene encoding the nonswitched isotype is typically the first CH gene downstream of the functionally rearranged VDJ gene. Isotype switching has been classified as classical or non-classical isotype switching. Classical isotype switching occurs through a recombination event involving at least one switch sequence region in the transgenic. Non-classical isotype switching can occur, for example, by homologous recombination between human σ μ and human ∑ μ (δ-related deletion). Other non-classical switching mechanisms, such as inter-transgenic and / or inter-chromosomal recombination, etc., may occur and effect isotype switching.

[0550] As used herein, the term "nucleic acid" refers to deoxyribonucleotides or ribonucleotides in single- or double-stranded form, and polymers thereof. Unless specifically restricted, the term encompasses nucleic acids containing known analogs of natural nucleotides that have similar binding properties as the reference nucleic acid and are metabolized in a manner similar to naturally occurring nucleotides. Unless otherwise indicated, a particular nucleic acid sequence also implicitly encompasses conservatively modified variants thereof (e.g., degenerate codon substitutions), and complementary sequences as well as the explicitly recited sequences. Specifically, degenerate codon substitutions may be achieved by generating sequences in which one or more selected (or all) codons' third positions are substituted with mixed bases and / or deoxyinosine residues (Batzer et al., Nucleic Acid Res. 19:5081, 1991; Ohtsuka et al., Biol. Chem. 260:2605-2608, 1985; and Cassol et al, 1992; Rossolini et al, Mol. Cell. Probes 8:91-98, 1994). For arginine and leucine, modifications at the second base may also be conservative. The term nucleic acid may be used interchangeably with gene, cDNA, and mRNA encoded by a gene.

[0551] The polynucleotides used herein can comprise any polyribonucleotides or polydeoxyribonucleotides, which can be unmodified RNA or DNA or modified RNA or DNA. For example, polynucleotides can comprise single-stranded and double-stranded DNA, DNA with a mixture of single-stranded and double-stranded regions, single-stranded and double-stranded RNA, and RNA with a mixture of single-stranded and double-stranded regions, hybrid molecules comprising DNA and RNA that can be single-stranded or more typically double-stranded or with a mixture of single-stranded and double-stranded regions. In addition, polynucleotides can comprise triple-stranded regions containing RNA or DNA or both RNA and DNA. Polynucleotides can also contain one or more modified bases or a DNA or RNA backbone modified for stability or other reasons. "Modified" bases include, for example, tritylated bases and unusual bases such as inosine. A variety of modifications can be made to DNA and RNA; thus, "polynucleotide" encompasses chemically, enzymatically, or metabolically modified forms.

[0552] When a nucleic acid is placed into a functional relationship with another nucleic acid sequence, it is "operably linked". For example, if a promoter or enhancer affects the transcription of a coding sequence, it is operably linked to that sequence. With respect to transcriptional regulatory sequences, operably linked means that the DNA sequences being linked are contiguous, and, in the case where two protein-coding regions are to be linked, contiguous and in the reading frame. For transposable sequences, operably linked means that the sequences are capable of effecting transposition recombination.

[0553] As used herein, the term "paratope", also known as "antigen-binding site", refers to a part of an antibody or an antigen-binding fragment thereof that recognizes and binds to an epitope on an antigen and comprises a set of complementarity-determining regions (CDRs) located within the variable heavy and light chains.

[0554] As used herein, "parenteral administration", "parenterally administered", and other grammatically equivalent phrases refer to modes of administration other than enteral and topical administration, typically by injection, and include, but are not limited to, intravenous, intranasal, intraocular, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, intratracheal, subcutaneous, subepidermal, intraarticular, subcapsular, subarachnoid, intraspinal, epidural, intracerebral, intracranial, intracarotid, and intrasternal injection and infusion.

[0555] As used herein, the term "patient" includes human and other mammalian subjects who are undergoing prophylactic or therapeutic treatment.

[0556] In the case of two or more nucleic acid or polypeptide sequences, the term "percent identity" refers to the percentage of identical nucleotide or amino acid residues between two or more sequences or subsequences when compared and aligned for maximum correspondence, determined using one of the following sequence comparison algorithms (e.g., BLASTP and BLASTN or other algorithms available to those of skill in the art) or by visual inspection. Depending on the application, the "percent identity" may exist over a region of the sequences being compared, such as a functional domain, or over the full length of the two sequences being compared. For sequence comparison, typically one sequence acts as a reference sequence to which the test sequence is compared. When using a sequence comparison algorithm, the test and reference sequences are entered into a computer, subsequence coordinates are designated if necessary, and sequence algorithm program parameters are designated. The sequence comparison algorithm then calculates the percent sequence identity of the test sequence relative to the reference sequence based on the designated program parameters.

[0557] Optimal alignment of sequences for comparison can be conducted, e.g., by the local homology algorithm of Smith & Waterman, Adv. Appl. Math. 2:482 (1981), by the homology alignment algorithm of Needleman & Wunsch, J. Mol. Biol. 48:443 (1970), by the similarity search method of Pearson & Lipman, Proc. Nat'l. Acad. Sci. USA 85:2444 (1988), by computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, Wis.), or by visual inspection (generally see Ausubel et al., supra).

[0558] An example of an algorithm suitable for determining percent sequence identity and sequence similarity is the BLAST algorithm, described in Altschul et al., J. Mol. Biol. 215:403-410 (1990). Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information website.

[0559] As used herein generally, "pharmaceutically acceptable" means those compounds, materials, compositions, and / or dosage forms that, within the scope of sound medical judgment, are suitable for use in contact with the tissues, organs, and / or bodily fluids of humans and animals without excessive toxicity, irritation, allergic response, or other problems or complications commensurate with a reasonable benefit / risk ratio.

[0560] As used herein, "pharmaceutically acceptable carrier" means and includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like that are physiologically compatible. The compositions can contain pharmaceutically acceptable salts, such as acid addition salts or base addition salts (see, e.g., Berge et al. (1977) J Pharm Sci 66 : 1-19).

[0561] As used herein, the terms "polypeptide", "peptide", and "protein" are used interchangeably to refer to polymers of amino acid residues. This term applies to amino acid polymers in which one or more of the amino acid residues are artificial chemical mimics of the corresponding naturally occurring amino acids, as well as to naturally occurring amino acid polymers and non-naturally occurring amino acid polymers.

[0562] As used herein, the term "prevent" when used in connection with a medical condition means that, relative to a subject who has not received the composition, administration of the composition reduces the frequency of symptoms of the medical condition in the subject or delays its onset.

[0563] As used herein, the terms "purified" or "isolated" as applied to any protein (antibody or fragment) described herein mean a polypeptide that has been separated or purified from the components that naturally accompany it (e.g., other proteins, lipids, and nucleic acids in a prokaryote that expresses the protein). Typically, a polypeptide is purified when it constitutes at least 60 (e.g., at least 65, 70, 75, 80, 85, 90, 92, 95, 97, or 99) % of the total protein weight in a sample.

[0564] As used herein, the term "rearranged" refers to the configuration of a heavy or light chain immunoglobulin locus in which a V segment is arranged adjacent to a D-J or J segment in a configuration that encodes substantially a complete V H or V L domain. Rearranged immunoglobulin loci can be identified by comparison to germline DNA; a rearranged locus will have at least one recombined heptamer / nonamer homology element.

[0565] As used herein, the term "receptor clustering" refers to a cellular process that results in the aggregation or local accumulation of a set of receptors at a specific cellular location, typically to induce or amplify a signaling response. Many protein receptors bind to cognate ligands and cluster, i.e., form dimers, trimers, oligomers, or polymers, after binding their cognate ligands. For example, the PDGF receptor and members of the TNF receptor superfamily form dimers and trimers, respectively, upon ligand binding. Cognate ligand-induced clustering (e.g., dimerization, polymerization) transduces signals through the receptor. Accordingly, the antibodies or antigen-binding fragments thereof of the present disclosure are capable of activating a receptor by binding to more than one receptor and inducing or stabilizing dimerization, trimerization, and / or polymerization, with or without cognate ligand binding.

[0566] TNFR signaling requires receptor clustering and polymerization (Wajant (2015) Cell Death Differ 22(11):1727-1741), and in particular TNFRSF activation. 4-1BB (CD137), CD40, GITR, CD27, DR3, DR5, and Fas are some TNFSF receptors known to require clustering to trigger downstream signaling. Experimental evidence that the 4-1BB receptor must be crosslinked for signaling comes from Rabu et al. These authors reported that the 1-trimer form of human 4-1BBL had no activating effect on human T cells, while crosslinking the protein into 2- or more trimers resulted in strong activation (Rabu et al., (2005) J Biol Chem 280:41472-41481). Thus, in some embodiments, an anti-CD137 agonist antibody induces the polymerization of 2 or more trimers of CD137.

[0567] As used herein, the term "recombinant host cell" (or simply "host cell") is intended to refer to a cell into which a recombinant expression vector has been introduced. It should be understood that such term is intended to refer not only to the particular subject cell but also to the progeny of such a cell. Since certain modifications may occur in progeny due to mutation or environmental influences, such progeny may in fact be different from the parental cell, but are still included within the scope of the term "host cell" as used herein.

[0568] As used herein, the term "recombinant human antibody" includes all human antibodies prepared, expressed, produced or isolated by recombinant means, e.g., (a) antibodies isolated from animals (e.g., mice) that are transgenic or transchromosomal for human immunoglobulin genes or from hybridomas prepared therefrom, (b) antibodies isolated from host cells transformed to express an antibody, e.g., from a transfectoma, (c) antibodies isolated from a recombinant combinatorial human antibody library, and (d) antibodies prepared, expressed, produced or isolated by any other means involving splicing of human immunoglobulin gene sequences to other DNA sequences. Such recombinant human antibodies contain variable and constant regions that utilize specific human germline immunoglobulin sequences encoded by germline genes, but include subsequent rearrangements and mutations that occur, e.g., during antibody maturation. As is known in the art (see, e.g., Lonberg (2005) Nature Biotech. 23(9):1117-1125), the variable region contains the antigen-binding domain, which is encoded by multiple genes that have rearranged to form an antibody specific for a foreign antigen. In addition to rearrangement, the variable region may be further modified by multiple single amino acid changes (termed somatic mutations or hypermutations) to increase the affinity of the antibody for the foreign antigen. The constant region will further vary in response to antigen (i.e., isotype switching). Thus, rearranged and somatically mutated nucleic acid molecules encoding light and heavy chain immunoglobulin polypeptides in response to antigen may not have sequence identity with the original nucleic acid molecules, but will be substantially the same or similar (i.e., have at least 80% identity).

[0569] As used herein, the term "reference antibody" (used interchangeably with "reference mAb") or "reference antigen-binding protein" refers to an antibody or antigen-binding fragment thereof that binds to a specific epitope on human CD137 and is used to establish a relationship between itself and one or more different antibodies. In some embodiments, the relationship is the binding of the reference antibody and one or more different antibodies to the same epitope on CD137. As used herein, this term means that an anti-CD137 antibody that can be used as a competitor in a test or assay such as those described herein (e.g., a competitive binding assay), where the assay can be used to discover, identify or develop one or more different antibodies that bind to the same epitope. The variable heavy chain (V H ) and light chain (V L ) amino acid sequences of an exemplary reference antibody (mAb1) are provided in Table 23 (V H 1, SEQ ID NO. 4; V H2, SEQ ID NO. 6). In some embodiments, this term refers to an anti-CD137 antibody that can be used as a comparator in a test or assay that can be used to distinguish characteristics of the antibody (e.g., hepatotoxicity, anti-tumor efficacy). In some embodiments, the reference antibody is usulerumab. In some embodiments, the reference antibody is utolumab.

[0570] As used herein, the terms "specifically bind," "selectively bind," "selectively binds to," and "specifically binds to" refer to the binding of an antibody to an epitope on a predetermined antigen. Typically, an antibody binds to an epitope with a specific binding affinity of less than 10% when measured by the following methods: -6 M, for example, less than about 10 -7 M, 10 -8 M, 10 -9 M or 10 -10 M or even lower equilibrium dissociation constant (K D ), the method uses surface plasmon resonance (SPR) technology in a BIACORE 2000 instrument, uses recombinant human CD137 as an analyte, and the antibody is used as a ligand and binds to a predetermined antigen with an affinity that is at least two times higher than its affinity for binding to a non-specific antigen (e.g., BSA, casein) other than the predetermined antigen or a closely related antigen. The phrases "antibodies that recognize antigens" and "antibodies specific for antigens" are used interchangeably herein with the term "antibodies that specifically bind to antigens."

[0571] As used herein, the term "switch sequence" refers to those DNA sequences responsible for switch recombination. The "switch donor" sequence, typically the μ switch region, will be located 5' (i.e., upstream) of the constructed region to be deleted during switch recombination. The "switch acceptor" region will be located between the constructed region to be deleted and the replacement constant region (e.g., γ, ε, etc.). Because there is no specific site where recombination always occurs, the final gene sequence is generally unpredictable from the construct.

[0572] As used herein, the term "subject" includes any human or non-human animal. For example, the methods and compositions of the present invention can be used to treat a subject suffering from an immune disorder. The term "non-human animal" includes all vertebrates, such as mammals and non-mammals, such as non-human primates, sheep, dogs, cows, chickens, amphibians, reptiles, etc.

[0573] For nucleic acids, the term "substantially homologous" means that, when optimally aligned and compared, two nucleic acids or their designated sequences are identical, having appropriate nucleotide insertions or deletions in at least about 80% of the nucleotides, usually at least about 90% to 95%, and more preferably at least about 98% to 99.5% of the nucleotides. Alternatively, substantial homology exists when a fragment hybridizes to the complement of the strand under selective hybridization conditions.

[0574] The percent identity between two sequences is a function of the number of identical positions shared by the sequences (i.e., % homology = number of identical positions / total number of positions x 100), taking into account the number of gaps that need to be introduced for the optimal alignment of the two sequences and the length of each gap. The comparison of sequences and the determination of the percent identity between two sequences can be accomplished using mathematical algorithms, as described in the following non-limiting examples.

[0575] The percent identity between two nucleotide sequences can be determined using the GAP program in the GCG software package (available from http: / / www.gcg.com), using the NWSgapdna.CMP matrix and gap weights of 40, 50, 60, 70, or 80 and length weights of 1, 2, 3, 4, 5, or 6. The algorithm of E. Meyers and W. Miller (CABIOS, 4:11-17 (1989)), incorporated into the ALIGN program (version 2.0), can also be used, using the PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4, to determine the percent identity between two nucleotide or amino acid sequences. In addition, the algorithm of Needleman and Wunsch (J. Mol. Biol. (48):444-453 (1970)), incorporated into the GAP program in the GCG software package (available from http: / / www.gcg.com), can be used, using the Blossum 62 matrix or the PAM250 matrix, and gap weights of 16, 14, 12, 10, 8, 6, or 4 and length weights of 1, 2, 3, 4, 5, or 6, to determine the percent identity between two amino acid sequences.

[0576] The nucleic acid and protein sequences of the present disclosure can further be used as a "query sequence" to search against public databases to identify related sequences, for example. Such searches can be performed using the NBLAST and XBLAST programs (version 2.0) of Altschul, et al. (1990) J. Mol. Biol. 215:403-10. A BLAST nucleotide search can be performed using the NBLAST program, score = 100, wordlength = 12, to obtain nucleotide sequences homologous to the nucleic acid molecules of the present invention. A BLAST protein search can be performed using the XBLAST program, score = 50, wordlength = 3, to obtain amino acid sequences homologous to the protein molecules of the present invention. To obtain gapped alignments for comparison purposes, gapped BLAST can be used as described in Altschul et al., (1997) Nucleic Acids Res. 25(17):3389-3402. When using the BLAST and gapped BLAST programs, the default parameters of the respective programs (e.g., XBLAST and NBLAST) can be used. See http: / / www.ncbi.nlm.nih.gov.

[0577] Nucleic acids can be present in whole cells, cell lysates, or in a partially purified or substantially pure form. A nucleic acid is "isolated" or "substantially pure" when purified from other cellular components or other contaminants (e.g., other cellular nucleic acids or proteins) by standard techniques including alkaline / SDS treatment, CsCl banding, column chromatography, agarose gel electrophoresis, and other methods known in the art. See F. Ausubel, et al., ed. Current Protocols in Molecular Biology, Greene Publishing and Wiley Interscience, New York (1987).

[0578] The nucleic acid compositions of the present disclosure from cDNA, genomic, or mixtures thereof, although typically in the native sequence (except for modified restriction sites, etc.), can also be mutated using standard techniques to provide gene sequences. For coding sequences, these mutations can affect the amino acid sequence as desired. Specifically, DNA sequences that are substantially homologous to or derived from the native V, D, J, constant, switch, and other such sequences described herein are encompassed (where "derived" means the sequence is the same as or modified from another sequence).

[0579] As used herein, the term "tumor microenvironment" (or "cancer microenvironment"; abbreviated as TME) refers to the cellular environment or milieu in which a tumor or neoplasm exists, including the surrounding blood vessels as well as non-cancer cells, including but not limited to immune cells, fibroblasts, bone marrow-derived inflammatory cells, and lymphocytes. Signaling molecules and the extracellular matrix are also included in the TME. The tumor is closely associated with and continuously interacts with the surrounding microenvironment. The tumor can affect the microenvironment by releasing extracellular signals, promoting tumor angiogenesis, and inducing peripheral immune tolerance, while immune cells in the microenvironment can affect the growth and evolution of tumor cells.

[0580] The term "T cell" refers to a type of white blood cell that can be distinguished from other white blood cells by the presence of a T cell receptor on its cell surface. There are multiple subsets of T cells, including but not limited to T helper cells (also known as T H cells or CD4 + T cells) and subtypes, including T H 1, T H 2, T H 3, T H 17, T H 9 and T FH cells, cytotoxic T cells (i.e., T C cells, CD8 + T cells, cytotoxic T lymphocytes, T killer cells, killer T cells), memory T cells and subtypes, including central memory T cells (T CM cells), effector memory T cells (T EM and T EMRA cells) and resident memory T cells (T RM cells), regulatory T cells (also known as T reg cells or suppressor T cells) and subtypes, including CD4 + FOXP3 + T reg cells, CD4 + FOXP3 - T reg cells, Tr1 cells, Th3 cells and T reg 17 cells, natural killer T cells (also known as NKT cells), mucosal associated invariant T cells (MAIT), and gamma delta T cells (γδT cells), including Vγ9 / Vδ2 T cells. Any one or more of the above-mentioned or unmentioned T cells can be the target cell type for the methods of the present invention.

[0581] As used herein, the term "T cell activation" or "activation of T cells" refers to a cellular process in which mature T cells expressing antigen-specific T cell receptors on their surface recognize their cognate antigen and respond by entering the cell cycle, secreting cytokines or lytic enzymes, and initiating or becoming capable of performing cell-based effector functions. T cell activation requires at least two signals for full activation. The first occurs after the T cell antigen-specific receptor (TCR) engages the antigen-major histocompatibility complex (MHC), and the second occurs upon engagement of subsequent costimulatory molecules (such as CD28). These signals are transmitted to the nucleus and result in clonal expansion of T cells, upregulation of cell surface activation markers, differentiation into effector cells, induction of cytotoxicity or cytokine secretion, induction of apoptosis, or a combination thereof.

[0582] As used herein, the term "T cell-mediated response" refers to any response mediated by T cells, including but not limited to effector T cells (such as CD8 + cells) and helper T cells (such as CD4 + cells). T cell-mediated responses include, for example, cytotoxicity and proliferation of T cells.

[0583] As used herein, the term "therapeutically effective amount" or "therapeutically effective dose" or similar terms used herein are intended to mean the amount of an agent (anti-CD137 antibody or antigen-binding fragment thereof) that will elicit the desired biological or medical response (such as improvement of one or more symptoms of cancer).

[0584] As used herein, the terms "treatment" and "therapy" refer to the therapeutic or prophylactic measures described herein. The method of "treatment" involves administering the human antibodies of the present disclosure to a subject in need of such treatment, for example, a subject in need of enhancing an immune response against a specific antigen, or a subject who may ultimately develop such a disorder, to prevent, cure, delay, reduce the severity of, or improve one or more symptoms of the disorder or recurrence of the disorder, or to extend the survival period of the subject beyond that expected in the absence of such treatment.

[0585] As used herein, the term "unrearranged" or "germline configuration" refers to a configuration in which the V segment has not been recombined to be adjacent to a D or J segment.

[0586] As used herein, the term "vector" is intended to refer to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked. One type of vector is a "plasmid", which refers to a circular double-stranded DNA loop into which additional DNA segments can be ligated. Another type of vector is a viral vector, into which additional DNA segments can be ligated into the viral genome. Certain vectors are capable of autonomous replication in the host cells into which they are introduced (e.g., bacterial vectors having a bacterial origin of replication and episomal mammalian vectors). Other vectors (e.g., non-episomal mammalian vectors) can integrate into the genome of the host cell after being introduced into the host cell and thereby replicate with the host genome. In addition, certain vectors are capable of directing the expression of genes operably linked thereto. Such vectors are referred to herein as "recombinant expression vectors" (or simply "expression vectors"). In general, expression vectors useful in recombinant DNA techniques are commonly in the form of plasmids. In the present specification, "plasmid" and "vector" are used interchangeably because plasmids are the most commonly used form of vectors. However, the present invention is intended to include such other forms of expression vectors, such as viral vectors (e.g., replication-defective retroviruses, adenoviruses, and adeno-associated viruses), which have equivalent functions.

[0587] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Preferred methods and materials are described below, although methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the methods and compositions of this disclosure. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety.

[0588] Anti-CD137 preparation

[0589] The present disclosure provides a preparation comprising an antibody that binds to human CD137 or an antigen-binding fragment thereof [hereinafter referred to as an "anti-CD137 antibody" or an "anti-human CD137 antibody"]. The anti-CD137 antibody preparation of the present disclosure maintains the antibody or its antigen-binding fragment under stable conditions, minimizes the formation of antibody aggregates (high molecular weight species) and particulates, reduces the percentage of charge variants, and maintains the structural integrity of the antibody.

[0590] On the one hand, the present disclosure provides, at least in part, various formulations of an anti-CD137 antibody or an antigen-binding fragment thereof. In some embodiments, the formulations of the present disclosure comprising an anti-CD137 or an antigen-binding fragment thereof further comprise: (i) a buffer (e.g., histidine), (ii) a disaccharide (e.g., sucrose), (iii) a non-ionic surfactant (e.g., polysorbate 80), and / or (iv) a salt (e.g., NaCl). In some embodiments, the formulations of the present disclosure have a pH of from about 5.0 to about 7.0. In some embodiments, the formulations of the present disclosure have a pH of from about 5.0 to about 7.4. In some embodiments, the pH of the formulation is from about 5.0 to about 8.0. In some embodiments, the formulations of the present disclosure further contain one or more solubilizers, diluents, binders, stabilizers, salts, lipophilic solvents, amino acids, chelating agents, or preservatives.

[0591] (i) Buffer

[0592] In some embodiments, a buffer is included in the antibody formulation to improve stability and / or control the pH of the formulation. As illustrated in the working examples described herein, the anti-CD137 antibody is stable at high concentrations and under forced degradation conditions when formulated in a histidine buffer.

[0593] In some embodiments, buffers that can be used in the formulations described herein include, for example, salts of citric acid, ascorbic acid, gluconic acid, carbonic acid, tartaric acid, succinic acid, acetic acid, or phthalic acid. In some embodiments, the buffer is a Tris-based buffer or a phosphate buffer.

[0594] In some embodiments, the formulations described herein comprise one or more amino acids that can provide buffering capacity and the like. Suitable amino acids for use in the formulations of the present disclosure include, for example, histidine, glycine, and serine. In some embodiments, the formulations of the present disclosure do not contain free amino acids as buffers. In some embodiments, the formulations of the present disclosure contain one free amino acid (e.g., histidine) as a buffer. In some embodiments, the formulations of the present disclosure contain two or more (e.g., two, three, four, five, six, or seven or more) different amino acids as buffers, such as serine and histidine.

[0595] Buffers are typically used at a concentration between about 10 mM and 100 mM, depending in part on the desired buffering capacity. In some embodiments, the formulations described herein contain a buffer at a concentration less than or about 100 mM (e.g., less than or about 90 mM, 80 mM, 70 mM, 60 mM, 50 mM, 40 mM, 30 mM, 25 mM, 20 mM, 15 mM, or 10 mM). In some embodiments, the formulations described herein contain a buffer at a concentration of at least 10 mM (e.g., at least 15 mM, 20 mM, 25 mM, 30 mM, 35 mM, 40 mM, 45 mM, 50 mM, 55 mM, 60 mM, 65 mM, 70 mM, 75 mM, 80 mM, 85 mM, 90 mM, 95 mM, 100 mM, or higher). In some embodiments, the formulations described herein contain a buffer at a concentration between about 10 mM and 20 mM, 15 mM and 20 mM, 10 mM and 25 mM, 15 mM and 25 mM, 20 mM and 25 mM, 10 mM and 30 mM, 15 mM and 30 mM, 20 mM and 30 mM, 25 mM and 30 mM, 10 mM and 40 mM, 15 mM and 40 mM, 20 mM and 40 mM, 25 mM and 40 mM, 30 mM and 40 mM, 10 mM and 50 mM, 15 mM and 50 mM, 20 mM and 50 mM, 25 mM and 50 mM, 30 mM and 50 mM, 40 mM and 50 mM, 10 mM and 100 mM, 15 mM and 100 mM, 20 mM and 100 mM, 25 mM and 100 mM, 30 mM and 100 mM, 40 mM and 100 mM, or 50 and 100 mM. It should be understood that in embodiments where the formulations of the present disclosure contain two or more (e.g., at least two, three, four, five, six, seven, eight, nine, or ten or more) different buffers, each of the two or more buffers can be present independently, e.g., at one of the above concentrations.

[0596] In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 10 - 100 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 15 - 100 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 20 - 100 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 25 - 100 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 30 - 100 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 40 - 100 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 50 - 100 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 10 - 50 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 15 - 50 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 20 - 50 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 25 - 50 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 30 - 50 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 40 - 50 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 10 - 40 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 15 - 40 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 20 - 40 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 25 - 40 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 30 - 40 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 10 - 30 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 15 - 30 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 20 - 30 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 25 - 30 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 10 - 25 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 15 - 25 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 20 - 25 mM.In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 10-20 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 15-20 mM.

[0597] In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 10 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 11 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 12 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 13 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 14 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 15 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 16 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 17 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 18 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 19 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 20 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 21 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 22 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 23 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 24 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 25 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 26 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 27 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 28 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 29 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 30 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 35 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 40 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 45 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 50 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 55 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 60 mM.In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 65 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 70 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 75 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 80 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 85 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 90 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 95 mM. In some embodiments, the formulations of the present disclosure comprise a buffer containing histidine at a concentration of about 100 mM.

[0598] (ii) Carbohydrate excipients

[0599] In some embodiments, a carbohydrate excipient is added to the antibody formulations of the present disclosure to improve stability. As provided in the working examples, it was found that the addition of sucrose to the anti-CD137 formulations of the present disclosure resulted in improved stability at elevated temperatures.

[0600] In some embodiments, the formulations described herein all contain a carbohydrate excipient. Suitable carbohydrate excipients are described, for example, in Katakam and Banga (1995) J Pharm Pharmacol 47(2) : 103-107; Andya et al. (2003) AAPS PharmSci 5(2):Article 10; and in Shire (2009) "Current Trends in Monoclonal Antibody Development and Manufacturing," Volume 11, Springer, 354 pages. Carbohydrate excipients suitable for the formulations described herein include, but are not limited to, monosaccharides such as fructose, maltose, galactose, glucose, D-mannose, and sorbose; disaccharides such as lactose, sucrose, trehalose, and cellobiose; polysaccharides such as maltodextrin, dextran, and starch; and sugar alcohols such as mannitol, xylitol, maltitol, lactitol, and sorbitol. In some embodiments, the carbohydrate excipient is a disaccharide (disaccharide or disaccharide sugar). In some embodiments, the disaccharide is sucrose. In some embodiments, the carbohydrate excipient is present in the formulations of the present disclosure at a concentration of at least or about 5% (e.g., at least or about 5.0%, 5.5%, 6.0%, 6.5%, 7.0%, 7.5%, 8.0%, 8.5%, 9.0%, 9.5%, 10.0%, 10.5%, 11.0%, 11.5%, 12.0%, 12.5%, 13.0%, 13.5%, 14.0%, 14.5%, 15.0% or higher) weight / volume (w / v). In embodiments where the formulations of the present disclosure contain two or more (e.g., at least two, three, four, five, six, seven, eight, nine, or ten or more) different carbohydrate excipients (e.g., sorbitol and mannitol), each excipient can be present independently at any of the above concentrations. In some embodiments of the present disclosure, the carbohydrate excipient is present in an amount of about 5-15% (w / v) (5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, or 15%). In some embodiments, the carbohydrate excipient is present in an amount of about 6% to about 15% (w / v). In some embodiments, the carbohydrate excipient is present in an amount of about 8% to about 15% (w / v). In some embodiments, the carbohydrate excipient is present in an amount of about 10% to about 15% (w / v). In some embodiments, the carbohydrate excipient is present in an amount of about 12% to about 15% (w / v). In some embodiments, the carbohydrate excipient is present in an amount of about 14% to about 15% (w / v). In some embodiments, the carbohydrate excipient is present in an amount of about 5% to about 12% (w / v). In some embodiments, the carbohydrate excipient is present in an amount of about 6% to about 12% (w / v). In some embodiments, the carbohydrate excipient is present in an amount of about 8% to about 12% (w / v). In some embodiments, the carbohydrate excipient is present in an amount of about 10% to about 12% (w / v).In some embodiments, the carbohydrate excipient is present in an amount of about 5% to about 10% (w / v). In some embodiments, the carbohydrate excipient is present in an amount of about 6% to about 10% (w / v). In some embodiments, the carbohydrate excipient is present in an amount of about 8% to about 10% (w / v). In some embodiments, the carbohydrate excipient is a disaccharide. In some embodiments, the carbohydrate excipient is sucrose.

[0601] In some embodiments, the carbohydrate excipient is present in an amount of about 5% (w / v). In some embodiments, the carbohydrate excipient is present in an amount of about 6% (w / v). In some embodiments, the carbohydrate excipient is present in an amount of about 7% (w / v). In some embodiments, the carbohydrate excipient is present in an amount of about 8% (w / v). In some embodiments, the carbohydrate excipient is present in an amount of about 9% (w / v). In some embodiments, the carbohydrate excipient is present in an amount of about 10% (w / v). In some embodiments, the carbohydrate excipient is present in an amount of about 11% (w / v). In some embodiments, the carbohydrate excipient is present in an amount of about 12% (w / v). In some embodiments, the carbohydrate excipient is present in an amount of about 13% (w / v). In some embodiments, the carbohydrate excipient is present in an amount of about 14% (w / v). In some embodiments, the carbohydrate excipient is present in an amount of about 15% (w / v). In some embodiments, the carbohydrate excipient is a disaccharide. In some embodiments, the carbohydrate excipient is sucrose. In some embodiments, the carbohydrate excipient is trehalose.

[0602] In some embodiments, the formulations of the present disclosure contain a disaccharide. In some embodiments, the formulations of the present disclosure contain sucrose. In some embodiments, the carbohydrate excipient is trehalose.

[0603] In some embodiments, the formulations of the present disclosure comprise sucrose in an amount of from about 5% to about 15% (w / v). In some embodiments, the formulations of the present disclosure comprise sucrose in an amount of from about 6% to about 15% (w / v). In some embodiments, the formulations of the present disclosure comprise sucrose in an amount of from about 8% to about 15% (w / v). In some embodiments, the formulations of the present disclosure comprise sucrose in an amount of from about 10% to about 15% (w / v). In some embodiments, the formulations of the present disclosure comprise sucrose in an amount of from about 12% to about 15% (w / v). In some embodiments, the formulations of the present disclosure comprise sucrose in an amount of from about 14% to about 15% (w / v). In some embodiments, the formulations of the present disclosure comprise sucrose in an amount of from about 5% to about 12% (w / v). In some embodiments, the formulations of the present disclosure comprise sucrose in an amount of from about 6% to about 12% (w / v). In some embodiments, the formulations of the present disclosure comprise sucrose in an amount of from about 8% to about 12% (w / v). In some embodiments, the formulations of the present disclosure comprise sucrose in an amount of from about 10% to about 12% (w / v). In some embodiments, the formulations of the present disclosure comprise sucrose in an amount of from about 5% to about 10% (w / v). In some embodiments, the formulations of the present disclosure comprise sucrose in an amount of from about 6% to about 10% (w / v). In some embodiments, the formulations of the present disclosure comprise sucrose in an amount of from about 8% to about 10% (w / v).

[0604] In some embodiments, the formulations of the present disclosure comprise about 5% (w / v) of sucrose. In some embodiments, the formulations of the present disclosure comprise about 6% (w / v) of sucrose. In some embodiments, the formulations of the present disclosure comprise about 7% (w / v) of sucrose. In some embodiments, the formulations of the present disclosure comprise about 8% (w / v) of sucrose. In some embodiments, the formulations of the present disclosure comprise about 9% (w / v) of sucrose. In some embodiments, the formulations of the present disclosure comprise about 10% (w / v) of sucrose. In some embodiments, the formulations of the present disclosure comprise about 11% (w / v) of sucrose. In some embodiments, the formulations of the present disclosure comprise about 12% (w / v) of sucrose. In some embodiments, the formulations of the present disclosure comprise about 13% (w / v) of sucrose. In some embodiments, the formulations of the present disclosure comprise about 14% (w / v) of sucrose. In some embodiments, the formulations of the present disclosure comprise about 15% (w / v) of sucrose.

[0605] (iii) Surfactant

[0606] In some embodiments, the antibody formulations of the present disclosure contain a surfactant. As used herein, the surfactant is an amphiphilic surfactant in nature. In some embodiments, the surfactant is added to the formulations herein to provide stability, reduce and / or prevent aggregation, or prevent and / or inhibit protein damage during processing conditions such as purification, filtration, lyophilization, transportation, storage, and delivery. In some embodiments of the present disclosure, the surfactant can be used to provide stability to the active ingredient.

[0607] In some embodiments, the formulations of the present disclosure contain a surfactant, such as an anionic, cationic, or nonionic surfactant. In some embodiments, the surfactant is a polyoxyethylene sorbitan fatty acid ester (polysorbate, sold under the trade name ), including polysorbate-20 (polyoxyethylene sorbitan monolaurate), polysorbate-40 (polyoxyethylene sorbitan monopalmitate), polysorbate-60 (polyoxyethylene sorbitan monostearate), and polysorbate-80 (polyoxyethylene sorbitan monooleate); polyoxyethylene alkyl ethers, such as 58 and 35; poloxamer (e.g., poloxamer 188); X-100 and X-114; NP40; Span20, Span40, Span60, Span65, Span80, and Span85; copolymers of ethylene and propylene glycol (e.g., series of nonionic surfactants, such as F68, 10R5, F108, F127, F38, L44, L62; and sodium dodecyl sulfate (SDS). In some embodiments, the surfactant is a nonionic surfactant. In some embodiments, the surfactant is a polysorbate. In some embodiments, the surfactant is polysorbate 80.

[0608] The amount of the surfactant included in the formulations of the present invention is an amount sufficient to perform the desired function, i.e., the minimum amount required to stabilize the active pharmaceutical ingredient (i.e., the anti-CD137 antibody or its antigen-binding fragment) in the formulation. All percentages regarding the surfactant are listed as w / v%.

[0609] In some embodiments, the formulations described herein contain a surfactant (e.g., any pharmaceutically acceptable surfactant described herein or known in the art) at a concentration of at least or about 0.01 (e.g., at least or about 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1 or higher) w / v%. In some embodiments, the formulations of the present disclosure contain no more than 0.1 (e.g., no more than 0.09, 0.08, 0.07, 0.06, 0.05, 0.04, 0.03, 0.02 or 0.01) w / v% of a pharmaceutically acceptable surfactant.

[0610] In some embodiments, the formulations of the present disclosure contain a surfactant at a concentration of about 0.01% to about 0.1% w / v. In some embodiments, the formulations of the present disclosure contain the following amounts of surfactant: about 0.01% to about 0.09% w / v; about 0.01% to about 0.08% w / v; about 0.01% to about 0.07% w / v; about 0.01% to about 0.06% w / v; about 0.01% to about 0.05% w / v; about 0.01% to about 0.04% w / v; about 0.01% to about 0.03% w / v, or about 0.01% to about 0.02% w / v.

[0611] In some embodiments, the formulations of the present disclosure contain a surfactant in an amount of about 0.01% w / v. In some embodiments, the formulations of the present disclosure contain a surfactant in an amount of about 0.02% w / v. In some embodiments, the formulations of the present disclosure contain a surfactant in an amount of about 0.025% w / v. In some embodiments, the formulations of the present disclosure contain a surfactant in an amount of about 0.03% w / v. In some embodiments, the formulations of the present disclosure contain a surfactant in an amount of about 0.035% w / v. In some embodiments, the formulations of the present disclosure contain a surfactant in an amount of about 0.04% w / v. In some embodiments, the formulations of the present disclosure contain a surfactant in an amount of about 0.05% w / v. In some embodiments, the formulations of the present disclosure contain a surfactant in an amount of about 0.06% w / v. In some embodiments, the formulations of the present disclosure contain a surfactant in an amount of about 0.07% w / v. In some embodiments, the formulations of the present disclosure contain a surfactant in an amount of about 0.08% w / v. In some embodiments, the formulations of the present disclosure contain a surfactant in an amount of about 0.09% w / v. In some embodiments, the formulations of the present disclosure contain a surfactant in an amount of about 0.1% w / v.

[0612] In some embodiments, the surfactant in the formulations of the present disclosure is polysorbate 80. In some embodiments, the formulations of the present disclosure contain polysorbate 80 in an amount of about 0.01% to about 0.1% w / v. In some embodiments, the formulations of the present disclosure contain polysorbate 80 in an amount of about 0.01% to about 0.09% w / v. In some embodiments, the formulations of the present disclosure contain polysorbate 80 in an amount of about 0.01% to about 0.08% w / v. In some embodiments, the formulations of the present disclosure contain polysorbate 80 in an amount of about 0.01% to about 0.07% w / v. In some embodiments, the formulations of the present disclosure contain polysorbate 80 in an amount of about 0.01% to about 0.06% w / v. In some embodiments, the formulations of the present disclosure contain polysorbate 80 in an amount of about 0.01% to about 0.05% w / v. In some embodiments, the formulations of the present disclosure contain polysorbate 80 in an amount of about 0.01% to about 0.04% w / v. In some embodiments, the formulations of the present disclosure contain polysorbate 80 in an amount of about 0.01% to about 0.03% w / v. In some embodiments, the formulations of the present disclosure contain polysorbate 80 in an amount of about 0.01% to about 0.02% w / v. In some embodiments, the formulations of the present disclosure contain polysorbate 80 in an amount of about 0.02% to about 0.1% w / v. In some embodiments, the formulations of the present disclosure contain polysorbate 80 in an amount of about 0.02% to about 0.09% w / v. In some embodiments, the formulations of the present disclosure contain polysorbate 80 in an amount of about 0.02% to about 0.08% w / v. In some embodiments, the formulations of the present disclosure contain polysorbate 80 in an amount of about 0.02% to about 0.07% w / v. In some embodiments, the formulations of the present disclosure contain polysorbate 80 in an amount of about 0.02% to about 0.06% w / v. In some embodiments, the formulations of the present disclosure contain polysorbate 80 in an amount of about 0.02% to about 0.05% w / v. In some embodiments, the formulations of the present disclosure contain polysorbate 80 in an amount of about 0.02% to about 0.04% w / v. In some embodiments, the formulations of the present disclosure contain polysorbate 80 in an amount of about 0.02% to about 0.035% w / v. In some embodiments, the formulations of the present disclosure contain polysorbate 80 in an amount of about 0.02% to about 0.03% w / v. In some embodiments, the formulations of the present disclosure contain polysorbate 80 in an amount of about 0.025% to about 0.05% w / v. In some embodiments, the formulations of the present disclosure contain polysorbate 80 in an amount of about 0.025% to about 0.04% w / v. In some embodiments, the formulations of the present disclosure contain polysorbate 80 in an amount of about 0.025% to about 0.035% w / v. In some embodiments, the formulations of the present disclosure contain polysorbate 80 in an amount of about 0.025% to about 0.03% w / v.In some embodiments, the formulations of the present disclosure comprise polysorbate 80 in an amount of from about 0.03% to about 0.1% w / v. In some embodiments, the formulations of the present disclosure comprise polysorbate 80 in an amount of from about 0.03% to about 0.09% w / v. In some embodiments, the formulations of the present disclosure comprise polysorbate 80 in an amount of from about 0.03% to about 0.08% w / v. In some embodiments, the formulations of the present disclosure comprise polysorbate 80 in an amount of from about 0.03% to about 0.07% w / v. In some embodiments, the formulations of the present disclosure comprise polysorbate 80 in an amount of from about 0.03% to about 0.06% w / v. In some embodiments, the formulations of the present disclosure comprise polysorbate 80 in an amount of from about 0.03% to about 0.05% w / v. In some embodiments, the formulations of the present disclosure comprise polysorbate 80 in an amount of from about 0.03% to about 0.04% w / v.

[0613] In some embodiments, the formulations of the present disclosure comprise polysorbate 80 in an amount of about 0.01% w / v. In some embodiments, the formulations of the present disclosure comprise polysorbate 80 in an amount of about 0.015% w / v. In some embodiments, the formulations of the present disclosure comprise polysorbate 80 in an amount of about 0.02% w / v. In some embodiments, the formulations of the present disclosure comprise polysorbate 80 in an amount of about 0.025% w / v. In some embodiments, the formulations of the present disclosure comprise polysorbate 80 in an amount of about 0.03% w / v. In some embodiments, the formulations of the present disclosure comprise polysorbate 80 in an amount of about 0.035% w / v. In some embodiments, the formulations of the present disclosure comprise polysorbate 80 in an amount of about 0.04% w / v. In some embodiments, the formulations of the present disclosure comprise polysorbate 80 in an amount of about 0.045% w / v. In some embodiments, the formulations of the present disclosure comprise polysorbate 80 in an amount of about 0.05% w / v. In some embodiments, the formulations of the present disclosure comprise polysorbate 80 in an amount of about 0.055% w / v. In some embodiments, the formulations of the present disclosure comprise polysorbate 80 in an amount of about 0.06% w / v. In some embodiments, the formulations of the present disclosure comprise polysorbate 80 in an amount of about 0.065% w / v. In some embodiments, the formulations of the present disclosure comprise polysorbate 80 in an amount of about 0.07% w / v. In some embodiments, the formulations of the present disclosure comprise polysorbate 80 in an amount of about 0.075% w / v. In some embodiments, the formulations of the present disclosure comprise polysorbate 80 in an amount of about 0.08% w / v. In some embodiments, the formulations of the present disclosure comprise polysorbate 80 in an amount of about 0.085% w / v. In some embodiments, the formulations of the present disclosure comprise polysorbate 80 in an amount of about 0.09% w / v. In some embodiments, the formulations of the present disclosure comprise polysorbate 80 in an amount of about 0.095% w / v. In some embodiments, the formulations of the present disclosure comprise polysorbate 80 in an amount of about 0.1% w / v.

[0614] (iv) salt

[0615] In some embodiments, salts can be included in the antibody formulations described herein to provide stability to the formulations. In some embodiments, the formulations described herein contain salts, such as sodium chloride, potassium chloride, or magnesium chloride. In some embodiments, the formulations described herein contain salts at a concentration of at least 50 (e.g., at least 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, or 200 or higher) mM. In some embodiments, the formulations of the present disclosure contain salts at a concentration of less than or about 200 (e.g., less than or about 195, 190, 185, 180, 175, 170, 165, 160, 155, 150, 145, 140, 135, 130, 125, 120, 115, 110, 105, 100, 95, 90, 85, 80, 75, 70, 65, 60, 55, or 50) mM. In some embodiments, the formulations of the present disclosure contain salts at a concentration between about 50 mM and 100 mM, 60 mM and 100 mM, 70 mM and 100 mM, 80 mM and 100 mM, 90 mM and 100 mM, 50 mM and 120 mM, 60 mM and 120 mM, 70 mM and 120 mM, 80 mM and 120 mM, 90 mM and 120 mM, 100 mM and 120 mM, 110 mM and 120 mM, 50 mM and 150 mM, 60 mM and 150 mM, 70 mM and 150 mM, 80 mM and 150 mM, 90 mM and 150 mM, 100 mM and 150 mM, 110 mM and 150 mM, 120 mM and 150 mM, 130 mM and 150 mM, 140 mM and 150 mM, 50 mM and 200 mM, 60 mM and 200 mM, 70 mM and 200 mM, 80 mM and 200 mM, 90 mM and 200 mM, 100 mM and 200 mM, 110 mM and 200 mM, 120 mM and 200 mM, 130 mM and 200 mM, 140 mM and 200 mM, 150 mM and 200 mM, 160 mM and 200 mM, 170 mM and 200 mM, 180 mM and 200 mM, or 190 mM and 200 mM.

[0616] It should be understood that in embodiments where the formulations of the present disclosure contain two or more (e.g., at least two, three, four, five, six, seven, eight, nine, or ten or more) different salts, each of the two or more salts can independently be present at, for example, one of the above concentrations.

[0617] In some embodiments, the formulations of the present disclosure contain NaCl. In some embodiments, the formulations of the present disclosure contain NaCl at a concentration of about 50 - 100 mM. In some embodiments, the formulations of the present disclosure contain NaCl at a concentration of about 60 - 100 mM. In some embodiments, the formulations of the present disclosure contain NaCl at a concentration of about 70 - 100 mM. In some embodiments, the formulations of the present disclosure contain NaCl at a concentration of about 80 - 100 mM. In some embodiments, the formulations of the present disclosure contain NaCl at a concentration of about 90 - 100 mM. In some embodiments, the formulations of the present disclosure contain NaCl at a concentration of about 90 - 110 mM. In some embodiments, the formulations of the present disclosure contain NaCl at a concentration of about 95 - 105 mM. In some embodiments, the formulations of the present disclosure contain NaCl at a concentration of about 50 - 120 mM. In some embodiments, the formulations of the present disclosure contain NaCl at a concentration of about 80 - 120 mM. In some embodiments, the formulations of the present disclosure contain NaCl at a concentration of about 100 - 120 mM. In some embodiments, the formulations of the present disclosure contain NaCl at a concentration of about 50 - 150 mM. In some embodiments, the formulations of the present disclosure contain NaCl at a concentration of about 80 - 150 mM. In some embodiments, the formulations of the present disclosure contain NaCl at a concentration of about 100 - 150 mM. In some embodiments, the formulations of the present disclosure contain NaCl at a concentration of about 120 - 150 mM. In some embodiments, the formulations of the present disclosure contain NaCl at a concentration of about 50 - 200 mM. In some embodiments, the formulations of the present disclosure contain NaCl at a concentration of about 80 - 200 mM. In some embodiments, the formulations of the present disclosure contain NaCl at a concentration of about 100 - 200 mM. In some embodiments, the formulations of the present disclosure contain NaCl at a concentration of about 120 - 200 mM. In some embodiments, the formulations of the present disclosure contain NaCl at a concentration of about 150 - 200 mM.

[0618] In some embodiments, the formulations of the present disclosure comprise NaCl at a concentration of about 50 mM. In some embodiments, the formulations of the present disclosure comprise NaCl at a concentration of about 60 mM. In some embodiments, the formulations of the present disclosure comprise NaCl at a concentration of about 70 mM. In some embodiments, the formulations of the present disclosure comprise NaCl at a concentration of about 75 mM. In some embodiments, the formulations of the present disclosure comprise NaCl at a concentration of about 80 mM. In some embodiments, the formulations of the present disclosure comprise NaCl at a concentration of about 85 mM. In some embodiments, the formulations of the present disclosure comprise NaCl at a concentration of about 90 mM. In some embodiments, the formulations of the present disclosure comprise NaCl at a concentration of about 95 mM. In some embodiments, the formulations of the present disclosure comprise NaCl at a concentration of about 100 mM. In some embodiments, the formulations of the present disclosure comprise NaCl at a concentration of about 105 mM. In some embodiments, the formulations of the present disclosure comprise NaCl at a concentration of about 110 mM. In some embodiments, the formulations of the present disclosure comprise NaCl at a concentration of about 115 mM. In some embodiments, the formulations of the present disclosure comprise NaCl at a concentration of about 120 mM. In some embodiments, the formulations of the present disclosure comprise NaCl at a concentration of about 125 mM. In some embodiments, the formulations of the present disclosure comprise NaCl at a concentration of about 130 mM. In some embodiments, the formulations of the present disclosure comprise NaCl at a concentration of about 140 mM. In some embodiments, the formulations of the present disclosure comprise NaCl at a concentration of about 150 mM. In some embodiments, the formulations of the present disclosure comprise NaCl at a concentration of about 160 mM. In some embodiments, the formulations of the present disclosure comprise NaCl at a concentration of about 170 mM. In some embodiments, the formulations of the present disclosure comprise NaCl at a concentration of about 180 mM. In some embodiments, the formulations of the present disclosure comprise NaCl at a concentration of about 190 mM. In some embodiments, the formulations of the present disclosure comprise NaCl at a concentration of about 200 mM.

[0619] (v) pH

[0620] It has been found that the pH of the anti-CD137 antibody formulations of the present disclosure affects the stability of the formulations. As provided in the working examples, improved stability was observed when the anti-CD137 antibody was formulated in a histidine buffer at pH 5.8 compared to formulations containing glutamate at pH 4.5 or Tris at pH 7.5.

[0621] In some embodiments, the formulations described herein include a buffer or a pH regulator. In some embodiments, any of the formulations described herein have or can be adjusted to have a physiologically acceptable pH. As used herein, "physiologically acceptable pH" is a pH that is between pH 5 and pH 7 and includes pH 5 and pH 7, and in some embodiments of the formulations described herein, is a pH that is between pH 5 and pH 8 and includes pH 5 and pH 8. Thus, as used herein, a physiologically acceptable pH includes specific pH values, such as 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, and 8.0. In some embodiments, a physiologically acceptable pH is at least pH 5 (e.g., at least pH 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, or 5.9), but less than pH 7 (e.g., less than pH 6.9, 6.8, 6.7, 6.6, 6.5, 6.4, 6.3, 6.2, or 6.1). That is, in some embodiments, a physiologically acceptable pH is, for example, at least pH 5.0, but less than pH 7.0. In some embodiments, a physiologically acceptable pH is between pH 5.0 and pH 7.0. In some embodiments, a physiologically acceptable pH is between pH 5.5 and pH 6.5. In some embodiments, a physiologically acceptable pH is, for example, pH 6. In some embodiments, the pH of the antibody formulations described herein is between about 5.0 and 7.0, including the end values (e.g., about 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, and 7.0). In some embodiments, a physiologically acceptable pH is at least pH 5 (e.g., at least pH 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, or 5.9), but less than pH 8 (e.g., less than pH 7.9, 7.8, 7.7, 7.6, 7.5, 7.4, 7.3, 7.2, or 7.1). That is, in some embodiments, a physiologically acceptable pH is, for example, at least pH 5.0, but less than pH 8.0. In some embodiments, a physiologically acceptable pH is between pH 5.0 and pH 8.0. In some embodiments, a physiologically acceptable pH is between pH 5.5 and pH 7.5. In some embodiments, a physiologically acceptable pH is, for example, pH 6. In some embodiments, a physiologically acceptable pH is, for example, pH 7.4.In some embodiments, the pH of the antibody formulations described herein is between about 5.0 and 8.0, inclusive of the end values (e.g., about 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, and 8.0).

[0622] In some embodiments, the formulations of the present disclosure have a pH of from about 5.0 to about 7.0. In some embodiments, the formulations of the present disclosure have a pH of from about 5.5 to about 7.0. In some embodiments, the formulations of the present disclosure have a pH of from about 6.0 to about 7.0. In some embodiments, the formulations of the present disclosure have a pH of from about 6.5 to about 7.0. In some embodiments, the formulations of the present disclosure have a pH of from about 5.0 to about 6.5. In some embodiments, the formulations of the present disclosure have a pH of from about 5.5 to about 6.5. In some embodiments, the formulations of the present disclosure have a pH of from about 6.0 to about 6.5. In some embodiments, the formulations of the present disclosure have a pH of from about 5.0 to about 6.0. In some embodiments, the formulations of the present disclosure have a pH of from about 5.5 to about 6.0. In some embodiments, the formulations of the present disclosure have a pH of from about 5.0 to about 5.5. In some embodiments, the formulations of the present disclosure have a pH of from about 5.8 to about 6.2.

[0623] In some embodiments, the formulations of the present disclosure have a pH of about 5.0. In some embodiments, the formulations of the present disclosure have a pH of about 5.1. In some embodiments, the formulations of the present disclosure have a pH of about 5.2. In some embodiments, the formulations of the present disclosure have a pH of about 5.3. In some embodiments, the formulations of the present disclosure have a pH of about 5.4. In some embodiments, the formulations of the present disclosure have a pH of about 5.5. In some embodiments, the formulations of the present disclosure have a pH of about 5.6. In some embodiments, the formulations of the present disclosure have a pH of about 5.7. In some embodiments, the formulations of the present disclosure have a pH of about 5.8. In some embodiments, the formulations of the present disclosure have a pH of about 5.9. In some embodiments, the formulations of the present disclosure have a pH of about 6.0. In some embodiments, the formulations of the present disclosure have a pH of about 6.1. In some embodiments, the formulations of the present disclosure have a pH of about 6.2. In some embodiments, the formulations of the present disclosure have a pH of about 6.3. In some embodiments, the formulations of the present disclosure have a pH of about 6.4. In some embodiments, the formulations of the present disclosure have a pH of about 6.5. In some embodiments, the formulations of the present disclosure have a pH of about 6.6. In some embodiments, the formulations of the present disclosure have a pH of about 6.7. In some embodiments, the formulations of the present disclosure have a pH of about 6.8. In some embodiments, the formulations of the present disclosure have a pH of about 6.9. In some embodiments, the formulations of the present disclosure have a pH of about 7.0.

[0624] In some embodiments, the formulations of the present disclosure have a pH of from about 5.0 to about 8.0. In some embodiments, the formulations of the present disclosure have a pH of from about 6.5 to about 8.0. In some embodiments, the formulations of the present disclosure have a pH of from about 6.0 to about 8.0. In some embodiments, the formulations of the present disclosure have a pH of from about 6.5 to about 8.0. In some embodiments, the formulations of the present disclosure have a pH of from about 5.0 to about 7.5. In some embodiments, the formulations of the present disclosure have a pH of from about 5.5 to about 7.5. In some embodiments, the formulations of the present disclosure have a pH of from about 6.0 to about 7.5. In some embodiments, the formulations of the present disclosure have a pH of from about 5.0 to about 7.0. In some embodiments, the formulations of the present disclosure have a pH of from about 5.5 to about 7.0. In some embodiments, the formulations of the present disclosure have a pH of from about 5.0 to about 7.5. In some embodiments, the formulations of the present disclosure have a pH of from about 5.8 to about 6.2. In some embodiments, the formulations of the present disclosure have a pH of from about 5.8 to about 7.4.

[0625] In some embodiments, the formulations of the present disclosure have a pH of about 5.0. In some embodiments, the formulations of the present disclosure have a pH of about 5.1. In some embodiments, the formulations of the present disclosure have a pH of about 5.2. In some embodiments, the formulations of the present disclosure have a pH of about 5.3. In some embodiments, the formulations of the present disclosure have a pH of about 5.4. In some embodiments, the formulations of the present disclosure have a pH of about 5.5. In some embodiments, the formulations of the present disclosure have a pH of about 5.6. In some embodiments, the formulations of the present disclosure have a pH of about 5.7. In some embodiments, the formulations of the present disclosure have a pH of about 5.8. In some embodiments, the formulations of the present disclosure have a pH of about 5.9. In some embodiments, the formulations of the present disclosure have a pH of about 6.0. In some embodiments, the formulations of the present disclosure have a pH of about 6.1. In some embodiments, the formulations of the present disclosure have a pH of about 6.2. In some embodiments, the formulations of the present disclosure have a pH of about 6.3. In some embodiments, the formulations of the present disclosure have a pH of about 6.4. In some embodiments, the formulations of the present disclosure have a pH of about 6.5. In some embodiments, the formulations of the present disclosure have a pH of about 6.6. In some embodiments, the formulations of the present disclosure have a pH of about 6.7. In some embodiments, the formulations of the present disclosure have a pH of about 6.8. In some embodiments, the formulations of the present disclosure have a pH of about 6.9. In some embodiments, the formulations of the present disclosure have a pH of about 7.0. In some embodiments, the formulations of the present disclosure have a pH of about 7.1. In some embodiments, the formulations of the present disclosure have a pH of about 7.2. In some embodiments, the formulations of the present disclosure have a pH of about 7.3. In some embodiments, the formulations of the present disclosure have a pH of about 7.4. In some embodiments, the formulations of the present disclosure have a pH of about 7.5. In some embodiments, the formulations of the present disclosure have a pH of about 7.6. In some embodiments, the formulations of the present disclosure have a pH of about 7.7. In some embodiments, the formulations of the present disclosure have a pH of about 7.8. In some embodiments, the formulations of the present disclosure have a pH of about 7.9. In some embodiments, the formulations of the present disclosure have a pH of about 8.0.

[0626] (vi) Concentration of the anti-CD137 antibody

[0627] In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulations of the present disclosure at a concentration of about 1 mg / ml to about 100 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 5 mg / ml to about 15 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 15 mg / ml to about 30 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 30 mg / ml to about 45 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 45 mg / ml to about 60 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 60 mg / ml to about 75 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 75 mg / ml to about 90 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 85 mg / ml to about 100 mg / ml.

[0628] In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulations of the present disclosure at a concentration of about 1 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 2 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 3 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 4 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 5 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 6 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 7 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 8 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 9 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 10 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 11 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 12 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 13 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 14 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 15 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 16 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 17 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 18 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 19 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 20 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 21 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 22 mg / ml.In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 23 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 24 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 25 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 30 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 35 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 40 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 45 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 50 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 55 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 60 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 65 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 70 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 75 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 80 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 85 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 90 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 95 mg / ml. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof is present in the formulation at a concentration of about 100 mg / ml.

[0629] (vii) Exemplary formulations

[0630] In some embodiments, the formulations of the present disclosure comprise the following: a buffer containing histidine and an anti-CD137 antibody or antigen-binding fragment thereof. In some embodiments, the formulation comprises from about 10 mM to about 100 mM histidine. In some embodiments, the formulation comprises an anti-CD137 antibody or antigen-binding fragment thereof at a concentration of from about 1 mg / ml to about 100 mg / ml.

[0631] In some embodiments, the formulations of the present disclosure comprise the following: a buffer containing histidine, a disaccharide, and an anti-CD137 antibody or an antigen-binding fragment thereof. In some embodiments, the formulation comprises from about 10 mM histidine to about 100 mM histidine. In some embodiments, the disaccharide is sucrose. In some embodiments, the formulation comprises from about 5-15% (w / v) of the disaccharide. In some embodiments, the formulation comprises an anti-CD137 antibody or an antigen-binding fragment thereof at a concentration of from about 1 mg / ml to about 100 mg / ml.

[0632] In some embodiments, the formulations of the present disclosure comprise the following: a buffer containing histidine and an anti-CD137 antibody or an antigen-binding fragment thereof. In some embodiments, the formulation has a pH of from about 5.0 to 7.0. In some embodiments, the formulation comprises from about 10 mM histidine to about 100 mM histidine. In some embodiments, the formulation comprises an anti-CD137 antibody or an antigen-binding fragment thereof at a concentration of from about 1 mg / ml to about 100 mg / ml.

[0633] In some embodiments, the formulations of the present disclosure comprise the following: a buffer containing histidine, a disaccharide, a nonionic surfactant, a salt, and an anti-CD137 antibody or an antigen-binding fragment thereof. In some embodiments, the formulation has a pH of from about 5.0 to 7.0. In some embodiments, the formulation comprises from about 10 mM histidine to about 100 mM histidine. In some embodiments, the disaccharide of the formulation is sucrose. In some embodiments, the formulation comprises from about 5-15% (w / v) of the disaccharide. In some embodiments, the nonionic surfactant of the formulation is polysorbate. In some embodiments, the polysorbate is polysorbate 80. In some embodiments, the formulation comprises from about 0.01% to about 0.1% (w / v) of the nonionic surfactant. In some embodiments, the salt of the formulation is NaCl. In some embodiments, the formulation comprises a salt at a concentration of from about 50 mM to about 200 mM. In some embodiments, the formulation comprises an anti-CD137 antibody or an antigen-binding fragment thereof at a concentration of from about 1 mg / ml to about 100 mg / ml.

[0634] In some embodiments, the formulations of the present disclosure comprise the following: a buffer comprising histidine, a disaccharide of about 5% to about 15% (w / v), a nonionic surfactant of about 0.01% to about 0.1% w / v, a salt of about 50 mM to about 200 mM, and an anti-CD137 antibody or an antigen-binding fragment thereof. In some embodiments, the formulation has a pH of about 5.0 to 7.0. In some embodiments, the formulation comprises about 10 mM histidine to about 100 mM histidine. In some embodiments, the disaccharide of the formulation is sucrose. In some embodiments, the disaccharide of the formulation is lactose. In some embodiments, the disaccharide of the formulation is maltose. In some embodiments, the disaccharide of the formulation is trehalose. In some embodiments, the nonionic surfactant of the formulation is polysorbate. In some embodiments, the polysorbate is polysorbate 80. In some embodiments, the salt of the formulation is NaCl. In some embodiments, the formulation comprises an anti-CD137 antibody or an antigen-binding fragment thereof at a concentration of about 1 mg / ml to about 100 mg / ml.

[0635] In some embodiments, the formulations of the present disclosure comprise the following: a buffer comprising about 10 mM to about 100 mM histidine, sucrose of about 5% to about 15% (w / v), polysorbate 80 of about 0.01% to about 0.1% w / v, NaCl of about 50 mM to about 200 mM, and an anti-CD137 antibody or an antigen-binding fragment thereof. In some embodiments, the formulation has a pH of about 5.0 to 7.0. In some embodiments, the formulation has a pH of about 5.0 to 7.4. In some embodiments, the formulation comprises an anti-CD137 antibody or an antigen-binding fragment thereof at a concentration of about 1 mg / ml to about 100 mg / ml.

[0636] In some embodiments, the formulations of the present disclosure comprise the following: a buffer containing about 20 mM histidine, about 10% (w / v) sucrose, about 0.03% w / v polysorbate 80, about 100 mM NaCl, and an anti-CD137 antibody or an antigen-binding fragment thereof. In some embodiments, the formulation has a pH of about 6.0. In some embodiments, the formulation comprises an anti-CD137 antibody or an antigen-binding fragment thereof at a concentration of about 1 mg / ml to about 100 mg / ml. In some embodiments, the formulation comprises an anti-CD137 antibody or an antigen-binding fragment thereof at a concentration of about 5 mg / ml to about 15 mg / ml. In some embodiments, the formulation comprises an anti-CD137 antibody or an antigen-binding fragment thereof at a concentration of about 15 mg / ml to about 30 mg / ml. In some embodiments, the formulation comprises an anti-CD137 antibody or an antigen-binding fragment thereof at a concentration of about 30 mg / ml to about 45 mg / ml. In some embodiments, the formulation comprises an anti-CD137 antibody or an antigen-binding fragment thereof at a concentration of about 45 mg / ml to about 60 mg / ml. In some embodiments, the formulation comprises an anti-CD137 antibody or an antigen-binding fragment thereof at a concentration of about 60 mg / ml to about 75 mg / ml. In some embodiments, the formulation comprises an anti-CD137 antibody or an antigen-binding fragment thereof at a concentration of about 75 mg / ml to about 90 mg / ml. In some embodiments, the formulation comprises an anti-CD137 antibody or an antigen-binding fragment thereof at a concentration of about 85 mg / ml to about 100 mg / ml. In some embodiments, the formulation comprises an anti-CD137 antibody or an antigen-binding fragment thereof at a concentration of about 5 mg / ml. In some embodiments, the formulation comprises an anti-CD137 antibody or an antigen-binding fragment thereof at a concentration of about 10 mg / ml. In some embodiments, the formulation comprises an anti-CD137 antibody or an antigen-binding fragment thereof at a concentration of about 15 mg / ml. In some embodiments, the formulation comprises an anti-CD137 antibody or an antigen-binding fragment thereof at a concentration of about 20 mg / ml.

[0637] The formulations of the present disclosure as described herein provide an anti-CD137 antibody or an antigen-binding fragment thereof with significant stability, minimizing the formation of antibody aggregates (high molecular weight species) and particulates, minimizing charge variants, and maintaining the structural integrity of the antibody.

[0638] (viii) Stability of the anti-CD137 formulation

[0639] In some embodiments, the formulations described herein are capable of maintaining the structural integrity of the anti-CD137 antibody or antigen-binding fragment thereof in solution for an extended period of time. In some embodiments, the anti-CD137 antibody in the formulations of the present disclosure is stored at about 2°C to 40°C (e.g., stored at, for example, 1°C, 2°C, 3°C, 4°C, 5°C, 6°C, 7°C, 8°C, 9°C, 10°C, 11°C, 12°C, 13°C, 14°C, 15°C, 16°C, 17°C, 18°C, 19°C, 20°C, 21°C, 22°C, 23°C, 24°C, 25°C, 26°C, 27°C, 28°C, 29°C, 30°C, 31°C, 32°C, 33°C, 34°C, 35°C, 36°C, 37°C, 38°C, 39°C, 40°C) for at least 4 weeks (e.g., at least five weeks, six weeks, seven weeks, eight weeks, nine weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, or 24 weeks, or at least one month, two months, three months, four months, five months, six months, seven months, eight months, nine months, 10 months, 11 months, 12 months, 13 months, 14 months, 15 months, 16 months, 17 months, 18 months, 19 months, 20 months, 21 months, 22 months, 23 months, 24 months or longer) and remains stable. In some embodiments, the anti-CD137 antibody in the formulations of the present disclosure is stored at about 2°C to 9°C for at least 4 weeks and remains stable. In some embodiments, the anti-CD137 antibody in the formulations of the present disclosure is stored at about 10°C to 19°C for at least 4 weeks and remains stable. In some embodiments, the anti-CD137 antibody in the formulations of the present disclosure is stored at about 20°C to 29°C for at least 4 weeks and remains stable. In some embodiments, the anti-CD137 antibody in the formulations of the present disclosure is stored at about 30°C to 40°C for at least 4 weeks and remains stable. In some embodiments, the anti-CD137 antibody in the formulations of the present disclosure is stored at about 4°C for at least 4 weeks and remains stable. In some embodiments, the anti-CD137 antibody in the formulations of the present disclosure is stored at about 25°C for at least 4 weeks and remains stable. In some embodiments, the anti-CD137 antibody in the formulations of the present disclosure is stored at about 40°C for at least 4 weeks and remains stable.

[0640] As illustrated in the working examples described herein, the inventors have provided a formulation suitable for maintaining approximately 10 mg / mL anti-CD137 antibody or antigen-binding fragment thereof in a predominantly monomeric form for at least 24 weeks at 4°C, at least 12 weeks at 25°C, and at least 4 weeks at 40°C. As used herein, the anti-CD137 antibody or antigen-binding fragment thereof in the formulations of the present disclosure is "predominantly monomeric", or "predominantly in monomeric form" if at least 95 (e.g., at least 95.1, 95.2, 95.3, 95.4, 95.5, 95.6, 95.7, 95.8, 95.9, 96, 96.1, 96.2, 96.3, 96.4, 96.5, 96.6, 96.7, 96.8, 96.9, 97, 97.1, 97.2, 97.3, 97.4, 97.5, 97.6, 97.7, 97.8, 97.9, 98, 98.1, 98.2, 98.3, 98.4, 98.5, 98.6, 98.7, 98.8, 98.9, 99, 99.1, 99.2, 99.3, 99.4, 99.5, 99.6, 99.7, 99.8 or 99.9 or higher) % monomer (e.g., determined using size exclusion chromatography (SEC)) of the antibody present in the solution. That is: less than 5 (e.g., less than 4.9, 4.8, 4.7, 4.6, 4.5, 4.4, 4.3, 4.2, 4.1, 4.0, 3.9, 3.8, 3.7, 3.6, 3.5, 3.4, 3.3, 3.2, 3.1, 3.0, 2.9, 2.8, 2.7, 2.6, 2.5, 2.4, 2.3, 2.2, 2.1, 2, 1.9, 1.8, 1.7, 1.6, 1.5, 1.4, 1.3, 1.2, 1.1, 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2 or 0.1) % of the antibody in the solution is oligomeric, aggregated and / or fragmented. As used herein, antibody fragmentation refers to improperly assembled components or degradation products of a full-length antibody that have a molecular weight lower than the full-length antibody. Such fragmented forms include, but are not limited to, free monomeric heavy chain polypeptides, dimeric heavy chain polypeptides (e.g., disulfide-linked heavy chain polypeptides), dimeric heavy chain polypeptides bound to one light chain polypeptide, monomeric heavy chain polypeptides bound to one light chain polypeptide, or further degradation products or fragments of light or heavy chain polypeptides.In some embodiments, less than 2 (e.g., less than 1.9, 1.8, 1.7, 1.6, 1.5, 1.4, 1.3, 1.2, 1.1, 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, or 0.1)% of the antibody aggregates after being stored at 2°C to 8°C for at least 4 weeks (e.g., at least five weeks, six weeks, seven weeks, eight weeks, nine weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, or 24 weeks, or at least one month, two months, three months, four months, five months, six months, seven months, eight months, nine months, 10 months, 11 months, 12 months, 13 months, 14 months, 15 months, 16 months, 17 months, 18 months, 19 months, 20 months, 21 months, 22 months, 23 months, 24 months, or longer). Methods for determining the amount of monomeric antibody of an anti-CD137 antibody or antigen-binding fragment thereof present in solution and the amount of oligomeric, aggregated, or fragmented forms are described herein and illustrated in the working examples. For example, one skilled in the art can use, for example, the following to determine the percentage of intact, fragmented, unfolded intermediates, and / or aggregated antibody species present in a given solution: size exclusion chromatography (SEC), size exclusion chromatography-high performance liquid chromatography (SEC-HPLC), static light scattering (SLS), Fourier transform infrared spectroscopy (FTIR), circular dichroism (CD), urea-induced protein unfolding techniques, intrinsic tryptophan fluorescence, non-reducing sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and differential scanning calorimetry (DSC).

[0641] Anti-CD137 antibody and antigen-binding fragment thereof

[0642] The present disclosure provides a formulation comprising an antibody and antigen-binding fragment thereof that specifically binds to and agonizes CD137. In some embodiments, the formulation of the present disclosure comprises any of the anti-CD137 antibodies or antigen-binding fragments thereof described herein.

[0643] In some aspects, the present disclosure provides a formulation comprising an anti-CD137 agonist antibody and antigen-binding fragment thereof that can be used to treat cancer. In some embodiments, the formulation of the present disclosure comprises an anti-CD137 agonist antibody and antigen-binding fragment thereof that induces cytokine production. In some embodiments, the formulation of the present disclosure comprises an anti-CD137 agonist antibody that increases the number of CD8+ T cells in the tumor microenvironment. In some embodiments, the formulation of the present disclosure comprises an anti-CD137 agonist antibody and antigen-binding fragment thereof that induces protective anti-tumor immunity. The present disclosure also provides a formulation comprising an anti-CD137 agonist antibody and antigen-binding fragment thereof that substantially does not increase the CD4+ and / or CD8+ T cell population in the spleen or liver when administered in vivo.

[0644] Human CD137 is a 255 - amino - acid transmembrane polypeptide (SEQ ID NO:3; accession number NM_001561; NP_001552) and is a member of the phylogenetically conserved tumor necrosis factor receptor (TNFR) superfamily. CD137 (or 4 - 1BB, TNFR superfamily 9) and its ligand (CD137L) are involved in regulating a wide range of immune activities. CD137 ligand cross - links with its receptor CD137 expressed on activated T cells and co - stimulates T - cell activity. CD137 is an activation - induced co - stimulatory molecule. Recent studies have shown that the anti - cancer effect mediated by CD137 is mainly based on its ability to activate T cells, especially its ability to induce cytotoxic T lymphocyte (CTL) responses and to induce cytokine production, especially large amounts of IFN - γ (Ye et al., (2014) Clin Cancer Res 20(1):44 - 55). CD137 ligand is a transmembrane protein on the cell surface, and it transmits signals into the cells on which it is expressed, a phenomenon called "reverse signaling" or "back signaling". CD137 ligand expression is found on most types of white blood cells and some non - immune cells. In monocytes (monocytes, macrophages, and DCs), CD137 ligand signaling induces activation, migration, survival, and differentiation.

[0645] Thus, in some embodiments, the isolated anti - CD137 agonist antibody or antigen - binding fragment thereof described herein binds to and agonizes CD137 and allows or promotes CD137L binding. In some embodiments, the isolated anti - CD137 agonist antibody or antigen - binding fragment thereof described herein binds to and agonizes CD137. In some embodiments, the anti - CD137 antibody provided by the present disclosure binds to and agonizes CD137 and co - stimulates the activation of T cells.

[0646] In some embodiments, the formulations of the present disclosure comprise the isolated anti - CD137 agonist antibody or antigen - binding fragment thereof described herein, which has one or more of the following properties or characteristics:

[0647] a) Specifically binds to human CD137;

[0648] b) Binds to human and cynomolgus monkey CD137; and

[0649] c) Binds to human and mouse CD137.

[0650] In some embodiments, the anti-CD137 agonist antibody or antigen-binding fragment thereof described herein binds to CD137 and co-stimulates T cell activity. In some embodiments, the anti-CD137 agonist antibody or antigen-binding fragment thereof described herein binds to CD137 and induces or enhances T cell activation, cytotoxic T lymphocyte (CTL) response, T cell proliferation, cytokine production, or a combination thereof. In some embodiments, the anti-CD137 agonist antibody or antigen-binding fragment thereof described herein binds to CD137 and induces or enhances T cell activation, cytotoxic T lymphocyte (CTL) response, T cell proliferation, cytokine production, or a combination thereof in the tumor microenvironment. In some embodiments, the anti-CD137 antibody or antigen-binding fragment thereof described herein does not significantly induce or enhance T cell activation and / or T cell proliferation in the liver and / or spleen. In some embodiments, the anti-CD137 antibody described herein binds to CD137 and induces the production of IFN-γ. In some embodiments, the antibodies provided by the present disclosure bind to CD137 and induce the production of IL-2, TNF-α, IL-13, or a combination thereof.

[0651] In some embodiments, the formulations of the present disclosure comprise the anti-CD137 antibody described herein, which specifically binds to CD137 and agonizes it. In some embodiments, the agonization of CD137 is assayed by determining the concentration of cytokines produced by immune cells. Methods for analyzing cytokine production are known in the art and are used in the Examples. In some embodiments, an increase in cytokines produced by immune cells indicates agonization of CD137. In some embodiments, the agonization of CD137 is assayed by analyzing T cell proliferation. In some embodiments, an increase in T cell proliferation indicates agonization of CD137. In some embodiments, the level of cell signaling is assayed by quantitative determination of phosphorylation of relevant molecules or expression of a gene reporter following a relevant promoter to assay the agonization of CD137. In some embodiments, an increase in cell signaling indicates agonization of CD137. In some embodiments, the agonization of CD137 is assayed by determining tumor volume. In some embodiments, a decrease in tumor volume indicates agonization of CD137.

[0652] In some embodiments, the formulations of the present disclosure comprise the anti-CD137 antibody described herein, which induces, increases, or stabilizes oligomerization, multimerization, or other higher-order clustering of CD137. In some embodiments, CD137 clustering on the cell surface is observed by fluorescence microscopy.

[0653] The present disclosure provides a formulation comprising an isolated monoclonal antibody or an antigen-binding fragment thereof that binds to and agonizes CD137. In some embodiments, the antibody or antigen-binding fragment thereof, (i) binds to human CD137 with an affinity (K D ) of about 30 - 100 nM (e.g., between about 30 nM and about 100 nM); (ii) binds to an epitope on human CD137 as described herein; and / or (iii) comprises a heavy chain CDR3 containing the amino acid sequence DXXXXLXXXXYXYYX (SEQ ID NO:126).

[0654] Affinity for CD137

[0655] In some embodiments, the formulation of the present disclosure comprises the isolated anti-CD137 agonist antibody or antigen-binding fragment thereof as described herein that binds to human CD137 with an affinity (K D ) of about 30 - 100 nM (e.g., between about 30 nM and about 100 nM or between about 40 nM and about 100 nM). In some embodiments, the anti-CD137 antibody has an affinity for human CD137 that is at least two-fold higher (e.g., at least three-fold, four-fold, five-fold, six-fold, seven-fold, eight-fold, nine-fold, or 10-fold) than the affinity of mAb10 for murine CD137. In some embodiments, the affinity of the anti-CD137 antibody is not greater than 500 nM, 450 nM, 400 nM, 350 nM, 300 nM, 250 nM, 200 nM, 250 nM, 200 nM, 175 nM, 150 nM, 125 nM, 110 nM, or 100 nM. In some embodiments, the anti-CD137 antibody has an affinity for human CD137 that is at least two-fold higher (e.g., at least three-fold, four-fold, five-fold, six-fold, seven-fold, eight-fold, nine-fold, or 10-fold) than the affinity of mAb10 for murine CD137, but is not greater than 500 nM, 450 nM, 400 nM, 350 nM, 300 nM, 250 nM, 200 nM, 250 nM, 200 nM, 175 nM, 150 nM, 125 nM, 110 nM, or 100 nM. The affinity of an antibody is the strength of binding to a single CD137 polypeptide. In some embodiments, the affinity is represented by the equilibrium dissociation constant (K D ). The value of K D is inversely proportional to the binding affinity of the antibody for the antigen. Thus, the smaller the value of K D , the greater the affinity of the antibody for its antigen.

[0656] Methods for determining the affinity of an antibody for its antigen are known in the art. Exemplary methods for determining binding affinity employ surface plasmon resonance. Surface plasmon resonance is an optical phenomenon that allows the analysis of real-time biospecific interactions by detecting changes in pr...

Claims

1. A preparation, comprising: (a) an anti-CD137 antibody at a concentration of 1 mg / ml to 100 mg / ml, wherein the anti-CD137 antibody comprises a heavy chain CDR1 shown in SEQ ID NO:48, a heavy chain CDR2 shown in SEQ ID NO:56, and a heavy chain CDR3 shown in SEQ ID NO:68, and a light chain CDR1 shown in SEQ ID NO:69, a light chain CDR2 shown in SEQ ID NO:78, and a light chain CDR3 shown in SEQ ID NO:89, and (b) a buffer containing histidine.

2. A preparation, comprising: (a) an anti-CD137 antibody at a concentration of 1 mg / ml to 100 mg / ml, wherein the anti-CD137 antibody comprises a heavy chain CDR1 shown in SEQ ID NO:48, a heavy chain CDR2 shown in SEQ ID NO:56, and a heavy chain CDR3 shown in SEQ ID NO:68, and a light chain CDR1 shown in SEQ ID NO:69, a light chain CDR2 shown in SEQ ID NO:78, and a light chain CDR3 shown in SEQ ID NO:89, (b) a buffer containing histidine; and (c) a disaccharide.

3. A preparation, comprising: (a) an anti-CD137 antibody at a concentration of 1 mg / ml to 100 mg / ml, wherein the anti-CD137 antibody comprises a heavy chain CDR1 shown in SEQ ID NO:48, a heavy chain CDR2 shown in SEQ ID NO:56, and a heavy chain CDR3 shown in SEQ ID NO:68, and a light chain CDR1 shown in SEQ ID NO:69, a light chain CDR2 shown in SEQ ID NO:78, and a light chain CDR3 shown in SEQ ID NO:89, and (b) a buffer containing histidine; wherein the preparation has a pH of 5.0 - 7.

0.

4. A preparation, comprising: (a) an anti-CD137 antibody at a concentration of 1 mg / ml to 100 mg / ml, wherein the anti-CD137 antibody comprises a heavy chain CDR1 shown in SEQ ID NO:48, a heavy chain CDR2 shown in SEQ ID NO:56, and a heavy chain CDR3 shown in SEQ ID NO:68, and a light chain CDR1 shown in SEQ ID NO:69, a light chain CDR2 shown in SEQ ID NO:78, and a light chain CDR3 shown in SEQ ID NO:89, and (b) a buffer containing histidine; wherein the preparation has a pH of 5.0 - 7.

4.

5. A preparation, comprising: (a) An anti-CD137 antibody at a concentration of 1 mg / ml to 100 mg / ml, wherein the anti-CD137 antibody comprises a heavy chain CDR1 shown in SEQ ID NO:48, a heavy chain CDR2 shown in SEQ ID NO:56, a heavy chain CDR3 shown in SEQ ID NO:68, a light chain CDR1 shown in SEQ ID NO:69, a light chain CDR2 shown in SEQ ID NO:78, and a light chain CDR3 shown in SEQ ID NO:89, (b) A buffer solution containing histidine, (c) A disaccharide, (d) A nonionic surfactant, and (e) A salt, wherein the pH of the preparation is 5.0 to 7.

0.

6. A preparation comprising: (a) An anti-CD137 antibody at a concentration of 1 mg / ml to 100 mg / ml, wherein the anti-CD137 antibody comprises a heavy chain CDR1 shown in SEQ ID NO:48, a heavy chain CDR2 shown in SEQ ID NO:56, a heavy chain CDR3 shown in SEQ ID NO:68, a light chain CDR1 shown in SEQ ID NO:69, a light chain CDR2 shown in SEQ ID NO:78, and a light chain CDR3 shown in SEQ ID NO:89, (b) A buffer solution containing histidine, (c) A disaccharide, (d) A nonionic surfactant, and (e) A salt, wherein the pH of the preparation is 5.0 to 7.

4.

7. A preparation comprising: (a) An anti-CD137 antibody at a concentration of 1 mg / ml to 100 mg / ml, wherein the anti-CD137 antibody comprises a heavy chain CDR1 shown in SEQ ID NO:48, a heavy chain CDR2 shown in SEQ ID NO:56, a heavy chain CDR3 shown in SEQ ID NO:68, a light chain CDR1 shown in SEQ ID NO:69, a light chain CDR2 shown in SEQ ID NO:78, and a light chain CDR3 shown in SEQ ID NO:89, (b) A buffer solution containing histidine, (c) A disaccharide at a weight / volume of 5% to 15%, (d) A nonionic surfactant at a weight / volume (w / v) of from 0.01% to 0.1%, and (e) A salt at 50 mM to 200 mM, wherein the pH of the preparation is 5.0 to 7.

0.

8. A preparation comprising: (a) An anti-CD137 antibody at a concentration of 1 mg / ml to 100 mg / ml, wherein the anti-CD137 antibody comprises a heavy chain CDR1 shown in SEQ ID NO:48, a heavy chain CDR2 shown in SEQ ID NO:56, a heavy chain CDR3 shown in SEQ ID NO:68, a light chain CDR1 shown in SEQ ID NO:69, a light chain CDR2 shown in SEQ ID NO:78, and a light chain CDR3 shown in SEQ ID NO:89, (b) A buffer solution containing histidine, (c) A disaccharide at a weight / volume of 5% to 15%, (d) A non-ionic surfactant with a weight / volume (w / v) of 0.01% to 0.1%, and (e) A salt with a concentration of 50 mM to 200 mM, wherein the pH of the formulation is 5.0 to 7.

4.

9. A formulation comprising: (a) An anti-CD137 antibody with a concentration of 1 mg / ml to 100 mg / ml, wherein the anti-CD137 antibody comprises a heavy-chain CDR1 shown in SEQ ID NO: 48, a heavy-chain CDR2 shown in SEQ ID NO: 56, a heavy-chain CDR3 shown in SEQ ID NO: 68, a light-chain CDR1 shown in SEQ ID NO: 69, a light-chain CDR2 shown in SEQ ID NO: 78, and a light-chain CDR3 shown in SEQ ID NO: 89, (b) A buffer containing 10 mM to 100 mM histidine, (c) Sucrose with a weight / volume of 5% to 15%, (d) Polysorbate-80 with a weight / volume (w / v) of 0.01% to 0.1%, and (e) 50 mM to 200 mM of NaCl, wherein the pH of the formulation is 5.0 to 7.

0.

10. A formulation comprising: (a) An anti-CD137 antibody with a concentration of 1 mg / ml to 100 mg / ml, wherein the anti-CD137 antibody comprises a heavy-chain CDR1 shown in SEQ ID NO: 48, a heavy-chain CDR2 shown in SEQ ID NO: 56, a heavy-chain CDR3 shown in SEQ ID NO: 68, a light-chain CDR1 shown in SEQ ID NO: 69, a light-chain CDR2 shown in SEQ ID NO: 78, and a light-chain CDR3 shown in SEQ ID NO: 89, (b) A buffer containing 10 mM to 100 mM histidine, (c) Sucrose with a weight / volume of 5% to 15%, (d) Polysorbate-80 with a weight / volume (w / v) of 0.01% to 0.1%, and (e) 50 mM to 200 mM of NaCl, wherein the pH of the formulation is 5.0 to 7.

4.

11. A formulation comprising: (a) An anti-CD137 antibody with a concentration of 1 mg / ml to 100 mg / ml, wherein the anti-CD137 antibody comprises a heavy-chain CDR1 shown in SEQ ID NO: 48, a heavy-chain CDR2 shown in SEQ ID NO: 56, a heavy-chain CDR3 shown in SEQ ID NO: 68, a light-chain CDR1 shown in SEQ ID NO: 69, a light-chain CDR2 shown in SEQ ID NO: 78, and a light-chain CDR3 shown in SEQ ID NO: 89, (b) A buffer containing 20 mM histidine, (c) Sucrose with a weight / volume (w / v) of 10%, (d) Polysorbate-80 with a weight / volume (w / v) of 0.03%, and (e) 100 mM of NaCl, wherein the pH of the formulation is 6.

0.

12. The preparation according to any one of claims 1 to 11, wherein the anti-CD137 antibody comprises a heavy chain and a light chain sequence each containing an amino acid sequence having at least 90% identity to SEQ ID NO: 4 and 6, respectively.

13. The preparation according to claim 12, wherein the anti-CD137 antibody comprises a heavy chain and a light chain sequence having the amino acid sequences shown in SEQ ID NO: 4 and 6, respectively.

14. The preparation according to any one of claims 1 to 11, wherein the anti-CD137 antibody comprises a heavy chain and a light chain sequence each containing an amino acid sequence having at least 90% identity to SEQ ID NO: 101 and 6, respectively.

15. The preparation according to claim 14, wherein the anti-CD137 antibody comprises a heavy chain and a light chain sequence having the amino acid sequences shown in SEQ ID NO: 101 and 6, respectively.

16. The preparation according to any one of claims 1 to 11, wherein the antibody comprises an IgG1 heavy chain constant region.

17. The preparation according to claim 16, wherein the IgG1 heavy chain constant region is a wild-type human IgG1 heavy chain constant region.

18. The preparation according to claim 16, wherein the IgG1 heavy chain constant region contains an amino acid substitution relative to the wild-type human IgG1 heavy chain constant region.

19. The preparation according to any one of claims 1 to 11, wherein the antibody comprises an IgG4 heavy chain constant region.

20. The preparation according to claim 19, wherein the IgG4 heavy chain constant region is a wild-type human IgG4 heavy chain constant region.

21. The preparation according to claim 19, wherein the IgG4 heavy chain constant region contains an amino acid substitution relative to the wild-type human IgG4 heavy chain constant region.

22. The preparation according to any one of claims 1 to 8, comprising 10 mM histidine to 100 mM histidine.

23. The preparation according to any one of claims 1 to 8, comprising 20 mM histidine.

24. The preparation according to any one of claims 3 to 4 and 9 to 10, further comprising a disaccharide.

25. The preparation according to any one of claims 2 and 5 to 8, wherein the disaccharide is selected from sucrose, lactose, maltose, and trehalose.

26. The preparation according to any one of claims 2 and 5 to 6, wherein the weight / volume of the disaccharide is 5% to 15%.

27. The preparation according to any one of claims 2 and 5 to 8, wherein the weight / volume of the disaccharide is 10%.

28. The preparation according to any one of claims 1 to 4 and 9 to 11, further comprising a salt.

29. The preparation according to any one of claims 1 to 4, further comprising NaCl.

30. The preparation according to claim 29, wherein the concentration of the salt is 50 mM to 200 mM.

31. The preparation according to claim 30, wherein the concentration of the salt is 100 mM.

32. The preparation according to claim 2, wherein the preparation has a pH of 5.0 - 7.

0.

33. The preparation according to claim 2, wherein the preparation has a pH of 5.0 - 7.

4.

34. The preparation according to claim 33, wherein the pH is 6.

0.

35. The preparation according to any one of claims 1 to 4, further comprising a non-ionic surfactant.

36. The preparation according to claim 35, wherein the non-ionic surfactant is polysorbate.

37. The preparation according to claim 36, wherein the polysorbate is polysorbate-80.

38. The preparation according to any one of claims 5 to 6 and 35 to 36, wherein the weight / volume (w / v) of the non-ionic surfactant is 0.01% to 0.1%.

39. The preparation according to any one of claims 5 to 8, wherein the weight / volume (w / v) of the non-ionic surfactant is 0.03%.

40. The preparation according to any one of claims 1 to 11, comprising the anti-CD137 antibody at a concentration of 5 mg / ml to 15 mg / ml.

41. The preparation according to any one of claims 1 to 11, comprising the anti-CD137 antibody at a concentration of 15 mg / ml to 30 mg / ml.

42. The preparation according to any one of claims 1 to 11, comprising the anti-CD137 antibody at a concentration of 30 mg / ml to 45 mg / ml.

43. The preparation according to any one of claims 1 to 11, comprising the anti-CD137 antibody at a concentration of 45 mg / ml to 60 mg / ml.

44. The preparation according to any one of claims 1 to 11, comprising the anti-CD137 antibody at a concentration of 60 mg / ml to 75 mg / ml.

45. The preparation according to any one of claims 1 to 11, comprising the anti-CD137 antibody at a concentration of 75 mg / ml to 90 mg / ml.

46. The preparation according to any one of claims 1 to 11, comprising the anti-CD137 antibody at a concentration of 85 mg / ml to 100 mg / ml.

47. The preparation according to any one of claims 1 to 11, comprising the anti-CD137 antibody at a concentration of 5 mg / ml.

48. The preparation according to any one of claims 1 to 11, comprising the anti-CD137 antibody at a concentration of 10 mg / ml.

49. The preparation according to any one of claims 1 to 11, comprising the anti-CD137 antibody at a concentration of 15 mg / ml.

50. The preparation according to any one of claims 1 to 11, comprising the anti-CD137 antibody at a concentration of 20 mg / ml.

51. Use of the preparation according to any one of claims 1 to 11 in the manufacture of a medicament for the treatment of cancer, wherein the preparation induces or enhances the dimerization of human CD137 trimers in a subject, and wherein the cancer is selected from the group consisting of melanoma, glioma, renal cancer, breast cancer, blood cancer and head and neck cancer. Use of a formulation according to any one of claims 1 to 11 in the preparation of a medicament for the treatment of cancer, wherein the formulation induces or enhances the multimerization of human CD137 trimers in a subject, and wherein the cancer is selected from the group consisting of: melanoma, glioma, renal cancer, breast cancer, blood cancer, and head and neck cancer. Use of a formulation according to any one of claims 1 to 11 in the preparation of a medicament for the treatment of cancer, wherein the formulation induces or enhances T cell activation in a subject, and wherein the cancer is selected from the group consisting of: melanoma, glioma, renal cancer, breast cancer, blood cancer, and head and neck cancer. Use according to claim 53, wherein the T cell activation occurs in the tumor microenvironment. Use of a formulation according to any one of claims 1 to 11 in the preparation of a medicament for the treatment of cancer, wherein the formulation induces or enhances a cytotoxic T cell response in a subject, and wherein the cancer is selected from the group consisting of: melanoma, glioma, renal cancer, breast cancer, blood cancer, and head and neck cancer. Use according to claim 55, wherein the cytotoxic T cell response occurs in the tumor microenvironment. Use of a formulation according to any one of claims 1 to 11 in the preparation of a medicament for the treatment of cancer, wherein the formulation induces or enhances cytokine production by immune cells in a subject, and wherein the cancer is selected from the group consisting of: melanoma, glioma, renal cancer, breast cancer, blood cancer, and head and neck cancer. Use according to claim 57, wherein the cytokines produced are IL-2, TNFα, IL-13, IFN-γ, or a combination thereof. Use according to claim 58, wherein the cytokine production occurs in the tumor microenvironment. Use of a formulation according to any one of claims 1 to 11 in the preparation of a medicament for the treatment of cancer, wherein the formulation induces or enhances T cell proliferation in a subject, and wherein the cancer is selected from the group consisting of: melanoma, glioma, renal cancer, breast cancer, blood cancer, and head and neck cancer. Use according to claim 60, wherein the T cell proliferation occurs in the tumor microenvironment. Use of a formulation according to any one of claims 1 to 11 in the preparation of a medicament for the treatment of cancer, wherein the formulation reduces or inhibits tumor growth, and wherein the cancer is selected from the group consisting of: melanoma, glioma, renal cancer, breast cancer, blood cancer, and head and neck cancer. Use of a formulation according to any one of claims 1 to 11 in the preparation of a medicament for the treatment of cancer, wherein the formulation increases the infiltration of immune cells into the tumor microenvironment after administration, and wherein the cancer is selected from the group consisting of: melanoma, glioma, renal cancer, breast cancer, blood cancer, and head and neck cancer. Use according to claim 63, wherein the immune cells express CD45. Use of a preparation according to any one of claims 1 to 11 in the manufacture of a medicament for the treatment of cancer, wherein the preparation reduces the number of regulatory T (Treg) cells in the tumor microenvironment after administration, and wherein the cancer is selected from the group consisting of: melanoma, glioma, renal cancer, breast cancer, blood cancer, and head and neck cancer.

66. The use according to claim 65, wherein the Treg cells express CD4, FOXP-3, and CD25.

67. Use of a preparation according to any one of claims 1 to 11 in the manufacture of a medicament for the treatment of cancer, wherein the preparation reduces the number of macrophages in the tumor microenvironment after administration of the antibody or its antigen-binding portion, and wherein the cancer is selected from the group consisting of: melanoma, glioma, renal cancer, breast cancer, blood cancer, and head and neck cancer.

68. The use according to claim 67, wherein the macrophages express CD45 and CD11b.

69. Use of a preparation according to any one of claims 1 to 11 in the manufacture of a medicament for the treatment of cancer, wherein the preparation reduces T cell exhaustion in the tumor microenvironment after administration of the antibody or its antigen-binding portion, and wherein the cancer is selected from the group consisting of: melanoma, glioma, renal cancer, breast cancer, blood cancer, and head and neck cancer, optionally, wherein the reduction of T cell exhaustion includes a decrease in the expression of TIGIT, PD-1, LAG-3, or a combination thereof.

70. Use of a preparation according to any one of claims 1 to 11 in the manufacture of a medicament for the treatment of cancer, wherein the preparation induces an anti-tumor memory immune response in a subject in need thereof, and wherein the cancer is selected from the group consisting of: melanoma, glioma, renal cancer, breast cancer, blood cancer, and head and neck cancer.

71. Use of the preparation according to claim 69 in the manufacture of a medicament for the treatment of cancer, wherein the anti-CD137 antibody binds to the Fcγ receptor, and wherein the cancer is selected from the group consisting of: melanoma, glioma, renal cancer, breast cancer, blood cancer, and head and neck cancer.

72. Use of a preparation according to any one of claims 1 to 11 in the manufacture of a medicament for the treatment of cancer, wherein depletion of CD4+ T cells, CD8+ T cells, natural killer cells, or a combination thereof reduces the efficacy of the preparation, and wherein the cancer is selected from the group consisting of: melanoma, glioma, renal cancer, breast cancer, blood cancer, and head and neck cancer.

73. A kit comprising a container containing a preparation according to any one of claims 1 to 11 and a package insert containing instructions for administering the preparation, wherein the preparation is a pharmaceutical preparation for treating or delaying the development of cancer in a subject in need thereof, or for reducing or inhibiting tumor growth in a subject in need thereof, and wherein the cancer is selected from the group consisting of: melanoma, glioma, renal cancer, breast cancer, blood cancer, and head and neck cancer.

74. A kit comprising a container containing the preparation according to any one of claims 1 to 11 and a package insert containing instructions for administering the preparation alone or in combination with another agent, wherein the preparation is a pharmaceutical preparation for treating or delaying the development of cancer in a subject in need thereof or for reducing or inhibiting tumor growth in a subject in need thereof, and wherein the cancer is selected from the group consisting of: melanoma, glioma, renal cancer, breast cancer, blood cancer, and head and neck cancer.

75. Use of the preparation according to any one of claims 1 to 11 in the manufacture of a medicament for treating cancer, wherein the preparation delays the development of cancer in a subject in need thereof, or reduces or inhibits tumor growth in a subject in need thereof, and wherein the cancer is selected from the group consisting of: melanoma, glioma, renal cancer, breast cancer, blood cancer, and head and neck cancer.

76. The preparation according to any one of claims 1 to 11 for use in the manufacture of a medicament for treating cancer, wherein the preparation delays the development of cancer in a subject in need thereof, or reduces or inhibits tumor growth in a subject in need thereof, and wherein the cancer is selected from the group consisting of: melanoma, glioma, renal cancer, breast cancer, blood cancer, and head and neck cancer.

77. The preparation according to any one of claims 1 to 11, wherein the preparation is a pharmaceutical preparation.

Images