Method for treating a tumor or cancer in an individual, binding agent, pharmaceutical composition, use of a binding agent, and kit

Description

METHOD FOR TREATING A TUMOR OR CANCER IN AN INDIVIDUAL, BINDING AGENT, PHARMACEUTICAL COMPOSITION, USE OF A BINDING AGENT, AND KIT TECHNICAL FIELD

[001] The present invention relates to therapy that uses a binding agent that binds to human PD-L1 and human CD137 to reduce or prevent tumor progression or to treat cancer. FUNDAMENTALS

[002] CD137 (4-1BB) is a member of the TNFR family and is a co-stimulatory molecule in CD8+ and CD4+ T cells, regulatory T cells (Tregs), natural killer (NK(T)) cells, B cells, and neutrophils. In T cells, CD137 is not constitutively expressed, but rather induced by activation of the T cell receptor (TCR) (e.g., in tumor-infiltrating lymphocytes (TILs) (Gros et al., J. Clin Invest 2014;124(5):2246-59)). Stimulation via its natural ligand 41BBL or agonist antibodies leads to signaling using TRAF-2 and TRAF-1 as adapters. Early signaling by CD137 involves K-63 polyubiquitination reactions that ultimately result in the activation of the nuclear factor (NF)-kB and mitogen-activated protein kinase (MAP) pathways. This signaling leads to increased T cell co-stimulation, proliferation, cytokine production, maturation, and prolonged survival of CD8+ T cells.Agonist antibodies against CD137 have been shown to promote antitumor control by T cells in several preclinical models (Murillo et al., Clin Cancer Res 2008;14(21):6895-906). Antibodies that stimulate CD137 can induce T cell survival and proliferation, thus potentiating the antitumor immune response. Antibodies that stimulate CD137 have been described in the state of the art and include urelumab, a human IgG4 antibody (AU 2004279877), and utomilumab, a human IgG2 antibody (Fisher et al., 2012, Cancer Immunol. Immunother. 61:). Petition 870250088654, dated 09 / 30 / 2025, page 16 / 329 / 254 1721-1733).

[003] Programmed death ligand 1 (PD-L1, PDL1, CD274, B7H1) is a 33 kDa single-pass type I membrane protein. Three isoforms of PD-L1 have been described, based on alternative splicing. PD-L1 belongs to the immunoglobulin (Ig) superfamily and contains an Ig-like C2 domain and an Ig-like V domain. Newly isolated T and B cells express negligible amounts of PD-L1, and a fraction (about 16%) of CD14+ monocytes constitutively express PD-L1. However, interferon-γ (IFNy) is known to increase PD-L1 expression in tumor cells.PD-L1 obstructs antitumor immunity by 1) rendering tumor-reactive T cells insensitive by binding to its receptor, programmed cell death protein 1 (PD-1) (CD279), on activated T cells; 2) rendering tumor cells resistant to lysis mediated by CD8+ T cells and FAS ligands through PD-1 signaling via PD-L1 expressed on tumor cells; 3) rendering T cells insensitive through reverse signaling via CD80 expressed on T cells (B7.1); and 4) promoting the development and maintenance of induced regulatory T cells. PD-L1 is expressed in many types of human cancer, including melanoma, ovarian, lung, and colon cancer (Latchman et al., 2004 Proc Natl Acad Sci USA 101, 10691-6). PD-L1 blocking antibodies have demonstrated clinical activity in several types of cancer that overexpress PD-L1 (including melanoma and NSCLC). For example, atezolizumab is a humanized IgG1 monoclonal antibody against PD-L1.Currently, it is in clinical trials as immunotherapy for several indications, including various types of solid tumors (see, for example, Rittmeyer et al., 2017 Lancet 389:255-265) and is approved for indications of non-small cell lung cancer and bladder cancer. Avelumab, a PD-L1 antibody (Kaufman et al. Lancet Oncol. 2016;17(10):1374-1385), was approved by the FDA for the treatment of adult and pediatric patients 12 years of age and older with carcinoma of... Petition 870250088654, dated 09 / 30 / 2025, page 17 / 329 / 254 metastatic Merkel cells, and is currently in clinical trials for several cancer indications, including bladder cancer, gastric cancer, head and neck cancer, mesothelioma, NSCLC, ovarian cancer, and renal cancer. Durvalumab, a PD-L1 antibody, is approved for indications of locally advanced or metastatic urothelial carcinoma and is in clinical development for multiple solid tumors and hematologic cancers (see, for example, Massard et al., 2016 J Clin Oncol. 34(26):3119-25). Other anti-PD-L1 antibodies have been described, for example, in WO2004004771.

[004] Horton et al (J Immunother Cancer. 2015; 3(Suppl 2): ​​​​O10) describe the combination of a 4-1BB agonist antibody with a PD-L1 neutralizing antibody. WO 2019 / 025545 provides binding agents, such as bispecific antibodies, that bind to human PD-L1 and bind to human CD137. GEN1046 (DuoBody®-PD-L1x4-1BB) is a bispecific PD-L1x4-1BB antibody that targets PD-L1 and 4-1BB.

[005] However, despite these advances in technique, there is a considerable need for improved therapies for the treatment of cancer. SUMMARY

[006] The present inventors have surprisingly found that a binding agent that binds to human PD-L1 and binds to human CD137 can be used to treat tumors or cancers that exhibit high microsatellite instability (MSI-H) or deficiency in nonspecific binding repair (dMMR) in humans.

[007] Thus, in a first aspect, the present description provides a method for treating a tumor or cancer in an individual, said method comprising administering to said individual a binding agent comprising a first binding region that binds to CD137 and a second binding region that binds to PD-L1, wherein said tumor or cancer Petition 870250088654, dated 09 / 30 / 2025, page 18 / 329 / 254, presents high microsatellite instability (MSI-H) or deficiency in nonspecific link repair (dMMR).

[008] In a second aspect, the present description provides a binding agent for use in a method for treating a tumor or cancer in an individual, said method comprising administering to said individual the binding agent comprising a first binding region that binds to CD137 and a second binding region that binds to PD-L1, wherein said tumor or cancer exhibits high microsatellite instability (MSI-H) or deficiency in nonspecific binding repair (dMMR).

[009] In a third aspect, the present description provides a pharmaceutical composition for use in a method for treating a tumor or cancer in an individual, comprising a binding agent comprising a first binding region that binds to CD137 and a second binding region that binds to PD-L1 and optionally a pharmaceutically acceptable carrier, wherein said tumor or cancer exhibits high microsatellite instability (MSI-H) or deficiency in nonspecific binding repair (dMMR).

[0010] In a fourth aspect, the present description provides a use of a binding agent for the manufacture of a medicament for treating a tumor or cancer in an individual, wherein said tumor or cancer exhibits high microsatellite instability (MSI-H) or deficiency in nonspecific binding repair (dMMR), wherein the binding agent comprises a first binding region that binds to CD137 and a second binding region that binds to PD-L1.

[0011] In a fifth aspect, the present description provides a kit for use in a method for treating a tumor or cancer in an individual, wherein said tumor or cancer exhibits high microsatellite instability (MSI-H) or deficiency in nonspecific binding repair (dMMR), comprising a) a binding agent comprising a first binding region that Petition 870250088654, dated 09 / 30 / 2025, page 19 / 329 / 254 links to CD137 and a second binding region that binds to PD-L1, and b) a PD1 inhibitor. BRIEF DESCRIPTION OF THE FIGURES

[0012] Figure 1 shows the change over time in target lesions for all 40 individuals with endometrial cancer measured with GEN1046 in the expansion cohort 4 of the GCT1046-01 Trial (spider plot).

[0013] Figure 2 shows the best overall change in target lesions for all 40 individuals with endometrial cancer measured with GEN1046 in the expansion cohort 4 of the GCT1046-01 Trial (cascade plot).

[0014] Figure 3 shows the best overall change in target lesions for 33 individuals with MSS tumors measured with GEN1046 in the expansion cohort 4 of the GCT1046-01 Trial (cascade plot).

[0015] Figure 4 shows the best overall change in target lesions for 7 individuals with MSI-H tumors measured with GEN1046 in the expansion cohort 4 of the GCT1046-01 Trial (cascade plot).

[0016] Figure 5 shows a schematic representation of the anticipated mode of action of the bispecific CD137xPD-L1 antibodies. (A) PD-L1 is expressed on antigen-presenting cells (APCs) as well as on tumor cells. The binding of PD-L1 to T cells expressing the negative regulatory molecule PD-1 effectively abolishes T cell activation signals and eventually leads to T cell inhibition. (B) After the addition of a bispecific CD137xPD-L1 antibody, the inhibitory PD-1:PD-L1 interaction is blocked via the PD-L1-specific arm and, at the same time, the bispecific antibody, through cell-cell interaction, provides agonistic signaling to CD137 expressed on T cells, resulting in strong T cell co-stimulation.

[0017] Figure 6 shows the MC38 syngeneic tumor model, established by subcutaneous inoculation of 1 χ¹⁰⁶ MC38 cells into Petition 870250088654, dated 09 / 30 / 2025, page 20 / 329 / 254 C57BL / 6 mice. When tumors reached a mean volume of 64 mm3, mice were randomized and treated with mbsIgG2a-PDL1*4-1BB (5 mg / kg), a mouse anti-PD-1 antibody (anti-mPD-1; 10 mg / kg), alone or in combination, or PBS (all 2QW*3). A. The data presented are the median tumor volume per treatment group (n=10), with data transferred to animals that met the termination criteria. Growth curves were stopped when <50% of animals in a treatment group remained alive (PBS, mbsIgG2aPD-L1x4-1BB, anti-mPD-1) or until Day 35 (combination of mbsIgG2a-PDL1x4-1BB with anti-mPD-1). Arrows indicate treatment days. B. Progression-free survival, defined as the percentage of mice with a tumor volume less than 500 mm3, is presented as a Kaplan-Meier curve.The Mantel-Cox analysis was used to compare survival between treatment groups on day 45 (Table 9).

[0018] Figure 7 shows the dose-response analysis of proliferation of GEN1046, anti-PD-1 antibody Nivolumab or anti-PD-1 antibody Pembrolizumab in an antigen-specific T cell assay with an active PD1 / PD-L1 axis. T cells labeled with CFSE electroporated with a claudin-6-specific TCR and a PD-1-IVT-RNA were incubated with immature dendritic cells electroporated with claudin-6-IVT-RNA in the presence of (A) GEN1046 (in 3-fold serial dilutions from 1 to 0.00015 pg / mL), (B) nivolumab (in 4-fold serial dilutions from 0.8 to 0.00005 pg / mL), or (C) pembrolizumab (in 4-fold serial dilutions from 0.8 to 0.00005 pg / mL) for five days. CD8+ T cell proliferation was measured by flow cytometry. The data presented are expansion indices as a function of antibody concentration. Error bars (SD) indicate variation in the experiment (n = 3 replicates in (A); n = 2 replicates in (B) and (C), using cells from a representative donor).The curves were fitted using a 4-parameter logarithmic fit, and the EC50 values ​​and Hill coefficients were used. Petition 870250088654, dated 09 / 30 / 2025, page 21 / 329 / 254 (shown in Table 10-12) were determined using GraphPad Prism v9.0 software.

[0019] Figure 8 shows the release of PD-1 / PD-L1-mediated T cell inhibition and additional co-stimulation of CD8+ T cell proliferation by GEN1046 in the absence or presence of the anti-PD-1 antibody Nivolumab or the anti-PD-1 antibody Pembrolizumab. T cells labeled with CFSE electroporated with a claudin-6 specific TCR and an in vitro translated RNA (IVT) for PD-1 were incubated with immature dendritic cells electroporated with claudin-6 IVT RNA in the presence of 0.2 pg / mL, 0.0067 pg / mL, or 0.0022 pg / mL of GEN1046 in combination with a fixed concentration of 1.6 pg / mL of nivolumab, in combination with a fixed concentration of 0.8 pg / mL of pembrolizumab, or 0.8 pg / mL of non-binding IgG1-ctrl control antibody for five days (n = 2 technical replicates per condition, using cells from n = 3 individual donors). Medium only, only 0.8 pg / mL of IgG1-ctrl.1.6 pg / mL of nivolumab alone and 0.8 pg / mL of pembrolizumab alone were used to determine basal proliferation in the absence of GEN1046. CD8+ T cell proliferation was measured by flow cytometry. Bar graphs represent the mean ± standard deviation of expansion indices by indicated condition, calculated using FlowJo v10.7.1 software. The dashed line represents basal proliferation in the presence of the anti-PD-1 antibody nivolumab. The dotted line represents basal proliferation in the presence of the anti-PD-1 antibody pembrolizumab.

[0020] Figure 9 shows the binding of IgG1-PD1 to PD-1 from different species. Transiently transfected CHO-S cells with PD-1 from different species were incubated with IgG1-PD1, pembrolizumab, or non-binding control antibodies IgG1-ctrl-FERR and IgG4-ctrl, and binding was analyzed using flow cytometry. Non-transfected CHO-S cells incubated with IgG1-PD1 were included as a negative control. AB. The Petition 870250088654, dated 09 / 30 / 2025, page 22 / 329 / 254. Data shown are the geometric mean fluorescence intensities (gMFI) ± SD of duplicate wells from a representative experiment out of four experiments. CD. Data shown are the geometric mean fluorescence intensities (gMFI) ± SD of duplicate wells from a representative experiment out of two experiments. E. Data shown are the geometric mean fluorescence intensities (gMFI) ± SD of duplicate wells from a representative experiment out of four experiments. Abbreviations: gMFI = geometric mean fluorescence intensity; PD-1 = programmed cell death protein 1; PE = R-Phycoerythrin.

[0021] Figure 10 shows the competitive binding of IgG1-PD1 with PD-L1 and PD-L2 to human PD-1. Transiently transfected CHO-S cells with human PD-1 were incubated with 1 μg / mL of biotinylated recombinant human PD-L1 (A) or PD-L2 (B) in the presence of IgG1-PD1 or pembrolizumab. IgG1-ctrl-FERR was included as a negative control. Cells were stained with streptavidin-allophycocyanin, and the percentage of cells that bound to biotinylated PD-L1 or PD-L2 was determined by measuring the percentage of streptavidin-allophycocyanin+ cells by flow cytometry. The percentage of streptavidin-allophycocyanin+ cells in the antibody-free control and in an untransfected sample is indicated by dashed lines. The data presented are from single replicates of a representative experiment out of three separate experiments.Abbreviations: Ab = antibody; CHO-S = Chinese hamster ovary, suspension; ctrl = control; FERR = L234F / L235E / G236R-K409R; PD 1 = programmed cell death protein 1; PD-L1 = programmed cell death ligand 1; PD-L2 = programmed cell death ligand 2.

[0022] Figure 11 shows the functional inhibition of the PD-1 / PD-L1 checkpoint by IgG1 PD1. Blockade of the PD-1 / PD-L1 axis was tested using a cell-based bioluminescent PD-1 / PD-L1 blocking reporter assay. Data presented are mean luminescence ± standard deviation. Petition 870250088654, dated 09 / 30 / 2025, page 23 / 329 / 254 standard deviation (SD) of duplicated wells in a representative experiment of five (pembrolizumab and IgG1-PD1), three (IgG1-ctrl-FERR) or two (nivolumab). Abbreviations: FERR = L234F / L235E / G236R-K409R; PD1 = programmed cell death protein 1; PD-L1 = programmed cell death ligand 1; URL = relative light units; SD = standard deviation.

[0023] Figure 12 shows the increase in CD8+ T cell proliferation by IgG1-PD1 in an antigen-specific T cell proliferation assay. Human CD8+ T cells were electroporated with RNA encoding a CLDN6-specific TCR and RNA encoding PD-1 and labeled with CFSE. The T cells were then co-cultured with iDCs electroporated with CLDN6-encoding RNA in the presence of IgG1-PD1, pembrolizumab, nivolumab, or IgG1-ctrl-FERR. CFSE dilution in T cells was analyzed by flow cytometry after 4 days and used to calculate the expansion index. Data from one representative donor (26268_B) of four donors evaluated in three independent experiments are presented. Error bars represent the standard deviation (SD) of duplicate wells. Curves were fitted by 4-parameter logarithmic fitting using GraphPad Prism.Abbreviations: CFSE = carboxyfluorescein succinimidyl ester; FERR = L234F / L235E / G236R-K409R; PD1 = programmed cell death protein 1; SD = standard deviation.

[0024] Figure 13 shows IgG1-PD1-induced IFNγ secretion in an allogeneic MLR assay. Three unique pairs of allogeneic human mDC donors and CD8+ T cells were co-cultured in the presence of IgG1-PD1 or pembrolizumab for 5 days. IgG1-ctrl-FERR and an IgG4 isotype control were included as negative controls. IFNγ secretion was analyzed in the supernatant using an IFNγ-specific immunoassay. Data presented are the mean ± standard error of mean concentration (SEM) for three unique pairs of allogeneic donors. Abbreviations: FERR = L234F / L235E / G236R-K409R; IFN = interferon; IgG = Petition 870250088654, dated 09 / 30 / 2025, page 24 / 329 / 254 immunoglobulin G; mDC = mature dendritic cell; MLR = mixed lymphocyte reaction; SEM = standard error of the mean.

[0025] Figure 14 shows IgG1PD1-induced cytokine secretion in an allogeneic MLR assay. Three unique pairs of allogeneic human mDC donors and CD8+ T cells were co-cultured in the presence of 1 μg / mL of IgG1-PD1 or pembrolizumab for 5 days. IgG1-ctrl-FERR was included as a negative control. Cytokine secretion was analyzed in the supernatant using Luminex. (A) Cytokine levels are represented as the mean variation relative to cytokine levels measured in untreated co-cultures. (B) Cytokine production levels of three unique pairs of allogeneic donors are shown, with horizontal lines indicating the mean, upper, and lower limits.Abbreviations: FC = variation relative to double; FERR = L234F / L235E / G236R-K409R; GM-CSF = granulocyte-macrophage colony-stimulating factor; IgG = immunoglobulin G; IL = interleukin; MCP-1 = monocyte chemotactic protein 1; mDC = mature dendritic cell; MLR = lymphocyte mixed reaction; TNF = tumor necrosis factor.

[0026] Figure 15 shows the binding of C1q to membrane-bound IgG1-PD1. The binding of C1q to IgG1-PD1 was analyzed using stimulated human CD8+ T cells. After incubation with IgG1-PD1, IgG1 ctrlFERR, IgG1-ctrl, or positive control antibody IgG1-CD52-E430G (without inertia mutations and with a mutation that enhances hexamerization), the cells were incubated with human serum as a source of C1q. C1q binding was detected with a rabbit anti-C1q antibody conjugated with FITC. The data presented are the geometric mean fluorescence intensities (gMFI) ± standard deviation (SD) of duplicate wells from a representative donor among seven donors in three comparable experiments. Abbreviations: FITC = fluorescein isothiocyanate; gMFI = geometric mean fluorescence intensity; PE = R-phycoerythrocyanin. Petition 870250088654, dated 09 / 30 / 2025, page 25 / 329 / 254

[0027] Figure 16 shows the binding of IgG1-PD1 by FcyR. The binding of IgG1-PD1 to immobilized recombinant human FcyR constructs was analyzed by SPR in a qualified assay (n = 1). FcYRIa (A), FcYRIIa-H131 (B), FcYRIIa-R131 (C), FcYRIIb (D), FcyRIIIa-F158 (E), and FcYRIIIa-V158 (F) bind to IgG1-PD1. The IgG1-ctrl antibody (without the FER inertia mutations) was included as a positive control for binding. Abbreviations: Ctrl = control; FcyR = Fc gamma receptor; IgG = immunoglobulin G; PD-1 = programmed cell death protein 1; UR = resonance units.

[0028] Figure 17 shows the binding of IgG1-PD1 and several other anti-PD-1 antibodies by FcyR. The binding of IgG1-PD1, nivolumab, pembrolizumab, dostarlimab, and cemiplimab to immobilized recombinant human FcyR constructs was analyzed by SPR (n = 3). FcYRIa (A), FcYRIIa-H131 (B), FcYRIIa-R131 (C), FcYRIIb (D), FcYRIIIaF158 (E), and FcYRIIIa-V158 (F) were identified as binding test antibodies. IgG1-ctrl and IgG4-ctrl antibodies were included as positive controls for the binding of IgG1 and IgG4 molecules by FcyR to wild-type Fc regions. The binding response ± standard deviation (SD) of three separate experiments is shown. Abbreviations: Ctrl = control; FcyR = Fc gamma receptor; IgG = immunoglobulin G; PD-1 = programmed cell death protein 1; UR = resonance units.

[0029] Figure 18 shows the binding of IgG1-PD1 and several other anti-PD-1 antibodies by FcYRIa. The binding of IgG1-PD1, nivolumab, pembrolizumab, dostarlimab, and cemiplimab to CHO-S cells transiently expressing human FcYRIa was analyzed by flow cytometry. IgG1-ctrl and IgG1-ctrl-FERR were included as positive and negative controls, respectively. Abbreviations: Ctrl = control; FcyR = Fc gamma receptor; FERR = L234F / L235E / G236R-K409R; huIgG = human immunoglobulin G; PD-1 = programmed cell death protein 1; PE = R-phycoerythrin. Petition 870250088654, dated 09 / 30 / 2025, page 26 / 329 / 254

[0030] Figure 19 shows total human IgG in mouse plasma samples. Mice were intravenously injected with 1 or 10 mg / kg of IgG1-PD1 at t = 0 and serial plasma samples were collected at 10 min, 4 h, 1 d, 2 d, 8 d, 14 and 21 d after injection. Total huIgG in plasma samples was determined by ECLIA for each mouse. Data are represented as mean huIgG concentration ± SD of three individual mice. Dashed lines indicate wild-type (wt) plasma huIgG concentrations predicted by a two-compartment model based on IgG clearance in humans (Bleeker et al., 2001, Blood. 98(10):3136-42). Dashed lines indicate the LLOQ and ULOQ. Abbreviations: huIgG = human IgG; IgG = immunoglobulin G; LLOQ = lower limit of quantification; PD-1 = programmed cell death protein 1; SD = standard deviation; ULOQ = upper limit of quantification.

[0031] Figure 20 shows the antitumor activity of IgG1-PD1 in human PD-1 knock-in mice. The MC38 colon cancer syngeneic tumor model was established by SC implantation in hPD-1 KI mice. Mice received 0.5, 2, or 10 mg / kg of IgG1-PD1 or pembrolizumab or 10 mg / kg of IgG1-ctrl-FERR 2QW*3 (9 mice per group). (A) Mean tumor volume ± SEM in each group, up to the last time point when the group was completed. (B) Tumor volumes of the different groups on the last day when all groups were completed (Day 11). The data shown are the tumor volumes in individual mice in each treatment group, as well as the mean tumor volume ± SEM per treatment group. Mann-Whitney analysis was used to compare tumor volumes of the treatment groups with the group treated with IgG1-ctrl-FERR, with *p<0.05, **p<0.01 and ***p<0.001. C.Progression-free survival, defined as the percentage of mice with a tumor volume less than 500 mm3, is shown as a Kaplan-Meier curve. The analysis excluded one mouse from the group. Petition 870250088654, dated 09 / 30 / 2025, page 27 / 329 / 254 IgG1-PD1 of 2 mg / kg that was found dead due to undetermined cause on day 16, before the tumor volume had exceeded 500 mm3. Abbreviations: 2QW*3 = twice a week for three weeks; Ctrl = control; FERR = L234F / L235E / G236R / K409R mutations; IgG = immunoglobulin G; KI = knock-in; PD-1 = programmed cell death protein 1; SC = subcutaneous; SEM = standard error of the mean.

[0032] Figure 21 shows IL-2 secretion induced by IgG1-PD1 in combination with GEN1046 in an allogeneic MLR assay. Two unique pairs of allogeneic human mDC donors and CD8+ T cells were co-cultured for 5 days in the presence of IgG1-PD1 (1 pg / mL), pembrolizumab (research grade, 1 pg / mL), GEN1046 (0.001 to 30 pg / mL), or the combination of pembrolizumab or IgG1-PD1 and GEN1046. IgG1-ctrlFERR (100 pg / mL), IgG4 (100 pg / mL), bsIgG1-PD-L1xctrl (30 pg / mL), bsIgG1-ctrlx4-1BB (30 pg / mL), and IgG1-ctrl-FEAL (30 pg / mL) were included as control antibodies. IL-2 secretion was analyzed in the supernatant by Luminex. The data shown are the mean IL-2 levels ± SEM of 2 pairs of single allogeneic donors.Abbreviations: bsIgG1 = bispecific immunoglobulin G1; ctrl = control; FERR = L234F / L235E / G236R, K409R mutations; FEAL = L234F / L235E / D265A, F405L mutations; IL = interleukin; IgG = immunoglobulin G; mDCs = mature dendritic cells; MLR = mixed lymphocyte reaction; PD1 = programmed cell death protein 1; PD-L1 = programmed cell death ligand 1; SEM = standard error of the mean.

[0033] Figure 22 shows the increased proliferation of CD8+ T cells by IgG1-PD1 in combination with GEN1046 in an antigen-specific T cell stimulation assay. Human CD8+ T cells were electroporated with RNA encoding a CLDN6-specific TCR and RNA encoding PD1 and labeled with CFSE. The T cells were then co-cultured with CLDN6-electroporated iDCs in the presence of 0.8 pg / mL Petition 870250088654, dated 09 / 30 / 2025, page 28 / 329 / 254 of IgG1-PD1, pembrolizumab, or IgG1-ctrl-FERR, alone or in combination with the indicated concentrations of GEN1046. The dilution of CFSE in T cells was analyzed by flow cytometry after 4 days and used to calculate the expansion index. Data from one representative donor among four donors evaluated in two independent experiments are presented. Error bars represent the standard deviation (SD) of duplicate wells. The dashed line indicates the expansion index of CD8+ T cells co-cultured with sham-electroporated iDCs (i.e., not expressing CLDN6).Abbreviations: CFSE = carboxyfluorescein succinimidyl ester; CLDN6 = claudin 6; ctrl = control; FERR = L234F / L235E / G236R, K409R mutations; iDCs = immature dendritic cells; IgG1 = immunoglobulin G1; PD1 = programmed cell death protein 1; PD-L1 = programmed cell death ligand 1; RNA = ribonucleic acid; DP = standard deviation; TCR = T-cell receptor.

[0034] Figure 23 shows the increased cytokine secretion by IgG1-PD1 in combination with GEN1046 after antigen-specific stimulation of CD8+ T cells. Human CD8+ T cells expressing a TCR specific for CLDN6 and PD1 were co-cultured with iDCs expressing CLDN6 in the presence of 0.8 pg / mL of IgG1-PD1, pembrolizumab, or IgG1-ctrl-FERR, alone or in combination with the indicated concentrations of GEN1046. Cytokine concentrations in the culture supernatants were determined after 4 days by multiplexed electrochemiluminescence immunoassay. Data from one representative donor among four donors evaluated in two independent experiments are shown. Error bars represent the standard deviation (SD) of duplicate wells.Abbreviations: CLDN6 = claudin 6; ctrl = control; FERR = L234F / L235E / G236R, K409R mutations; GM-CSF = granulocyte / macrophage colony-stimulating factor; iDCs = immature dendritic cells; IgG1 = immunoglobulin G1; IFN = interferon; IL = interleukin; PD1 =. Petition 870250088654, dated 09 / 30 / 2025, page 29 / 329 / 254 programmed cell death protein 1; PD-L1 = programmed cell death ligand 1; RNA = ribonucleic acid; DP = standard deviation; TCR = T cell receptor.

[0035] Figure 24 shows the MC38 colon cancer model established by subcutaneous inoculation of 1 χ¹⁰⁶ MC38 cells into C57BL / 6 mice. When tumors reached a mean volume of 60 mm³, mice were randomized and treated with the indicated antibodies or combinations thereof (all 2QWx3). A. Data presented are the median tumor volume per treatment group (n = 10), with data transferred to animals that met termination criteria. Growth curves were stopped when <50% of animals in a treatment group remained alive (mIgG2a-ctrlAAKR, mbsIgG2a-PD-L1x4-1BB, mouse anti-PD-1 antibody [antimPD-1]) or until Day 69 (combination of mbsIgG2a-PD-L1*4-1BB with antimPD-1). Downward-pointing triangles indicate treatment days. B. Progression-free survival, defined as the percentage of mice with a tumor volume less than 500 mm3, is shown as a Kaplan-Meier curve.

[0036] Figure 25 shows the new challenge of mice with complete tumor regression after treatment and a control group of tumor-free mice. The mice were again challenged with 1 χ¹⁰⁶ MC38 tumor cells that were injected subcutaneously on Day 121 after the start of antibody treatment. The data presented are mean tumor volumes ± SEM.

[0037] Figure 26 shows the cytokine levels in the peripheral blood of C57BL / 6 mice bearing the MC38 tumor treated with mbsIgG2a-PD-L1x4-1BB, an anti-mPD-1 antibody, as single agents or in combination, or with the non-binding control antibody IgG2a-ctrl-AAKR. Peripheral blood samples were collected at the beginning of the study (one day Petition 870250088654, dated 09 / 30 / 2025, page 30 / 329 / 254 before treatment [Day -1], dotted line) and two days after each treatment (Day 2 and Day 5). Cytokine analysis was performed by ECLIA.

[0038] Figure 27 shows quantitative IHC and ISH data on immune and tumor cell markers expressed in resected tumor tissues from the MC38 colon cancer model. C57BL / 6 mice were inoculated with 1 χ¹⁰⁶ MC38 cells. When tumors reached a mean volume of 50-70 mm³, mice were randomized and treated with mbsIgG2α-PD-L1 χ¹⁰BB, anti-mPD-1, or a combination thereof. Tumors were resected on Day 7 (n = 5 per treatment group) or Day 14 (n = 5 per treatment group) after the start of treatment. Some of the resected tumor samples were too small for IHC analysis, resulting in the analysis of 4-5 tumors per treatment group. Sections of resected tumors (4 μm) were stained with anti-CD3, anti-CD4, anti-CD8, or anti-PD-L1 antibodies by immunohistochemistry (IHC), or were stained for 4-1BB or PD-L2 by in situ hybridization (ISH).IHC data are presented as the percentage of marker-positive cells out of the total cells counted on the slide, as well as the mean ± SEM per treatment group. ISH data are presented as the RNAscope H-score per slide, as well as the mean ± SEM per treatment group.

[0039] Figure 28 shows the expression of GzmB and Ki67 in CD8 T cell subsets from dissociated tumor tissue of the MC38 colon cancer model. C57BL / 6 mice were inoculated with 1 χ¹⁰⁶ MC38 cells. When tumors reached a mean volume of 50–70 mm³, mice were randomized and treated with mbsIgG2α-PDL1x4-1BB, anti-mPD-1, or a combination thereof. Tumors were resected on day 7 (n = 5 per treatment group) after the start of treatment, dissociated into single-cell suspensions, and analyzed by flow cytometry. The data presented are the percentage of cells Petition 870250088654, dated 09 / 30 / 2025, page 31 / 329 / 254 GzmB+ (A) or Ki67+(B) in the CD8+ T cell population of individual mice and the mean ± SEM by treatment group. Mann-Whitney statistical analysis was performed to compare the percentage of GzmB+ or Ki67+ cells in the CD8+ T cell population between treatment groups, with * p < 0.05 and ** p < 0.01.

[0040] Figure 29 shows the characterization of the depleted phenotype of CD3+ T cells after two rounds of CD3 / CD28 stimulation. (A) In vitro depleted CD3+ T cells or naive T cells were stimulated with CD3 / CD28 beads. IFNγ secretion was analyzed by ELISA. Data presented are mean ± standard deviation (SD) of duplicate wells from a representative donor pair. (B) The expression of TIM3, LAG3, PD-1, and 4-1BB in in vitro naive and depleted CD3+ T cells was determined by flow cytometry. Data presented are the median fluorescence intensity corrected for background fluorescence (ΔMFI). (C) Ki67 expression in in vitro naive and depleted CD3+ T cells was determined by flow cytometry.

[0041] Figure 30 shows IFNγ secretion induced by GEN1046 in combination with pembrolizumab in a mixed lymphocyte reaction (MLR) of mature dendritic cells (mDCs) and depleted CD3+ T cells in vitro (Tex). Tex were co-cultured with DCs matured in allogeneic LPS (at a DC:T cell ratio of 1:4) in the presence of GEN1046 (0.001 - 30 pg / mL) or pembrolizumab (1 pg / mL) alone or in combination for 5 days. Co-cultivars without antibody treatment (no antibody) or treated with bs!gG1-PD-L1xctrl (30 pg / mL), bs!gG1-ctrl*4-1BB (30 pg / mL), IgG4 isotype control (1 pg / mL) or IgG1-ctrl-FEAL (30 pg / mL) were included as controls. IFNγ secretion was analyzed by ELISA. The data presented are mean ± standard deviation (SD) of duplicate wells from a representative donor pair among four donor pairs tested.

[0042] Figure 31 shows the highest synergy scores of Petition 870250088654, dated 09 / 30 / 2025, page 32 / 329 / 254 single agent (HSA) for the combination of GEN1046 with pembrolizumab in a multidrug-resistant leprosy (MLR) of mDCs and Tex. Tex were co-cultured with DCs matured in allogeneic LPS (in a DC:T cell ratio of 1:4) in the presence of GEN1046 (0.001 - 30 pg / mL) or pembrolizumab (1 pg / mL) alone or in combination for 5 days. The data presented are HSA synergy scores from a representative donor pair among four donor pairs tested. Scores > 10 are indicative of synergy in this model. Table 1 - Sequences: References are made to the sequences and SEQ ID NOs shown, among others, in the sequence listing. References are also made to specific examples of antibodies of the invention described herein, but without limiting the present invention to them. These exemplary, but not limiting, antibodies of the invention are designated herein by reference to the antibody designation. In bold and underlined are F; E; G; A; L; R and G, corresponding to positions 234; 235; 236; 265; 405; 409 and 430, respectively, these positions being in accordance with EU numbering. In SEQ ID Nos. 83 and 84, the amino acids in bold represent the -AAKR or -AALT mutations required for controlled Fab arm switching. In variable regions, CDR regions that have been annotated according to IMGT definitions (unless otherwise stated or contradicted by the context) are underlined. SEQ ID NAME SEQUENCE Organism 1 VH_CD137-009-H7 EVQLVESGGGLVQPGRSLRLSCTASGFSLNDYWMSW vrqapgkglewvgyidvggslyyaasvkgrftisrd DSKSIAYLQMNSLKTEDTAVYYCARGGLTYGFDLW GQGTLVTVSS synthetic construction 2 VH_CD137-009-H7_CDR1 GFSLNDYW synthetic construction 3 VH_CD137-009-H7_CDR2 IDVGGSL synthetic construction 4 VH_CD137-009-H7_CDR3 ARGGLTYGFDL synthetic construction 5 VL_CD137-009-L2 DIVMTQSPSSLSASVGDRVTITCQASEDISSYLAWYQ QKPGKAPKRLIYGASDLASGVPSRFSASGSGTDYTFTI SSLQPEDIATYYCHYYATISGLGVAFGGGTKVEIK synthetic construction Petition 870250088654, dated 09 / 30 / 2025, page 33 / 329 / 254 6 VL_CD137-009- L2_CDR1 EDISSY synthetic construction 7 VL_CD137-009L2_CDR2 GAS synthetic construction 8 VL_CD137-009- L2_CDR3 HYYATISGLGVA synthetic construction 9 VH_CD137-009 QSLEESGGRLVTPGTPLTLTCTVSGFSLNDYWMSWV rqapgkglewigyidvggslyyaswakgrftisrtst TVDLKMTSLTTEDTATYFCARGGLTYGFDLWGPGTL VTVSS synthetic construction 10 VL_CD137-009 DIVMTQTPASVSEPVGGTVTINCQASEDISSYLAWYQ QKPGQRPKRLIYGASDLASGVPSRFSASGSGTEYALTI SDLESADAATYYCHYYATISGLGVAFGGGTEVVVK synthetic construction 11 VH-PD-L1-547 EVQLLEPGGGLVQPGGSLRLSCEASGSTFSTYAMSW VRQAPGKGLEWVSGFSGSGGFTFYADSVRGRFTISRD SSKNTLFLQMSSLRAEDTAVYYCAIPARGYNYGSFQ HWGQGTLVTVSS synthetic construction 12 VH- PD-L1-547- CDR1 GSTFSTYA synthetic construction 13 VH- PD-L1-547- CDR2 FSGSGGFT synthetic construction 14 VH- PD-L1-547- CDR3 AIPARGYNYGSFQH synthetic construction 15 VL- PD-L1-547 SYVLTQPPSVSVAPGQTARITCGGNNIGSKSVHWYQQ KPGQAPVLVVYDDNDRPSGLPERFSGSNSGNTATLTI SRVEAGDEADYYCQVWDSSSDHVVFGGGTKLTVL synthetic construction 16 VL- PD-L1-547- CDR1 NIGSKS synthetic construction 17 VL-PD-L1-547- CDR2 DDN synthetic construct 18 VL- PD-L1-547- CDR3 QVWDSSSDHVV synthetic construct 19 IgG1-Fc ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPV TVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP CPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDV SHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYR VVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTIS KAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKL TVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP GK synthetic construction 20 IgG1-Fc_F405L ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPV TVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP CPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDV SHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYR VVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTIS KAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFLLYSKL TVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP GK synthetic construction Petition 870250088654, dated 09 / 30 / 2025, page 34 / 329 / 254 21 IgG1-Fc_K409R ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPV TVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP CPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDV SHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYR VVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTIS KAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRL TVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP GK synthetic construction 22 IgG1-Fc_FEA ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPV TVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP CPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVA VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTI SKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGF YPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSK LTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP GK synthetic construction 23 IgGl-FEAR-Fc ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPV TVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP CPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVA VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGF YPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSR LTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP GK synthetic construction 24 IgGl-FEAL-Fc ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPV TVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP CPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVA VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTI SKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGF YPSDIAVEWESNGQPENNYKTTPPVLDSDGSFLLYSK LTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP GK synthetic construction 25 IgGl-Fc without C-terminal lysine ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPV TVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP CPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDV SHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYR VVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTIS KAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKL TVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP G synthetic construction Petition 870250088654, dated 09 / 30 / 2025, page 35 / 329 / 254 26 IgG1-Fc_F405L without C-terminal lysine ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPV TVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP CPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDV SHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYR VVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTIS KAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFLLYSKL TVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP G synthetic construction 27 IgG1-Fc_K409R without lysine C-terminal ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPV TVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP CPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDV SHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYR VVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTIS KAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRL TVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP G synthetic construct 28 IgG1-Fc_FEA without lysine C-terminal ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPV TVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP CPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVAVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTI SKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGF YPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSK LTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP G synthetic construction 29 IgG1-FEAR-Fc without C-terminal lysine ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPV TVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP CPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVA VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTI SKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGF YPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSR LTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP G synthetic construction 30 IgG1-FEAL-Fc without C-terminal lysine ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPV TVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP CPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVA VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTI SKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGF YPSDIAVEWESNGQPENNYKTTPPVLDSDGSFLLYSK LTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP G constructionsynthetic Petition 870250088654, dated 09 / 30 / 2025, p. 36 / 329 / 254 31 Cadeia pesada de CD137-009-H7 EVQLVESGGGLVQPGRSLRLSCTASGFSLNDYWMSW VRQAPGKGLEWVGYIDVGGSLYYAASVKGRFTISRD DSKSIAYLQMNSLKTEDTAVYYCARGGLTYGFDLW GQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGC LVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPK SCDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRT PEVTCVVVAVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVS LTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSD GSFFLYSRLTVDKSRWQQGNVFSCSVMHEALHNHYT QKS LSLSPG construção sintética 32 Cadeia leve de CD137-009-L2 DIVMTQSPSSLSASVGDRVTITCQASEDISSYLAWYQ QKPGKAPKRLIYGASDLASGVPSRFSASGSGTDYTFTI SSLQPEDIATYYCHYYATISGLGVAFGGGTKVEIKRT VAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKV QWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLS KADYEKHKVYACEVTHQGLSSPVTKSFNRGEC construção sintética 33 Cadeia pesada de PDL1-547 EVQLLEPGGGLVQPGGSLRLSCEASGSTFSTYAMSW VRQAPGKGLEWVSGFSGSGGFTFYADSVRGRFTISRD SSKNTLFLQMSSLRAED TAVYYCAIPARGYNYGSFQHWGQGTLVTVSSASTKG PSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQT YICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEF EGGPSVFLFPPKPKDTLMISRTPEVTCVVVAVSHEDP EVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVL TVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQ PREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAV EWESNGQPENNYKTTPPVLDSDGSFLLYSKLTVDKSR WQQGNVFSCSVMHEALHNHYTQKSLSLSPG synthetic construction 34 PDL1-547 light chain SYVLTQPPSVSVAPGQTARITCGGNNIGSKSVHWYQQ KPGQAPVLVVYDDNDRPSGLPERFSGSNSGNTATLTI SRVEAGDEADYYCQVWDSSSDHVVFGGGTKLTVLG QPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVT VAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLT PEQWKSHRSYSCQVTHEGSTVEKTVAPTECS synthetic construct 35 Kappa-C RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREA KVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC synthetic construct 36 Lambda-C GQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLS LTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS synthetic construct 37 CD137 of human (UniProtKB - Q07011; incl. signal peptide sequence: aa 1-23) MGNSCYNIVATLLLVLNFERTRSLQDPCSNCPAGTFCDNNRNQICSPCPPNSFSSAGGQRTCDICRQCKGVFRT RKECSSTSNAECDCTPGFHCLGAGCSMCEQDCKQGQ ELTKKGCKDCCFGTFNDQKRGICRPWTNCSLDGKSV LVNGTKERDVVCGPSPADLSPGASSVTPPAPAREPGH SPQIISFFLALTSTALLFLLFFLTLRFSVVKRGRKKLLY IFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCEL Homo sapiens Petition 870250088654, dated 09 / 30 / 2025, page 37 / 329 / 254 38 Human CD137 (UniProtKB - Q07011; mature sequence) LQDPCSNCPAGTFCDNNRNQICSPCPPNSFSSAGGQR TCDICRQCKGVFRTRKECSSTSNAECDCTPGFHCLGA GCSMCEQDCKQGQELTKKGCKDCCFGTFNDQKRGIC RPWTNCSLDGKSVLVNGTKERDVVCGPSPADLSPGA SSVTPPAPAREPGHSPQIISFFLALTSTALLFLLFFLTLR FSVVKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFP EEEEGGCEL Homo sapiens 39 Human PD-L1 (UniProtKB - Q9NZQ7; incl. signal peptide sequence: aa 1-18) MRIFAVFIFMTYWHLLNAFTVTVPKDLYVVEYGSNM TIECFPVEKQLDLAALIVYWEMEDKNIIQFVHGEEDL KVQHSSYRQRARLLKDQLSLGNAALQITDVKLQDAG VYRCMISYGGADYKRITVKVNAPYNKINQRILVVDP VTSEHELTCQAEGYPKAEVIWTSSDHQVLSGKTTTTN SKREEKLFNVTSTLRINTTTNEIFYCTFRRLDPEENHT AELVIPELPLAHPPNERTHLVILGAILLCLGVALTFIFR LRKGRMMDVKKCGIQDTNSKKQSDTHLEET Homo sapiens 40 human PD-L1 (UniProtKB - Q9NZQ7; mature sequence) FTVTVPKDLYVVEYGSNMTIECKFPVEKQLDLAALIV YWEMEDKNIIQFVHGEEDLKVQHSSYRQRARLLKDQ LSLGNAALQITDVKLQDAGVYRCMISYGGADYKRIT VKVNAPYNKINQRILVVDPVTSEHELTCQAEGYPKAE VIWTSSDHQVLSGKTTTTNSKREEKLFNVTSTLRINTT TNEIFYCTFRRLDPEENHTAELVIPELPLAHPPNERTHLVILGAILLCLGVALTFIFRLRKGRMMDVKKCGIQDT NSKKQSDTHLEET Homo sapiens 41 PD-1 de humano MQIPQAPWPVVWAVLQLGWRPGWFLDSPDRPWNPP TFSPALLVVTEGDNATFTCSFSNTSESFVLNWYRMSP SNQTDKLAAFPEDRSQPGQDCRFRVTQLPNGRDFHM SVVRARRNDSGTYLCGAISLAPKAQIKESLRAELRVT ERRAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLV LLVWVLAVICSRAARGTIGARRTGQPLKEDPSAVPVF SVDYGELDFQWREKTPEPPVPCVPEQTEYATIVFPSG MGTSSPARRGSADGPRSAQPLRPEDGHCSWPL Homo sapiens 42 CTLA-4 MACLGFQRHKAQLNLATRTWPCTLLFFLLFIPVFCKA MHVAQPAVVLASSRGIASFVCEYASPGKATEVRVTV LRQADSQVTEVCAATYMMGNELTFLDDSICTGTSSG NQVNLTIQGLRAMDTGLYICKVELMYPPPYYLGIGN GTQIYVIDPEPCPDSDFLLWILAAVSSGLFFYSFLLTA VSLSKMLKKRSPLTTGVYVKMPPTEPECEKQFQPYFI PIN Homo sapiens 43 VH_MAB-19-0618 QVQLVESGGGLVQPGTSLRLSCSVSGFSLYSYNMGW VRQAPGKGLEYIGIISGGTIGHYASWAKGRFTISRDTS KTTLYLQMNSLTTEDTATYFCARAFYDDYDYNVWG PGTLVTVSS construção sintética 44 VL_MAB-19-0618 AIQLTQSPSSLSASVGGTVTITCQSSQSVYGNNQLSW YQQKPGQPPKLLIYQASKLETGVPSRFRGSGSGTQFT LTISSLQSEDFATYYCAGGYSSSSDTTFGGGTEVVVK construção sintética 45 VH_MAB-19-0618 CDR1 GFSLYSYNSynthetic construction 46 VH_MAB-19-0618 CDR2 IISGGTIG Synthetic construction 47 VH_MAB-19-0618 CDR3 AFYDDYDYNV Synthetic construction 48 VL_MAB-19-0618 CDR1 QSVYGNNQ Synthetic construction 49 VL_MAB-19-0618 CDR2 QAS Synthetic construction Petition 870250088654, dated 09 / 30 / 2025, page 38 / 329 / 254 50 VL_MAB-19-0618 CDR3 AGGYSSSSDTT synthetic construct 51 VH_IgG1-PD1MDX1106-FEAL CDR1 GITFSNSG synthetic construct 52 VH_IgG1-PD1MDX1106-FEAL CDR2 IWYDGSKR synthetic construct 53 VH_IgG1-PD1MDX1106-FEAL CDR3 ATNDDY synthetic construct 54 VL_IgG1-PD1MDX1106-FEAL CDR1 QSVSSY synthetic construct 55 VL_IgG1-PD1MDX1106-FEAL CDR2 DAS synthetic construct 56 VL_IgG1-PD1MDX1106-FEAL CDR3 QQSSNWPRT synthetic construct 57 IgG1-PD1MDX1106FEAL_VH QVQLVESGGGVVQPGRSLRLDCKASGITFSNSGMHW VRQAPGKGLEWVAVIWYDGSKRYYADSVKGRFTISR DNSKNTLFLQMNSLRAEDTAVYYCATNDDYWGQGT LVTVSS synthetic construct 58 IgG1-PD1MDX1106-FEAL_VL EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQ QKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTI SSLEPEDFAVYYCQQSSNWPRTFGQGTKVEIK synthetic construct 59 Pembrolizumab VH CDR1 GYTFTNYY synthetic construct 60 Pembrolizumab VH CDR2 INPSNGGT synthetic construct 61 Pembrolizumab VH CDR3 ARRDYRFDMGFDY synthetic construct 62 Pembrolizumab VL CDR1 KGVSTSGYSY synthetic construction 63 VL CDR2Pembrolizumab LAS synthetic construct 64 VL CDR3 of Pembrolizumab QHSRDLPLT synthetic construct 65 VH of Pembrolizumab QVQLVQSGVEVKKPGASVKVSCKASGYTFTNYYMY WVRQAPGQGLEWMGGINPSNGGTNFNEKFKNRVTL TTDSSTTTAYMELKSLQFDDTAVYYCARRDYRFDMG FDYWGQGTTVTVSS synthetic construct 66 VL of Pembrolizumab EIVLTQSPATLSLSPGERATLSCRASKGVSTSGYSYLH WYQQKPGQAPRLLIYLASYLESGVPARFSGSGSGTDF TLTISSLEPEDFAVYYCQHSRDLPLTFGGGTKVEIK synthetic construct Petition 870250088654, dated 09 / 30 / 2025, page 39 / 329 / 254 67 Pembrolizumab Heavy Chain QVQLVQSGVEVKKPGASVKVSCKASGYTFTNYYMY WVRQAPGQGLEWMGGINPSNGGTNFNEKFKNRVTL TTDSSTTTAYMELKSLQFDDTAVYYCARRDYRFDMG FDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTA ALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS SGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDK RVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMIS RTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKT KPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVS NKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQ VSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLD SDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNH YTQKSLSLSLGK synthetic construct 68 Pembrolizumab light chain EIVLTQSPATLSLSPGERATLSCRASKGVSTSGYSYLH WYQQKPGQAPRLLIYLASYLESGVPARFSGSGSGTDF TLTISSLEPEDFAVYYCQHSRDLPLTFGGGTKVEIKRT VAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKV QWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLS KADYEKHKVYACEVTHQGLSSPVTKSFNRGEC synthetic construct 69 Nivolumab VH CDR1 GITFSNSG synthetic construct 70 Nivolumab VH CDR2 IWYDGSKR synthetic construct 71 Nivolumab VH CDR3 ATNDDY synthetic construct 72 Nivolumab VL CDR1 QSVSSY construct73 Nivolumab VL CDR2 synthetic construct 74 Nivolumab VL CDR3 synthetic construct 75 Nivolumab VH synthetic construct QVQLVESGGGVVQPGRSLRLDCKASGITFSNSGMHW VRQAPGKGLEWVAVIWYDGSKRYYADSVKGRFTISR DNSKNTLFLQMNSLRAEDTAVYYCATNDDYWGQGT LVTVSS synthetic construct 76 Nivolumab VL EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQ QKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTI SSLEPEDFAVYYCQQSSNWPRTFGQGTKVEIK synthetic construct 77 Nivolumab heavy chain QVQLVESGGGVVQPGRSLRLDCKASGITFSNSGMHW VRQAPGKGLEWVAVIWYDGSKRYYADSVKGRFTISR DNSKNTLFLQMNSLRAEDTAVYYCATNDDYWGQGT LVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKD YFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV VTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGP PCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCV VVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFN STYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIE KTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVK GFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSL SLGK synthetic construction Petition 870250088654, dated 09 / 30 / 2025, page 40 / 329 / 25478 Nivolumab light chain EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQ QKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTI SSLEPEDFAVYYCQQSSNWPRTFGQGTKVEIKRTVAA PSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWK VDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADY EKHKVYACEVTHQGLSSPVTKSFNRGEC synthetic construction 79 VH_IgG1-b12 QVQLVQSGAEVKKPGASVKVSCQASGYRFSNFVIHW VRQAPGQRFEWMGWINPYNGNKEFSAKFQDRVTFT ADTSANTAYMELRSLRSADTAVYYCARVGPYSWDD SPQDNYYMDVWGKGTTVIVSS synthetic construction 80 VL_IgG1-b12 EIVLTQSPGTLSLSPGERATFSCRSSHSIRSRRVAWYQ HKPGQAPRLVIHGVSNRASGISDRFSGSGSGTDFTLTI TRVEPEDFALYYCQVYGASSYTFGQGTKLERK synthetic construction 81 m4-1BB-3H3 VH EMQLVESGGGLVQPGRSMKLSCAGSGFTLSDYGVA WVRQAPKKGLEWVAYISYAGGTTYYRESVKGRFTIS RDNAKSTLYLQMDSLRSEDTATYYCTIDGYGGYSGS HWYFDFWGPGTMVTVSS synthetic construction 82 m4-1BB-3H3 VL DIQMTQSPSLLSASVGDRVTLNCRTSQNVYKNLAWY QQKLGEAPKLLIYNANSLQAGIPSRFSGSGSGTDFTLT ISSLQPEDVATYFCQQYYSGNTFGAGTNLELK synthetic construction 83 AALT AKTTAPSVYPLAPVCGDTTGSSVTLGCLVKGYFPEPV TLTWNSGSLSSGVHTFPAVLQSDLYTLSSSVTVTSSTWPSQSITCNVAHPASSTKVDKKIEPRGPTIKPCPPCKC PAPNAAGGPSVFIFPPKIKDVLMISLSPMVTCVVVDVS EDDPDVQISWFVNNVEVLTAQTQTHREDYNSTLRVV SALPIQHQDWMSGKEFKCKVNNKALPAPIERTISKPK GSVRAPQVYVLPPPEEEMTKKQVTLTCMVTDFMPED IYVEWTNNGKTELNYKNTEPVLDSDGSYLMYSKLTV EKKNWVERNSYSCSVVHEGLHNHHTTKSFSRTPGK synthetic construction 84 AAKR AKTTAPSVYPLAPVCGDTTGSSVTLGCLVKGYFPEPV TLTWNSGSLSSGVHTFPAVLQSDLYTLSSSVTVTSST WPSQSITCNVAHPASSTKVDKKIEPRGPTIKPCPPCKC PAPNAAGGPSVFIFPPKIKDVLMISLSPMVTCVVVDVS EDDPDVQISWFVNNVEVLTAQTQTHREDYNSTLRVV SALPIQHQDWMSGKEFKCKVNNKALPAPIERTISKPK GSVRAPQVYVLPPPEEEMTKKQVTLTCMVKDFMPE DIYVEWTNNGKTELNYKNTEPVLDSDGSYFMYSRLR VEKKNWVERNSYSCSVVHEGLHNHHTTKSFSRTPGK synthetic construction 85 constant region kappa LC of mouse RADAAPTVSIFPPSSEQLTSGGASVVCFLNNFYPKDIN VKWKIDGSERQNGVLNSWTDQDSKDSTYSMSSTLTL TKDEYERHNSYTCEATHKTSTSPIVKSFNRNEC synthetic construction 86 MPDL3280A VH EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHW YRQAPGKGLEWYAWISPYGGSTYYADSVKGRFTISA DTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDY WGQGTLVTVSS synthetic construction 87 MPDL3280A VLDIQMTQSPSSLSASVGDRVTITCRASQDVSTAVAWY QQKPGKAPKLLIYSASFLYSGVPSRFSGSGSGTDFTLT ISSLQPEDFATYYCQQYLYHPATFGQGTKVEIK synthetic construction 88 VH IgG1-PD1 QVQLVESGGGLVQPGTSLRLSCSVSGFSLYSYNMGW VRQAPGKGLEYIGIISGGTIGHYASWAKGRFTISRDTS KTTLYLQMNSLTTEDTATYFCARAFYDDYDYNVWG PGTLVTVSS synthetic construction Petition 870250088654, dated 09 / 30 / 2025, page 41 / 329 / 254 89 VL IgG1-PD1 AIQLTQSPSSLSASVGGTVTITCQSSQSVYGNNQLSW YQQKPGQPPKLLIYQASKLETGVPSRFRGSGSGTQFT LTISSLQSEDFATYYCAGGYSSSSDTTFGGGTEVVVK synthetic construct 90 human constant region________HC IgG1m(f)-L234FL235E-G236R ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPV TVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP CPAPEFERGPSVFLFPPKPKDTLMISRTPEVTCVVVD VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTI SKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGF YPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSK LTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP G synthetic construction 91 human constant region________HC IgG1m(f)-L234F- L235E-G236R- K409R ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPV TVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP CPAPEFERGPSVFLFPPKPKDTLMISRTPEVTCVVVD VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTI SKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGF YPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSR LTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP G constructionSynthetic 92 Human HC Constant Region IgGd1m(f) ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPV TVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP CPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDV SHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYR VVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTIS KAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKL TVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP G Synthetic construct 93 Human HC Constant Region IgG1m(f)-L234FL235E-G236R and variants ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPV TVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP CPAPEX1X2X3GPSVFLFPPKPKDTLMISRTPEVTCVVV DVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNST YRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKT ISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGF YPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSK LTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP G where: Xi = A, V, L, I, P, F, M or W, preferably I, P, F, M or W, more preferably F; X2 = L, D or E, preferably D or E, more preferably E; X3 = not G,preferably K, R or H, more preferably R; above all preferably X1X2X3 = FER synthetic construction Petition 870250088654, dated 09 / 30 / 2025, page 42 / 329 / 254 94 constant region of human HC IgG1m(f)-L234FL235E-G236RK409R ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPV TVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP CPAPEFERGPSVFLFPPKPKDTLMISRTPEVTCVVVD VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTI SKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGF YPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSR LTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP G synthetic construction 95 Human constant region HC IgG1m(f)-E430G ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPV TVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP CPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDV SHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYR VVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTIS KAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKL TVDKSRWQQGNVFSCSVMHGALHNHYTQKSLSLSP G synthetic construct 96 Human constant region HC IgG4 ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVT VSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSS LGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPSCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQE DPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVS VLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAK GQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDI AVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVD KSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK synthetic construct 97 human kappa constant region LC RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREA KVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC synthetic construct 98 Human IgG1 heavy chain - LALA ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPV TVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPP CPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVD VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTI SKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGF YPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSK LTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP G synthetic construction 99 HCDR 3 intersection of Kabat and IMGT (= Kabat) AFYDDYDYNV synthetic construction 100 HCDR 3 IMGT ARAFYDDYDYNV synthetic construction 101 HCDR 2 intersection of Kabat and IMGT (= IMGT) ISGGTIGsynthetic construction 102 HCDR 2 Kabat IISGGTIGHYASWAKG synthetic construction Petition 870250088654, dated 09 / 30 / 2025, page 43 / 329 / 254 HCDR 1 intersection of Kabat and IMGT SYN synthetic construction 103 HCDR 1 Kabat SYNMG synthetic construction 104 HCDR 1 IMGT GFSLYSYN synthetic construction 105 LCDR 3 intersection = Kabat = IMGT AGGYSSSDTT synthetic construction LCDR2 intersection of Kabat and IMGT (= IMGT) QAS synthetic construction 106 LCDR2 Kabat QASKLET synthetic construction 107 LCDR1 intersection of Kabat and IMGT (= IMGT) QSVYGNNQ synthetic construction 108 LCDR1 Kabat QSSQSVYGNNQLS synthetic construction 109 VH MAB-19-0202 QSVEESGGRLVTPGTPLTLTCTVSGFSLYSYNMGWV RQAPGKGLEYIGIISGGTIGHYASWAKGRFTISKTSST TVDLKMTSLTTEDTATYFCARAFYDDYDYNVWGPG TLVTVSS synthetic construct 110 VL MAB-19-0202 AAVLTQTPSPVSAAVGGTVTISCQSSQSVYGNNQLS WYQQKPGQPPKLLIYQASKLETGVPSRFKGSGSGTQF TLTISDLESDDAATYYCAGGYSSSSDTTFGGGTEVVV K synthetic construct 111 VH IgG1-PD1 (H5 derived from MAB-19-0202) QVQLVESGGGLVQPGTSLRLSCSVSGFSLYSYNMGW VRQAPGKGLEYIGIISGGTIGHYASWAKGRFTISRDTS KTTLYLQMNSLTTEDTATYFCARAFYDDYDYNVWG PGTLVTVSS synthetic construct 112 VL IgG1-PD1 (L4 derived fromMAB-19-0202) AIQLTQSPSSLSASVGGTVTITCQSSQSVYGNNQLSW YQQKPGQPPKLLIYQASKLETGVPSRFRGSGSGTQFT LTISSLQSEDFATYYCAGGYSSSSDTTFGGGTEVVVK synthetic construction 113 PD-1 of complete human MQIPQAPWPVVWAVLQLGWRPGWFLDSPDRPWNPP TFSPALLVVTEGDNATFTCSFSNTSESFVLNWYRMSP SNQTDKLAAFPEDRSQPGQDCRFRVTQLPNGRDFHM SVVRARRNDSGTYLCGAISLAPKAQIKESLRAELRVT ERRAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLV LLVWVLAVICSRAARGTIGARRTGQPLKEDPSAVPVF SVDYGELDFQWREKTPEPPVPCVPEQTEYATIVFPSG MGTSSPARRGSADGPRSAQPLRPEDGHCSWPL Homo sapiens 114 Complete human PD-1 extracellular domain FLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSE SFVLNWYRMSPSNQTDKLAAFPEDRSQPGQDCRFRV TQLPNGRDFHMSVVRARRNDSGTYLCGAISLAPKAQ IKESLRAELRVTERRAEVPTAHPSPSPRPAGQFQTLV Homo sapiens Petition 870250088654, dated 09 / 30 / 2025, page 44 / 329 / 254 115 Human PD-1 nucleic acid agtttcccttccgctcacctcgcctgagcagtggagaaggcggcactctggtggg gctgctccaggcat gcagatcccacggcgccctcgtcgtctgggcggcgct acaactgggctggcggcggcggcctccccc ccccccaccttctccccagccctgctcgtggtgaccgaaggggacaacgccacctt cacctgcagcttctccaacacatcggagagctcgtgctaactggtaccgcatgag ccccagcaaccagacggacaagct cggccaggactgccgcttccgtgtcacacaactgcccaacgggcgtgacttccaca tgagcgtggtcgggcccggcgcaatgacagcggcacctaccctgtggggccat ctccctggcccccaaggcgcagatcaagagagcctgcgggcgagctcaggt gacagagagaagggcagaagtgcccacagcccaccccagcccctcacccaggcgcggccagttccaaaccctggttggtgtcgtgggcggctgctgggcgc ctggtgctgctagtgggtcctggccgtctctccgggcgcgcgcgc aataggagccaggcgcaccggccagccctgaaggacccctcagccgtgcc tgtgttctctgtgtggactatggggctggattccagtggcgagagaccccgga gccccgtgccctgtgtccctgagcacggagtatgccaccattgtctttcctagc gcccagccactgaggcctgaggatggacactgctcttggcccctccttggccaccagtgttctgcagaccctccaccatgagcccgggtcagcgcatttcct caggagaagcaggcaggctgcaggccattgcaggccgtccaggggctgagctgc ctggggcgaccggggctccagcctgcacctgcaccaggcacagccccaccaca ggactcatgtctcaatgcccacagtgagcccaggcagcaggtgtcaccgtccccta caggagggccagatgcagtcactgcttcaggtcctgccagcacagagctgcctg cgtccagctccctgaatctctgctgctgctgctgctgctgctgctgctgcctgcggcc cggggctgaaggcgccgtggccctgcctgacgccccggagcctcctgcctgaact tggggctggttggagatggccttggagcagccaaggtgcccctggcagtggcat cccgaaacgccctggaggcaggccccaagaactgggcacaggaggtgggaggtac atggggctggggactccccaggagttatctgctccctgcaggcctagagaagtttca gggaaggtcagaagagctcctggctgtggtgggcagggcaggaaacccctccac ctttacacatgcccaggcaggcacctcaggccctttgtggggcagggaagctgaggc agtaagcgggcaggcagagctggaggcctttcaggcccagccagcactctggcct cctgccgccgcattccacccagcccctcacaccactcgggagagggacatccta cggtcccaaggtcaggagggcagggctggggttgactcaggccctcccagctgt ggccacctgggttgggaggcagaagtgcaggcacctagggccccatgtg cccaccctgggagctctccttggaacccattcctgaaattatttaaaggggttggccgggctcccaccagggcctgggtgggaaggtacaggcgttcccccggggcctagtac ccccgccgtggcctatccactcctcacatccacacactgcaccccccactcctgggg cagggccaccagcatccaggcggccagcaggcacctgagtggctgggacaagg gatcccccttccctgtggttctattatattataattataattaaatatgagagcatgctaag gaaaa Homo sapiens 116 VH________CD52- CAMPATH-1H QVQLQESGPGLVRPSQTLSLTCTVSGFTFTDFYMNW VRQPPGRGLEWIGFIRDKAKGYTTEYNPSVKGRVTM LVDTSKNQFSLRLSSVTAADTAVYYCAREGHTAAPF DYWGQGSLVTVSS synthetic construction 117 VH CD52- CAMPATH-1H CDR1 GFTFTDFY synthetic construction 118 VH CD52- CAMPATH-1H CDR2 IRDKAKGYTT synthetic construction 119 VH CD52- CAMPATH-1H CDR3 AREGHTAAPFDY synthetic construction 120 VL_________CD52- CAMPATH-1H DIQMTQSPSSLSASVGDRVTITCKASQNIDKYLNWYQ QKPGKAPKLLIYNTNNLQTGVPSRFSGSGSGTDFTFTI SSLQPEDIATYYCLQHISRPRTFGQGTKVEIK synthetic construction Petition 870250088654, dated 09 / 30 / 2025, page 45 / 329 / 254 121 VL CD52- CAMPATH-1H CDR1 QNIDKY synthetic construction VL CD52- CAMPATH-1H CDR2 NTN synthetic construction 122 VL CD52- CAMPATH-1H CDR3 LQHISRPRT synthetic construction 123 VH gp120-b12 QVQLVQSGAEVKKPGASVKVSCQASGYRFSNFVIHW VRQAPGQRFEWMGWINPYNGNKEFSAKFQDRVTFT ADTSANTAYMELRSLRSADTAVYYCARVGPYSWDD SPQDNYYMDVWGKGTTVIVSS synthetic construction 124 VH gp120-b12 CDR1 GYRFSNFV synthetic construction 125 VH gp120-b12 CDR2 INPYNGNK synthetic construction 126 VH gp120-b12 CDR3 ARVGPYSWDDSPQDNYYMDV synthetic construction 127 VL gp120-b12 EIVLTQSPGTLSLSPGERATFSCRSSHSIRSRRVAWYQ HKPGQAPRLVIHGVSNRASGISDRFSGSGSGTDFTLTI TRVEPEDFALYYCQVYGASSYTFGQGTKLERK synthetic construction 128 VL gp120-b12 CDR1 HSIRSRR synthetic construction VL gp120-b12 CDR2 GVS synthetic construction 129 VL gp120-b12-CDR3 QVYGASSYT synthetic construction 130 MAB-19-0202-HC FR1 QSVEESGGRLVTPGTPLTLTCTVSGFSLY synthetic construction 131 MAB-19-0202-HC FR2 WVRQAPGKGLEYIG synthetic construction 132 MAB-19-0202-HC FR3 RFTISKTSSTTVDLKMTSLTTEDTATYFCAR constructionsynthetic construction 133 MAB-19-0202-HC FR4 WGPGTLVTVSS synthetic construction 134 MAB-19-0202-LC FR1 AAVLTQTPSPVSAAVGGTVTISC synthetic construction 135 MAB-19-0202-LC FR2 WYQQKPGQPPKLLIY synthetic construction 136 MAB-19-0202-LC FR3 GVPSRFKGSGSGTQFTLTISDLESDDAATYYC synthetic construction 137 MAB-19-0202-LC FR4 FGGGTEVVVK synthetic construction 138 MAB-19-0202-H5 FR1 QVQLVESGGGLVQPGTSLRLSCSVSGFSLY synthetic construction 139 MAB-19-0202-H5 FR2 WVRQAPGKGLEYIG synthetic construction Petition 870250088654, dated 09 / 30 / 2025, p. 46 / 329 / 254 140 MAB-19-0202-H5 FR3 RFTISRDTSKTTLYLQMNSLTTEDTATYFCAR synthetic construct 141 MAB-19-0202-H5 FR4 WGPGTLVTVSS synthetic construct 142 MAB-19-0202-L4 FR1 AIQLTQSPSSLSASVGGTVTITC synthetic construct 143 MAB-19-0202-L4 FR2 WYQQKPGQPPKLLIY synthetic construct 144 MAB-19-0202-L4 FR3 GVPSRFRGSGSGTQFTLTISSLQSEDFATYYC synthetic construct 145 MAB-19-0202-L4 FR4 FGGGTEVVVK synthetic construct 146 Pembrolizumab VH CDR1 (Kabat numbering) NYYMY synthetic construct 147 Pembrolizumab VH CDR2 Pembrolizumab (Kabat numbering) GINPSNGGTNFNEKFKN synthetic construct 148 Pembrolizumab VH CDR3 (Kabat numbering) RDYRFDMGFDY synthetic construct 149 Pembrolizumab VL CDR1 (Kabat numbering) RASKGVSTSGYSYLH synthetic construct 150 Pembrolizumab VL CDR2 (Kabat numbering) LASYLES synthetic construct 151 Pembrolizumab VL CDR3 (Kabat numbering) QHSRDLPLT synthetic construct 152 IgG1-PD1 heavy chain QVQLVESGGGLVQPGTSLRLSCSVSGFSLYSYNMGW VRQAPGKGLEYIGIISGGTIGHYASWAKGRFTISRDTSKTTLYLQMNSLTTEDTATYFCARAFYDDYDYNVWG PGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLV KDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLS SVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSC DKTHTCPPCPAPEFERGPSVFLFPPKPKDTLMISRTPE VTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPRE EQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKAL PAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGS FFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQ KSLSLSPG synthetic construction 153 Lightweight chain IgG1PD1 AIQLTQSPSSLSASVGGTVTITCQSSQSVYGNNQLSW YQQKPGQPPKLLIYQASKLETGVPSRFRGSGSGTQFT LTISSLQSEDFATYYCAGGYSSSSDTTFGGGTEVVVK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREA KVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC synthetic construction DETAILED DESCRIPTION OF THE INVENTION

[0043] Although the present description is described in more detail to Petition 870250088654, dated 09 / 30 / 2025, p. 47 / 329 / 254, it should be understood that this description is not limited to the specific methodologies, protocols, and reagents described herein, as these may vary. It should also be understood that the terminology used herein is intended only to describe particular embodiments and is not intended to limit the scope of the present description, which will be limited only by the appended claims. Unless otherwise defined, all technical and scientific terms used herein have the same meanings commonly understood by those skilled in the art.

[0044] The elements of this description will be described in more detail below. These elements are listed with particular modalities; however, it should be understood that they can be combined in any way and in any number to create additional modalities. The examples and preferred modalities described in various ways should not be interpreted as limiting this description to only the modalities explicitly described. This description should be understood as supporting and including modalities that combine the explicitly described modalities with any number of the described and / or preferred elements. Furthermore, any permutations and combinations of all the elements described in this application should be considered described by the description of this application, unless the context indicates otherwise.For example, if in a preferred embodiment of the linking agent used herein the first heavy chain comprises or consists essentially of an amino acid sequence presented in SEQ ID NO: 23 or 29 [IgG1Fc_FEAR], and in another preferred embodiment of the linking agent used herein the second heavy chain comprises or consists essentially of an amino acid sequence presented in SEQ ID NO: 24 or 30 [IgG1Fc_FEAL], then in a further preferred embodiment of the linking agent used herein the first heavy chain comprises or consists essentially of an amino acid sequence presented in SEQ ID. Petition 870250088654, dated 09 / 30 / 2025, p. 48 / 329 / 254 NO: 23 or 29 [IgG1-Fc_FEAR], and the second heavy chain comprises or consists essentially of an amino acid sequence presented in SEQ ID NO: 24 or 30 [IgG1-Fc_FEAL].

[0045] Preferably, the terms used here are defined in the manner described in “A multilingual glossary of biotechnological terms: (IUPAC Recommendations)”, HGW Leuenberger, B. Nagel and H. Kolbl, Eds., Helvetica Chimica Acta, CH-4010 Basel, Switzerland, (1995).

[0046] The practice described herein will employ, unless otherwise indicated, conventional chemistry, biochemistry, cell biology, immunology, and recombinant DNA techniques as explained in the literature in the field (compare, for example, Organikum, Deutscher Verlag der Wissenschaften, Berlin 1990; Streitwieser / Heathcook, “Organische Chemie”, VCH, 1990; Beyer / Walter, “Lehrbuch der Organischen Chemie”, S. Hirzel Verlag Stuttgart, 1988; Carey / Sundberg, “Organische Chemie”, VCH, 1995; March, “Advanced Organic Chemistry”, John Wiley & Sons, 1985; Rompp Chemie Lexikon, Falbe / Regitz (Hrsg.), Georg Thieme Verlag Stuttgart, New York, 1989; Molecular Cloning: A Laboratory Manual, 2nd Edition, J. Sambrook et al. al. eds., Cold Spring Harbor Laboratory Press, Cold Spring Harbor 1989.

[0047] All methods described herein may be carried out in any suitable order unless otherwise indicated or clearly contradicted by the context. The use of any and all examples, or exemplary language (e.g., “such as”), provided herein is merely to better illustrate the present description and does not represent a limitation on the scope of the present description otherwise claimed. No language in the descriptive report should be construed as indicating any unclaimed element essential to the practice of the present description.

[0048] The citation of value ranges here is intended merely to Petition 870250088654, dated 09 / 30 / 2025, page 49 / 329 / 254, functions as an abbreviated method of individual reference to each separate value in the range. Unless otherwise indicated herein, each individual value is incorporated into the descriptive report as if it were individually cited herein.

[0049] Several documents are cited throughout the text of the same descriptive report. Each of the documents cited herein (including all patents, patent applications, scientific publications, manufacturer specifications, instructions, etc.), whether above or below, is incorporated herein by reference in its entirety. Nothing contained herein should be construed as an admission that the invention does not have the right to precede such description by virtue of a prior invention. Definitions

[0050] Definitions that apply to all aspects of this description will be provided below. The following terms have the following meanings, unless otherwise indicated. Any undefined terms have their meanings recognized in the art.

[0051] Throughout the same descriptive report and the attached claims, unless the context requires otherwise, the word “comprise” and variations such as “comprises” and “comprising” shall be understood as implying the inclusion of a member, whole number or step or group of members, whole numbers or steps, but not the exclusion of any other member, whole number or step or group of members, whole numbers or steps. The term “essentially consisting of” means the exclusion of other members, whole numbers or steps of any essential meaning. The term “comprising” includes the term “essentially consisting of” which, in turn, includes the term “consisting of”. Thus, in each occurrence in the present application, the term “comprising” may be replaced by the term “essentially consisting of” or “consisting of”. Similarly, in each Petition 870250088654, dated 09 / 30 / 2025, p. 50 / 329 / 254 occurrence in the present request, the term “consisting essentially of” can be replaced by the term “consisting of”.

[0052] The terms “a”, “an”, “the” and “the” and similar references used in the context of the description of the present description (especially in the context of the claims) should be interpreted as including both the singular and the plural, unless otherwise indicated herein or clearly contradicted by the context.

[0053] When used herein, “and / or” should be interpreted as a specific description of each of the two specified features or components, with or without the other. For example, “X and / or Y” should be interpreted as a specific description of each of (i) X, (ii) Y and (iii) X and Y, as if each were presented individually herein.

[0054] In the context of the present description, the term “approximately” indicates a range of precision that a professional with average knowledge will understand still guarantees the technical effect of the characteristic in question. The term typically indicates a deviation from the stated numerical value of ±5%, ±4%, ±3%, ±2%, ±1%, ±0.9%, ±0.8%, ±0.7%, ±0.6%, ±0.5%, ±0.4%, ±0.3%, ±0.2%, ±0.1%, ±0.05%, and, for example, ±0.01%. As will be understood by those skilled in the art, the specific deviation for a numerical value for a given technical effect will depend on the nature of the technical effect. For example, a natural or biological technical effect may generally exhibit a greater deviation than an artificial or engineered technical effect.

[0055] The term “binding agent”, in the context of this description, refers to any agent capable of binding to desired antigens. In certain embodiments of this description, the binding agent is an antibody, antibody fragment or construct thereof. The binding agent may also comprise synthetic, modified or unnaturally occurring fractions, in particular non-peptidic fractions. Such fractions may, for example, link functionalities or binding regions to desired antigens, Petition 870250088654, dated 09 / 30 / 2025, page 51 / 329 / 254 as antibodies or antibody fragments. In one embodiment, the binding agent is a synthetic construct comprising CDRs or antigen-binding variable regions.

[0056] As used herein, “immune checkpoint” refers to regulators of the immune system and, in particular, co-stimulatory and inhibitory signals that regulate the amplitude and quality of T cell receptor recognition of an antigen. In certain embodiments, the immune checkpoint is an inhibitory signal. In certain embodiments, the inhibitory signal is the interaction between PD-1 and PD-L1 and / or PD-L2. In certain embodiments, the inhibitory signal is the interaction between CTLA-4 and CD80 or CD86 to displace CD28 binding. In certain embodiments, the inhibitory signal is the interaction between LAG-3 molecules and MHC class II. In certain embodiments, the inhibitory signal is the interaction between TIM-3 and one or more of its ligands, such as galectin 9, PtdSer, HMGB1, and CEACAM1. In certain embodiments, the inhibitory signal is the interaction between one or more KIRs and their ligands. In certain modalities, the inhibitory signal is the interaction between TIGIT and one or more of its ligands, PVR, PVRL2, and PVRL3.In certain modalities, the inhibitory signal is the interaction between CD94 / NKG2A and HLA-E. In certain modalities, the inhibitory signal is the interaction between VISTA and its binding partner(s). In certain modalities, the inhibitory signal is the interaction between one or more Siglecs and their ligands. In certain modalities, the inhibitory signal is the interaction between GARP and one or more of its ligands. In certain modalities, the inhibitory signal is the interaction between CD47 and SIRPa. In certain modalities, the inhibitory signal is the interaction between PVRIG and PVRL2. In certain modalities, the inhibitory signal is the interaction between CSF1R and CSF1. In certain modalities, the inhibitory signal is the interaction between BTLA and HVEM. In certain modalities, the inhibitory signal is part of the adenosinergic pathway, for example, the interaction between A2AR and / or A2BR and adenosine, produced by CD39 and CD73. In certain modalities, the inhibitory signal is the interaction between... Petition 870250088654, dated 09 / 30 / 2025, page 52 / 329 / 254 B7-H3 and its receptor and / or B7-H4 and its receptor. In certain modalities, the inhibitory signal is mediated by IDO, CD20, NOX, or TDO.

[0057] The terms “checkpoint inhibitor” (CPI) and “immunological checkpoint inhibitor (ICP)” are used herein synonymously. The terms refer to molecules, such as binding agents, that reduce, inhibit, interfere totally or partially with, or negatively modulate one or more checkpoint proteins, or that reduce, inhibit, interfere totally or partially with, or negatively modulate the expression of one or more checkpoint proteins, such as molecules, such as binding agents, that inhibit an immunological checkpoint, in particular, that inhibit the inhibitory signal of an immunological checkpoint. In one embodiment, the immunological checkpoint inhibitor binds to one or more checkpoint proteins. In another embodiment, the immunological checkpoint inhibitor binds to one or more molecules that regulate checkpoint proteins.In one embodiment, the immune checkpoint inhibitor binds to precursors of one or more checkpoint proteins, for example, at the DNA or RNA level. Any agent that functions as a checkpoint inhibitor according to the present description may be used. The term “partially” as used herein means at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% at the level of, for example, inhibition of a checkpoint protein.

[0058] In one embodiment, the checkpoint inhibitor can be any compound, such as any binding agent, that inhibits the inhibitory signal of an immune checkpoint, wherein the inhibitory signal is selected from the group consisting of: the interaction between PD-1 and PDL1 and / or PD-L2; the interaction between CTLA-4 and CD80 or CD86 to displace the binding of CD28; the interaction between LAG-3 molecules and MHC class II;Petition 870250088654, dated 09 / 30 / 2025, p. 53 / 329 / 254 interaction between TIM-3 and one or more of its ligands, such as galectin 9, PtdSer, HMGB1 and CEACAM1; the interaction between one or more KIRs and their ligands; the interaction between TIGIT and one or more of its ligands, PVR, PVRL2 and PVRL3; the interaction between CD94 / NKG2A and HLA-E; the interaction between VISTA and its binding partner(s); the interaction between one or more Siglecs and their ligands; the interaction between GARP and one or more of its ligands; the interaction between CD47 and SIRPa; the interaction between PVRIG and PVRL2; the interaction between CSF1R and CSF1; the interaction between BTLA and HVEM; part of the adenosinergic pathway, for example, the interaction between A2AR and / or A2BR and adenosine, produced by CD39 and CD73; the interaction between B7-H3 and its receptor and / or B7-H4 and its receptor; an inhibitory signal mediated by IDO, CD20, NOX or TDO.In one embodiment, the checkpoint inhibitor is at least one selected from the group consisting of PD-1 inhibitors, PD-L1 inhibitors, PD-L2 inhibitors, CTLA-4 inhibitors, TIM-3 inhibitors, KIR inhibitors, LAG-3 inhibitors, TIGIT inhibitors, VISTA inhibitors, and GARP inhibitors. In one embodiment, the checkpoint inhibitor may be a blocking antibody, such as a PD-1 blocking antibody, a CTLA4 blocking antibody, a PD-L1 blocking antibody, a PD-L2 blocking antibody, a TIM-3 blocking antibody, a KIR blocking antibody, a LAG-3 blocking antibody, a TIGIT blocking antibody, a VISTA blocking antibody, or a GARP blocking antibody. Examples of a PD-1 blocking antibody include pembrolizumab, nivolumab, cemiplimab, and espartalizumab. Examples of CTLA4-blocking antibodies include ipilimumab and tremelimumab.Examples of PD-L1 blocking antibodies include atezolizumab, durvalumab, and avelumab.

[0059] In one embodiment, the anti-PD-1 antibody or antigen-binding fragment thereof comprises a variable chain region. Petition 870250088654, dated 09 / 30 / 2025, page 54 / 329 / 254 heavy (VH) comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NO: 43, and a variable light chain (VL) region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NO: 44.

[0060] In one embodiment, the anti-PD-1 antibody or antigen-binding fragment thereof comprises a variable heavy chain region and a variable light chain region, wherein the variable heavy chain region comprises: (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 45; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 46; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 47; and wherein the variable light chain region comprises: (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 48; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 49; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 50.

[0061] In one embodiment of the anti-PD-1 antibodies described herein, the variable heavy chain domain comprises the amino acid sequence of SEQ ID NO: 43 and the variable light chain domain comprises the amino acid sequence of SEQ ID NO: 44.

[0062] The term “immunoglobulin” refers to proteins of the immunoglobulin superfamily, preferably to antigen receptors such as antibodies or the B cell receptor (BCR). Immunoglobulins are distinguished by a structural domain, namely the immunoglobulin domain, with a characteristic immunoglobulin (Ig) folding pattern. Petition 870250088654, dated 09 / 30 / 2025, page 55 / 329 / 254 The term includes membrane-bound immunoglobulins as well as soluble immunoglobulins. Membrane-bound immunoglobulins are also called surface immunoglobulins or membrane immunoglobulins, which are generally part of the BCR. Soluble immunoglobulins are generally called antibodies.

[0063] The structure of immunoglobulins has been well distinguished. See, for example, Fundamental Immunology Chapter 7 (Paul, W., ed., 2nd ed. Raven Publishers, NY (1989)). In summary, immunoglobulins generally comprise several chains, typically two identical heavy chains and two identical light chains, which are linked by disulfide bonds. These chains are mainly composed of immunoglobulin domains or regions, such as the VL or VL domain / region (variable light chain), CL or CL domain / region (constant light chain), VH or VH domain / region (variable heavy chain), and the CH or CH domains / regions (constant heavy chain) CH1 (CH1), CH2 (CH2), CH3 (CH3) and CH4 (CH4). The constant heavy chain region typically comprises three domains, CH1, CH2 and CH3. The hinge region is the region between the CH1 and CH2 domains of the heavy chain and is highly flexible.Disulfide bonds in the hinge region are part of the interactions between two heavy chains in an IgG molecule. Each light chain is typically comprised of a VL and a CL. The constant light chain region is typically comprised of a domain, CL. The VH and VL regions can be further subdivided into hypervariability regions (or hypervariable regions that may be hypervariable in sequence and / or form of structurally defined loops), also called complementarity-determining regions (CDRs), interspersed with more conserved regions, called framework regions (FRs). Each VH and VL is typically composed of three CDRs and four FRs, arranged from the amino-terminal to the carboxy-terminal in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4 (see also Chothia and Lesk J. Mol. Biol. 196). Petition 870250088654, dated 09 / 30 / 2025, page 56 / 329 / 254 901-917 (1987)). Unless otherwise stated or contradicted by the context, CDR sequences are identified herein according to IMGT rules using DomainGapAlign (Lefranc MP., Nucleic Acids Research 1999;27:209-212 and Ehrenmann F., Kaas Q. and Lefranc M.-P. Nucleic Acids Res., 38, D301-307 (2010); see also the internet address www.imgt.org. Unless otherwise stated or contradicted by the context, reference to amino acid positions in the regions described herein is in accordance with EU numbering (Edelman et al., Proc Natl Acad Sci USA. May 1969;63(1):78-85; Kabat et al., Sequences of Proteins of Immunological Interest, Fifth Edition. 1991 NIH Publication Number 91-3242).

[0064] There are five types of mammalian immunoglobulin heavy chains, namely α, δ, ε, γ, and μ, which represent the different classes of antibodies, namely IgA, IgD, IgE, IgG, and IgM. Unlike soluble immunoglobulin heavy chains, membrane or surface immunoglobulin heavy chains comprise a transmembrane domain and a short cytoplasmic domain at their carboxy terminus. In mammals, there are two types of light chains, namely lambda and kappa. Immunoglobulin chains comprise a variable region and a constant region. The constant region is essentially conserved across different immunoglobulin isotypes, while the variable part is highly diverse and represents antigen recognition.

[0065] The terms “amino acid” and “amino acid residue” may be used interchangeably herein and should not be understood as limiting. Amino acids are organic compounds containing amine (-NH2) and carboxyl (-COOH) functional groups, along with a side chain (R group) specific to each amino acid. In the context of the present description, amino acids can be classified based on their structure and chemical characteristics. Thus, classes of amino acids can be reflected in one or Petition 870250088654, dated 09 / 30 / 2025, page 57 / 329 / 254, both from the following tables: Table 2: Main classification based on the structure and general chemical characterization of the R group. Amino Acid Class: Acidic Residues D and E; Basic Residues K, R, and H; Uncharged Hydrophilic Residues S, T, N, and Q; Uncharged Aliphatic Residues G, A, V, L, and I; Uncharged Nonpolar Residues C, M, and P; Aromatic Residues F, Y, and W. Table 3: Alternative physical and functional classifications of amino acid residues Amino Acid Class Hydroxyl Group-Containing Residues S and T Aliphatic Residues I, L, V, and M Cycloalkenyl-Associated Residues F, H, W, and Y Hydrophobic Residues A, C, F, G, H, I, L, M, R, T, V, W, and Y Negatively Charged Residues D and E Polar Residues C, D, E, H, K, N, Q, R, S, and T Positively Charged Residues H, K, and R Small Residues A, C, D, G, N, P, S, T, and V Very Small Residues A, G, and S Residues Involved in Coil Formation A, C, D, E, G, H, K, N, Q, R, S, P, and T Flexible Residues Q, T, K, S, G, P, D, E, and R

[0066] For the purposes of this description, “variants” of an amino acid sequence (peptide, protein or polypeptide) comprise amino acid insertion variants, amino acid addition variants, amino acid deletion variants and / or amino acid substitution variants. The term “variant” includes all mutants, splicing variants, post-translationally modified variants, conformations, isoforms, allelic variants, species variants, species homologs, in particular those that are naturally occurring. The term “variant” includes, in particular, fragments of an amino acid sequence.

[0067] Amino acid insertion variants comprise insertions Petition 870250088654, dated 09 / 30 / 2025, p. 58 / 329 / 254 of one or two or more amino acids in a particular amino acid sequence. In the case of amino acid sequence variants with insertion, one or more amino acid residues are inserted into a specific site in the amino acid sequence, although random insertion with proper screening of the resulting product is also possible.

[0068] Amino acid addition variants comprise amino and / or carboxy-terminal fusions of one or more amino acids, such as 1, 2, 3, 5, 10, 20, 30, 50 or more amino acids.

[0069] Amino acid deletion variants are distinguished by the removal of one or more amino acids from the sequence, such as the removal of 1, 2, 3, 5, 10, 20, 30, 50 or more amino acids. Deletions can occur at any position in the protein. Amino acid deletion variants that comprise deletion at the N-terminal and / or C-terminal end of the protein are also called N-terminal and / or C-terminal truncation variants.

[0070] Amino acid substitution variants are distinguished by the removal of at least one residue in the sequence and the insertion of another residue in its place. The substitution of one amino acid for another can be classified as a conservative or non-conservative substitution. Preference is given to modifications in positions in the amino acid sequence that are not conserved between homologous proteins or peptides and / or to the substitution of amino acids for others with similar properties. Preferably, amino acid alterations in peptide and protein variants are conservative amino acid alterations, i.e., amino acid substitutions with or without similar charge. A conservative amino acid alteration involves the substitution of an amino acid from a family of amino acids that are related in their side chains.In the context of this description, a "conservative substitution" is the replacement of one amino acid with another amino acid that has similar structural and / or functional characteristics. Petition 870250088654, dated 09 / 30 / 2025, page 59 / 329 / 254 similar chemical reactions, such as the substitution of one amino acid residue for another amino acid residue of the same class, in the manner defined in either of the two previous tables: for example, leucine can be substituted for isoleucine, since both are aliphatic and hydrophobic branched. Similarly, aspartic acid can be substituted for glutamic acid, since both are small residues with a negative charge. Naturally occurring amino acids can also be broadly divided into four families: acidic (aspartate, glutamate), basic (lysine, arginine, histidine), nonpolar (alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), and uncharged polar (glycine, asparagine, glutamine, cysteine, serine, threonine, tyrosine). Phenylalanine, tryptophan, and tyrosine are sometimes collectively classified as aromatic amino acids.In one embodiment, conservative amino acid substitutions include substitutions in the following groups: - glycine, alanine; - valine, isoleucine, leucine; - aspartic acid, glutamic acid; - asparagine, glutamine; - serine, threonine; - lysine, arginine; and - phenylalanine, tyrosine.

[0071] The term “amino acid corresponding to position...” and similar expressions, as used herein, refer to an amino acid position number in a human IgG1 heavy chain. The corresponding amino acid positions in other immunoglobulins can be found by alignment with human IgG1. Thus, an amino acid or segment in one sequence that “corresponds to” an amino acid or segment in another sequence is one that aligns with the other amino acid or segment using an alignment program. Petition 870250088654, dated 09 / 30 / 2025, page 60 / 329 / 254 standard sequence, such as ALIGN, ClustalW or similar, typically in standard configurations and exhibiting at least 50%, at least 80%, at least 90%, or at least 95% identity with a human IgG1 heavy chain. It is considered well known in the art to align a sequence or segment within a sequence and thus determine the corresponding position in a sequence to an amino acid position, according to the present description.

[0072] The term “antibody” (Ab) in the context of this description refers to an immunoglobulin molecule, a fragment of an immunoglobulin molecule, or a derivative of any of them, which has the ability to bind specifically to an antigen (in particular an epitope on an antigen) under typical physiological conditions, preferably with a half-life of significant time periods, such as at least about 30 minutes, at least about 45 minutes, at least about one hour, at least about two hours, at least about four hours, at least about 8 hours, at least about 12 hours, about 24 hours or more, about 48 hours or more, about 3, 4, 5, 6, 7 or more days, etc., or any other functionally relevant defined period (such as sufficient time to induce, promote, intensify and / or modulate a physiological response associated with antibody binding to the antigen, and / or sufficient time for the antibody to recruit effector activity). In particular, the term “antibody” refers to a glycoprotein comprising at least two heavy chains (H) and two light chains (L) interconnected by disulfide bonds. The term “antibody” includes monoclonal antibodies, recombinant antibodies, human antibodies, humanized antibodies, chimeric antibodies and combinations of any of the foregoing. Each heavy chain comprises a variable heavy chain region (VH) and a constant heavy chain region (CH). Each light chain comprises a variable light chain region (VL) and a constant heavy chain region (CH). Petition 870250088654, dated 09 / 30 / 2025, page 61 / 329 / 254 constant light chain (LC) region. The variable regions and constant regions are also referred to here as variable domains and constant domains, respectively. The VH and VL regions can be further subdivided into hypervariability regions, called complementarity-determining regions (CDRs), interspersed with more conserved regions, called framework regions (FRs). Each VH and VL is composed of three CDRs and four FRs, arranged from amino-terminal to carboxy-terminal in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable chains (CDRs) of a hepatitis V virus (HV) are designated HCD1, HCD2, and HCD3 (or CDR-H1, HCD-H2, and HCD-H3), while the variable chains of a leishmaniasis (LV) are designated LCDR1, LCDR2, and LCDR3 (or CDR-L1, LCD-L2, and LCD-L3). The variable regions of the heavy and light chains contain a binding domain that interacts with an antigen.The constant regions of an antibody comprise the heavy chain constant region (CH) and the light chain constant region (CL), where CH can be further subdivided into the CH1 constant domain, a hinge region, and the CH2 and CH3 constant domains (arranged from amino-terminal to carboxy-terminal in the following order: CH1, CH2, CH3). The constant regions of antibodies can mediate the binding of immunoglobulin to host tissues or factors, including various cells of the immune system (e.g., effector cells) and components of the complement system, such as C1q. Antibodies can be intact immunoglobulins derived from natural or recombinant sources, and can be immunoactive portions of intact immunoglobulins. Antibodies are typically tetramers of immunoglobulin molecules.Antibodies can exist in a variety of forms including, for example, polyclonal antibodies, monoclonal antibodies, Fv, Fab and F(ab)2, as well as single-chain antibodies and humanized antibodies.

[0073] The variable regions of the heavy and light chains of the immunoglobulin molecule contain a binding domain that interacts with a Petition 870250088654, dated 09 / 30 / 2025, page 62 / 329 / 254 antigen. The terms “binding region” and “antigen-binding region” are used interchangeably herein and refer to the region that interacts with the antigen and comprises both a VH region and a VL region. An antibody as used herein comprises not only monospecific antibodies but also multispecific antibodies comprising multiple, such as two or more, for example, three or more, different antigen-binding regions.

[0074] As indicated above, the term antibody here, unless otherwise indicated or clearly contradicted by the context, includes fragments of an antibody that are antigen-binding fragments, that is, retain the ability to bind specifically to the antigen. It has been shown that the antigen-binding function of an antibody can be performed by fragments of a full-length antibody.Examples of antigen-binding fragments included in the term “antibody” include (i) a Fab' or Fab fragment, a monovalent fragment consisting of the VL, VH, CL and CH1 domains, or a monovalent antibody as described in WO 2007 / 059782 (Genmab); (ii) F(ab')2 fragments, bivalent fragments comprising two Fab fragments linked by a disulfide bridge in the hinge region; (iii) an Fd fragment consisting essentially of the VH and CH1 domains; (iv) an Fv fragment consisting essentially of the VL and VH domains of a single arm of an antibody; (v) a dAb fragment (Ward et al., Nature 341, 544-546 (1989)), consisting essentially of a VH domain and also termed antibody domains (Holt et al; Trends Biotechnol. 2003 Nov;21(11):484-90); (vi) camelid molecules or Nanobody (Revets et al; Expert Opin Biol Ther. 2005 Jan;5(1):111-24); and (vii) an isolated complementarity determination region (CDR).Furthermore, although the two domains of the Fv fragment, VL and VH, are encoded by separate genes, these can be joined, using recombinant methods, by a synthetic linker that... Petition 870250088654, dated 09 / 30 / 2025, pp. 63 / 329 / 254, allows them to be prepared as a single-chain protein in which the VL and VH regions pair to form monovalent molecules (known as single-chain antibodies or single-chain Fv (scFv), see, for example, Bird et al., Science 242, 423-426 (1988) and Huston et al., PNAS USA 85, 5879-5883 (1988)). Such single-chain antibodies are included in the term antibody unless otherwise noted or clearly indicated by the context. Although such fragments are generally included in the meaning of antibody, they are collectively and each independently unique features of the present description, exhibiting different biological properties and utility. These and other antibody fragments useful in the context of the present description, as well as the bispecific formats of such fragments, are discussed in more detail here.It should also be understood that the term antibody, unless otherwise specified, also includes polyclonal antibodies, monoclonal antibodies (mAbs), antibody-like polypeptides such as chimeric antibodies and humanized antibodies, and antibody fragments that retain the ability to bind specifically to the antigen (antigen-binding fragments) provided by any known technique, such as enzymatic cleavage, peptide synthesis, and recombinant techniques.

[0075] A generated antibody may present any isotype. As used herein, the term “isotype” refers to the class of immunoglobulins (e.g., IgG (such as IgG1, IgG2, IgG3, IgG4), IgD, IgA (such as IgA1, IgA2), IgE, IgM, or IgY) that is encoded by genes in the constant region of the heavy chain. When a specific isotype, for example, IgG1, is mentioned herein, the term is not limited to a specific sequence of the isotype, for example, a specific IgG1 sequence, but is used to indicate that the antibody is more closely sequenced to that isotype, for example, IgG1, than to other isotypes. Thus, for example, an IgG1 antibody described herein may be a sequence variant of an IgG1 antibody of Petition 870250088654, dated 09 / 30 / 2025, p. 64 / 329 / 254 natural occurrence, including variations in the constant regions.

[0076] IgG1 antibodies can exist in multiple polymorphic variants called allotypes (reviewed in Jefferis and Lefranc 2009. mAbs Vol. 1 issue 4 1-7), any of which is suitable for use in some of the embodiments described herein. Common allotypic variants in human populations are those designated by the letters a, f, n, z, or combinations thereof. In any of the embodiments described herein, the antibody may comprise a heavy chain Fc region comprising a human IgG Fc region. In additional embodiments, the human IgG Fc region comprises a human IgG1.

[0077] The term “multispecific antibody,” in the context of this description, refers to an antibody with at least two different antigen-binding regions, defined by different antibody sequences. In some embodiments, the different antigen-binding regions bind to different epitopes on the same antigen.However, in preferred embodiments, the different antigen-binding regions bind to different target antigens. In one embodiment, the multispecific antibody is a “bispecific antibody” or “bs”. A multispecific antibody, like a bispecific antibody, can have any shape, including any of the bispecific or multispecific antibody shapes described below.

[0078] The term “full length”, when used in the context of an antibody, indicates that the antibody is not a fragment, but contains all the specific isotype domains normally found for that isotype in nature, for example, the VH, CH1, CH2, CH3, hinge, VL and CL domains for an IgG1 antibody.

[0079] The term “human antibody,” as used herein, is intended to include antibodies with variable and framework regions derived from human germline immunoglobulin sequences. Petition 870250088654, dated 09 / 30 / 2025, page 65 / 329 / 254 and a constant domain of human immunoglobulin. The human antibodies described herein may include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations, insertions, or deletions introduced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo). However, the term "human antibody," as used herein, is not intended to include antibodies in which germline-derived CDR sequences from another non-human species, such as a mouse, have been grafted onto human structural sequences.

[0080] The term “chimeric antibody,” as used herein, refers to an antibody in which the variable region is derived from a non-human species (e.g., derived from rodents) and the constant region is derived from a different species, such as humans. Chimeric antibodies can be generated by antibody engineering. “Antibody engineering” is a term used generically for different types of antibody modifications, and the processes for antibody engineering are well known to the expert. In particular, a chimeric antibody can be generated using standard DNA techniques, as described in Sambrook et al., 1989, Molecular Cloning: A Laboratory Manual, New York: Cold Spring Harbor Laboratory Press, Ch. 15. Thus, the chimeric antibody can be a genetically or enzymatically modified recombinant antibody.It is known to those skilled in the art that a chimeric antibody can be generated, and therefore, the generation of the chimeric antibody can be performed by methods other than those described here. Chimeric monoclonal antibodies for therapeutic applications in humans are developed to reduce the anticipated immunogenicity of non-human antibodies, for example, rodent antibodies. These may typically contain non-human variable regions (e.g., murine or rabbit), which are specific for the antigen of interest, and constant heavy and light chain domains. Petition 870250088654, dated 09 / 30 / 2025, page 66 / 329 / 254 human antibodies. The terms “variable region” or “variable domain”, as used in the context of chimeric antibodies, refer to a region comprising the CDRs and the structural regions of the heavy and light chains of an immunoglobulin, as described below.

[0081] The term “humanized antibody,” as used herein, refers to a genetically modified non-human antibody containing constant domains of human antibodies and non-human variable domains modified to contain a high level of sequence homology with human variable domains. This can be achieved by grafting the six complementarity-determining regions (CDRs) of non-human antibodies, which together form the antigen-binding site, onto a homologous human acceptor structural region (FR) (see WO 92 / 22653 and EP 0 629 240). To fully reconstitute the binding affinity and specificity of the parental antibody, it may be necessary to replace structural residues of the parental antibody (i.e., the non-human antibody) in the human structural regions (reverse mutations).Structural homology modeling can help identify amino acid residues in structural regions that are important for antibody binding properties. Thus, a humanized antibody may comprise non-human CDR sequences, primarily human structural regions, optionally comprising one or more reverse amino acid mutations in the non-human amino acid sequence, and fully human constant regions. Optionally, additional amino acid modifications, which are not necessarily reverse mutations, may be applied to obtain a humanized antibody with preferred characteristics, such as affinity and biochemical properties.

[0082] As used here, a protein that is “derived from” another protein, for example, a parent protein, means that one or more amino acid sequences of the protein are identical or similar to one or more of the parent protein's amino acids. Petition 870250088654, dated 09 / 30 / 2025, page 67 / 329 / 254 more amino acid sequences in the other protein or in the parental protein. For example, in an antibody, binding arm, antigen-binding region, constant region, or similar that is derived from another antibody, binding arm, antigen-binding region, or constant region or from a parental antibody, one or more amino acid sequences are identical or similar to those of the other antibody, binding arm, antigen-binding region, or constant region or of the parental antibody. Examples of one or more of these amino acid sequences include, but are not limited to, those of the VH and VL CDRs and / or one or more or all of the scaffold regions, VH, VL, CL, hinge, or CH regions.For example, a humanized antibody can be described here as “derived from” a non-human parental antibody, meaning that at least the VL and VH CDR sequences are identical or similar to the VH and VL CDR sequences of said non-human parental antibody. A chimeric antibody can be described here as being “derived from” a non-human parental antibody, meaning that typically the VH and VL sequences may be identical or similar to those of the non-human parental antibody. Another example is a binding arm or antigen-binding region that can be described here as being “derived from” a specific parental antibody, meaning that said binding arm or antigen-binding region typically comprises identical or similar VH and / or VL CDRs, or VH and / or VL sequences to the binding arm or antigen-binding region of said parental antibody.As described elsewhere in the same document, however, amino acid modifications, such as mutations, can be made to CDRs, constant regions, or other parts of the antibody, binding arm, antigen-binding region, or similar structures to introduce desired characteristics. When used in the context of one or more sequences derived from a primary protein or parental protein, a “similar” amino acid sequence preferably exhibits identity. Petition 870250088654, dated 09 / 30 / 2025, p. 68 / 329 / 254 sequence of at least about 50%, such as at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95% or at least about 97%, 98% or 99%.

[0083] Non-human antibodies can be generated in several different species, such as mice, rabbits, chickens, guinea pigs, llamas, and goats.

[0084] Monoclonal antibodies can be produced by a variety of techniques, including conventional monoclonal antibody methodology, for example, the standard somatic cell hybridization technique of Kohler and Milstein, Nature 256: 495 (1975). Other techniques for the production of monoclonal antibodies can be employed, for example, viral or oncogenic transformation of B lymphocytes or phage display techniques using antibody gene libraries, and such methods are well known to those skilled in the art.

[0085] Hybridoma production in these non-human species is a well-established procedure. Immunization protocols and techniques for isolating splenocytes from immunized non-human animals / species for fusion are known in the art. Fusion partners (e.g., murine myeloma cells) and fusion procedures are also known.

[0086] When used herein, unless contradicted by context, the term “Fab arm” or “arm” refers to a heavy-chain-light chain pair and is used interchangeably with “half molecules” herein.

[0087] The term “binding arm comprising an antigen-binding region” means an antibody molecule or fragment comprising an antigen-binding region. Thus, a binding arm may comprise, for example, the six CDR sequences VH and VL, the VH and VL sequences, a Fab or Fab' fragment, or a Fab arm.

[0088] When used in this document, unless contradicted by context, the term “Fc region” refers to an antibody region consisting of Petition 870250088654, dated 09 / 30 / 2025, page 69 / 329 / 254 in the two Fc sequences of the heavy chains of an immunoglobulin, wherein said Fc sequences comprise at least one hinge region, a CH2 domain, and a CH3 domain. In one embodiment, the term “Fc region,” as used herein, refers to a region comprising, in the direction from the N-terminal to the C-terminal end of the antibody, at least one hinge region, a CH2 region, and a CH3 region. An Fc region of the antibody may mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (such as effector cells) and components of the complement system.

[0089] In the context of the present description, the term “inducing Fc-mediated effector function to a lesser degree”, used in relation to an antibody, including a multispecific antibody, means that the antibody induces Fc-mediated effector functions, such function being, in particular, selected from the list of binding to the IgG Fc receptor (FcgammaR, FcyR), binding to C1q, ADCC or CDC, to a lesser degree compared with a human IgG1 antibody comprising (i) the same CDR sequences, in particular comprising the same first and second antigen-binding regions, as said antibody, and (ii) two heavy chains comprising the hinge regions, CH2 and CH3 of human IgG1.

[0090] Fc-mediated effector function can be measured by binding to FcyRs, binding to C1q, or induction of Fc-mediated crosslinking via FcyRs.

[0091] The term “hinge region,” as used herein, refers to the hinge region of an immunoglobulin heavy chain. Thus, for example, the hinge region of a human IgG1 antibody corresponds to amino acids 216–230, according to the EU numbering established in Kabat (Kabat, EA et al., Sequences of proteins of immunological interest. 5th Edition – U.S. Department of Health and Human Services, NIH publication number 91-3242, pp. 662, 680, 689 (1991)). However, the hinge region can also be any of the Petition 870250088654, dated 09 / 30 / 2025, p. 70 / 329 / 254 other subtypes described here.

[0092] The term “CH1 region” or “CH1 domain”, as used herein, refers to the CH1 region of an immunoglobulin heavy chain. Thus, for example, the CH1 region of a human IgG1 antibody corresponds to amino acids 118-215, according to the EU numbering established in Kabat (ibid). However, the CH1 region may also be any of the other subtypes described herein.

[0093] The term “CH2 region” or “CH2 domain”, as used herein, refers to the CH2 region of an immunoglobulin heavy chain. Thus, for example, the CH2 region of a human IgG1 antibody corresponds to amino acids 231-340, according to the EU numbering, as established in Kabat (ibid). However, the CH2 region can also be any of the other subtypes, as described herein.

[0094] The term “CH3 region” or “CH3 domain,” as used herein, refers to the CH3 region of an immunoglobulin heavy chain. Thus, for example, the CH3 region of a human IgG1 antibody corresponds to amino acids 341–447, according to the EU numbering, as established in Kabat (ibid). However, the CH3 region may also be any of the other subtypes, as described herein.

[0095] The term “monovalent antibody” means, in the context of the present description, that an antibody molecule is capable of binding to only one molecule of the antigen and, therefore, is not capable of cross-linking with the antigen.

[0096] A “CD137 antibody” or “anti-CD137 antibody” is an antibody, as described above, that binds specifically to the CD137 antigen.

[0097] A “CD137xPD-L1 antibody” or “antiCD137xPD-L1 antibody” is a bispecific antibody comprising two regions of Petition 870250088654, dated 09 / 30 / 2025, p. 71 / 329 / 254 different antigen binding, one of which binds specifically to the CD137 antigen and the other binds specifically to the PD-L1 antigen.

[0098] The term “biosimilar” (e.g., of an approved reference product / biological medicinal product), as used herein, refers to a biological product that is similar to the reference product based on data from (a) analytical studies demonstrating that the biological product is highly similar to the reference product despite minor differences in clinically inactive components; (b) animal studies (including toxicity assessment); and / or (c) one or more clinical studies (including immunogenicity and pharmacokinetic or pharmacodynamic assessment) that are sufficient to demonstrate safety, purity and potency under one or more appropriate conditions of use for which the reference product is approved and intended to be used and for which approval is sought (e.g., that there are no clinically significant differences between the biological product and the reference product in terms of product safety, purity and potency).In some embodiments, the biosimilar biological product and the reference product utilize the same mechanism or mechanisms of action for the prescribed, recommended, or suggested condition or conditions of use in the proposed labeling, but only to the extent that the mechanism or mechanisms of action are known for the reference product. In some embodiments, the prescribed, recommended, or suggested condition or conditions of use in the proposed labeling for the biological product have been previously approved for the reference product. In some embodiments, the route of administration, pharmaceutical form, and / or concentration of the biological product are the same as those of the reference product. A biosimilar may be, for example, a currently known antibody with the same primary amino acid sequence as a marketed antibody, but may be produced in different cell types or by different production, purification, or formulation methods. Petition 870250088654, dated 09 / 30 / 2025, p. 72 / 329 / 254

[0099] As used herein, the terms “binding” or “capable of binding” in the context of antibody binding to a predetermined antigen or epitope typically refer to binding with an affinity corresponding to a Kd of about 10-7 M or less, such as about 10-8 M or less, such as about 10-9 M or less, about 10-10 M or less, or about 10-11 M or even less, when determined using Biolayer Interferometry (BLI) or, for example, when determined using surface plasmon resonance (SPR) technology on a BIAcore 3000 instrument using the antigen as the ligand and the antibody as the analyte. The antibody binds to the predetermined antigen with an affinity corresponding to a KD that is at least ten times smaller, such as at least 100 times smaller, for example, at least 1,000 times smaller, such as at least 10,000 times smaller, for example, at least 100.000 times smaller than its KD for binding to a non-specific antigen (e.g., BSA, casein) other than the predetermined antigen or a closely related antigen. The degree to which the affinity is greater depends on the antibody's KD, so that when the antibody's KD is very low (i.e., the antibody is highly specific), the degree to which the affinity for the antigen is less than the affinity for a non-specific antigen can be at least 10,000 times.

[00100] The term “kd” (sec-1), as used here, refers to the dissociation rate constant of a specific antibody-antigen interaction. This value is also called the koff value.

[00101] The term “KD” (M), as used herein, refers to the dissociation equilibrium constant of a specific antibody-antigen interaction.

[00102] Two antibodies exhibit the “same specificity” if they bind to the same antigen and the same epitope. If an antibody being tested recognizes the same epitope as a given binding antibody to Petition 870250088654, dated 09 / 30 / 2025, p. 73 / 329 / 254 antigen, that is, antibodies that bind to the same epitope, can be tested by different methods well known to those skilled in the art.

[00103] Competition between antibodies can be detected by a cross-blocking assay. For example, a competitive ELISA assay can be used as a cross-blocking assay. For example, the target antigen can be coated into the wells of a microtiter plate, and the antigen-binding antibody and the candidate competing test antibody can be added. The amount of antigen-binding antibody bound to the well correlates indirectly with the binding capacity of the candidate competing test antibody that competes with it for binding to the same epitope. Specifically, the higher the affinity of the candidate competing test antibody for the same epitope, the smaller the amount of antigen-binding antibody bound to the antigen-coated well. The amount of antigen-binding antibody bound to the well can be measured by labeling the antibody with detectable or measurable labeling substances.

[00104] An antibody that competes for binding to an antigen with another antibody, for example, an antibody comprising variable heavy and light chain regions, as described herein, or an antibody that has specificity for an antigen of another antibody, for example, an antibody comprising variable heavy and light chain regions, as described herein, may be an antibody comprising variants of said variable heavy and / or light chain regions, as described herein, for example, modifications in the CDRs and / or a certain degree of identity, as described herein.

[00105] A “single multispecific antibody”, as used herein, refers to a multispecific antibody that is substantially free of other antibodies with different antigenic specificities (e.g., a single bispecific antibody that binds specifically to CD137 and PD). Petition 870250088654, dated 09 / 30 / 2025, page 74 / 329 / 254 L1 is substantially free of monospecific antibodies that bind specifically to CD137 or PD-L1.

[00106] The term “monoclonal antibody,” as used herein, refers to a preparation of antibody molecules of unique molecular composition. A monoclonal antibody composition exhibits unique specificity and binding affinity for a specific epitope.

[00107] When used herein, the term “heterodimeric interaction between the first and second CH3 regions” refers to the interaction between the first CH3 region and the second CH3 region in a first CH3 / second CH3 heterodimeric antibody.

[00108] When used herein, the term “homodimeric interactions of the first and second CH3 regions” refers to the interaction between a first CH3 region and another first CH3 region in a first CH3 / first CH3 homodimeric antibody and to the interaction between a second CH3 region and another second CH3 region in a second CH3 / second CH3 homodimeric antibody.

[00109] When used herein, the term “homodimeric antibody” refers to an antibody comprising two initial Fab arms or half-molecules, in which the amino acid sequence of said Fab arms or half-molecules is the same.

[00110] When used herein, the term “heterodimeric antibody” refers to an antibody comprising a first and a second Fab arm or half-molecule, wherein the amino acid sequence of said first and second Fab arms or half-molecules is different. In particular, the CH3 region, or the antigen-binding region, or the CH3 region and the antigen-binding region of said first and second Fab arms / half-molecules are different.

[00111] The term “reducing conditions” or “reducing environment” refers to a condition or environment in which a substrate, such as a residue of Petition 870250088654, dated 09 / 30 / 2025, page 75 / 329 / 254 cysteine ​​in the hinge region of an antibody is more likely to be reduced than oxidized.

[00112] The present description also describes multispecific antibodies, such as bispecific antibodies, comprising functional variants of the VL regions, VH regions, or one or more CDRs of the bispecific antibodies in the examples. A functional variant of a VL, VH, or CDR used in the context of a bispecific antibody still allows each antigen-binding region of the bispecific antibody to retain at least a substantial proportion (at least about 50%, 60%, 70%, 80%, 90%, 95% or more) of the affinity and / or specificity / selectivity of the original bispecific antibody and, in some cases, such a bispecific antibody may be associated with higher affinity, selectivity, and / or specificity than the original bispecific antibody.

[00113] Such functional variants typically retain significant sequence identity with the original bispecific antibody. The percent identity between two sequences is a function of the number of identical positions shared by the sequences (i.e., % homology = # of identical positions / # total of positions x 100), taking into account the number of gaps and the length of each gap, which need to be introduced for optimal alignment of the two sequences. The percent identity between two nucleotide or amino acid sequences can, for example, be determined using the algorithm of E. Meyers and W. Miller, Comput. Appl. Biosci 4, 11-17 (1988), which was incorporated into the ALIGN program (version 2.0), using a PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4.Furthermore, the percent identity between two amino acid sequences can be determined using the Needleman and Wunsch algorithm, J. Mol. Biol. 48, 444-453 (1970).

[00114] In the context of the present description, unless otherwise stated Petition 870250088654, dated 09 / 30 / 2025, page 76 / 329 / 254. As indicated, the following notations are used to describe a mutation: i) the substitution of an amino acid at a given position is written as, for example, K409R, which means a substitution of a lysine at position 409 of the protein by an arginine; and ii) for specific variants, specific three-letter or one-letter codes are used, including the codes Xaa and X to indicate any amino acid residue. Thus, the substitution of lysine by arginine at position 409 is designated as: K409R, and the substitution of lysine by any amino acid residue at position 409 is designated as K409X. In the case of a lysine deletion at position 409, it is indicated by K409*.

[00115] Exemplary variants include those that differ from the VH and / or VL and / or CDR sequences of the parental sequences primarily by conservative substitutions; for example, 12, such as 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1, of the substitutions in the variant are conservative amino acid residue substitutions.

[00116] In the context of the present description, conservative substitutions can be defined as substitutions in the amino acid classes, in the manner defined in tables 2 and 3.

[00117] The term “CD137”, as used herein, refers to CD137 (4-1BB), also known as member 9 of the tumor necrosis factor receptor superfamily (TNFRSF9), which is the receptor for the TNFSF9 / 4-1BBL ligand. CD137 (4-1BB) is believed to be involved in T cell activation. Other synonyms for CD137 include, but are not limited to, receptor ligand 4-1BB, CD137, 4-1BB T cell antigen homolog, and T cell antigen ILA. In one embodiment, CD137 (4-1BB) is human CD137 (4-1BB), with UniProt accession number Q07011. The sequence of human CD137 is also shown in SEQ ID NO: 37. Amino acids 1 to 23 of SEQ ID NO: 37 correspond to the human CD137 signal peptide; while amino acids 24 to 186 of SEQ ID NO: 37 Petition 870250088654, dated 09 / 30 / 2025, page 77 / 329 / 254 correspond to the extracellular domain of human CD137; and the remainder of the protein, i.e., amino acids 187 to 213 and 214 to 255 of SEQ ID NO: 37, are transmembrane and cytoplasmic domains, respectively.

[00118] The “Programmed Death-1 (PD-1)” receptor refers to an immunoinhibitory receptor belonging to the CD28 family. PD-1 (also known as CD279 or SLEB2) is predominantly expressed on previously activated T cells in vivo and binds to two ligands, PD-L1 (also known as B7-H1 or CD274) and PD-L2 (also known as B7-DC or CD273). The term “PD-1”, as used herein, includes human PD1 (hPD-1), variants, isoforms and species homologs of hPD-1, and analogs that have at least one epitope in common with hPD-1, in particular a protein that has the amino acid sequence (NCBI Reference Sequence: NP_005009.2) as set forth in SEQ ID NO: 113 of the sequence listing, or a protein preferably encoded by a nucleic acid sequence (NCBI Reference Sequence: NM_005018.2) as set forth in SEQ ID NO: 115.“Programmed Death Ligand-1 (PD-L1)” is one of two cell surface glycoprotein ligands for PD-1 (the other being PD-L2) that negatively regulates T cell activation and cytokine secretion by binding to PD-1.

[00119] The term “PD-L1”, as used herein, includes human PD-L1 (hPD-L1), variants, isoforms and species homologs of hPD-L1, such as monkey PD-L1 (cynomolgus macaque), African elephant, wild boar and mouse (see, for example, Genbank accession numbers NP_054862.1, XP_005581836, XP_003413533, XP_005665023 and NP_068693, respectively), and analogs that possess at least one common epitope with hPD-L1. The human PD-L1 sequence is also shown in SEQ ID NO: 40 (mature sequence) and in SEQ ID NO: 39, where amino acids 1 to 18 are predicted to be a signal peptide. The term “PDL2”, as used herein, includes human PD-L2 (hPD-L2), variants, Petition 870250088654, dated 09 / 30 / 2025, page 78 / 329 / 254 isoforms and homologs of hPD-L2 species, and analogs that possess at least one common epitope with hPD-L2. PD-1 ligands (PD-L1 and PD-L2) are expressed on the surface of antigen-presenting cells, such as dendritic cells or macrophages, and other immune cells. The binding of PD-1 to PD-L1 or PD-L2 results in the downregulation of T cell activation. Cancer cells expressing PD-L1 and / or PD-L2 are able to deactivate T cells expressing PD-1, resulting in the suppression of the anti-cancer immune response. The interaction between PD-1 and its ligands results in a decrease in tumor-infiltrating lymphocytes, a decrease in T cell receptor-mediated proliferation, and immune evasion by cancer cells. Immunosuppression can be reversed by inhibiting the local interaction of PD-1 with PD-L1, and the effect is additive when the interaction of PD-1 with PD-L2 is also blocked.

[00120] The term “dysfunctional,” as used herein, refers to an immune cell that is in a state of reduced immunological responsiveness to antigenic stimulation. Dysfunctional includes absence of response to antigen recognition and impaired ability to translate antigen recognition into downstream T cell effector functions, such as proliferation, cytokine production (e.g., IL-2), and / or target cell killing.

[00121] The term “anergy,” as used herein, refers to the state of non-response to antigenic stimulation resulting from incomplete or insufficient signals transmitted by the T cell receptor (TCR). T cell anergy can also occur after antigen stimulation in the absence of co-stimulation, resulting in the cell becoming refractory to subsequent activation by the antigen, even in the context of co-stimulation. The non-responsive state can often be overridden by the presence of IL-2. Anergic T cells do not undergo clonal expansion and / or acquire effector functions. Petition 870250088654, dated 09 / 30 / 2025, page 79 / 329 / 254

[00122] The term “exhaustion,” as used herein, refers to the exhaustion of immune cells, such as T cell exhaustion, as a state of T cell dysfunction arising from sustained TCR signaling, which occurs during many chronic infections and cancer. It is distinguished from anergy in that it does not arise from incomplete or deficient signaling, but from sustained signaling. Exhaustion is defined by deficient effector function, sustained expression of inhibitory receptors, and a transcriptional state distinct from that of functional or memory effector T cells. Exhaustion prevents optimal control of diseases (e.g., infections and tumors). Exhaustion can result from both extrinsic negative regulatory pathways (e.g., immunoregulatory cytokines) and intrinsic negative regulatory pathways of the cell (immune checkpoint inhibitory pathways, such as those described herein).

[00123] “Enhancing T cell function” means inducing, causing, or stimulating a T cell to exhibit sustained or amplified biological function, or renewing or reactivating depleted or inactive T cells. Examples of enhancing T cell function include increased interferon-γ secretion by CD8+ T cells, increased proliferation, and increased response to antigens (e.g., tumor clearance) relative to pre-intervention levels. In a given modality, the intensification level is at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 110%, 120%, 130%, 140%, 150%, 200% or more. The methods for measuring this intensification are known to those skilled in the art.

[00124] The term “inhibitory nucleic acid” or “inhibitory nucleic acid molecule,” as used herein, refers to a nucleic acid molecule, for example, DNA or RNA, that reduces, inhibits, interferes with, or negatively modulates, wholly or partially, one or more PD-1 proteins. Inhibitory nucleic acid molecules include, without limitation, oligonucleotides, Petition 870250088654, dated 09 / 30 / 2025, p. 80 / 329 / 254 siRNA, shRNA, antisense DNA or RNA molecules and aptamers (e.g., DNA or RNA aptamers).

[00125] The term “oligonucleotide,” as used herein, refers to a nucleic acid molecule capable of decreasing protein expression, in particular the expression of a PD-1 protein, such as the PD-1 proteins described herein. Oligonucleotides are short molecules of DNA or RNA, typically comprising 2 to 50 nucleotides. Oligonucleotides can be single-stranded or double-stranded. A PD-1 inhibitory oligonucleotide can be an antisense oligonucleotide.

[00126] Antisense oligonucleotides are single-stranded DNA or RNA molecules that are complementary to a specific sequence, in particular to a nucleic acid sequence (or a fragment thereof) of a PD-1 protein. Antisense RNA is typically used to prevent protein translation of mRNA, for example, mRNA encoding a PD-1 protein, by binding to that mRNA. Antisense DNA is typically used to target a specific complementary RNA (coding or non-coding). If binding occurs, this DNA / RNA hybrid can be degraded by the enzyme RNase H. Furthermore, morpholino antisense oligonucleotides can be used for gene knockdowns in vertebrates. For example, Kryczek et al., 2006 (J Exp Med, 203:871-81) developed B7-H4-specific morpholinos that specifically blocked B7H4 expression in macrophages, resulting in increased T cell proliferation and reduced tumor volume in mice with tumor-associated antigen (TAA)-specific T cells.

[00127] The terms “siRNA” or “small interfering RNA” or “small inhibitory RNA” are used interchangeably here and refer to a double-stranded RNA molecule with a typical length of 20 to 25 base pairs that interferes with the expression of a specific gene, such as a gene encoding a PD-1 protein, with a nucleotide sequence Petition 870250088654, dated 09 / 30 / 2025, page 81 / 329 / 254 supplementary. In one embodiment, siRNA interferes with mRNA, thus blocking translation, for example, the translation of a PD-1 protein. Transfection with exogenous siRNA can be used for gene knockdown; however, the effect may be only transient, especially in rapidly dividing cells. Stable transfection can be achieved, for example, by RNA modification or by using an expression vector. Useful modifications and vectors for stable cell transfection with siRNA are known in the art. siRNA sequences can also be modified to introduce a short loop between the two strands, resulting in a “small hairpin RNA” or “shRNA”. The shRNA can be processed into a functional siRNA by Dicer. The shRNA exhibits a relatively low rate of degradation and turnover. Consequently, the PD-1 inhibitor can be an shRNA.

[00128] The term “aptamer,” as used herein, refers to a single-stranded nucleic acid molecule, such as DNA or RNA, typically 25 to 70 nucleotides long, capable of binding to a target molecule, such as a polypeptide. In one embodiment, the aptamer binds to an immune PD-1 protein, such as the PD-1 checkpoint proteins described herein. For example, an aptamer, as described, may specifically bind to a PD-1 protein or polypeptide, or to a molecule in a signaling pathway that modulates the expression of a PD-1 protein or polypeptide. The generation and therapeutic use of aptamers are well known in the art (see, for example, US 5,475,096).

[00129] The terms “small molecule inhibitor” or “small molecule” are used interchangeably here and refer to a low molecular weight organic compound, generally up to 1,000 daltons, that reduces, inhibits, interferes with, or negatively modulates, totally or partially, one or more PD-1 proteins, in the manner described above. These small molecule inhibitors are generally synthesized by organic chemistry, but Petition 870250088654, dated 09 / 30 / 2025, page 82 / 329 / 254 can also be isolated from natural sources, such as plants, fungi, and microbes. The low molecular weight allows a small molecule inhibitor to diffuse rapidly through cell membranes. For example, several A2AR antagonists known in the art are organic compounds with a molecular weight of less than 500 daltons.

[00130] The term “cell-based therapy” refers to the transplantation of cells (e.g., T lymphocytes, dendritic cells, or stem cells) that express an immune inhibitor of PD-1 into an individual with the aim of treating a disease or disorder (e.g., cancer).

[00131] The term “oncolytic virus,” as used herein, refers to a virus capable of selectively replicating and retarding the growth or inducing the death of a cancerous or hyperproliferative cell, in vitro or in vivo, without any or minimal effect on normal cells. An oncolytic virus for the administration of a PD-1 inhibitor comprises an expression cassette that may encode a PD-1 inhibitor that is an inhibitory nucleic acid molecule, such as an siRNA, shRNA, an oligonucleotide, antisense DNA or RNA, an aptamer, an antibody or a fragment thereof, or a soluble PD-1 protein or fusion. The oncolytic virus is preferably replication-competent and the expression cassette is under the control of a viral promoter, for example, a synthetic early / late poxvirus promoter.Exemplary oncolytic viruses include vesicular stomatitis virus (VSV), rhabdoviruses (e.g., picornaviruses such as Seneca Valley virus; SVV-001), coxsackievirus, parvovirus, Newcastle disease virus (NDV), herpes simplex virus (HSV; OncoVEX GMCSF), retroviruses (e.g., influenza virus), measles virus, reovirus, Sinbis virus, vaccinia virus, as exemplarily described in WO 2017 / 209053 (including Copenhagen, Western Reserve, Wyeth strains) and adenoviruses (e.g., Delta-24, Delta-24-RGD, ICOVIR-5, ICOVIR-7, Onyx-015, ColoAd1, H101, AD5 / 3-D24-GMCSF). The generation of viruses. Petition 870250088654, dated 09 / 30 / 2025, page 83 / 329 / 254 recombinant oncolytic viruses comprising a soluble form of a PD-1 inhibitor and methods for their use are described in document WO 2018 / 022831, which is incorporated herein by reference in its entirety. Oncolytic viruses can be used as attenuated viruses.

[00132] “Treatment cycle” is defined here as the period of time in which the effects of separate doses of the binding agent are added due to the pharmacodynamics of the binding agent, or in other words, the period of time after the individual’s body has essentially eliminated the administered binding agent. Multiple small doses in a short time window, for example, in 2 to 24 hours, such as 2 to 12 hours or on the same day, may be equivalent to a single larger dose.

[00133] In the present context, the term “treatment”, “treating” or “therapeutic intervention” refers to the management and care of an individual for the purpose of combating a condition, such as a disease or disorder. The term is intended to include the entire spectrum of treatments for a given condition from which the subject suffers, such as the administration of a therapeutically effective compound to alleviate symptoms or complications, slow the progression of the disease, disorder or condition, relieve or alleviate symptoms and complications and / or cure or eliminate the disease, disorder or condition, as well as prevent the condition, with prevention being understood as the management and care of an individual for the purpose of combating the disease, condition or disorder and includes the administration of active compounds to prevent the appearance of symptoms or complications.In one embodiment, "treatment" refers to the administration of an effective amount of a therapeutically active binding agent, such as a therapeutically active antibody, as described herein, for the purpose of relieving, improving, interrupting, or eradicating (curing) symptoms or disease states.

[00134] Response to treatment, as well as resistance, failure to Petition 870250088654, dated 09 / 30 / 2025, page 84 / 329 / 254. Response and / or relapse of treatment with a binding agent of the present description may be determined according to the Response Evaluation Criteria in Solid Tumors; version 1.1 (RECIST Criteria v1.1). The RECIST Criteria are presented in the following table (LD: longest dimension). Table 4: Response Definition (RECIST v1.1 Criteria) Category Criteria Based on Target Lesions Complete Response (CR) Disappearance of all target lesions. Any pathological lymph nodes must show a reduction in the short axis to <10 mm. Partial Response (PR) > 30% reduction in the sum of target lesion lymph nodes, taking the baseline sum of target lymph nodes as a reference. Stable Disease (SD) Neither sufficient reduction to qualify for PR, nor sufficient increase to qualify for PD, taking as a reference the lowest sum of target lymph nodes recorded since the beginning of treatment. Progressive Disease (PD) > 20% increase in the sum of target lesion lymph nodes, taking as a reference the lowest sum of target lymph nodes recorded since the beginning of treatment or the appearance of one or more new lesions. Based on Non-Target Lesions CR Disappearance of all non-target lesions and normalization of tumor marker levels. All lymph nodes must be non-pathological in size (< 10 mm in the short axis).SD: Persistence of one or more non-target lesions and / or maintenance of tumor marker levels above normal limits. PD: Appearance of one or more new lesions and / or unequivocal progression of existing non-target lesions.

[00135] The “best overall response” is the best recorded response from the start of treatment until disease progression / recurrence (the lowest recorded measures since the start of treatment will be used as a reference for PD). Individuals with CR or PR are considered to have an objective response. Individuals with CR, PR, or SD are considered to be in disease control. Individuals with NE are counted as non-responders. The best overall response is the best recorded response from the start of treatment until disease progression / recurrence (the lowest recorded measures since the start of treatment will be used as a reference for PD). Individuals with CR, PR, or SD are considered to be in disease control. Individuals with NE are counted as non-responders.

[00136] The “Duration of response (DOR)” applies only to individuals Petition 870250088654, dated 09 / 30 / 2025, page 85 / 329 / 254, whose best confirmed overall response is CR or PR and is defined as the time from the first documented objective tumor response (CR or PR) to the date of the first PD or death from the underlying cancer.

[00137] “Progression-free survival (PFS)” is defined as the number of days from Day 1 of Cycle 1 until the first documented progression or death from any cause.

[00138] “Overall survival (OS)” is defined as the number of days from Day 1 of Cycle 1 until death from any cause. If it is not known that an individual has died, the OS will be censored at the most recent date on which it was known that the individual was alive (at or before the cutoff date).

[00139] In the context of this description, the term “treatment regimen” refers to a structured treatment plan designed to improve and maintain health.

[00140] The term “effective amount” or “therapeutically effective amount” refers to an effective quantity, at the dosages and for the time periods required, to achieve a desired therapeutic result. A therapeutically effective amount of a binding agent, such as an antibody, whether a multispecific antibody or a monoclonal antibody, may vary according to factors such as the disease state, age, sex, and weight of the individual, and the ability of the binding agent to induce a desired response in the individual. A therapeutically effective amount is also one in which any toxic or harmful effects of the binding agent, or a fragment thereof, are offset by the therapeutically beneficial effects. If the response in a patient is insufficient with an initial dose, higher doses (or effectively larger doses obtained by a different and more localized route of administration) may be used.If undesirable side effects occur in a patient with a given dose, lower doses (or effectively lower doses obtained by a different, more localized route of administration) may be used. Petition 870250088654, dated 09 / 30 / 2025, page 86 / 329 / 254

[00141] As used herein, the term “cancer” includes a disease distinguished by aberrantly regulated cell growth, proliferation, differentiation, adhesion, and / or migration. A “cancer cell” is understood to be an abnormal cell that grows through rapid and uncontrolled cell proliferation and continues to grow after the stimuli that initiated the new growth have ceased.

[00142] The term “cancer,” as described herein, also includes cancerous metastases. By “metastasis,” we mean the spread of cancerous cells from their site of origin to another part of the body. Metastasis formation is a very complex process and depends on the detachment of malignant cells from the primary tumor, invasion of the extracellular matrix, penetration of endothelial basement membranes to enter the body cavity and vessels, and then, after transport by the blood, infiltration of target organs. Finally, the growth of a new tumor, that is, a secondary or metastatic tumor, at the target site depends on angiogenesis. Tumor metastasis frequently occurs even after removal of the primary tumor, as tumor cells or components may remain and develop metastatic potential.In one embodiment, the term "metastasis," as described herein, refers to "distant metastasis," which refers to metastasis distant from the primary tumor and the regional lymph node system.

[00143] Terms such as “reduce”, “inhibit”, “interfere” and “negatively modulate”, as used herein, mean the ability to cause an overall reduction, for example, of about 5% or more, about 10% or more, about 15% or more, about 20% or more, about 25% or more, about 30% or more, about 40% or more, about 50% or more, or about 75% or more, in the level. The term “inhibit” or similar phrases includes complete or essentially complete inhibition, that is, a reduction to zero or essentially to zero. Petition 870250088654, dated 09 / 30 / 2025, p. 87 / 329 / 254

[00144] Terms such as “increase” or “intensification”, in a modality, refer to an increase or intensification of at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 80% or at least about 100%.

[00145] The “physiological pH”, as used here, refers to a pH of 7.5 or around 7.5.

[00146] As used in this description, “% by weight” refers to the percentage by weight, which is a unit of concentration that measures the amount of a substance in grams (g), expressed as a percentage of the total weight of the composition in grams (g).

[00147] The term “freezing” refers to the solidification of a liquid, usually with the removal of heat.

[00148] The term “lyophilize” or “lyophilization” refers to the freeze-drying of a substance by means of freezing and then reducing the surrounding pressure (e.g., below 15 Pa, such as below 10 Pa, below 5 Pa, or 1 Pa or less) to allow the frozen medium in the substance to sublime directly from the solid phase to the gaseous phase. Thus, the terms “lyophilize” and “lyophilization” are used interchangeably here.

[00149] The term “recombinant,” in the context of this description, means “made by means of genetic engineering.” In one embodiment, a “recombinant object,” in the context of this description, does not occur naturally.

[00150] The term “natural occurrence,” as used herein, refers to the fact that an object can be found in nature. For example, a peptide or nucleic acid present in an organism (including viruses) that can be isolated from a source in nature and has not been intentionally modified by humans in a laboratory, occurs naturally. The term Petition 870250088654, dated 09 / 30 / 2025, page 88 / 329 / 254 “found in nature” means “present in nature” and includes known objects as well as objects that have not yet been discovered and / or isolated from nature, but which may be discovered and / or isolated in the future from a natural source.

[00151] According to the present description, the term “peptide” comprises oligo- and polypeptides and refers to substances comprising about two or more, about 3 or more, about 4 or more, about 6 or more, about 8 or more, about 10 or more, about 13 or more, about 16 or more, about 20 or more, and up to about 50, about 100, or about 150 consecutive amino acids linked together by peptide bonds. The term “protein” refers to large peptides, in particular peptides with at least about 151 amino acids, but the terms “peptide” and “protein” are used herein generally as synonyms.

[00152] A “therapeutic protein” has a positive or beneficial effect on an individual’s disease condition or state when provided to him or her in a therapeutically effective amount. In one embodiment, a therapeutic protein has curative or palliative properties and may be administered to improve, alleviate, attenuate, reverse, delay the onset of, or lessen the severity of one or more symptoms of a disease or disorder. A therapeutic protein may have prophylactic properties and may be used to delay the onset of a disease or to lessen the severity of such a disease or pathological condition. The term “therapeutic protein” includes whole proteins or peptides and may also refer to therapeutically active fragments thereof. It may also include therapeutically active variants of a protein. Examples of therapeutically active proteins include, but are not limited to, antigens for vaccination and immunostimulants such as cytokines.

[00153] The term “portion” refers to a fraction. With respect to a specific structure, such as an amino acid sequence or protein, the Petition 870250088654, dated 09 / 30 / 2025, p. 89 / 329 / 254 the term “portion” can designate a continuous or discontinuous fraction of that structure.

[00154] The terms “part” and “fragment” are used interchangeably here and refer to a continuous element. For example, a part of a structure, such as an amino acid sequence or a protein, refers to a continuous element of that structure. When used in the context of a composition, the term “part” means a portion of the composition. For example, a part of a composition can be any portion from 0.1% to 99.9% (such as 0.1%, 0.5%, 1%, 5%, 10%, 50%, 90% or 99%) of said composition.

[00155] “Fragment”, with reference to an amino acid sequence (peptide or protein), refers to a part of an amino acid sequence, that is, a sequence that represents the amino acid sequence shortened at the N-terminal and / or C-terminal end. A C-terminal fragment (N-terminal fragment) can be obtained, for example, by translating a truncated open reading frame that does not have the 3' end of the open reading frame. A C-terminal fragment (C-terminal fragment) can be obtained, for example, by translating a truncated open reading frame that does not have the 5' end of the open reading frame, provided that the truncated open reading frame contains a start codon that serves to initiate translation.A fragment of an amino acid sequence comprises, for example, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% of the amino acid residues of an amino acid sequence. A fragment of an amino acid sequence preferably comprises at least 6, in particular at least 8, at least 12, at least 15, at least 20, at least 30, at least 50, or at least 100 consecutive amino acids of an amino acid sequence.

[00156] According to the present description, a part or fragment Petition 870250088654, dated 09 / 30 / 2025, page 90 / 329 / 254 of a peptide or protein preferably possesses at least one functional property of the peptide or protein from which it was derived. Such functional properties include pharmacological activity, interaction with other peptides or proteins, enzymatic activity, interaction with antibodies, and selective binding of nucleic acids. For example, a pharmacologically active fragment of a peptide or protein possesses at least one of the pharmacological activities of the peptide or protein from which the fragment was derived. A part or fragment of a peptide or protein preferably comprises a sequence of at least 6, in particular, at least 8, at least 10, at least 12, at least 15, at least 20, at least 30, or at least 50 consecutive amino acids of the peptide or protein.A part or fragment of a peptide or protein preferably comprises a sequence of up to 8, in particular, up to 10, up to 12, up to 15, up to 20, up to 30 or up to 55 consecutive amino acids of the peptide or protein.

[00157] By “variant,” we mean here an amino acid sequence that differs from an original amino acid sequence by virtue of at least one amino acid modification. The original amino acid sequence may be a naturally occurring or wild-type (WT) amino acid sequence, or it may be a modified version of a wild-type amino acid sequence. Preferably, the variant amino acid sequence has at least one amino acid modification compared to the original amino acid sequence, for example, from 1 to about 20 amino acid modifications, and preferably from 1 to about 10 or from 1 to about 5 amino acid modifications compared to the original.

[00158] By “wild type” or “WT” or “natural” we mean here an amino acid sequence found in nature, including allelic variations. An amino acid sequence, peptide or protein of type Petition 870250088654, dated 09 / 30 / 2025, p. 91 / 329 / 254 wild type has an amino acid sequence that has not been intentionally modified.

[00159] Preferably, the degree of similarity, preferably of identity, between a given amino acid sequence and an amino acid sequence that is a variant of said amino acid sequence will be at least about 60%, 70%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%. The degree of similarity or identity is preferably given for an amino acid region that is at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or about 100% of the entire length of the reference amino acid sequence.For example, if the reference amino acid sequence consists of 200 amino acids, the degree of similarity or identity is preferably given for at least about 20, at least about 40, at least about 60, at least about 80, at least about 100, at least about 120, at least about 140, at least about 160, at least about 180, or about 200 amino acids, in some embodiments, continuous amino acids. In some embodiments, the degree of similarity or identity is given for the entire length of the reference amino acid sequence. The alignment to determine sequence similarity, preferably sequence identity, can be done with tools known in the art, preferably using best sequence alignment, for example, using Align, using default settings, preferably EMBOSS::needle, Matrix: Blosum62, Open spacing 10.0, Spacing extension 0.5.

[00160] “Sequence similarity” indicates the percentage of amino acids that are identical or that represent conservative substitutions. Petition 870250088654, dated 09 / 30 / 2025, p. 92 / 329 / 254 of amino acids. “Sequence identity” between two amino acid sequences indicates the percentage of amino acids that are identical between the sequences. “Sequence identity” between two nucleic acid sequences indicates the percentage of nucleotides that are identical between the sequences.

[00161] The terms “% identical” and “% identity” or similar terms refer, in particular, to the percentage of nucleotides or amino acids that are identical in an optimal alignment between the sequences to be compared. This percentage is purely statistical, and the differences between the two sequences may be, but not necessarily, randomly distributed throughout the length of the sequences to be compared. Comparisons of two sequences are generally performed by comparing them, after optimal alignment, with respect to a segment or “comparison window” in order to identify local regions of corresponding sequences. Optimal alignment for a comparison can be performed manually or with the aid of the local homology algorithm of Smith and Waterman, 1981, Ads App. Math. 2, 482, with the aid of the local homology algorithm of Needleman and Wunsch, 1970, J. Mol. Biol.48, 443, with the aid of the Pearson and Lipman similarity search algorithm, 1988, Proc. Natl Acad. Sci. USA 88, 2444, or with the aid of computer programs that use said algorithms (GAP, BESTFIT, FASTA, BLAST P, BLAST N and TFASTA in the Wisconsin Genetics Software package, Genetics Computer Group, 575 Science Drive, Madison, Wisconsin). In some embodiments, the percent identity of two sequences is determined using the BLASTN or BLASTP algorithm, available on the website of the United States National Center for Biotechnology Information (NCBI) (e.g., at blast.ncbi.nlm.nih.gov / Blast.cgi). In some embodiments, the algorithm parameters used for the BLASTN algorithm on the NCBI website include: (i) Expected Limit set to 10; (ii). Petition 870250088654, dated 09 / 30 / 2025, page 93 / 329 / 254 Word size set to 28; (iii) Maximum number of matches in a query range set to 0; (iv) Match / Mismatch Scores set to 1, -2; (v) Spacing Costs set to Linear; and (vi) filtering for low-complexity regions being used. In some embodiments, the algorithm parameters used for the BLASTP algorithm on the NCBI website include: (i) Expected Limit set to 10; (ii) Word Size set to 3; (iii) Maximum number of matches in a query range set to 0; (iv) Matrix set to BLOSUM62; (v) Spacing Costs set to Existence: 11 Extension: 1; and (vi) adjustment of the conditional compositional scoring matrix.

[00162] Percent identity is obtained by determining the number of identical positions to which the sequences to be compared correspond, dividing that number by the number of positions compared (for example, the number of positions in the reference sequence) and multiplying that result by 100.

[00163] In some embodiments, the degree of similarity or identity is given for a region that is at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or about 100% of the entire length of the reference sequence. For example, if the reference amino acid sequence consists of 200 amino acid residues, the degree of identity is given for at least about 100, at least about 120, at least about 140, at least about 160, at least about 180, or about 200 amino acid residues, in some embodiments, contiguous amino acid residues. In some embodiments, the degree of similarity or identity is given for the entire length of the reference sequence.

[00164] Homologous amino acid sequences show, according to the present description, at least 40%, in particular at least 50%, Petition 870250088654, dated 09 / 30 / 2025, page 94 / 329 / 254 at least 60%, at least 70%, at least 80%, at least 90% and, preferably, at least 95%, at least 98% or at least 99% identity of amino acid residues.

[00165] The amino acid sequence variants described herein can be readily prepared by a person skilled in the art, for example, by recombinant DNA manipulation. The manipulation of DNA sequences for the preparation of peptides or proteins with substitutions, additions, insertions, or deletions is described in detail in Sambrook et al. (1989), for example. Furthermore, the peptides and amino acid variants described herein can be readily prepared using known peptide synthesis techniques, such as solid-phase synthesis and similar methods.

[00166] In one embodiment, a fragment or variant of an amino acid sequence (peptide or protein) is preferably a “functional fragment” or “functional variant.” The term “functional fragment” or “functional variant” of an amino acid sequence refers to any fragment or variant that exhibits one or more functional properties identical or similar to those of the amino acid sequence from which it is derived, i.e., it is functionally equivalent. With respect to antigens or antigenic sequences, a specific function is one or more immunogenic activities exhibited by the amino acid sequence from which the fragment or variant is derived.The term “functional fragment” or “functional variant,” as used herein, refers in particular to a variant molecule or sequence comprising an amino acid sequence that is altered by one or more amino acids compared to the amino acid sequence of the original molecule or sequence and that is still capable of performing one or more functions of the original molecule or sequence, for example, inducing an immune response. In one embodiment, the modifications to the amino acid sequence of the original molecule or sequence do not significantly affect or alter the... Petition 870250088654, dated 09 / 30 / 2025, page 95 / 329 / 254 characteristics of the molecule or sequence. In different embodiments, the function of the functional fragment or functional variant may be reduced, but still significantly present; for example, the immunogenicity of the functional variant may be at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% of the original molecule or sequence. However, in other embodiments, the immunogenicity of the functional fragment or functional variant may be increased compared to the original molecule or sequence.

[00167] An amino acid sequence (peptide, protein, or polypeptide) “derived from” a designated amino acid sequence (peptide, protein, or polypeptide) refers to the origin of the first amino acid sequence. Preferably, the amino acid sequence derived from a specific amino acid sequence has an amino acid sequence identical, essentially identical, or homologous to that specific sequence or a fragment thereof. Amino acid sequences derived from a specific amino acid sequence may be variants of that specific sequence or a fragment thereof. For example, it will be understood by those skilled in the art that antigens suitable for use herein may be altered so that their sequence varies relative to the natural or native sequences from which they were derived, while maintaining the desired activity of the native sequences.

[00168] “Isolated” means altered or removed from its natural state. For example, a nucleic acid or peptide naturally present in a living animal is not “isolated,” but the same nucleic acid or peptide partially or completely separated from the coexisting materials in its natural state is “isolated.” An isolated nucleic acid or protein may exist in a substantially purified form or may exist in a non-natural environment, such as, for example, a host cell. In a preferred embodiment, the binding agent used in the present description is in a form Petition 870250088654, dated 09 / 30 / 2025, page 96 / 329 / 254 substantially purified.

[00169] The term “genetic modification” or simply “modification” includes the transfection of cells with nucleic acid. The term “transfection” refers to the introduction of nucleic acids, in particular RNA, into a cell. For the purposes of this description, the term “transfection” also includes the introduction of a nucleic acid into a cell or the uptake of a nucleic acid by such a cell, wherein the cell may be present in an individual, for example, a patient. Thus, according to this description, a cell for transfection of a nucleic acid described herein may be present in vitro or in vivo, for example, the cell may be part of an organ, tissue and / or organism of a patient. According to this description, transfection may be transient or stable. For some applications of transfection, it is sufficient that the transfected genetic material is expressed only transiently.RNA can be transfected into cells to transiently express its encoded protein. Because the nucleic acid introduced in the transfection process is generally not integrated into the nuclear genome, the foreign nucleic acid will be diluted by mitosis or degraded. Cells that allow episomal amplification of nucleic acids significantly reduce the dilution rate. If it is desired that the transfected nucleic acid remain in the genome of the cell and its daughter cells, a stable transfection must occur. Such a stable transfection can be achieved using virus-based systems or transposon-based systems for transfection. Generally, the nucleic acid encoding the antigen is transiently transfected into the cells. RNA can be transfected into cells to transiently express its encoded protein.

[00170] According to the present description, a peptide or protein analogue is a modified form of said peptide or protein from which it was derived and possesses at least one functional property of said peptide or protein. Petition 870250088654, dated 09 / 30 / 2025, page 97 / 329 / 254 peptide or protein. For example, a pharmacologically active analog of a peptide or protein possesses at least one of the pharmacological activities of the peptide or protein from which the analog was derived. Such modifications include any chemical modification and comprise single or multiple substitutions, deletions, and / or additions of any molecules associated with the protein or peptide, such as carbohydrates, lipids, and / or proteins or peptides. In one embodiment, “analogues” of proteins or peptides include those modified forms resulting from glycosylation, acetylation, phosphorylation, amidation, palmitoylation, myristoylation, isoprenylation, lipidation, alkylation, derivatization, introduction of protective / blocking groups, proteolytic cleavage, or binding to an antibody or other cellular ligand. The term "analogue" also extends to all functional chemical equivalents of said proteins and peptides.

[00171] “Activation” or “stimulation,” as used herein, refers to the state of an immune effector cell, such as a T cell, that has been sufficiently stimulated to induce detectable cell proliferation. Activation may also be associated with the initiation of signaling pathways, the induced production of cytokines, and detectable effector functions. The term “activated immune effector cells” refers, among other things, to immune effector cells that are in the process of cell division.

[00172] The term “priming” refers to a process in which an immune effector cell, such as a T cell, has its first contact with its specific antigen and causes differentiation into effector cells, such as effector T cells.

[00173] The term “clonal expansion” or “expansion” refers to a process in which a specific entity is multiplied. In the context of the present description, the term is preferably used in the context of an immune response in which immune effector cells are stimulated by an antigen, proliferate, and the specific immune effector cell that recognizes the Petition 870250088654, dated 09 / 30 / 2025, page 98 / 329 / 254 said antigen is amplified. Preferably, clonal expansion leads to the differentiation of immune effector cells.

[00174] An “antigen,” according to the present description, includes any substance that induces an immune response and / or any substance against which an immune response or an immune mechanism, such as a cellular response, is directed. This also includes situations where the antigen is processed into antigenic peptides and an immune response or immune mechanism is directed against one or more antigenic peptides, particularly if presented in the context of MHC molecules. In particular, an “antigen” refers to any substance, preferably a peptide or protein, that reacts specifically with antibodies or T lymphocytes (T cells). According to the present description, the term “antigen” includes any molecule containing at least one epitope, such as a T cell epitope.Preferably, an antigen, in the context of the present description, is a molecule that, optionally after processing, induces an immune reaction, which is preferably specific to the antigen (including cells expressing the antigen). In one embodiment, an antigen is a disease-associated antigen, such as a tumor antigen, a viral antigen, or a bacterial antigen, or an epitope derived from such an antigen.

[00175] The term “epitope” refers to an antigenic determinant in a molecule, such as an antigen, that is, a part or fragment of the molecule that is recognized by the immune system, for example, by T-cell or B-cell antibodies, particularly when presented in the context of MHC molecules. In one embodiment, “epitope” means a protein determinant capable of specifically binding to an antibody. Epitopes generally consist of surface clusters of molecules, such as amino acids or sugar side chains, and usually possess specific three-dimensional structural features, as well as Petition 870250088654, dated 09 / 30 / 2025, page 99 / 329 / 254 specific charge characteristics. Conformational and non-conformational epitopes are distinguished by the fact that binding to the former, but not the latter, is lost in the presence of denaturing solvents. The epitope may comprise amino acid residues directly involved in binding and other amino acid residues that are not directly involved in binding, such as amino acid residues that are effectively blocked or covered by the peptide that specifically binds to the antigen (in other words, the amino acid residue is in the coverage area of ​​the peptide that specifically binds to the antigen).

[00176] An epitope of a protein preferably comprises a continuous or discontinuous portion of said protein and preferably has between about 5 and about 100, preferably between about 5 and about 50, more preferably between about 8 and about 0, even more preferably between about 10 and about 25 amino acids in length. For example, the epitope may preferably have 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25 amino acids in length. It is particularly preferred that the epitope, in the context of the present description, be a T-cell epitope.

[00177] The term “optional” or “optionally,” as used herein, means that the event, circumstance, or condition subsequently described may or may not occur, and that the description includes instances where said event, circumstance, or condition occurs and instances where it does not occur.

[00178] In the manner used here, the terms “linked”, “fused” or “fusion” are used interchangeably. These terms refer to the union of two or more elements, components or domains.

[00179] The term “disease” (also referred to as “disorder” here) refers to an abnormal condition affecting an individual’s body. A disease is often interpreted as a medical condition associated with specific symptoms and signs. A disease can be caused by factors Petition 870250088654, dated 09 / 30 / 2025, page 100 / 329 / 254 originating from an external source, such as an infectious disease, or may be caused by internal dysfunctions, such as autoimmune diseases. In humans, "disease" is often used more broadly to refer to any condition that causes pain, dysfunction, suffering, social problems, or death to the affected individual, or similar problems for those in contact with the individual. In this broader sense, it sometimes includes injuries, disabilities, disorders, syndromes, infections, isolated symptoms, deviant behaviors, and atypical variations in structure and function, while in other contexts and for other purposes, these categories may be considered distinguishable. Diseases generally affect individuals not only physically but also emotionally, as contracting and living with many diseases can alter a person's outlook on life and personality.

[00180] The term “therapeutic treatment” refers to any treatment that improves the state of health and / or prolongs (increases) the life expectancy of an individual. Such treatment may eliminate the disease in an individual, stop or delay the development of a disease in an individual, inhibit or delay the development of a disease in an individual, decrease the frequency or severity of symptoms in an individual, and / or decrease the recurrence in an individual who currently has or has had a disease.

[00181] The terms “prophylactic treatment” or “preventive treatment” refer to any treatment aimed at preventing the occurrence of a disease in an individual. The terms “prophylactic treatment” or “preventive treatment” are used interchangeably here. Similarly, the term “method for prevention,” in the context of disease progression, such as the progression of a tumor or cancer, refers to any method aimed at preventing the progression of the disease in an individual.

[00182] The terms “individual” and “subject” are used here Petition 870250088654, dated 09 / 30 / 2025, page 101 / 329 / 254 indifferently. They refer to a human being or another mammal (for example, mouse, rat, rabbit, dog, cat, cattle, pig, sheep, horse or primate) or any other non-mammalian animal, including birds (chickens), fish or any other animal species that may be affected by or susceptible to a disease or disorder (for example, cancer). Unless otherwise indicated, the terms “individual” and “subject” do not denote a specific age and therefore include adults, the elderly, children and newborns. In embodiments of the present description, the “individual” or “subject” is a “patient”.

[00183] The term “patient” means an individual or subject for treatment, in particular an individual or subject who is ill.

[00184] The term “microsatellite instability (MSI)” refers to the form of genomic instability associated with defective DNA mismatch repair (MMR) in tumors. See Boland et al., Cancer Research 58, 5258-5257, 1998. Based on the extent of instability, MSI can be classified as microsatellite unstable-high (MSI-H), microsatellite unstable-low (MSI-L), and microsatellite stable (MSS). In one embodiment, MSI analysis can be performed using the five microsatellite markers recommended by the National Cancer Institute (NCI): BAT25 (GenBank accession number: 9834508), BAT26 (GenBank accession number: 9834505), D5S346 (GenBank accession number: 181171), D2S123 (GenBank accession number: 187953), and D17S250 (GenBank accession number: 177030). In another embodiment, MSI analysis can be performed using a panel of five poly-A mononucleotide repeat markers (BAT-25, BAT-26, NR-21, NR-24, NR-27).The microsatellite high instability (MSI-H) status can be determined if two or more of the five previously indicated NCI markers exhibit instability or > 30% of the total markers in other marker panels demonstrate instability (i.e., Petition 870250088654, dated 09 / 30 / 2025, page 102 / 329 / 254; exhibit insertion / deletion mutations); low microsatellite instability (MSI-L) status can be determined if one of the five previously indicated NCI markers exhibits instability or less than 30% of the total markers in other marker panels demonstrate instability (i.e., exhibit insertion / deletion mutations); and microsatellite stability (MSS) status can be determined if none of the five previously indicated NCI markers or other marker panels exhibit instability (i.e., exhibit insertion / deletion mutations). Commercially available tests to determine MMR status include, but are not limited to, the VENTANA MMR RxDx Panel.Proficient mismatch repair (pMMR) status refers to the normal expression status of MMR proteins (MLH1, PMS2, MSH2, and MSH6) in tumor specimens by IHC, and deficient mismatch repair (dMMR) status refers to the low or no expression status, such as loss of nuclear expression, of one or more MMR proteins (MLH1, PMS2, MSH2, and MSH6) in a tumor specimen by IHC. MSI-H status is generally consistent with dMMR status, while MSI-L and MSS statuses are generally consistent with pMMR status. Techniques for determining MSI or MMR status (such as MSIH, MSS, pMMR, dMMR) in tumors are within the knowledge of the specialist and are not limited to the modalities described here. Examples of such techniques are well known in the art, such as, but not limited to, those described in Gilson et al., Cancers (Basel), 2021 Mar 24;13(7):1491.Commercially available tests for MSI or MMR analysis include, but are not limited to, for example, the Promega® MSI multiplex PCR assay, FoundationOne® CDx (F1CDx), Guardant360® CDx, Idylla® MSI test, and the VENTANA MMR RxDx Panel. In some modalities, the VENTANA MMR RxDx Panel can be used to determine MSI or MMR status. In certain modalities, MSI or MMR status is determined by... Petition 870250088654, dated 09 / 30 / 2025, page 103 / 329 / 254 results of microsatellite mismatch repair (MMR) / MSI tests using immunohistochemistry (IHC), polymerase chain reaction (PCR) or next-generation sequencing (NGS) performed with a test approved by the Food and / or Drug Administration (FDA) / with Conformité Européenne (CE) marking. Many types of cancer or tumors have been found to be associated with microsatellite instability, such as colorectal cancer, gastric cancer, endometrial cancer, ovarian cancer, hepatobiliary tract cancer, pancreatic cancer, urinary tract cancer, bladder cancer, thyroid cancer, breast cancer, prostate cancer, ovarian cancer, central nervous system (CNS) cancer, and skin cancer, such as melanoma (Han et al., Front Genet. 2022 Dec 1;13:933475). Aspects and modalities of the present description

[00185] In a first aspect, the present description provides a method for treating a tumor or cancer in an individual, said method comprising administering to said individual a binding agent comprising a first binding region that binds to CD137 and a second binding region that binds to PD-L1, wherein said tumor or cancer exhibits high microsatellite instability (MSI-H) or deficiency in nonspecific binding repair (dMMR). Liaison agent that binds to CD137 and PD-L1

[00186] In one embodiment, CD137 is human CD137, in particular human CD137 comprising the sequence presented in SEQ ID NO: 38. In one embodiment, PD-L1 is human PD-L1, in particular human PD-L1 comprising the sequence presented in SEQ ID NO: 40. In one embodiment, CD137 is human CD137 and PD-L1 is human PD-L1. In one embodiment, CD137 is human CD137 comprising the sequence presented in SEQ ID NO: 38, and PD-L1 is human PD-L1 comprising the sequence presented in SEQ ID NO: 40.

[00187] In a liaison agent modality according to Petition 870250088654, dated 09 / 30 / 2025, page 104 / 329 / 254 first aspect, a) the first binding region that binds to human CD137 comprises a variable heavy chain (VH) region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NO: 1 or 9, and a variable light chain (VL) region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NO: 5 or 10; and b) the second antigen-binding region that binds to human PD-L1 comprises a variable heavy chain (VH) region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NO: 11, and a variable light chain (VL) region comprising the CDR1, CDR2 and CDR3 sequences of SEQ ID NO: 15.

[00188] In a liaison agent embodiment according to the first aspect, a) The first binding region that binds to human CD137 comprises a variable heavy chain (VH) region comprising the CDR1, CDR2, and CDR3 sequences presented in: SEQ ID NO: 2, 3, and 4, respectively, and a variable light chain (VL) region comprising the CDR1, CDR2, and CDR3 sequences presented in: SEQ ID NO: 6, 7, and 8, respectively; and b) The second antigen-binding region that binds to human PD-L1 comprises a variable heavy chain (VH) region comprising the CDR1, CDR2, and CDR3 sequences presented in: SEQ ID NO: 12, 13, and 14, respectively, and a variable light chain (VL) region comprising the CDR1, CDR2, and CDR3 sequences presented in: SEQ ID NO: 16, 17, and 18, respectively.

[00189] In a first-aspect-based embodiment of the binding agent, the first binding region that binds to human CD137 Petition 870250088654, dated 09 / 30 / 2025, page 105 / 329 / 254 comprises a variable heavy chain (VH) region comprising an amino acid sequence with at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with SEQ ID NO: 1 or 9 and a variable light chain (VL) region comprising an amino acid sequence with at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with SEQ ID NO: 5 or 10.

[00190] In a further embodiment of the binding agent according to the first aspect, the second binding region that binds to human PD-L1 comprises a variable heavy chain (VH) region comprising an amino acid sequence with at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with SEQ ID NO: 11 and a variable light chain (VL) region comprising an amino acid sequence with at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with SEQ ID NO: 15.

[00191] In a liaison agent embodiment according to the first aspect, (a) the first binding region that binds to human CD137 comprises a variable heavy chain (VH) region comprising an amino acid sequence with at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with SEQ ID NO: 1 or 9 and a variable light chain (VL) region comprising an amino acid sequence with at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with SEQ ID NO: 5 or 10; and b) the second binding region that binds to human PD-L1 comprises a variable heavy chain (VH) region comprising an amino acid sequence with at least 90%, at least 95%, by Petition 870250088654, dated 09 / 30 / 2025, page 106 / 329 / 254 less than 97%, at least 99%, or 100% sequence identity with SEQ ID NO: 11 and a variable light chain (VL) region comprising an amino acid sequence with at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with SEQ ID NO: 15.

[00192] In one embodiment of the binding agent according to the first aspect, the first binding region that binds to human CD137 comprises a variable heavy chain (VH) region comprising the amino acid sequence presented in SEQ ID NO: 1 or 9 and a variable light chain (VL) region comprising the amino acid sequence presented in SEQ ID NO: 5 or 10.

[00193] In a further embodiment of the binding agent according to the first aspect, the second binding region that binds to human PD-L1 comprises a variable heavy chain (VH) region comprising the amino acid sequence presented in SEQ ID NO: 11 and a variable light chain (VL) region comprising the amino acid sequence presented in SEQ ID NO: 15.

[00194] In a liaison agent embodiment according to the first aspect, a) the first binding region that binds to human CD137 comprises a variable heavy chain (VH) region comprising the amino acid sequence presented in SEQ ID NO: 1 or 9 and a variable light chain (VL) region comprising the amino acid sequence presented in SEQ ID NO: 5 or 10; and b) The second binding region that binds to human PD-L1 comprises a variable heavy chain (VH) region comprising the amino acid sequence presented in SEQ ID NO: 11 and a variable light chain (VL) region comprising the amino acid sequence Petition 870250088654, dated 09 / 30 / 2025, page 107 / 329 / 254 submitted in SEQ ID NO: 15.

[00195] In a liaison agent embodiment according to the first aspect, a) The first binding region that binds to human CD137 comprises a variable heavy chain (VH) region comprising the amino acid sequence presented in SEQ ID NO: 1 and a variable light chain (VL) region comprising the amino acid sequence presented in SEQ ID NO: 5; and b) the second binding region that binds to human PD-L1 comprises a variable heavy chain (VH) region comprising the amino acid sequence presented in SEQ ID NO: 11 and a variable light chain (VL) region comprising the amino acid sequence presented in SEQ ID NO: 15.

[00196] The binding agent may be in particular an antibody, such as a multispecific antibody, for example, a bispecific antibody. Likewise, the binding agent may be in the form of a full-length antibody or an antibody fragment.

[00197] It is additionally preferable that the binding agent be a human antibody or a humanized antibody.

[00198] Each variable region may comprise three complementarity determination regions (CDR1, CDR2, and CDR3) and four framework regions (FR1, FR2, FR3, and FR4).

[00199] Complementary determination regions (CDRs) and framework regions (FRs) can be arranged from amino-terminal to carboxy-terminal in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.

[00200] In one embodiment of the first aspect, the linking agent comprises: Petition 870250088654, dated 09 / 30 / 2025, page 108 / 329 / 254 i) a polypeptide comprising said first variable heavy chain region (VH) and a first constant heavy chain region (CH), and ii) a polypeptide comprising said second variable heavy chain region (VH) and a second constant heavy chain region (CH).

[00201] In one embodiment of the first aspect, the linking agent comprises: i) a polypeptide comprising said first variable light chain (VL) region and further comprising a first constant light chain (CL) region, and ii) a polypeptide comprising said second variable light chain (VL) region and further comprising a second constant light chain (CL) region.

[00202] In one embodiment of the first aspect, the binding agent is an antibody comprising a first binding arm and a second binding arm, wherein the first binding arm comprises: i) a polypeptide comprising said first variable heavy chain region (VH) and said first constant heavy chain region (CH), and ii) a polypeptide comprising said first variable light chain region (VL) and said first constant light chain region (CL); and the second connecting arm comprises: iii) a polypeptide comprising said second variable heavy chain region (VH) and said second constant heavy chain region (CH), and iv) a polypeptide comprising said second variable light chain region (VL) and said second constant light chain region. Petition 870250088654, dated 09 / 30 / 2025, page 109 / 329 / 254 (CL).

[00203] In one embodiment of the first aspect, the binding agent comprises i) a first heavy chain and light chain comprising said antigen-binding region capable of binding to CD137, the first heavy chain comprising a first constant region of heavy chain and the first light chain comprising a first constant region of light chain; and ii) a second heavy chain and light chain comprising said antigen-binding region capable of binding to PDL1, the second heavy chain comprising a second constant region of heavy chain and the second light chain comprising a second constant region of light chain.

[00204] Each of the first and second constant heavy chain (CH) regions may comprise one or more of a constant heavy chain 1 (CH1) region, a hinge region, a constant heavy chain 2 (CH2) region and a constant heavy chain 3 (CH3) region, preferably at least one hinge region, a CH2 region and a CH3 region.

[00205] Each of the first and second constant heavy chain (CH) regions may comprise a CH3 region, wherein the two CH3 regions comprise asymmetric mutations. Asymmetric mutations mean that the sequences of said first and second CH3 regions contain amino acid substitutions at non-identical positions. For example, one of said first and second CH3 regions contains a mutation at the position corresponding to position 405 in a human IgG1 heavy chain according to EU numbering, and the other of said first and second CH3 regions contains a mutation at the position corresponding to position 409 in a human IgG1 heavy chain according to EU numbering.

[00206] In the said first constant region of heavy chain (CH) by Petition 870250088654, dated 09 / 30 / 2025, p. 110 / 329 / 254 less one of the amino acids at a position corresponding to a position selected from the group consisting of T366, L368, K370, D399, F405, Y407, and K409 in a human IgG1 heavy chain according to EU numbering may have been substituted, and in said second constant region of heavy chain (CH) at least one of the amino acids at a position corresponding to a position selected from the group consisting of T366, L368, K370, D399, F405, Y407, and K409 in a human IgG1 heavy chain according to EU numbering may have been substituted. In particular forms, the first and second heavy chains are not substituted at the same positions (i.e., the first and second heavy chains contain asymmetric mutations).

[00207] In one embodiment of the linking agent according to the first aspect, (i) the amino acid at the position corresponding to F405 in a human IgG1 heavy chain according to EU numbering is L in said first heavy chain constant region (CH), and the amino acid at the position corresponding to K409 in a human IgG1 heavy chain according to EU numbering is R in said second heavy chain constant region (CH), or (ii) the amino acid at the position corresponding to K409 in a human IgG1 heavy chain according to EU numbering is R in said first heavy chain, and the amino acid at the position corresponding to F405 in a human IgG1 heavy chain according to EU numbering is L in said second heavy chain.

[00208] In one embodiment of the first aspect, the binding agent induces Fc-mediated effector function to a lesser extent compared to another antibody comprising the same first and second antigen-binding regions and two heavy chain constant (CH) regions comprising the hinge regions, CH2 and CH3 of human IgG1.

[00209] In a specific embodiment of the linking agent, according to the first aspect, the said first and second constant regions Petition 870250088654, dated 09 / 30 / 2025, p. 111 / 329 / 254 of the heavy chain (HCs) are modified so that the antibody induces Fc-mediated effector function to a lesser degree compared with an identical antibody, except for comprising unmodified first and second constant regions of the heavy chain (HCs). In particular, each or both of the so-called unmodified first and second constant regions of the heavy chain (HCs) may comprise, consist of, or consist essentially of the amino acid sequence shown in SEQ ID NO: 19 or 25.

[00210] Fc-mediated effector function can be determined by measuring the binding of the binding agent to Fcy receptors, the binding to C1q, or the Fc-mediated crosslinking induction of Fcy receptors. In particular, Fc-mediated effector function can be determined by measuring the binding of the binding agent to C1q.

[00211] The first and second constant regions of the heavy chain of the binding agent may have been modified so that the binding of C1q to said antibody is reduced compared to a wild-type antibody, preferably reduced by at least 70%, at least 80%, at least 90%, at least 95%, at least 97% or 100%, wherein the binding of C1q is preferably determined by ELISA.

[00212] In an embodiment of the binding agent according to the first aspect, in at least one of said first and second constant regions of the heavy chain (CH), one or more amino acids in the positions corresponding to the L234, L235, D265, N297, and P331 positions in a human IgG1 heavy chain according to EU numbering, are not L, L, D, N, and P, respectively.

[00213] In one embodiment of the binding agent according to the first aspect, the positions corresponding to positions L234 and L235 on a human IgG1 heavy chain according to EU numbering may be F and E, respectively, on said first and second chains. Petition 870250088654, dated 09 / 30 / 2025, p. 112 / 329 / 254 heavy.

[00214] In particular, the positions corresponding to positions L234, L235, and D265 in a human IgG1 heavy chain according to EU numbering may be F, E, and A, respectively, in said first and second constant regions of the heavy chain.

[00215] In an embodiment of the binding agent according to the first aspect, the positions corresponding to positions L234 and L235 in a human IgG1 heavy chain according to the EU numbering of both the first and second heavy chain constant regions are F and E, respectively, wherein (i) the position corresponding to F405 in a human IgG1 heavy chain according to the EU numbering of the first heavy chain constant region is L, and the position corresponding to K409 in a human IgG1 heavy chain according to the EU numbering of the second heavy chain is R, or (ii) the position corresponding to K409 in a human IgG1 heavy chain according to the EU numbering of the first heavy chain constant region is R, and the position corresponding to F405 in a human IgG1 heavy chain according to the EU numbering of the second heavy chain is L.

[00216] In an embodiment of the binding agent according to the first aspect, the positions corresponding to positions L234, L235, and D265 in a human IgG1 heavy chain according to the EU numbering of both the first and second constant regions of the heavy chain are F, E, and A, respectively, wherein (i) the position corresponding to F405 in a human IgG1 heavy chain according to the EU numbering of the first constant region of the heavy chain is L, and the position corresponding to K409 in a human IgG1 heavy chain according to the EU numbering of the second constant region of the heavy chain is R, or (ii) the position corresponding to K409 in a human IgG1 heavy chain according to the EU numbering of the first heavy chain is R, and the position that Petition 870250088654, dated 09 / 30 / 2025, p. 113 / 329 / 254 corresponds to F405 in a human IgG1 heavy chain according to the EU numbering of the second heavy chain is L.

[00217] In one embodiment of the linking agent according to the first aspect, the constant region of said first and / or second heavy chain comprises an amino acid sequence selected from the group consisting of a) the sequence shown in SEQ ID NO: 19 or SEQ ID NO: 25 [IgG1-FC]; b) a subsequence of the sequence in a), such as a subsequence in which 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 consecutive amino acids have been deleted, starting from the N-terminus or C-terminus of the sequence defined in a); and c) a sequence with at most 10 substitutions, such as at most 9 substitutions, at most 8, at most 7, at most 6, at most 5, at most 4, at most 3, at most 2, or at most 1 substitution compared to the amino acid sequence defined in a) or b).

[00218] In one embodiment of the linking agent according to the first aspect, the constant region of said first or second heavy chain, such as the second heavy chain, comprises or consists essentially of, or consists of, an amino acid sequence selected from the group consisting of: a) the sequence shown in SEQ ID NO: 20 or SEQ ID NO: 26 [IgG1-F405L]; b) a subsequence of the sequence in a), such as a subsequence in which 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 consecutive amino acids have been deleted, starting from the N-terminus or C-terminus of the sequence defined in a); and c) a sequence with at most 9 substitutions, such as Petition 870250088654, dated 09 / 30 / 2025, p. 114 / 329 100 / 254 maximum 8, maximum 7, maximum 6, maximum 5, maximum 4, maximum 3, maximum 2 or maximum 1 substitution compared to the amino acid sequence defined in a) or b).

[00219] In one embodiment of the linking agent according to the first aspect, the constant region of said first or second heavy chain, such as the first heavy chain, comprises or consists essentially of, or consists of an amino acid sequence selected from the group consisting of: a) the sequence shown in SEQ ID NO: 21 or 27 [IgG1F409R]; b) a subsequence of the sequence in a), such as a subsequence in which 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 consecutive amino acids have been deleted, starting from the N-terminus or C-terminus of the sequence defined in a); and c) a sequence with at most 10 substitutions, such as at most 9 substitutions, at most 8, at most 7, at most 6, at most 5, at most 4 substitutions, at most 3, at most 2, or at most 1 substitution compared to the amino acid sequence defined in a) or b).

[00220] In one embodiment of the linking agent according to the first aspect, the constant region of said first and / or second heavy chain comprises or consists essentially of, or consists of, an amino acid sequence selected from the group consisting of: a) the sequence shown in SEQ ID NO: 22 or SEQ ID NO: 28 [IgG1-Fc_FEA]; b) a subsequence of the sequence in a), such as a subsequence in which 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 consecutive amino acids have been deleted, starting from the N-terminus or C-terminus of the sequence defined in a); and Petition 870250088654, dated 09 / 30 / 2025, p. 115 / 329 101 / 254 c) a sequence with at most 7 substitutions, such as at most 6 substitutions, at most 5, at most 4, at most 3, at most 2, or at most 1 substitution compared to the amino acid sequence defined in a) or b).

[00221] In one embodiment of the linking agent according to the first aspect, the constant region of said first and / or second heavy chain, such as the second heavy chain, comprises or consists essentially of, or consists of, an amino acid sequence selected from the group consisting of: a) the sequence shown in SEQ ID NO: 24 or SEQ ID NO: 30 [IgG1-Fc_FEAL]; b) a subsequence of the sequence in a), such as a subsequence in which 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 consecutive amino acids have been deleted, starting from the N-terminus or C-terminus of the sequence defined in a); and c) a sequence with at most 6 substitutions, such as at most 5 substitutions, at most 4 substitutions, at most 3, at most 2, or at most 1 substitution compared to the amino acid sequence defined in a) or b).

[00222] In one embodiment of the linking agent according to the first aspect, the constant region of said first and / or second heavy chain, such as the first heavy chain, comprises or consists essentially of, or consists of, an amino acid sequence selected from the group consisting of: a) the sequence shown in SEQ ID NO: 23 or SEQ ID NO: 29 [IgG1-Fc_FEAR]; b) a subsequence of the sequence in a), such as a subsequence in which 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 consecutive amino acids have been deleted, starting from the N-terminus or C-terminus of the sequence. Petition 870250088654, dated 09 / 30 / 2025, pp. 116 / 329 102 / 254 defined in a); and c) a sequence with at most 6 substitutions, such as at most 5 substitutions, at most 4, at most 3, at most 2, or at most 1 substitution compared to the amino acid sequence defined in a) or b).

[00223] In one embodiment of the first aspect, the linking agent comprises a constant region of kappa (κ) light chain.

[00224] In one embodiment of the first aspect, the linking agent comprises a constant region of lambda (λ) light chain.

[00225] In one embodiment of the linking agent according to the first aspect, the first light chain constant region is either a kappa (κ) light chain constant region or a lambda (λ) light chain constant region.

[00226] In one embodiment of the linking agent according to the first aspect, the second light chain constant region is either a lambda (λ) light chain constant region or a kappa (κ) light chain constant region.

[00227] In one embodiment of the linking agent according to the first aspect, the first constant light chain region is a kappa (κ) constant light chain region and the second constant light chain region is a lambda (λ) constant light chain region or the first constant light chain region is a lambda (λ) constant light chain region and the second constant light chain region is a kappa (κ) constant light chain region.

[00228] In one embodiment of the linking agent according to the first aspect, the kappa (κ) light chain comprises an amino acid sequence selected from the group consisting of a) the sequence presented in SEQ ID NO: 35; b) a subsequence of the sequence in a), such as a Petition 870250088654, dated 09 / 30 / 2025, page 117 / 329 103 / 254 subsequence in which 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 consecutive amino acids have been deleted, starting from the N-terminus or C-terminus of the sequence defined in a); and c) a sequence with at most 10 substitutions, such as at most 9 substitutions, at most 8, at most 7, at most 6, at most 5, at most 4 substitutions, at most 3, at most 2, or at most 1 substitution, compared to the amino acid sequence defined in a) or b).

[00229] In one embodiment of the linking agent according to the first aspect, the lambda (λ) light chain comprises an amino acid sequence selected from the group consisting of: a) the sequence shown in SEQ ID NO: 36; b) a subsequence of the sequence in a), such as a subsequence in which 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 consecutive amino acids have been deleted, starting from the N-terminus or C-terminus of the sequence defined in a); and c) a sequence with at most 10 substitutions, such as at most 9 substitutions, at most 8, at most 7, at most 6, at most 5, at most 4 substitutions, at most 3, at most 2, or at most 1 substitution, compared to the amino acid sequence defined in a) or b).

[00230] The linking agent (in particular, antibody) according to the first aspect is of an isotype selected from the group consisting of IgG1, IgG2, IgG3 and IgG4. In particular, the linking agent may be a full-length IgG1 antibody. In preferred embodiments of the first aspect, the linking agent (in particular, antibody) is of the IgG1m(f) allotype.

[00231] In a preferred embodiment of the linking agent according to the first aspect, the linking agent comprises: i) a first heavy chain and a light chain comprising Petition 870250088654, dated 09 / 30 / 2025, pp. 118 / 329 104 / 254 said antigen-binding region capable of binding to CD137, wherein the first heavy chain comprises the sequence presented in SEQ ID NO: 31, and the first light chain comprises the sequence presented in SEQ ID NO: 32; ii) a second heavy chain and light chain comprising said antigen-binding region capable of binding to PD-L1, wherein the second heavy chain comprises the sequence presented in SEQ ID NO: 33, and the second light chain comprises the sequence presented in SEQ ID NO: 34.

[00232] The linking agent for use according to the first aspect may be in particular acasunlimab or a biosimilar thereof.

[00233] In currently preferred embodiments, the amount of binding agent administered in each dose and / or in each treatment cycle is: a) approximately 0.3-5 mg / kg of body weight or approximately 25-400 mg in total; and / or b) about 2.1 x 10-9- 3.4 x 10-8mol / kg body weight or about 1.7 x 10-7- 2.7 x 10-6mol in total.

[00234] According to these modalities, the dose defined in mg / kg can be converted into a fixed dose, and vice versa, based on the body weight of the individuals to whom the binding agent is administered, being 80 kg.

[00235] The amount of binding agent administered in each dose and / or in each treatment cycle may be in particular about 0.3-4.0 mg / kg of body weight or about 25-320 mg in total; and / or about 2.1 x 10-9- 2.7 x 10-8 mol / kg of body weight or about 1.7 x 10-7- 2.2 x 10-6 mol in total.

[00236] The amount of binding agent administered in each dose and / or in each treatment cycle may be in particular about 0.38-4.0 mg / kg of body weight or about 30-320 mg in total; and / or about 2.6 x 10-9- 2.7 x 10-8 mol / kg of body weight or Petition 870250088654, dated 09 / 30 / 2025, pp. 119 / 329 105 / 254 about 2.4 x 10-7- 2.2 x 10-6mol in total.

[00237] The amount of binding agent administered in each dose and / or in each treatment cycle may be in particular about 0.5-3.3 mg / kg of body weight or about 40-260 mg in total; and / or about 3.4 x 10-9- 2.2 x 10-8 mol / kg of body weight or about 2.7 x 10-7- 1.8 x 10-6 mol in total.

[00238] The amount of binding agent administered in each dose and / or in each treatment cycle may be in particular about 0.6-2.5 mg / kg of body weight or about 50-200 mg in total; and / or about 4.3 x 10-9- 1.7 x 10-8 mol / kg of body weight or about 3.4 x 10-7- 1.4 x 10-6 mol in total.

[00239] The amount of binding agent administered in each dose and / or in each treatment cycle may be in particular about 0.8-1.8 mg / kg of body weight or about 60-140 mg in total; and / or about 5.1 x 10-9- 1.2 x 10-8 mol / kg of body weight or about 4.1 x 10-7- 9.5 x 10-7 mol in total.

[00240] The amount of binding agent administered in each dose and / or in each treatment cycle may be in particular about 0.9-1.8 mg / kg of body weight or about 70-140 mg in total; and / or about 6.0 x 10-9- 1.2 x 10-8 mol / kg of body weight or about 4.8 x 10-7- 9.5 x 10-7 mol in total.

[00241] The amount of binding agent administered in each dose and / or in each treatment cycle may be in particular about 1-1.5 mg / kg of body weight or about 80-120 mg in total; and / or about 6.8 x 10-9- 1.0 x 10-8 mol / kg of body weight or about 5.5 x 10-7- 8.2 x 10-7 mol in total.

[00242] The amount of binding agent administered in each dose and / or in each treatment cycle may be in particular about 1.1-1.4 mg / kg of body weight or about 90-110 mg in total; and / or Petition 870250088654, dated 09 / 30 / 2025, pp. 120 / 329 106 / 254 about 7.7 x 10-9- 9.4 x 10-9mol / kg body weight or about 6.1 x 10-7- 7.5 x 10-7mol in total.

[00243] The amount of binding agent administered in each dose and / or in each treatment cycle may be in particular about 1.2-1.3 mg / kg of body weight or about 95-105 mg in total; and / or about 6.8 x 10-9- 8.9 x 10-9 mol / kg of body weight or about 6.5 x 10-7- 7.2 x 10-7 mol in total.

[00244] The amount of binding agent administered in each dose and / or in each treatment cycle may be in particular about 0.8-1.5 mg / kg of body weight or about 65-120 mg in total; and / or about 5.5 x 10-9- 1.0 x 10-8 mol / kg of body weight or about 4.4 x 10-7- 8.2 x 10-7 mol in total.

[00245] The amount of binding agent administered in each dose and / or in each treatment cycle may be in particular about 0.9-1.3 mg / kg of body weight or about 70-100 mg in total; and / or about 6.0 x 10-9- 8.5 x 10-9 mol / kg of body weight or about 4.8 x 10-7- 6.8 x 10-7 mol in total.

[00246] approximately 0.9-1.1 mg / kg of body weight or approximately 75-90 mg in total; and / or approximately 6.4 x 10-9- 7.7 x 10-9 mol / kg of body weight or approximately 5.1 x 10-7- 6.1 x 10-7 mol in total.

[00247] Additionally, the amount of binding agent administered in each dose and / or in each treatment cycle may be in particular 0.3-4.0 mg / kg of body weight or 25-320 mg in total; and / or 2.1 x 10-9 - 2.7 x 10-8 mol / kg of body weight or 1.7 x 10-7 2.2 x 10-6 mol in total.

[00248] The amount of binding agent administered in each dose and / or in each treatment cycle may be in particular 0.38-4.0 mg / kg of body weight or 30-320 mg in total; and / or Petition 870250088654, dated 09 / 30 / 2025, pp. 121 / 329 107 / 254 2.6 x 10-9 - 2.7 x 10-8 mol / kg of body weight or 2.4 x 10-7 2.2 x 10-6 mol in total.

[00249] The amount of binding agent administered in each dose and / or in each treatment cycle may be in particular 0.5-3.3 mg / kg of body weight or 40-260 mg in total; and / or 3.4 x 10-9 - 2.2 x 10-8 mol / kg of body weight or 2.7 x 10-7 1.8 x 10-6 mol in total.

[00250] The amount of binding agent administered in each dose and / or in each treatment cycle may be in particular 0.6-2.5 mg / kg of body weight or 50-200 mg in total; and / or 4.3 x 10-9 - 1.7 x 10-8 mol / kg of body weight or 3.4 x 10-7 1.4 x 10-6 mol in total.

[00251] The amount of binding agent administered in each dose and / or in each treatment cycle may be in particular 0.8-1.8 mg / kg of body weight or 60-140 mg in total; and / or 5.1 x 10-9 - 1.2 x 10-8 mol / kg of body weight or 4.1 x 10-7 9.5 x 10-7 mol in total.

[00252] The amount of binding agent administered in each dose and / or in each treatment cycle may be in particular 0.9-1.8 mg / kg of body weight or 70-140 mg in total; and / or 6.0 x 10-9 - 1.2 x 10-8 mol / kg of body weight or 4.8 x 10-7 9.5 x 10-7 mol in total.

[00253] The amount of binding agent administered in each dose and / or in each treatment cycle may be in particular 1-1.5 mg / kg of body weight or 80-120 mg in total; and / or 6.8 x 10-9 - 1.0 x 10-8 mol / kg of body weight or 5.5 x 10-7 8.2 x 10-7 mol in total.

[00254] The amount of binding agent administered in each dose and / or in each treatment cycle may be in particular 1.1-1.4 mg / kg of Petition 870250088654, dated 09 / 30 / 2025, pp. 122 / 329 108 / 254 body weight or 90-110 mg total; and / or 7.7 x 10-9 - 9.4 x 10-9 mol / kg of body weight or 6.1 x 10-7 7.5 x 10-7 mol in total.

[00255] The amount of binding agent administered in each dose and / or in each treatment cycle may be in particular 1.2-1.3 mg / kg of body weight or 95-105 mg in total; and / or 6.8 x 10-9 - 8.9 x 10-9 mol / kg of body weight or 6.5 x 10-7 7.2 x 10-7 mol in total.

[00256] The amount of binding agent administered in each dose and / or in each treatment cycle may be in particular 0.8-1.5 mg / kg of body weight or 65-120 mg in total; and / or 5.5 x 10-9 - 1.0 x 10-8 mol / kg of body weight or 4.4 x 10-7 8.2 x 10-7 mol in total.

[00257] The amount of binding agent administered in each dose and / or in each treatment cycle may be in particular 0.9-1.3 mg / kg of body weight or 70-100 mg in total; and / or 6.0 x 10-9 - 8.5 x 10-9 mol / kg of body weight or 4.8 x 10-7 6.8 x 10-7 mol in total.

[00258] The amount of binding agent administered in each dose and / or in each treatment cycle may be in particular 0.9-1.1 mg / kg of body weight or 75-90 mg in total; and / or 6.4 x 10-9 - 7.7 x 10-9 mol / kg of body weight or 5.1 x 10-7 6.1 x 10-7 mol in total.

[00259] The amount of binding agent administered in each dose and / or in each treatment cycle may be: a) approximately 1.1 mg / kg of body weight or approximately 80 mg in total; and / or b) around 6.8 x 10-9mol / kg of body weight or around 5.5 x 10-7mol in total. Petition 870250088654, dated 09 / 30 / 2025, pp. 123 / 329 109 / 254

[00260] The amount of binding agent administered in each dose and / or in each treatment cycle may be: a) 1.1 mg / kg of body weight or 80 mg in total; and / or b) 6.8 x 10⁻⁹ mol / kg of body weight or 5.5 x 10⁻⁷ mol in total.

[00261] Currently, it is preferable that the amount of binding agent administered in each dose and / or in each treatment cycle be: a) approximately 1.25 mg / kg of body weight or approximately 100 mg in total; and / or b) around 8.5 x 10-9mol / kg of body weight or around 6.8 x 10-7mol in total.

[00262] It is equally preferable that the amount of binding agent administered in each dose and / or in each treatment cycle be: a) 1.25 mg / kg of body weight or 100 mg in total; and / or b) 8.5 x 10⁻⁹ mol / kg of body weight or 6.8 x 10⁻⁷ mol in total.

[00263] The binding agent can be administered in any manner and by any route known in the art. In a preferred embodiment, the binding agent is administered systemically, such as parenterally, in particular intravenously.

[00264] The binding agent may be administered in the form of any suitable pharmaceutical composition, as described herein. In a preferred embodiment, the binding agent is administered in the form of an infusion.

[00265] The binding agent for use according to the invention can be administered using intravenous (IV) infusion, such as by intravenous infusion for at least 30 minutes, or for at least 60 minutes, for example, using intravenous infusion for 30 to 120 minutes. Preferably, the binding agent for use according to the invention is Petition 870250088654, dated 09 / 30 / 2025, pp. 124 / 329 110 / 254 administered using intravenous (IV) infusion over 30 minutes.

[00266] The binding agent may be administered before, simultaneously with, or after administration of the PD-1 inhibitor.

[00267] In one embodiment, the binding agent is administered before the administration of the PD-1 inhibitor. For example, the interval between the end of administration of the binding agent and the beginning of administration of the PD-1 inhibitor may be at least about 10 min, such as at least about 15 min, at least about 20 min, at least about 25 min, at least about 30 min, at least about 35 min, at least about 40 min, at least about 45 min, at least about 50 min, at least about 55 min, at least about 60 min, at least about 90 min, or at least about 120 min, and up to about 14 days (up to about 2 weeks), as well as up to about 13 days, up to about 12 days, up to about 11 days, up to about 10 days, up to about 9 days, up to about 8 days, up to about 7 days (up to about 1 week), up to about 6 days, up to about 5 days, up to about 4 days, up to about 3 days, up to about 2 days, up to about 1 day (up to about 24 hours),until approximately 6 PM, until approximately 12 PM, until approximately 6 AM, until approximately 5 AM, until approximately 4 AM, until approximately 3 AM, until approximately 2.5 AM, or until approximately 2 AM.

[00268] In one embodiment, the binding agent is administered after the administration of the PD-1 inhibitor. For example, the interval between the end of PD-1 inhibitor administration and the start of binding agent administration may be at least about 10 min, such as at least about 15 min, at least about 20 min, at least about 25 min, at least about 30 min, at least about 35 min, at least about 40 min, at least about 45 min, at least about 50 min, at least about 55 min, at least about 60 min, at least about 90 min, or at least about 120 min, and up to about 14 days (up to about 2 weeks), such as up to about 13 days, up to about 12 days, up to about 11 days, up to about 10 days, up to about 9 days, up to about 8 days, up to about 7 days (up to Petition 870250088654, dated 09 / 30 / 2025, pp. 125 / 329 111 / 254 approximately 1 week), up to approximately 6 days, up to approximately 5 days, up to approximately 4 days, up to approximately 3 days, up to approximately 2 days, up to approximately 1 day (up to approximately 24 h), up to approximately 18 h, up to approximately 12 h, up to approximately 6 h, up to approximately 5 h, up to approximately 4 h, up to approximately 3 h, up to approximately 2.5 h, or up to approximately 2 h.

[00269] In one embodiment, the binding agent is administered simultaneously with the PD-1 inhibitor. For example, the binding agent and the PD-1 inhibitor may be administered using a composition comprising both drugs. Alternatively, the binding agent may be administered at one end of the individual, and the PD-1 inhibitor may be administered at the other end of the individual. PD-1 inhibitor (also known as programmed cell death protein 1, PD1, CD279)

[00270] In one embodiment, the PD-1 inhibitor prevents inhibitory signals associated with PD-1. In one embodiment, the PD-1 inhibitor is an antibody, or fragment thereof, that interrupts or inhibits inhibitory signaling associated with PD-1. In one embodiment, the PD-1 inhibitor is a small molecule inhibitor that interrupts or inhibits inhibitory signaling. In one embodiment, the PD-1 inhibitor is a peptide-based inhibitor that interrupts or inhibits inhibitory signaling. In one embodiment, the PD-1 inhibitor is an inhibitory nucleic acid molecule that interrupts or inhibits inhibitory signaling.

[00271] Inhibition or blocking of PD-1 signaling, as described herein, results in the prevention or reversal of immunosuppression and the establishment or enhancement of T-cell immunity against cancerous cells. In one embodiment, inhibition of PD-1 signaling, as described herein, reduces or inhibits immune system dysfunction. In one embodiment, inhibition of PD-1 signaling, as described herein, renders dysfunctional immune cells less dysfunctional. In one embodiment, inhibition of PD-1 signaling, as described herein, renders Petition 870250088654, dated 09 / 30 / 2025, pp. 126 / 329 112 / 254 a less dysfunctional T cell.

[00272] In one embodiment, the PD-1 inhibitor prevents the interaction between PD-1 and PD-L1.

[00273] The PD-1 inhibitor may be an antibody, an antigen-binding fragment thereof, or a construct thereof comprising an antibody moiety with an antigen-binding fragment of the required specificity. Antibodies or antigen-binding fragments thereof are as described herein. Antibodies or antigen-binding fragments thereof that are PD-1 inhibitors include, in particular, antibodies or antigen-binding fragments thereof that bind to PD-1. Antibodies or antigen-binding fragments thereof that are PD-1 inhibitors also include antibodies or antigen-binding fragments thereof that bind to PD-L1. Antibodies or antigen-binding fragments may also be conjugated to additional moieties as described herein. In particular, antibodies or antigen-binding fragments thereof are chimerized, humanized, or human antibodies.

[00274] In a preferred embodiment, an antibody that is a PD-1 inhibitor is a single antibody.

[00275] In one embodiment, the PD-1 inhibitor is an antibody, a fragment or a construct thereof that prevents the interaction between PD-1 and PD-L1.

[00276] The PD-1 inhibitor can be an inhibitory nucleic acid molecule, such as an oligonucleotide, siRNA, shRNA, an antisense DNA or RNA molecule, and an aptamer (e.g., DNA or RNA aptamer), in particular an antisense oligonucleotide. In one embodiment, the PD-1 checkpoint inhibitor, being an siRNA, interferes with mRNA, thus blocking translation, for example, the translation of a PD-1 protein.

[00277] In one embodiment, the PD-1 inhibitor is an antibody, a Petition 870250088654, dated 09 / 30 / 2025, pp. 127 / 329 113 / 254 antigen-binding moiety or a construct thereof that disrupts or inhibits the interaction between the PD-1 receptor and one or more of its ligands, PD-L1 and / or PD-L2. Antibodies that bind to PD-1 and disrupt or inhibit the interaction between PD-1 and one or more of its ligands are known in the art. In certain embodiments, the antibody, antigen-binding moiety, or a construct thereof binds specifically to PD-1.

[00278] In other preferred embodiments, the PD-1 inhibitor is an antibody that binds to PD-1, such as a PD-1 blocking antibody. Without being tied to any theory, it is believed that combining a binding agent comprising a first CD137 binding region and a second PD-L1 binding region with a PD-1 bound antibody increases the response rate and leads to an improved duration of response in individuals receiving combination therapy, as combination therapy leads to complete blockade of the PD-1 pathway with simultaneous conditional activation of 41BB. A PD-1 blocking antibody blocks the interaction with PD-L1 and PD-L2. It is further believed that combination therapy with a PD-1 bound antibody makes larger amounts of PD-L1 available to be bound by the binding agent.

[00279] Exemplary PD-1 inhibitors include, without limitation, anti-PD-1 antibodies such as BGB-A317 (BeiGene; see US 8,735,553, WO 2015 / 35606 and US 2015 / 0079109), lambrolizumab (e.g., described as hPD109A and its humanized derivatives h409A1, h409A16 and h409A17 in WO2008 / 156712), AB137132 (Abcam), EH12.2H7 and RMP1-14 (#BE0146; Bioxcell Lifesciences Pvt. LTD.), MIH4 (Affymetrix eBioscience), nivolumab (OPDIVO, BMS-936558; Bristol Myers Squibb; see US Patent 8,008,449; WO 2013 / 173223; WO 2006 / 121168), pembrolizumab (KEYTRUDA; MK-3475; Merck; see WO 2008 / 156712), pidilizumab (CT-011; CureTech; see Hardy et al., 1994, Cancer Res., 54(22):5793-6 and WO 2009 / 101611), PDR001 (Novartis; see WO Petition 870250088654, dated 09 / 30 / 2025, pp. 128 / 329 114 / 254 2015 / 112900), MEDI0680 (AMP-514; AstraZeneca; vide WO 2012 / 145493), TSR-042 (vide WO 2014 / 179664), cemiplimabe (REGN-2810; Regeneron; H4H79; cf. USN798). 2015 / 0203579 and WO 2015 / 112800), JS001 (TAIZHOU JUNSHI PHARMA; vide Si-Yang Liu et al., 2007, J. Hematol. Oncol. 70: 136), AMP-224 (GSK-26; cf. 2016, Li et al., 2016; cf. Int J Mol Sci 17(7):1151 e WO 2010 / 027827 e WO 2011 / 066342), PF-06801591 (Pfizer), tislelizumabe (BGB-A317; BeiGene; vide WO 2015 / 356, Patent 6969.69 US. 2015 / 0079109), BI 754091, SHR-1210 (vide WO 2015 / 085847) and antibodies 17D8, 2D3, 4H1, 4A11, 7D3 and 5F4 as described in WO 2006 / 0079109, 2018, 2016 INHRCJ Hengrui Medicine also known as SHR1210 vide WO 2015 / 085847), TSR-042 (Tesaro Biopharmaceutical; also known as ANB011; vide W02014 / 179664), GLS-010 (WO / Pharmaceuticals; known as WBP3055 vide Si-Yang et al., 2017, J. Hematol Oncol.70:136), STI-1110 (Sorrento Therapeutics; pursuant to WO 2014 / 194302), AGEN2034 (Agenus; pursuant to WO 2017 / 040790), MGA012 (Macrogenics; pursuant to WO 2017 / 19846), IBI308 (Innovent; pursuant to WO 2017 / 19846). 2017 / 024465, WO 2017 / 025016, WO 2017 / 132825, and WO 2017 / 133540), cetrelimabe (JNJ-63723283; JNJ-3283; vide Calvo et al., J. Clin. Oncol. 36, no. 5_suppl (2018) 58), genolimzumab (CBT-501; vide Patel et al., J. ImmunoTher. Cancer, 2017, 5(Supplement 2):P242), sasanlimab (PF06801591; vide Youssef et al., Proc. Am. Assoc. Cancer Res. Ann. Meeting 2017; Cancer Res 2017;77(13 Suppl):Abstract), toripalimab (JS-001; vide US 2016 / 0272708), camrelizumab (SHR-1210; INCSHR-1210; vide US 2016 / 376367; Huang et al., Clin. Cancer Res. 2018; 24(6):1296-1304), espartalizumab (PDR001; vide WO 2017 / 106656; Naing et al., J. Clin. Oncol. 34, no.15_suppl (2016) 3060-3060), BCD-100 (JSC BIOCAD, Russia; vide WO 2018 / 103017), balstilimabe (AGEN2034; vide WO 2017 / 040790), syntilimabe (IBI-308; vide WO 2017 / 024465). and WO 2017 / 133540), for nonlife (BI-754091; vide US 2017 / 334995; Johnson et al. Petition 870250088654, 09 / 30 / 2025, p. 129 / 3 115 / 254 al., J. Clin. Oncol. 36, no. 5_suppl (2018) 212-212), zimberelimab (GLS010; see WO 2017 / 025051), LZM-009 (see US 2017 / 210806), AK-103 (see WO 2017 / 071625, WO 2017 / 166804 and WO 2018 / 036472), retifanlimab (MGA-012; see WO 2017 / 019846), Sym-021 (see WO 2017 / 055547), CS1003 (see CN107840887), Dostarlimab (JEMPERLI®, GlaxoSmithKline LLC, Philadelphia, PA), anti-PD-1 antibodies as described, for example, in US 7,488,802, US 8,008,449, US 8,168,757, WO 03 / 042402, WO 2010 / 089411 (further describing anti-PD-L1 antibodies), WO 2010 / 036959, WO 2011 / 159877 (further describing antibodies against TIM-3), WO 2011 / 082400, WO 2011 / 161699, WO 2009 / 014708, WO 03 / 099196, WO 2009 / 114335, WO 2012 / 145493 (further describing antibodies against PD-L1), WO 2015 / 035606, WO 2014 / 055648 (further describing anti-KIR antibodies), US 2018 / 0185482 (further describing anti-PD-L1 and anti-TIGIT antibodies), US 8,008,449, US 8,779,105, US 6,808,710, US 8,168.757, US 2016 / 0272708 and US 8,354,509, small molecule antagonists for the PD-1 signaling pathway, as described, for example, in Shaabani et al., 2018, Expert Op Ther Pat., 28(9):665-678 and Sasikumar and Ramachandra, 2018, BioDrugs, 32(5):481-497, PD-1-targeted siRNAs, as described, for example, in WO 2019 / 000146 and WO 2018 / 103501, soluble PD-1 proteins, as described in WO 2018 / 222711 and oncolytic viruses comprising a soluble form of PD-1, as described, for example, in WO 2018 / 022831. Exemplary PD-1 inhibitors also include, but are not limited to, PD-L1 inhibitors such as Atezolizumab, Avelumab, Durvalumab, Envafolimab, Cosibelimab, AUNP12, CA-170, and BMS-986189.

[00280] In a certain embodiment, the PD-1 inhibitor is nivolumab (OPDIVO; BMS-936558) or a biosimilar thereof, pembrolizumab (KEYTRUDA; MK-3475) or a biosimilar thereof, pidilizumab (CT Petition 870250088654, dated 09 / 30 / 2025, pp. 130 / 329 116 / 254 011), PDR001, MEDI0680 (AMP-514) or a biosimilar thereof, TSR042, REGN2810, JS001, AMP-224 (GSK-2661380), PF-06801591, BGBA317, BI 754091, or SHR-1210.

[00281] The PD-1 inhibitor may be, in particular, pembrolizumab or a biosimilar thereof. Alternatively, the antibody may be nivolumab or a biosimilar thereof.

[00282] In certain embodiments, the PD-1 inhibitor is selected from the group consisting of Pembrolizumab, Nivolumab, Cemiplimab, Dostarlimab, Vopratelimab, Espartalizumab, Camrelizumab, Sintilimab, Tislelizumab, Toripalimab, Retifanlimab, Pidilizumab, AMP-224, AMP514, Atezolizumab, Avelumab, Durvalumab, Envafolimab, Cosibelimab, AUNP12, CA-170, BMS-986189, or a respective biosimilar thereof.

[00283] In certain embodiments, the PD-1 inhibitor is an anti-PD-1 antibody, anti-PD-L1 antibody or antigen-binding fragment thereof comprising the complementarity-determining regions (CDRs) of one of the anti-PD-1 antibodies or antigen-binding fragments described above, such as the CDRs of an anti-PD-1 antibody or antigen-binding fragment selected from the group consisting of Pembrolizumab, Nivolumab, Cemiplimab, Dostarlimab, Espartalizumab, Camrelizumab, Sintilimab, Tislelizumab, Toripalimab, Retifanlimab, Pidilizumab, AMP-514, Atezolizumab, Avelumab, Durvalumab, Envafolimab and Cosibelimab, or a respective biosimilar thereof.

[00284] In some embodiments, the CDRs of the anti-PD-1 antibody or the anti-PD-L1 antibody are delineated using the Kabat numbering scheme (Kabat, EA, et al. (1991) Sequences of Proteins Immunological Interest, Fifth Edition, US Department of Health and Human Services, publication number NTH 91-3242). Petition 870250088654, dated 09 / 30 / 2025, pp. 131 / 329 117 / 254

[00285] In certain embodiments, the PD-1 inhibitor is an anti-PD-1 antibody, an anti-PD-L1 antibody or antigen-binding fragment thereof comprising the variable heavy chain region and the variable light chain region of one of the anti-PD-1 antibodies or antigen-binding fragments described above, such as the variable heavy chain region and the variable light chain region of an anti-PD-1 antibody or antigen-binding fragment selected from the group consisting of Pembrolizumab, Nivolumab, Cemiplimab, Dostarlimab, Espartalizumab, Camrelizumab, Sintilimab, Tislelizumab, Toripalimab, Retifanlimab, Pidilizumab, AMP-514, Atezolizumab, Avelumab, Durvalumab, Envafolimab, Cosibelimab, or a respective biosimilar thereof.

[00286] In certain embodiments, the PD-1 inhibitor is an anti-PD-1 antibody, an anti-PD-L1 antibody or an antigen-binding fragment thereof selected from the group consisting of Pembrolizumab, Nivolumab, Cemiplimab, Dostarlimab, Spartalizumab, Camrelizumab, Sintilimab, Tislelizumab, Toripalimab, Retifanlimab, Pidilizumab, AMP-514, Atezolizumab, Avelumab, Durvalumab, Envafolimab, Cosibelimab or a respective biosimilar thereof.

[00287] In certain embodiments, the PD-1 inhibitor is an anti-PD-1 antibody or its antigen-binding fragment selected from the group consisting of Pembrolizumab, Nivolumab, Cemiplimab, Dostarlimab, Spartalizumab, Camrelizumab, Sintilimab, Tislelizumab, Toripalimab, Retifanlimab, Pidilizumab, AMP-514 or a respective biosimilar thereof.

[00288] The pembrolizumab CDR sequences are identified here by SEQ ID NOs: 59-61 (VH 1, 2 and 3 CDRs, respectively) and by SEQ ID NOs: 62-64 (VL 1, 2 and 3 CDRs, respectively). The VH and VL sequences are identified by SEQ ID NOs: 65 and 66. Petition 870250088654, dated 09 / 30 / 2025, pp. 132 / 329 118 / 254 respectively, and the heavy and light chain sequences are identified by SEQ ID NOs: 67 and 68, respectively. Therefore, in one embodiment, the PD-1 inhibitor is an antibody comprising a variable heavy chain (VH) region comprising the CDR1, CDR2, and CDR3 sequences presented in SEQ ID NOs: 59, 60, and 61, respectively, and a variable light chain (VL) region comprising the CDR1, CDR2, and CDR3 sequences presented in SEQ ID NOs: 62, 63, and 64, respectively.

[00289] In a further embodiment, the PD-1 inhibitor is an antibody comprising a variable heavy chain (VH) region comprising or consisting of, or consisting essentially of, the sequence presented in SEQ ID NO: 65, and a variable light chain (VL) region comprising, consisting of, or consisting essentially of, the sequence presented in SEQ ID NO: 66. The PD-1 inhibitor may in particular be an antibody comprising a heavy chain comprising, consisting of, or consisting essentially of, the amino acid sequence presented in SEQ ID NO: 67, and a light chain comprising, consisting of, or consisting essentially of, the amino acid sequence presented in SEQ ID NO: 68.

[00290] The CDR sequences of nivolumab are identified here by SEQ ID NOs: 69-71 (VH 1, 2 and 3 CDRs, respectively) and SEQ ID NOs: 72-74 (VL 1, 2 and 3 CDRs, respectively). The VH and VL sequences are identified by SEQ ID NOs: 75 and 76, respectively, and the heavy and light chain sequences are identified by SEQ ID NOs: 77 and 78, respectively. Therefore, in one embodiment, the PD-1 inhibitor is an antibody comprising a variable heavy chain (VH) region comprising the CDR1, CDR2, and CDR3 sequences presented in SEQ ID NOs: 69, 70, and 71, respectively, and a variable light chain (VL) region comprising the CDR1, CDR2, and CDR3 sequences presented in SEQ ID NOs: 72, 73, and 74, respectively. Petition 870250088654, dated 09 / 30 / 2025, pp. 133 / 329 119 / 254

[00291] In a further embodiment, the PD-1 inhibitor is an antibody comprising a variable heavy chain (VH) region comprising or consisting of, or consisting essentially of, the sequence presented in SEQ ID NO: 75, and a variable light chain (VL) region comprising, consisting of, or consisting essentially of, the sequence presented in SEQ ID NO: 76. The PD-1 inhibitor may in particular be an antibody comprising a heavy chain comprising, consisting of, or consisting essentially of, the amino acid sequence presented in SEQ ID NO: 77, and a light chain comprising, consisting of, or consisting essentially of, the amino acid sequence presented in SEQ ID NO: 78.

[00292] Anti-PD-1 antibodies described herein are preferably monoclonal, and may be multispecific, human, humanized, or chimeric antibodies, single-stranded, Fab fragments, F(ab') fragments, fragments produced by a Fab expression library, and PD-1-binding fragments of any of the foregoing. In some embodiments, an anti-PD-1 antibody described herein binds specifically to PD-1 (e.g., human PD-1). The immunoglobulin molecules described herein may be of any isotype (e.g., IgG, IgE, IgM, IgD, IgA, and IgY), class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2), or subclass of immunoglobulin molecule.

[00293] In certain embodiments of the description, anti-PD-1 antibodies are antigen-binding fragments (e.g., human antigen-binding fragments) as described herein and include, among others, Fab, Fab' and F(ab')2, Fd, single-chain Fvs (scFv), single-chain antibodies, disulfide-linked Fvs (sdFv), and fragments comprising a VL or VH domain. Antigen-binding fragments, including single-chain antibodies, may comprise the variable region(s) alone or in combination with all or part of the following: hinge region, Petition 870250088654, dated 09 / 30 / 2025, pp. 134 / 329 120 / 254 CH1, CH2, CH3 and CL domains. Also included in this description are antigen-binding fragments comprising any combination of variable region(s) with a hinge region, CH1, CH2, CH3 and CL domains. In some embodiments, the anti-PD-1 antibodies or their antigen-binding fragments are human, murine (e.g., mouse and rat), donkey, sheep, rabbit, goat, guinea pig, camelid, horse or chicken.

[00294] The anti-PD-1 antibodies described herein may be monospecific, bispecific, trispecific, or of greater multispecificity. Multispecific antibodies may be specific for different epitopes of PD-1 or may be specific for both PD-1 and a heterologous protein. See, for example, publications PCT WO 93 / 17715; WO 92 / 08802; WO 91 / 00360; WO 92 / 05793; Tutt et al., 1991, J. Immunol. 147:60-69; US Patents 4,474,893; 4,714,681; 4,925,648; 5,573,920; 5,601,819; Kostelny et al., 1992, J. Immunol. 148:1547 1553.

[00295] The anti-PD-1 antibodies described herein may be described or specified in terms of the particular CDRs they comprise. The precise amino acid sequence boundaries of a given CDR or FR may be readily determined using any of several well-known schemes, including those described by Kabat et al. (1991) “Protein Sequences of Immunological Interest,” 5th Edition. Public Health Service, National Institutes of Health, Bethesda, MD (numbering scheme “Kabat”); Al-Lazikani et al., (1997) JMB 273, 927-948 (numbering scheme “Chothia”); MacCallum et al., J. Mol. Biol. 262:732-745 (1996), “Antigen-antigen interactions: contact analysis and topography of binding sites,” J. Mol. Biol. 262, 732-745.” (numbering scheme for “Contact”); Lefranc MP et al., “IMGT unique numbering for immunoglobulin and T cell receptor variable domains and Ig superfamily V-like domains,” Dev Comp Immunol, 2003; 27(1):55-77 (numbering scheme for “IMGT”); Honegger A and Petition 870250088654, dated 09 / 30 / 2025, pp. 135 / 329 121 / 254 Plückthun A, “Yet another numbering scheme for immunoglobulin variable domains: an automatic modeling and analysis tool,” J Mol Biol, 2001;309(3):657-70, (numbering scheme “Aho”); and Martin et al., “Modeling antibody hypervariable loops: a combined algorithm,” PNAS, 1989, 86(23):9268-9272, (numbering scheme “AbM”). The boundaries of a given CDR may vary depending on the scheme used for identification. In some embodiments, a specified individual CDR or CDRs (e.g., CDR-H1, CDR-H2, CDR-H3) of a given antibody or region thereof (e.g., variable region thereof) should be understood as including a CDR (or the specific one) in the manner defined by any of the schemes previously mentioned.For example, when it is stated that a specific CDR (e.g., a CDR-H3) contains the amino acid sequence of a corresponding CDR in a given amino acid sequence of the VH or VL region, it is understood that such CDR possesses a sequence of the corresponding CDR (e.g., CDR-H3) in the variable region, in the manner defined by any of the previously mentioned schemes. The scheme for identifying one or more specific CDRs can be specified, such as the CDR in the manner defined by the Kabat, Chothia, AbM, or IMGT methods.

[00296] In some embodiments, the numbering of amino acid residues in CDR sequences of anti-PD-1 antibodies or their antigen-binding fragments provided herein is in accordance with the IMGT numbering scheme, as described in Lefranc, MP et al., Dev. Comp. Immunol., 2003, 27, 55-77.

[00297] In some embodiments, the anti-PD-1 antibodies described herein comprise the CDRs of the nivolumab antibody. See WO 2006 / 121168. In some embodiments, the CDRs of the nivolumab antibody are delineated using the Kabat numbering scheme (Kabat, EA et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, US Petition 870250088654, dated 09 / 30 / 2025, pp. 136 / 329 122 / 254 Department of Health and Human Services, publication number NTH 913242). The present description includes an anti-PD-1 antibody or a derivative thereof, comprising a variable heavy or light chain domain, wherein said variable domain comprises (a) a set of three CDRs, wherein said set of CDRs is from the monoclonal antibody nivolumab, and (b) a set of four structural regions, wherein said set of structural regions differs from the set of structural regions of the monoclonal antibody nivolumab, and wherein said anti-PD-1 antibody or a derivative thereof binds to PD-1. In certain embodiments, the anti-PD-1 antibody is nivolumab.

[00298] Anti-PD-1 antibodies described herein may also be described or specified in terms of their binding affinity to PD-1 (e.g., human PD-1).Preferred binding affinities include those with a dissociation constant or Kd less than 5 x 10⁻² M, 10⁻² M, 5 x 10⁻³ M, 10⁻³ M, 5 x 10⁻⁴ M, 10⁻⁴ M, 5 x 10⁻⁵ M, 10⁻⁵ M, 5 x 10⁻⁶ M, 10⁻⁶ M, 5 x 10⁻⁷ M, 10⁻⁷ M, 5 x 10⁻⁸ M, 10⁻⁸ M, 5 x 10⁻⁹ M, 10⁻⁹ M, 5 x 10⁻¹⁰ M, 10⁻¹⁰ M, 5 x 10⁻¹⁰ M, 10⁻¹⁰ M, 5 x 10⁻¹¹ M, 10⁻¹¹ M, 5 x 10⁻¹³ M, 10⁻¹³ M, 5 x 10⁻¹⁴ M, 10⁻¹⁴ M. 5x10-15M, or 10-15M.

[00299] Anti-PD-1 antibodies also include derivatives and constructs that are modified, i.e., by the covalent attachment of any type of molecule to the antibody, so that the covalent attachment does not prevent the antibody from binding to PD-1. For example, but not limited to, derivatives of anti-PD-1 antibodies include antibodies that have been modified, for example, by glycosylation, acetylation, PEGylation, phosphorylation, amidation, derivatization by known protective / blocking groups, proteolytic cleavage, binding to a cellular ligand or other protein, etc. Any of the numerous chemical modifications can be performed by known techniques, including, but not limited to, specific chemical cleavage, acetylation, formylation, metabolic synthesis of tunicamycin, etc. In addition, the derivative or construct may contain one or more non-classical amino acids. Petition 870250088654, dated 09 / 30 / 2025, pp. 137 / 329 123 / 254

[00300] In a preferred embodiment, the PD-1 inhibitor is an antibody, in particular an antagonist or blocking antibody, that interrupts or inhibits the PD-1 pathway (interaction of PD-1 with one or more of its ligands (such as PD-L1 and / or PD-L2)). In a preferred embodiment, the PD-1 inhibitor is an antibody, in particular an antagonist or blocking antibody, that interrupts or inhibits the interaction between PD-1 and PD-L1.

[00301] PD-1 inhibitors can be administered in the form of nucleic acid, such as DNA or RNA molecules encoding a PD-1 inhibitor, for example, an inhibitory nucleic acid molecule or an antibody or fragment thereof. For example, antibodies can be administered encoded in expression vectors, as described herein. Nucleic acid molecules can be administered as such, for example, in the form of a plasmid or mRNA molecule, or complexed with an administration vehicle, for example, a liposome, lipoplex, or nucleic acid lipid particles. PD-1 inhibitors can also be administered via an oncolytic virus comprising an expression cassette encoding the PD-1 inhibitor. PD-1 can also be administered via the administration of endogenous or allogeneic cells capable of expressing a PD-1 inhibitor, for example, in the form of cell therapy.

[00302] Preferably, the PD-1 inhibitor is administered in an adequate amount. The amount of PD-1 inhibitor administered in each dose and / or treatment cycle may be in particular within a range, wherein more than 5%, preferably more than 10%, more preferably more than 15%, even more preferably more than 20%, even more preferably more than 25%, even more preferably more than 30%, even more preferably more than 35%, even more preferably more than 40%, even more preferably more than 45%, above all preferably more than 50% of said PD-1 inhibitors bind to PD-1. Petition 870250088654, dated 09 / 30 / 2025, pp. 138 / 329 124 / 254

[00303] In certain embodiments, the PD-1 inhibitor is pembrolizumab or a biosimilar thereof and the amount of PD-1 inhibitor administered, for example, in each dose and / or in each treatment cycle, is approximately 10 - approximately 1,000 mg in total, such as approximately 100 - approximately 600 mg in total, for example, approximately 150 - approximately 600 mg in total, approximately 150 - approximately 500 mg in total, approximately 175 - approximately 500 mg in total, approximately 175 - approximately 450 mg in total, approximately 200 - approximately 450 mg in total or such as approximately 200 - approximately 400 mg in total.

[00304] In certain embodiments, the PD-1 inhibitor is pembrolizumab or a biosimilar thereof and the amount of PD-1 inhibitor administered, for example, in each dose and / or in each treatment cycle, is 10 - 1,000 mg in total such as 100 - 600 mg in total, for example, 150 - 600 mg in total, 150 - 500 mg in total, 175 - 500 mg in total, 175 - 450 mg in total, 200 - 450 mg in total or such as 200 - 400 mg in total.

[00305] In certain embodiments, the PD-1 inhibitor is pembrolizumab or a biosimilar thereof and the amount of PD-1 inhibitor administered, for example, in each dose and / or in each treatment cycle, is approximately 100 - 600 mg in total; and / or approximately 6.84 x 10-7 - 4.11 x 10-7 mol in total.

[00306] In certain embodiments, the PD-1 inhibitor is pembrolizumab or a biosimilar thereof and the amount of PD-1 inhibitor administered, for example, in each dose and / or in each treatment cycle, is approximately 100 - 400 mg in total; and / or approximately 6.84 x 10-7- 2.73 x 10-6 mol in total, such as 100 - 400 mg in total; and / or 6.84 x 10-7- 2.73 x 10-6 mol in total.

[00307] In certain embodiments, the PD-1 inhibitor is pembrolizumab or a biosimilar thereof and the amount of PD-1 inhibitor administered, for example, in each dose and / or in each treatment cycle, is approximately 200–400 mg in total; and / or approximately 6.84 x 10⁻⁷ mol in the Petition 870250088654, dated 09 / 30 / 2025, pp. 139 / 329 125 / 254 total, such as 200 - 400 mg in total; and / or 6.84 x 10-7 - 2.73 x 10-6 mol in total.

[00308] In certain modalities, the amount of PD-1 inhibitor administered, for example, in each dose and / or in each treatment cycle, is approximately 200 mg or approximately 1.37 x 10-6 mol in total, such as 200 mg or 1.37 x 10-6 mol in total.

[00309] In certain embodiments, the PD-1 inhibitor is pembrolizumab or a biosimilar thereof and the amount of PD-1 inhibitor administered, for example, in each dose and / or in each treatment cycle, is approximately 200 mg or approximately 1.37 x 10-6 mol in total, such as 200 mg or 1.37 x 10-6 mol in total.

[00310] In certain modalities, the amount of PD-1 inhibitor administered, for example, in each dose and / or in each treatment cycle, is approximately 400 mg in total or approximately 2.73 x 10-6 in total, such as 400 mg in total or 2.73 x 10-6 in total.

[00311] In certain embodiments, the PD-1 inhibitor is pembrolizumab or a biosimilar thereof and the amount of PD-1 inhibitor administered, for example, in each dose and / or in each treatment cycle, is approximately 400 mg in total or approximately 2.73 x 10-6 in total, such as 400 mg in total or 2.73 x 10-6 in total.

[00312] PD-1 inhibitors can be administered in any manner and by any route known in the art. The mode and route of administration will depend on the type of PD-1 inhibitor to be used. In a preferred embodiment, the PD-1 inhibitor is administered systemically, such as parenterally, particularly intravenously.

[00313] PD-1 inhibitors can be administered in the form of any suitable pharmaceutical composition, as described herein. In a preferred embodiment, the PD-1 inhibitor is administered as an infusion, such as an intravenous infusion. Petition 870250088654, dated 09 / 30 / 2025, pp. 140 / 329 126 / 254

[00314] The antibody that binds to PD-1 may comprise a variable heavy chain (VH) region comprising an HCDR1, HCDR2, and HCDR3 sequence and a variable light chain (VL) region comprising an LCDR1, LCDR2, and LCDR3 sequence, wherein the HCDR1, HCDR2, and HCDR3 sequence comprises or presents the sequence in the manner presented in SEQ ID NO: 104, SEQ ID NO: 101, and SEQ ID NO: 100, respectively, and the LCDR1, LCDR2, and LCDR3 sequence comprises or presents the sequence in the manner presented in SEQ ID NO: 107, QAS, and SEQ ID NO: 105, respectively. A specific, but not limiting, example of such an antibody is MAB-19-0202.

[00315] The terms “variable heavy chain region” (also referred to as “VH”) and “variable light chain region” (also referred to as “VL”) are used here in their most general meaning and comprise any sequences capable of comprising complementarity-determining regions (CDRs), interspersed with other regions, also referred to as scaffold regions (FRs). The scaffold regions, among other things, space the CDRs so that they are able to form the antigen-binding site, particularly after the folding and pairing of VH and VL. Preferably, each VH and VL is composed of three CDRs and four FRs, arranged from the amino terminus to the carboxy terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.In other words, the terms “variable heavy chain region” and “variable light chain region” should not be interpreted as limited to sequences found in a native antibody or in VH and VL sequences, as exemplified here (SEQ ID NOs: 109 to 112 of the sequence listing). These terms include any sequences capable of properly understanding and positioning CDRs, for example, sequences derived from VL and VH regions of native antibodies or derived from the sequences established in SEQ ID NOs: 109 to 112 of the sequence listing. Those skilled in the art will understand that, in particular, the... Petition 870250088654, dated 09 / 30 / 2025, pp. 141 / 329 127 / 254 sequences of the structural regions can be modified (including variants with respect to amino acid substitutions and variants with respect to sequence length, i.e., insertion or deletion variants) without losing the characteristics of the VH and VL, respectively. In a preferred embodiment, any modification is limited to the structural regions. However, those skilled in the art will also be aware of the fact that CDRs, hypervariable and variable regions can be modified without losing the ability to bind to PD-1. For example, the CDR regions will be identical or highly homologous to the regions specified herein. By "highly homologous" it is understood that 1 to 5, preferably 1 to 4, such as 1 to 3 or 1 or 2 substitutions, can be made in the CDRs. Furthermore, the hypervariable and variable regions can be modified so that they exhibit substantial homology with the regions specifically described herein.

[00316] In the antibody binding to PD-1, CDRs, as specified here, were identified using two different CDR identification methods. The first numbering scheme used here is that of Kabat (Wu and Kabat, 1970; Kabat et al., 1991), and the second scheme is the IMGT numbering (Lefranc, 1997; Lefranc et al., 2005). In a third approach, the intersection of both identification schemes was used.

[00317] The antibody that binds to PD-1 may comprise one or more CDRs, a set of CDRs, or a combination of sets of CDRs, as described herein, comprising said CDRs together with their intervening scaffold regions (also referred to as the scaffold region or FR herein) or with portions of said scaffold regions. Preferably, the portion will include at least about 50% of one or both of the first and fourth scaffold regions, the 50% being the C-terminal 50% of the first scaffold region and the N-terminal 50% of the fourth scaffold region. The construction of antibodies by recombinant DNA techniques may result in the introduction of N- or C-terminal residues into the regions Petition 870250088654, dated 09 / 30 / 2025, pp. 142 / 329 128 / 254 variables encoded by linkers introduced to facilitate cloning or other manipulation steps, including the introduction of linkers to join variable regions of the description to other protein sequences, including immunoglobulin heavy chains, other variable domains (e.g., in the production of diabodies) or protein markers.

[00318] The antibody that binds to PD-1 may comprise a variable heavy chain (VH) region comprising a sequence with at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% identity to the amino acid sequence of the VH sequence as presented in any of SEQ ID NO: 111. In one embodiment, the antibody comprises a variable heavy chain (VH) region, wherein the VH comprises the sequence as presented in any of SEQ ID NO: 111. In one embodiment, the antibody comprises a variable light chain (VL) region comprising a sequence with at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% identity with the amino acid sequence of the VL sequence as presented in any of SEQ ID NO: 112.In one embodiment, the antibody comprises a variable light chain (VL) region, wherein the VL comprises the sequence in the manner presented in any of SEQ ID NO: 112.

[00319] The antibody that binds to PD-1 may comprise a variable heavy chain (VH) region and a variable light chain (VL) region, wherein the VH comprises or presents the sequence in the manner shown in SEQ ID NO: 111 and the VL comprises or presents the sequence in the manner shown in SEQ ID NO: 112, or respective variants of these sequences. Another example of an antibody that binds to PD-1 may comprise a VH comprising or presenting the sequence in the manner shown Petition 870250088654, dated 09 / 30 / 2025, pp. 143 / 329 129 / 254 in SEQ ID NO: 111, or a variant thereof, and a VL comprising or presenting the sequence in the manner presented in SEQ ID NO: 112, or a variant thereof. A specific, but not limiting, example of such an antibody is MAB-19-0618. The antibody MAB-19-0618 is derived from MAB-19-0202. Also included by this description are variants of said variable heavy chain (VH) regions and said variable light chain (VL) regions, and the respective combinations of the same VH and VL variants.

[00320] The antibody that binds to PD-1 may comprise a heavy chain and a light chain, wherein the heavy chain comprises a constant heavy chain region comprising or presenting the sequence in the manner presented in SEQ ID NO: 93 or 90 and a variable heavy chain (VH) region comprising or presenting the sequence in the manner presented in SEQ ID NO: 111, and wherein the light chain comprises a constant light chain region comprising or presenting the sequence in the manner presented in SEQ ID NO: 97 and a variable light chain (VL) region comprising or presenting the sequence in the manner presented in SEQ ID NO: 112.

[00321] The antibody that binds to PD-1 may comprise a heavy chain and a light chain, wherein the heavy chain comprises a constant region of heavy chain comprising or presenting the sequence in the manner presented in SEQ ID NO: 93 or 90 and a variable region of heavy chain (VH) comprising the CDR1, CDR2, and CDR3 sequences of the sequence in the manner presented in SEQ ID NO: 111, and wherein the light chain comprises a constant region of light chain comprising or presenting the sequence in the manner presented in SEQ ID NO: 97 and a variable region of light chain comprising the CDR1, CDR2, and CDR3 sequences of the sequence in the manner presented in SEQ ID NO: 112. For example, the CDR1, CDR2, and CDR3 sequences are in the manner shown here. Petition 870250088654, dated 09 / 30 / 2025, page 144 / 329 130 / 254 specified.

[00322] The antibody that binds to PD-1 can be a monoclonal, chimeric, or humanized monoclonal antibody, or a fragment of such an antibody. Antibodies can be complete antibodies or antigen-binding fragments thereof, including, for example, bispecific antibodies.

[00323] In the antibody that binds to PD-1, one or more, preferably both, constant regions of the heavy chain may be modified in such a way that the binding of C1q to said antibody is reduced, compared to a wild-type antibody, preferably reduced by at least 70%, at least 80%, at least 90%, at least 95%, at least 97% or 100%. In one embodiment, the binding to C1q may be determined by ELISA.

[00324] By “wild type”, or “WT”, or “native” is meant here an amino acid sequence found in nature, including allelic variations. A wild amino acid, peptide or protein sequence has an amino acid sequence that has not been intentionally modified.

[00325] In the antibody binding to PD-1, one or more, preferably both, constant regions of the heavy chain may have been modified so that the binding to one or more IgG Fc-gamma receptors to the antibody is reduced compared to a wild-type antibody, preferably by at least 70%, at least 80%, at least 90%, at least 95%, at least 97%, or 100%. In one embodiment, one or more IgG Fc-gamma receptors are selected from at least one of the Fc-gamma RI, Fc-gamma RII, and Fc-gamma RIII receptors. In another embodiment, the IgG Fc-gamma receptor is the Fc-gamma RI receptor.

[00326] In one embodiment, the antibody that binds to PD-1 is not capable of inducing Fc-gamma RI-mediated effector functions or in which the Petition 870250088654, dated 09 / 30 / 2025, pp. 145 / 329 131 / 254 Fc-gamma RI-mediated effector functions induced are reduced compared with a wild-type antibody, preferably by at least 70%, at least 80%, at least 90%, at least 95%, at least 97% or 100%.

[00327] In one embodiment, the antibody that binds to PD-1 is not capable of inducing at least one of the following conditions: complement-dependent cytotoxicity-mediated lysis (CDC), antibody-dependent cellular cytotoxicity-mediated lysis (ADCC), apoptosis, homotypic adhesion and / or phagocytosis, or in which at least one of the following conditions: complement-dependent cytotoxicity-mediated lysis (CDC), antibody-dependent cellular cytotoxicity-mediated lysis (ADCC), apoptosis, homotypic adhesion and / or phagocytosis is induced to a reduced extent, preferably reduced by at least 70%, at least 80%, at least 90%, at least 95%, at least 97% or 100%.

[00328] Antibody-dependent cell-mediated cytotoxicity is also referred to as “ADCC” herein. ADCC describes the ability of effector cells, in the manner described herein, to kill cells, particularly lymphocytes, which preferably requires that the target cell be marked by an antibody.

[00329] ADCC occurs preferentially when antibodies bind to antigens on tumor cells and the Fc domains of the antibody connect to Fc receptors (FcRs) on the surface of immune effector cells. Several families of Fc receptors have been identified, and specific cell populations characteristically express defined Fc receptors. ADCC can be viewed as a mechanism to directly induce a variable degree of immediate tumor destruction, leading to antigen presentation and the induction of tumor-directed T cell responses. Preferably, in vivo induction of ADCC will lead to both tumor-directed T cell responses and host-derived antibody responses. Petition 870250088654, dated 09 / 30 / 2025, pp. 146 / 329 132 / 254

[00330] Complement-dependent cytotoxicity is also referred to as “CDC” here. CDC is another method of cell destruction that can be targeted by antibodies. IgM is the most effective isotype for complement activation. IgG1 and IgG3 are also very effective in targeting CDC via the classical complement activation pathway. Preferably, in this cascade, the formation of antigen-antibody complexes results in the unblocking of multiple closely located C1q binding sites in the CH2 domains of participating antibody molecules, such as IgG molecules (C1q is one of the three subcomponents of complement C1). Preferably, these unblocked C1q binding sites convert the previously low-affinity C1q-IgG interaction into a high-avidity one, which triggers a cascade of events involving a number of other complement proteins and leads to the proteolytic release of the chemotactic / activating agents C3a and C5a from the effector cell.Preferably, the complement cascade ends in the formation of a membrane attack complex, which creates pores in the cell membrane that facilitate the free passage of water and solutes into and out of the cell, potentially leading to apoptosis.

[00331] In one embodiment, antibody binding to PD-1 has reduced or depleted effector functions. In one embodiment, the antibody does not mediate ADCC or CDC, or both.

[00332] In one embodiment, one or more, preferably both, constant regions of the PD-1-binding antibody heavy chain have been modified such that the binding of the neonatal Fc receptor (FcRn) to the antibody is not affected, compared to a wild-type antibody.

[00333] In one embodiment, the PD-1 to which the antibody is capable of binding is human PD-1. In one embodiment, PD-1 presents or comprises the amino acid sequence in the manner presented in SEQ ID NO: 113 or SEQ ID NO: 114, or the amino acid sequence of PD-1 Petition 870250088654, dated 09 / 30 / 2025, pp. 147 / 329 133 / 254 exhibits at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% identity with the amino acid sequence as presented in SEQ ID NO: 113 or SEQ ID NO: 114, or is an immunogenic fragment thereof. In one embodiment, the antibody exhibits the ability to bind to a natural PD-1 epitope present on the surface of living cells.

[00334] In one embodiment, the antibody that binds to PD-1 comprises a heavy chain constant region, wherein the heavy chain constant region comprises an aromatic or nonpolar amino acid at the position corresponding to position 234 in a human IgG1 heavy chain according to EU numbering and an amino acid other than glycine at the position corresponding to position 236 in a human IgG1 heavy chain according to EU numbering.

[00335] The term “amino acid corresponding to position...” and similar expressions as used herein refer to an amino acid position number in a human IgG1 heavy chain. Corresponding amino acid positions in other immunoglobulins can be observed by alignment with human IgG1. Thus, an amino acid or segment in a sequence that “corresponds to” an amino acid or segment in another sequence is one that aligns with the other amino acid or segment using a standard sequence alignment program such as ALIGN, ClustalW, or similar, typically in standard settings, and exhibits at least 50%, at least 80%, at least 90%, or at least 95% identity with a human IgG1 heavy chain. It is considered well known in the art to align a sequence or segment within a sequence, and thereby determine the corresponding position in a sequence for an amino acid position according to the present description. Petition 870250088654, dated 09 / 30 / 2025, pp. 148 / 329 134 / 254

[00336] With reference, for example, to the amino acid sequence according to SEQ ID NO. 93 of the sequence listing in this description, the amino acid positions corresponding to positions 234 to 236 in a human IgG1 heavy chain according to EU numbering are amino acid positions 117 to 119 of SEQ ID NO. 93, with F being positioned at position 117 (corresponding to positions 234 in a human IgG1 heavy chain according to EU numbering), E being positioned at position 118 (corresponding to positions 235 in a human IgG1 heavy chain according to EU numbering), and R being positioned at position 119 (corresponding to positions 236 in a human IgG1 heavy chain according to EU numbering). In the following manner, the amino acid sequence FER is underlined and shown in bold letters. ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTS GVHTFPAVLQSS 60 GLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHT CPPCPAPEFERG 120 PSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEV HNAKTKPREEQYN 180 STYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPR EPQVYTLPPSREE 240 MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSF FLYSKLTVDKSRW 300 QQGNVFSCSVMHEALHNHYTQKSLSLSPG 329

[00337] Unless otherwise indicated or otherwise clearly contradicted by the context, all references to amino acid positions in constant regions of antibody heavy chains throughout this description refer to the corresponding positions in a human IgG1 heavy chain, in accordance with the Petition 870250088654, dated 09 / 30 / 2025, pp. 149 / 329 135 / 254 EU numbering established in Kabat (described in Kabat, EA et al., Sequences of proteins of immunological interest. 5th Edition - US Department of Health and Human Services, NIH publication number 91-3242, pp. 662, 680, 689 (1991)).

[00338] In one embodiment, the PD-1-binding antibody comprises a heavy chain constant region that has reduced or depleted Fc-mediated effector function or that induces Fc-mediated effector function to a lesser degree compared with another antibody comprising the same antigen-binding regions and heavy chain constant regions (HCs), comprising the hinge, CH2, and CH3 regions of human IgG1.

[00339] In a specific embodiment, the said constant region of the heavy chain (HCs) in the antibody that binds to PD-1 is modified so that the antibody induces Fc-mediated effector function to a lesser extent compared to an identical antibody, except for comprising unmodified constant regions of the heavy chain (HCs).

[00340] The term “Fc-mediated effector function”, as used herein, refers to such functions, in particular, being selected from the list of binding to the IgG Fc receptor (FcgammaR, FcyR), binding to C1q, ADCC, CDC and any combination thereof.

[00341] In the context of the present description, the term “exhibits reduced or depleted Fc-mediated effector function” used in relation to an antibody, including a multispecific antibody, means that the antibody causes an overall decrease in Fc-mediated effector functions, such function in particular being selected from the list of IgG Fc receptor binding (FcgammaR, FcyR), C1q binding, ADCC or CDC, preferably by 5% or more, 10% or more, 20% or more, more preferably by 50% or more and most preferably by 75% or more, at the level compared to a human IgG1 antibody comprising (i) the same CDR sequences, in Petition 870250088654, dated 09 / 30 / 2025, pp. 150 / 329 136 / 254 particularly comprising the same first and second antigen-binding regions as said antibody and (ii) two heavy chains comprising the hinge regions, CH2 and CH3 of human IgG1. A “depleted Fc-mediated effector function” or similar phrases includes complete or essentially complete inhibition, i.e., a reduction to zero or essentially to zero.

[00342] In the context of the present description, the term “inducing Fc-mediated effector function to a lesser extent”, used in relation to an antibody, including a multispecific antibody, means that the antibody induces Fc-mediated effector functions, such function being, in particular, selected from the list of binding to the IgG Fc receptor (FcgammaR, FcyR), binding to C1q, ADCC or CDC, to a lesser extent compared with a human IgG1 antibody comprising (i) the same CDR sequences, in particular comprising the same first and second antigen-binding regions, as said antibody and (ii) two heavy chains comprising the hinge regions, CH2 and CH3 of human IgG1.

[00343] Fc-mediated effector function can be determined by measuring the binding of the binding agent to Fcy receptors, binding to C1q, or Fc-mediated crosslinking induction of Fcy receptors. In particular, Fc-mediated effector function can be determined by measuring the binding of the binding agent to C1q and / or FC-gamma RI of IgG.

[00344] In an embodiment related to the use of the antibody that binds to PD-1, the amino acid at the position corresponding to position 236 in a human IgG1 heavy chain, according to EU numbering, is a basic amino acid.

[00345] The terms “amino acid” and “amino acid residue” may be used interchangeably here and should not be understood as limiting. Amino acids are organic compounds that contain amine (-NH2) and carboxyl (COOH) functional groups, along with a side chain (R group). Petition 870250088654, dated 09 / 30 / 2025, pp. 151 / 329 137 / 254 specific to each amino acid. In the context of this description, amino acids can be classified based on their structure and chemical characteristics.

[00346] In the present description, amino acid residues are expressed using the following abbreviations. In addition, unless explicitly indicated otherwise, amino acid sequences of peptides and proteins are identified from the N-terminal to the C-terminal (left terminal to right terminal), with the N-terminal identified as a first residue. Amino acids are designated by their 3-letter abbreviation, 1-letter abbreviation, or full name, as follows: Ala: A: alanine; Asp: D: aspartic acid; Glu: E: glutamic acid; Phe: F: phenylalanine; Gly: G: glycine; His: H: histidine; Ile: I: isoleucine; Lys: K: lysine; Leu: L: leucine; Met: M: methionine; Asn: N: asparagine; Pro: P: proline; Gln: Q: glutamine; Arg: R: arginine; Ser: S: serine; Thr: T: threonine; Val: V: valine; Trp: W: tryptophan; Tyr: Y: tyrosine; Cys: C: cysteine.

[00347] Naturally occurring amino acids can also be broadly divided into four families: acidic (aspartate, glutamate), basic (lysine, arginine, histidine), nonpolar (alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), and uncharged polar (glycine, asparagine, glutamine, cysteine, serine, threonine, tyrosine). Phenylalanine, tryptophan, and tyrosine are sometimes collectively classified as aromatic amino acids.

[00348] In an embodiment related to the use of an antibody that binds to PD-1, the basic amino acid at the position corresponding to position 236 in a human IgG1 heavy chain, according to EU numbering, is selected from the group consisting of lysine, arginine, and histidine. In one embodiment, the basic amino acid at the position corresponding to position 236 in a human IgG1 heavy chain, according to EU numbering, is arginine (G236R). Such amino acid substitution is also referred to herein. Petition 870250088654, dated 09 / 30 / 2025, pp. 152 / 329 138 / 254 as G236R. The term “G236R” indicates that at position 236 in a human IgG1 heavy chain, according to EU numbering, the amino acid glycine (G) is replaced by arginine (R). In the present description, similar terms are used for other amino acid positions and amino acids. Unless otherwise indicated, the amino acid position referenced in these terms is the amino acid position in a human IgG1 heavy chain, according to EU numbering.

[00349] In one embodiment related to the use of an antibody that binds to PD-1, the amino acid at the position corresponding to position 234 in a human IgG1 heavy chain, according to EU numbering, is an aromatic amino acid. In one embodiment, the aromatic amino acid at that position is selected from the group consisting of phenylalanine, tryptophan, and tyrosine.

[00350] In an embodiment related to the use of an antibody that binds to PD-1, the amino acid at the position corresponding to position 234 in a human IgG1 heavy chain, according to EU numbering, is a nonpolar amino acid. In one embodiment, the nonpolar amino acid at this position is selected from the group consisting of alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, and tryptophan. In another embodiment, the nonpolar amino acid at this position is selected from the group consisting of isoleucine, proline, phenylalanine, methionine, and tryptophan.

[00351] In an embodiment related to the use of an antibody that binds to PD-1, the amino acid at the position corresponding to position 234 in a human IgG1 heavy chain, according to EU numbering, is phenylalanine (L234F).

[00352] Exemplary combinations of possible amino acids at positions 234 and 236 in a human IgG1 heavy chain, according to EU numbering, are presented in the following table: Petition 870250088654, dated 09 / 30 / 2025, pp. 153 / 329 139 / 254 Table 5: Amino acid position 234 Amino acid position 236 Phenylalanine (F) Arginine (R) Tryptophan (W) Arginine (R) Tyrosine (Y) Arginine (R) Alanine (A) Arginine (R) Valine (V) Arginine (R) Leucine (L) Arginine (R) Isoleucine (I) Arginine (R) Proline (P) Arginine (R) Methionine (M) Arginine (R) Phenylalanine (F) Lysine (K) Tryptophan (W) Lysine (K) Tyrosine (Y) Lysine (K) Alanine (A) Lysine (K) Valine (V) Lysine (K) Leucine (L) Lysine (K) Isoleucine (I) Lysine (K) Proline (P) Lysine (K) Methionine (M) Lysine (K) Phenylalanine (F) Histidine (H) Tryptophan (W) Histidine (H) Tyrosine (Y) Histidine (H) Alanine (A) Histidine (H) Valine (V) Histidine (H) Leucine (L) Histidine (H) Isoleucine (I) Histidine (H) Proline (P) Histidine (H) Methionine (M) Histidine (H)

[00353] For example, at the positions corresponding to positions 234 and 236 in a human IgG1 heavy chain according to EU numbering, in particular, the following amino acids may be present in the constant region of the antibody heavy chain that binds to PD-1: 234F / 236R, 234W / 236R, 234Y / 236R, 234A / 236R, 234L / 236R, 234F / 236K, 234W / 236K, Petition 870250088654, dated 09 / 30 / 2025, pp. 154 / 329 140 / 254 234Y / 236K, 234A / 236K, 234L / 236K, 234F / 236H, 234W / 236H, 234Y / 236H, 234A / 236H, or 234L / 236H.

[00354] The amino acids or amino acid substitutions mentioned in positions 234 and 236 may be present only in one heavy chain of the antibody that binds to PD-1 or in both heavy chains of the antibody that binds to PD-1. The respective amino acids present in the first and second heavy chains of the antibody may be selected independently of each other.

[00355] For example, at least one heavy chain of the antibody that binds to PD-1 may comprise the following sequence (SEQ ID NO: 93): ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTS GVHTFPAVLQSS 60 GLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHT CPPCPAPEFLRG 120 PSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEV HNAKTKPREEQYN 180 STYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPR EPQVYTLPPSREE 240 MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSF FLYSKLTVDKSRW 300 QQGNVFSCSVMHEALHNHYTQKSLSLSPG 329

[00356] In an embodiment related to antibody binding to PD1, said heavy chain in which the amino acids at the position corresponding to positions 234 and 236 in a human IgG1 heavy chain, according to EU numbering, are as specified above; furthermore, the amino acid at the position corresponding to position 235 in a human IgG1 heavy chain, according to EU numbering, is an acidic amino acid. In one embodiment, the acidic amino acid at this position is selected from aspartate or glutamate. In another embodiment, the amino acid at the position Petition 870250088654, dated 09 / 30 / 2025, page 155 / 329 141 / 254 corresponding to position 235 in a human IgG1 heavy chain, according to EU numbering, is glutamate (L235E).

[00357] In an embodiment related to antibody binding to PD1, in the constant region of the heavy chain, the amino acids at positions 234, 235, and 236 in a human IgG1 heavy chain, according to EU numbering, are a nonpolar or aromatic amino acid at position 234, an acidic amino acid at position 235, and a basic amino acid at position 236.

[00358] Exemplary combinations of possible amino acids at positions 234, 235, and 236 in a human IgG1 heavy chain, according to EU numbering, are presented in the following table: Table 6: Amino acid position 234 Amino acid position 235 Amino acid position 236 Phenylalanine (F) Aspartate (D) or Glutamate (E) Arginine (R) Tryptophan (W) Aspartate (D) or Glutamate (E) Arginine (R) Tyrosine (Y) Aspartate (D) or Glutamate (E) Arginine (R) Alanine (A) Aspartate (D) or Glutamate (E) Arginine (R) Valine (V) Aspartate (D) or Glutamate (E) Arginine (R) Leucine (L) Aspartate (D) or Glutamate (E) Arginine (R) Isoleucine (I) Aspartate (D) or Glutamate (E) Arginine (R) Proline (P) Aspartate (D) or Glutamate (E) Arginine (R) Methionine (M) Aspartate (D) or Glutamate (E) Arginine (R) Phenylalanine (F) Aspartate (D) or Glutamate (E) Lysine (K) Tryptophan (W) Aspartate (D) or Glutamate (E) Lysine (K) Tyrosine (Y) Aspartate (D) or Glutamate (E) Lysine (K) Alanine (A) Aspartate (D) or Glutamate (E) Lysine (K) Valine (V) Aspartate (D) or Glutamate (E) Lysine (K) Leucine (L) Aspartate (D) or Glutamate (E) Lysine (K) Isoleucine (I) Aspartate (D) or Glutamate (E) Lysine (K) Proline (P)Aspartate (D) or Glutamate (E) Lysine (K) Methionine (M) Aspartate (D) or Glutamate (E) Lysine (K) Petition 870250088654, dated 09 / 30 / 2025, pp. 156 / 329 142 / 254 Phenylalanine (F) Aspartate (D) or Glutamate (E) Histidine (H) Tryptophan (W) Aspartate (D) or Glutamate (E) Histidine (H) Tyrosine (Y) Aspartate (D) or Glutamate (E) Histidine (H) Alanine (A) Aspartate (D) or Glutamate (E) Histidine (H) Valine (V) Aspartate (D) or Glutamate (E) Histidine (H) Leucine (L) Aspartate (D) or Glutamate (E) Histidine (H) Isoleucine (I) Aspartate (D) or Glutamate (E) Histidine (H) Proline (P) Aspartate (D) or Glutamate (E) Histidine (H) Methionine (M) Aspartate (D) or Glutamate (E) Histidine (H)

[00359] For example, at positions 234 and 236 on a human IgG1 heavy chain, according to EU numbering, the following amino acids may be present in the constant region of the antibody heavy chain that binds to PD-1: 234F / 235E / 236R, 234W / 235E / 236R, 234Y / 235E / 236R, 234A / 235E / 236R, 234L / 235E / 236R, 234F / 235D / 236R, 234W / 235D / 236R, 234Y / 235D / 236R, 234A / 235D / 236R, 234L / 235D / 236R, 234F / 235L / 236R, 234W / 235L / 236R, 234Y / 235L / 236R, 234A / 235L / 236R, 234L / 235L / 236R, 234F / 235A / 236R, 234W / 235A / 236R, 234Y / 235A / 236R, 234A / 235A / 236R, 234L / 235A / 236R, 234F / 235E / 236K, 234W / 235E / 236K, 234Y / 235E / 236K, 234A / 235E / 236K, 234L / 235E / 236K, 234F / 235D / 236K, 234W / 235D / 236K, 234Y / 235D / 236K, 234A / 235D / 236K, 234L / 235D / 236K, 234F / 235L / 236K, 234W / 235L / 236K, 234Y / 235L / 236K, 234A / 235L / 236K, 234L / 235L / 236K, 234F / 235A / 236K, 234W / 235A / 236K, 234Y / 235A / 236K, 234A / 235A / 236K, 234L / 235A / 236K, 234F / 235E / 236H, 234W / 235E / 236H, 234Y / 235E / 236H, 234A / 235E / 236H, 234L / 235E / 236H,234F / 235D / 236H, 234W / 235D / 236H, 234Y / 235D / 236H, 234A / 235D / 236H, 234L / 235D / 236H, 234F / 235L / 236H, 234W / 235L / 236H, 234Y / 235L / 236H, 234A / 235L / 236H, 234L / 235L / 236H, 234F / 235A / 236H, 234W / 235A / 236H, 234Y / 235A / 236H, 234A / 235A / 236H, or 234L / 235A / 236H.,

[00360] The amino acids or amino acid substitutions previously mentioned at positions 234 and 236 may be present only in one heavy chain of the antibody that binds to PD-1 or in both. Petition 870250088654, dated 09 / 30 / 2025, pp. 157 / 329 143 / 254 the antibody heavy chains that bind to PD-1. The respective amino acids present in the first and second antibody heavy chains can be selected independently of each other.

[00361] For example, at least one heavy chain of the antibody that binds to PD-1 may comprise the following sequence (SEQ ID NO: 93): ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTS GVHTFPAVLQSS 60 GLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHT CPPCPAPEFERG 120 PSVFLFPKPKDTLMISRTPEVTVCVVDVSHEDPEVKFNWYVDGVEV HNAKTKPREEQYN 180 STYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAGQPR EPQVYTLPPSREE 240 MTKNQVSLTCLVKGGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSF FLYSKLTVDKSRW 300 QQGNVFSCSVMHEALHNHYTQKSLSLSPG 329

[00362] Any permutations and combinations of all amino acid substitutions described in headings 234, 236 and 235, if applicable, in this application, for example, in the manner shown in Tables 5 and 6, shall be considered as described by the description of this application, unless otherwise indicated by the context. For example, in an embodiment of the antibody the first heavy chain comprises the amino acids FER at the position corresponding to headings 234 to 236 in a human IgG1 heavy chain according to EU numbering, or the first heavy chain comprises or consists essentially of, or consists of, an amino acid sequence presented in SEQ ID NO: 93, and the second heavy chain of said antibody comprises other amino acids, for example, the amino acids AAG or LLG at the headings corresponding to headings 234 to 236 in a human IgG1 heavy chain according to EU numbering. Petition 870250088654, dated 09 / 30 / 2025, pp. 158 / 329 144 / 254 with EU numbering or comprises or the second heavy chain of said antibody comprises or essentially consists of, or consists of an amino acid sequence presented in SEQ ID NO: 92 or 98. In another embodiment of the antibody, the first and second heavy chains comprise the same amino acids at the position corresponding to positions 234 to 236 in a human IgG1 heavy chain according to EU numbering, i.e., the same aromatic or nonpolar amino acid at the position corresponding to position 234 in a human IgG1 heavy chain according to EU numbering, for example F, and the same amino acid other than glycine at the position corresponding to position 236 in a human IgG1 heavy chain according to EU numbering, for example R, such as the specific combination of FER or FLR.

[00363] In one embodiment, the antibody that binds to PD-1 comprises at least one or two constant heavy chain regions, wherein the amino acid corresponding to position 234 is phenylalanine, the amino acid corresponding to position 235 is glutamate, and the amino acid corresponding to position 236 is arginine (L234F / L235E / G236R = FER).

[00364] In one embodiment, the antibody that binds to PD-1 comprises one or more heavy chain (HC) constant regions comprising a sequence with at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% identity to the amino acid sequence of the heavy chain constant region sequence as presented in SEQ ID NO: 93.

[00365] In one embodiment, the antibody that binds to PD-1 comprises one or more, for example, two heavy chain constant (CH) regions, wherein the heavy chain constant region comprises the sequence in the manner presented in SEQ ID NO: 93.

[00366] The antibody is preferably of the IgG1 isotype. Petition 870250088654, dated 09 / 30 / 2025, pp. 159 / 329 145 / 254

[00367] As used herein, the term “isotype” refers to the class of immunoglobulins encoded by genes in the constant region of the heavy chain. When the IgG1 isotype is mentioned herein, the term is not limited to a specific isotype sequence, for example, a particular IgG1 sequence, but is used to indicate that the antibody is more closely related in sequence to that isotype, for example, IgG1, than to other isotypes. Thus, for example, an IgG1 antibody described herein may be a sequence variant of a naturally occurring IgG1 antibody, including variations in the constant regions.

[00368] IgG1 antibodies can exist in multiple polymorphic variants called allotypes (reviewed in Jefferis and Lefranc 2009. mAbs Vol 1 Publication 4 1-7), any of which is suitable for use in some of the embodiments described herein. Common allotypic variants in human populations are those designated by the letters a, f, n, z, or combinations thereof. In any of the embodiments described herein, the antibody may comprise an Fc heavy chain region comprising a human IgG Fc region. In other embodiments, the human IgG Fc region comprises a human IgG1.

[00369] In mammals, there are two types of light chains, namely lambda and kappa. Immunoglobulin chains comprise a variable region and a constant region. The constant region is essentially conserved in the different isotypes of immunoglobulins, while the variable part is highly diverse and responsible for antigen recognition.

[00370] For example, or in one embodiment, an antibody, preferably a monoclonal antibody, used in accordance with the present invention is an IgG1,κ isotype or an isotype λ, preferably comprising human IgG1 / κ or human IgG1 / λ constant parts, or the antibody, preferably the monoclonal antibody, is derived from an IgG1,λ (lambda) or IgG1,κ (kappa) antibody, preferably from an IgG1,λ antibody. Petition 870250088654, dated 09 / 30 / 2025, pp. 160 / 329 146 / 254 (lambda) human or IgG1, κ (kappa) human.

[00371] In one embodiment, the PD-1-binding antibody comprises a light chain with a constant light chain (LC) region comprising a sequence with at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% identity with the amino acid sequence of the LC sequence as presented in SEQ ID NO: 97. In another embodiment, the antibody comprises a light chain with a constant light chain (LC) region comprising the sequence as presented in SEQ ID NO: 97.

[00372] In one embodiment, the PD-1-binding antibody comprises a heavy chain comprising a sequence with at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% identity to the amino acid sequence as presented in SEQ ID NO: 152 and a light chain comprising a sequence with at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% identity to the amino acid sequence as presented in SEQ ID NO: 153. In another embodiment, the PD-1-binding antibody comprises a heavy chain comprising the amino acid sequence as presented in SEQ ID NO: 152 and a light chain comprising the amino acid sequence as presented in SEQ ID NO: 153.

[00373] In one embodiment of the invention, the antibody that binds to PD-1 is a full-length IgG1 antibody, for example, IgG1,κ. In another embodiment of the invention, the binding agent is a full-length human IgG1 antibody, for example, IgG1,κ.

[00374] In one embodiment, the antibody that binds to PD-1 can be derivatized, linked, or co-expressed with other binding specificities. In Petition 870250088654, dated 09 / 30 / 2025, pp. 161 / 329 147 / 254 In another embodiment, the antibody may be derivatized, linked or co-expressed with another functional molecule, for example, another peptide or protein (e.g., a Fab' fragment). For example, the antibody may be functionally linked (e.g., by chemical coupling, gene fusion, non-covalent association or otherwise) to one or more other molecular entities, such as another antibody (e.g., to produce a bispecific or multispecific antibody).

[00375] The antibody that binds to PD-1 may be a human antibody. The term “human antibody,” as used herein, is intended to include antibodies with variable and constant regions derived from human germline immunoglobulin sequences. The human antibody that binds to PD-1 may include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo).

[00376] The present description includes the use of bispecific and multispecific molecules comprising at least a first binding specificity for PD-1 and a second binding specificity (or other binding specificities) for a second target epitope (or for other target epitopes).

[00377] In one embodiment, the first antigen-binding region of the multispecific antibody that binds to PD-1 comprises the variable region of the heavy chain (VH) and / or the variable region of the light chain (VL), as presented herein.

[00378] In an embodiment related to the use of a multispecific antibody that binds to PD-1, the antibody comprises first and second binding arms derived from full-length antibodies, such as full-length IgG1, λ (lambda) or IgG1, κ (kappa) antibodies, in the manner previously mentioned. In one embodiment, the first and Petition 870250088654, dated 09 / 30 / 2025, page 162 / 329 148 / 254 second linking arms are derived from monoclonal antibodies. For example, or in a preferred embodiment, the first and / or second linking arm is derived from an IgG1, κ or λ isotype, preferably comprising human IgG1 / K or human IgG1^ constant parts.

[00379] Said first antigen-binding region that binds to PD1 of the multispecific or bispecific antibody used according to the present invention may comprise variable heavy and light chain regions of an antibody that competes for PD-1 binding with PD-L1 and / or PDL2. In an embodiment related to the use of the multispecific or bispecific antibody, the first antigen-binding region that binds to PD-1 comprises the variable heavy chain (VH) region and / or the variable light chain (VL) region, as presented herein.

[00380] As used herein, the term “effector cell” refers to an immune cell that is involved in the effector phase of an immune response, as opposed to the cognitive and activation phases of an immune response. Exemplary immune cells include cells of myeloid or lymphoid origin, for example, lymphocytes (e.g., B cells and T cells, including cytotoxic T cells (CTLs), killer cells, natural killer cells, macrophages, monocytes, eosinophils, neutrophils, polymorphonuclear cells, granulocytes, mast cells, and basophils).

[00381] “Target cell” means any unwanted cell in an individual (e.g., a human or animal) that can be targeted by an antibody. In preferred embodiments, the target cell is a tumor cell. Individual and tumor or cancer to be treated.

[00382] The individual to be treated according to the present description is preferably a human individual.

[00383] In preferred modalities, the tumor or cancer exhibits high microsatellite instability (MSI-H) or impaired ligand repair. Petition 870250088654, dated 09 / 30 / 2025, page 163 / 329 149 / 254 nonspecific (dMMR).

[00384] In a preferred embodiment, the tumor or cancer to be treated is a solid tumor or cancer. The tumor or cancer may be a metastatic tumor or cancer. The tumor or cancer may be an unresectable tumor or cancer. The tumor or cancer may be a recurrent tumor or cancer.

[00385] In one form, the tumor or cancer is leukemia, such as acute myeloid leukemia (AML).

[00386] The tumor or cancer may be selected from the group consisting of melanoma, ovarian cancer, lung cancer (e.g., non-small cell lung cancer (NSCLC)), colorectal cancer, head and neck cancer, gastric cancer, breast cancer, renal cancer, urothelial cancer, bladder cancer, esophageal cancer, pancreatic cancer, liver cancer, thymoma and thymic carcinoma, brain cancer, glioma, adrenocortical carcinoma, thyroid cancer, other skin cancers, sarcoma, multiple myeloma, leukemia, lymphoma, myelodysplastic syndromes, endometrial cancer, prostate cancer, penile cancer, cervical cancer, Hodgkin lymphoma, non-Hodgkin lymphoma, Merkel cell carcinoma, and mesothelioma. More preferably, the tumor or cancer is selected from the group consisting of melanoma, lung cancer, colorectal cancer, pancreatic cancer, and head and neck cancer.

[00387] Preferably, the tumor or cancer is selected from the group consisting of colorectal cancer, gastric cancer, endometrial cancer, ovarian cancer, hepatobiliary tract cancer, pancreatic cancer, urinary tract cancer, bladder cancer, thyroid cancer, breast cancer, prostate cancer, ovarian cancer, central nervous system (CNS) cancer, and skin cancer such as melanoma, and more preferably selected from the group consisting of colon cancer, gastric cancer, and endometrial cancer.

[00388] In particular modalities, the tumor or cancer is endometrial cancer. Endometrial cancer is one of the most common gynecological neoplasms. Petition 870250088654, dated 09 / 30 / 2025, page 164 / 329 Endometrial cancer is the most common cancer worldwide, with over 400,000 new cases and over 97,000 deaths in 2020 (Globocan, 2020). The highest incidence rates are found in North America and Northern, Central, and Eastern Europe (Globocan, 2020). The global burden of disease caused by endometrial cancer is also increasing, sharply in North America and European regions (Zhang et al., 2019). Approximately two-thirds of women with endometrial cancer have early-stage disease confined to the uterus, which is usually treated surgically with or without radiotherapy, with excellent results. However, women with recurrent and / or distant metastatic disease are incurable and have a 5-year survival rate of <20% and limited treatment options (SEER database, 2021).For many years, first-line therapy for treatment-naive patients with unresectable and / or metastatic endometrial cancer consisted of dual or triple chemotherapy, with response rates around 40% to 50% and a median survival in the range of 15 months (McMeekin et al., 2007; Miller et al., 2020), without significant advances in identifying subgroups with potentially different treatment needs. Endometrial cancers have historically been classified as type I or type II cancers. Type I cancers comprise approximately 85% of endometrial cancers and generally present with low- to intermediate-grade endometrioid histology. Type II cancers include non-endometrioid cases, most commonly with serous papillary or clear cell histology.Importantly, studies have evaluated the variation in response rates for different histological subtypes and found similar efficacy of SoC chemotherapy in serous and endometrioid tumors, both with an ORR of ~45% (McMeekin et al., 2007). New insights into the heterogeneity of endometrial tumors emerged from the genomic and transcriptomic profile of a large cohort of treatment-naive endometrial tumors using TCGA, leading to classification. Petition 870250088654, dated 09 / 30 / 2025, page 165 / 329 151 / 254 molecular classification of 4 subtypes of endometrial cancer, each with distinct molecular characteristics (Masood and Singh, 2021; Talhouk et al., 2015): POLE-mutated, an ultramutated subtype (<10% of all endometrial cancers); dMMR or MSI-H, a hypermutated subtype (25% to 30% of all endometrial cancers); pMMR or MSS, low CN subtype (30% to 40% of all endometrial cancers); pMMR or MSS, high CN subtype (25% to 30% of all endometrial cancers). PD-1 inhibitors, dostarlimab and pembrolizumab, are now FDA and EMA approved as second-line treatment for adult patients with recurrent or advanced endometrial cancer that is dMMR or MSI-H, as determined by an FDA-approved trial.

[00389] In one modality, the tumor is a PD-L1 positive tumor. In certain modalities, it is preferable that PD-L1 be expressed in >1% of cancer cells or tumor cells. In another modality, the tumor is a PD-L1 negative tumor. PD-L1 expression can be determined using techniques known to the specialist in the field and can, for example, be evaluated by immunohistochemistry (IHC).

[00390] In one embodiment, the individual has experienced progression during or after at least one prior line of treatment for the unresectable and / or metastatic tumor or cancer. According to a preferred embodiment, the treatment regimen is systemic chemotherapy such as platinum-based chemotherapy. According to this embodiment, the tumor or cancer is preferably endometrial cancer.

[00391] In one embodiment, the said individual has received prior treatment with a checkpoint inhibitor. The checkpoint inhibitor is, for example, a PD-1 inhibitor or a PD-L1 inhibitor, such as an anti-PD-1 antibody or an anti-PD-L1 antibody. The PD-1 inhibitor or PD-L1 inhibitor is administered as monotherapy or as part of a combination therapy. In a particular embodiment, the said individual Petition 870250088654, dated 09 / 30 / 2025, page 166 / 329 152 / 254 showed progression after treatment with a PD-1 inhibitor or a PD-L1 inhibitor, such as an anti-PD-1 antibody or an anti-PD-L1 antibody. According to this modality, the tumor or cancer is preferably endometrial cancer.

[00392] In a further embodiment, the said individual has not received prior treatment with a checkpoint inhibitor, such as an anti-PD-1 antibody, an anti-PD-L1 antibody, an anti-CTLA-4 antibody, an anti-LAG3 antibody, or an anti-TIGIT antibody. According to this embodiment, the tumor or cancer is preferably endometrial cancer. Treatment regimen

[00393] The binding agent and PD-1 inhibitor can be administered by any suitable route, such as intravenous, intra-arterial, subcutaneous, intradermal, intramuscular, intranodal or intratumoral.

[00394] In one embodiment of the first aspect, the binding agent is administered to the individual by systemic administration. Preferably, the binding agent is administered to the individual by intravenous injection or infusion. In one embodiment, the binding agent is administered in at least one treatment cycle.

[00395] In one embodiment, the PD-1 inhibitor is administered to the individual by systemic administration. Preferably, the PD-1 inhibitor is administered to the individual by intravenous injection or infusion. In one embodiment, the PD-1 inhibitor is administered in at least one treatment cycle.

[00396] In one embodiment, the binding agent and the PD-1 inhibitor are administered to the individual by systemic administration. Preferably, the binding agent and the PD-1 inhibitor are administered to the individual by intravenous injection or infusion. In one embodiment, the binding agent and the PD-1 inhibitor are administered in at least one treatment cycle.

[00397] In one modality, each treatment cycle is approximately two Petition 870250088654, dated 09 / 30 / 2025, page 167 / 329 153 / 254 weeks (14 days), three weeks (21 days) or four weeks (28 days), five weeks (35 days) or six weeks (42 days). In preferred modalities, each treatment cycle is three weeks (21 days). In other preferred modalities, each treatment cycle is six weeks (42 days).

[00398] In particular embodiments, a dose of the binding agent is administered or infused every two weeks (1Q2W), every three weeks (1Q3W) or every four weeks (1Q4W), every five weeks (1Q5W), every six weeks (1Q6W), preferably every three weeks (1Q3W) or every six weeks (1Q6W).

[00399] In particular embodiments, a dose of the binding agent and a dose of the PD-1 inhibitor are administered or infused every two weeks (1Q2W), every three weeks (1Q3W) or every four weeks (1Q4W), every five weeks (1Q5W), every six weeks (1Q6W), preferably every three weeks (1Q3W) or every six weeks (1Q6W).

[00400] In some embodiments, one dose or each dose is administered or infused on day 1 of each treatment cycle. For example, a dose of the binding agent and a dose of the PD-1 inhibitor may be administered on day 1 of each treatment cycle.

[00401] In some embodiments, a dose of 100 mg (or about 1.25 mg / kg of body weight) of the binding agent is administered every three weeks (1Q3W).

[00402] In some embodiments, a dose of 100 mg (or approximately 1.25 mg / kg of body weight) of the binding agent is administered every three weeks (1Q3W) for one or more treatment cycles, followed by the administration of a dose of 500 mg (or approximately 6.25 mg / kg of body weight) of the binding agent every six weeks (1Q6W) for one or more treatment cycles.

[00403] In some embodiments, a dose of 100 mg (or approximately 1.25 mg / kg of body weight) of the binding agent, preferably Petition 870250088654, dated 09 / 30 / 2025, pp. 168 / 329 154 / 254 acasunlimab or a biosimilar thereof is administered every three weeks (1Q3W) for two treatment cycles, followed by administration of a 500 mg dose (or approximately 6.25 mg / kg body weight) of the binding agent every six weeks (1Q6W) for one or more treatment cycles, preferably until complete tumor regression or disease progression.

[00404] In some embodiments, a 100 mg dose (or approximately 1.25 mg / kg body weight) of the binding agent is administered every six weeks (1Q6W).

[00405] In some embodiments, a dose of 100 mg (or about 1.25 mg / kg of body weight) of the binding agent and a dose of 200 mg of the PD-1 inhibitor are administered every three weeks (1Q3W).

[00406] In some embodiments, a dose of 100 mg (or about 1.25 mg / kg of body weight) of the binding agent and a dose of 400 mg of the PD-1 inhibitor are administered every six weeks (1Q6W).

[00407] In particular embodiments, a 100 mg dose (or approximately 1.25 mg / kg of body weight) of the binding agent, which is acasunlimab or a biosimilar thereof, and a 200 mg dose of the PD-1 inhibitor, which is pembrolizumab or a biosimilar thereof, are administered every three weeks (1Q3W), such as on day one of each three-week treatment cycle.

[00408] In particular embodiments, a 100 mg dose (or approximately 1.25 mg / kg of body weight) of the binding agent, which is acasunlimab or a biosimilar thereof, and a 400 mg dose of the PD-1 inhibitor, which is pembrolizumab or a biosimilar thereof, are administered every six weeks (1Q6W), such as on day one of each six-week treatment cycle.

[00409] The PD-1 inhibitor may be administered first, followed by the binding agent. Alternatively, the binding agent is administered first, followed by the PD-1 inhibitor. Petition 870250088654, dated 09 / 30 / 2025, pp. 169 / 329 155 / 254

[00410] Each dose may be administered or infused over a minimum of 30 minutes, as well as a minimum of 60 minutes, a minimum of 90 minutes, a minimum of 120 minutes, or a minimum of 240 minutes.

[00411] The binding agent can be administered in particular using intravenous (IV) infusion over 30 minutes, or for a minimum of 40 minutes, a minimum of 50 minutes, or a minimum of 60 minutes.

[00412] The PD-1 inhibitor can be administered in particular as an intravenous infusion over 30 minutes, or for a minimum of 40 minutes, a minimum of 50 minutes, or a minimum of 60 minutes.

[00413] The binding agent and the PD-1 inhibitor can be administered simultaneously. In a preferred alternative embodiment, the binding agent and the PD-1 inhibitor are administered separately.

[00414] The binding agent and the PD-1 inhibitor may be administered in any suitable form (e.g., pure as such). However, it is preferable that the binding agent and the PD-1 inhibitor be administered in the form of any suitable pharmaceutical composition, as described herein. In one embodiment, at least the binding agent and the PD-1 inhibitor are administered in the form of separate pharmaceutical compositions (i.e., a pharmaceutical composition for the binding agent and a pharmaceutical composition for the PD-1 inhibitor), preferably the binding agent and the PD-1 inhibitor are administered in the form of separate pharmaceutical compositions (i.e., a pharmaceutical composition for the binding agent and a pharmaceutical composition for the PD-1 inhibitor).

[00415] A pharmaceutical composition or compound may be formulated with a carrier, excipient and / or diluent, as well as any other components suitable for pharmaceutical compositions, including known adjuvants, in accordance with conventional techniques, such as those described in Remington: The Science and Practice of Pharmacy, 19th Edition, Petition 870250088654, dated 09 / 30 / 2025, page 170 / 329 156 / 254 Gennaro, Ed., Mack Publishing Co., Easton, PA, 1995. Pharmaceutically acceptable carriers or diluents, as well as any known adjuvants and excipients, must be suitable for the binding agent and / or PD-1 inhibitor and the chosen route of administration. The suitability of carriers and other components of pharmaceutical compositions is determined based on the absence of a significant negative impact on the desired biological properties of the chosen compound or pharmaceutical composition (e.g., impact less than a substantial impact [relative inhibition of 10% or less, relative inhibition of 5% or less, etc.] on antigen binding).

[00416] A composition, in particular the pharmaceutical composition of the binding agent and the pharmaceutical composition of the PD-1 inhibitor, may include diluents, fillers, salts, buffers, detergents (e.g., a non-ionic detergent such as Tween-20 or Tween-80), stabilizers (e.g., sugars or protein-free amino acids), preservatives, solubilizers and / or other materials suitable for inclusion in a pharmaceutical composition.

[00417] Pharmaceutically acceptable carriers, excipients or diluents for therapeutic use are well known in the pharmaceutical field and are described, for example, in Remington's Pharmaceutical Sciences, Mack Co. Publisher (A. R Gennaro edition 1985).

[00418] Pharmaceutical carriers, excipients, or diluents may be selected in relation to the intended route of administration and standard pharmaceutical practice.

[00419] Pharmaceutically acceptable carriers include any and all solvents, dispersing media, coatings, antibacterial and antifungal agents, isotonic agents, antioxidants and suitable absorption retardants, and the like, that are physiologically compatible with the active compound, in particular an agent Petition 870250088654, dated 09 / 30 / 2025, page 171 / 329 157 / 254 binding and the PD-1 inhibitor.

[00420] Examples of suitable aqueous and non-aqueous carriers that can be used in (pharmaceutical) compositions include water, saline solution, phosphate-buffered saline solution, ethanol, dextrose, polyols (such as glycerol, propylene glycol, polyethylene glycol and the like) and suitable mixtures thereof, vegetable oils such as olive oil, corn oil, peanut oil, cottonseed oil and sesame oil, colloidal solutions of carboxymethylcellulose, tragacanth gum and injectable organic esters such as ethyl oleate, and / or various buffers. Other carriers are well known in the pharmaceutical field.

[00421] Pharmaceutically acceptable carriers include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. The use of such media and agents for pharmaceutically active substances is known in the art. Except to the extent that any conventional media or agent is incompatible with the active compound, their use in (pharmaceutical) compositions is contemplated.

[00422] The term “excipient”, as used herein, refers to a substance that may be present in a (pharmaceutical) composition of the present description, but which is not an active ingredient. Examples of excipients include, without limitation, carriers, binders, diluents, lubricants, thickeners, surfactants, preservatives, stabilizers, emulsifiers, buffers, flavoring agents or colorings.

[00423] The term “diluent” refers to a diluting agent and / or diluent. In addition, the term “diluent” includes any one or more of the following fluids, liquids or solids, suspensions and / or mixing media. Examples of suitable diluents include ethanol, glycerol and water.

[00424] A (pharmaceutical) composition may also comprise pharmaceutically acceptable antioxidants, for example, (1) water-soluble antioxidants such as ascorbic acid, cysteine ​​hydrochloride, bisulfate Petition 870250088654, dated 09 / 30 / 2025, page 172 / 329 158 / 254 sodium, sodium metabisulfite, sodium sulfite and the like; (2) oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol and the like; and (3) metal chelating agents, such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid and the like.

[00425] A (pharmaceutical) composition may also comprise isotonic agents, such as sugars, polyalcohols such as mannitol, sorbitol, glycerol or sodium chloride in the composition.

[00426] A (pharmaceutical) composition may also contain one or more adjuvants appropriate for the chosen route of administration, such as preservatives, wetting agents, emulsifying agents, dispersing agents, preservatives or buffers, which may increase the shelf life or effectiveness of the composition. The composition, as used in this document, may be prepared with carriers that will protect the compound against rapid release, such as a controlled-release formulation, including implants, transdermal patches and microencapsulated delivery systems. Such carriers may include gelatin, glyceryl monostearate, glyceryl distearate, biodegradable and biocompatible polymers such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters and polylactic acid, alone or with a wax, or other materials well known in the art.The methods for preparing such formulations are generally known to those skilled in the art; see, for example, Sustained and Controlled Release Drug Delivery Systems, J.R. Robinson, ed., Marcel Dekker, Inc., New York, 1978.

[00427] “Pharmaceutically acceptable salts” include, for example, acid addition salts which may, for example, be formed using a pharmaceutically acceptable acid, such as hydrochloric acid, sulfuric acid, fumaric acid, maleic acid, succinic acid, acetic acid, Petition 870250088654, dated 09 / 30 / 2025, page 173 / 329 159 / 254 benzoic acid, citric acid, tartaric acid, carbonic acid or phosphoric acid. In addition, suitable pharmaceutically acceptable salts may include alkali metal salts (e.g., sodium or potassium salts); alkaline earth metal salts (e.g., calcium or magnesium salts); ammonium (NH4+); and salts formed with suitable organic ligands (e.g., quaternary ammonium and amine cations formed using counter-anions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, alkyl sulfonate and aryl sulfonate). Illustrative examples of pharmaceutically acceptable salts include, but are not limited to, acetate, adipate, alginate, arginate, ascorbate, aspartate, benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate, bromide, butyrate, calcium edetate, camphorate, camphorsulfonate, camsylate, carbonate, chloride, citrate, clavulanate, cyclopentanopropionate, digluconate, dihydrochloride, dodecyl sulfate, edetate, edisylate, estolate, esilate,ethanesulfonate, formate, fumarate, galactate, galacturonate, gluceptate, glucoheptonate, gluconate, glutamate, glycerophosphate, glycolylarsanilate, hemisulfate, heptanoate, hexanoate, hexylresorcinate, hydrabamine, hydrobromide, hydrochloride, hydrogen iodide, 2-hydroxyethanesulfonate, hydroxynaphthoate, iodide, isobutyrate, isothionate, lactate, lactobionate, laurate, lauryl sulfate, malate, maleate, malonate, mandelate, mesylate, methanesulfonate, methylsulfate, mucate, 2-naphthalenesulfonate, napsylate, nicotinate, nitrate, ammonium salt of N-methylglucamine, oleate, oxalate, pamoate (embonate), palmitate, pantothenate, pectinate, persulfate, 3phenylpropionate, phosphate / diphosphate, phthalate, picrate, pivalate, polygalacturonate, propionate, salicylate, stearate, sulfate, suberate, succinate, tannate, tartrate, theoclate, tosylate, triethiodide, undecanoate, valerate and the like (see, for example, SM Berge et al., “Pharmaceutical Salts”, J. Pharm. Sci., 66,pp. 119 (1977)). Salts that are not pharmaceutically acceptable may be used to prepare pharmaceutically acceptable salts and are included in this description. Petition 870250088654, dated 09 / 30 / 2025, page 174 / 329 160 / 254

[00428] In one embodiment, the binding agent and PD-1 inhibitor used herein can be formulated to ensure adequate distribution in vivo. Pharmaceutically acceptable carriers for parenteral administration include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. The use of such means and agents for pharmaceutically active substances is known in the art. Except to the extent that any conventional means or agent is incompatible with the active compound, its use in the compositions is contemplated. Other active or therapeutic compounds may also be incorporated into the compositions.

[00429] Pharmaceutical compositions for injection should typically be sterile and stable under manufacturing and storage conditions. The composition may be formulated as a solution, microemulsion, liposome, or other ordered structure suitable for high drug concentrations. The vehicle may be an aqueous or non-aqueous solvent or dispersion medium containing, for example, water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like) and suitable mixtures thereof, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Adequate flowability may be maintained, for example, by the use of a coating such as lecithin, by maintaining the required particle size in the case of dispersion, and by the use of surfactants. In many cases, it will be preferable to include isotonic agents, for example, sugars, polyalcohols such as glycerol, mannitol, sorbitol, or sodium chloride in the composition.Prolonged absorption of injectable compositions can be achieved by including an absorption-retardant agent in the composition, for example, monostearate salts and gelatin. Sterile injectable solutions can be prepared by incorporating the active compound in the required quantity into a suitable solvent with one or a combination of ingredients, for example, in the manner enumerated. Petition 870250088654, dated 09 / 30 / 2025, page 175 / 329 161 / 254 previously, as needed, followed by microfiltration for sterilization. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle containing a basic dispersion medium and other necessary ingredients, for example, those listed previously. In the case of sterile powders for the preparation of sterile injectable solutions, examples of preparation methods are vacuum drying and lyophilization (freeze-drying), which produce a powder of the active ingredient plus any desired additional ingredient from a previously sterile filtered solution.

[00430] Sterile injectable solutions can be prepared by incorporating the active compounds in the required quantity into an appropriate solvent with one or a combination of ingredients listed above, as needed, followed by microfiltration for sterilization. Generally, dispersions are prepared by incorporating the ac...

Claims

1. A method for treating a tumor or cancer in an individual, characterized in that said method comprises administering to said individual a binding agent comprising a first binding region that binds to CD137 and a second binding region that binds to PD-L1, wherein said tumor or cancer exhibits high microsatellite instability (MSI-H) or deficiency in nonspecific binding repair (dMMR).

2. Method according to claim 1, characterized in that PD-L1 is human PD-L1, in particular human PD-L1 comprising the sequence presented in SEQ ID NO: 40; and / or CD137 is human CD137, in particular human CD137 comprising the sequence presented in SEQ ID NO:

38.

3. Method according to claim 1 or 2, characterized in that a) the first binding region of the binding agent comprises a variable heavy chain (VH) region comprising the CDR1, CDR2 and CDR3 sequences presented in: SEQ ID NO: 2, 3 and 4, respectively, and a variable light chain (VL) region comprising the CDR1, CDR2 and CDR3 sequences presented in: SEQ ID NO: 6, GAS and SEQ ID NO: 8, respectively; (b) the second binding region of the binding agent comprises a variable heavy chain (VH) region comprising the CDR1, CDR2 and CDR3 sequences presented in: SEQ ID NO: 12, 13, and 14, respectively, and a variable light chain (VL) region comprising the CDR1, CDR2 and CDR3 sequences presented in: SEQ ID NO: 16, DDN and SEQ ID NO: 18, respectively.

4. Method according to any of the preceding claims, characterized in that a) the first binding region of the binding agent comprises a variable heavy chain (VH) region comprising the amino acid sequence presented in SEQ ID NO: 1 and a variable light chain (VL) region comprising the amino acid sequence presented in SEQ ID NO: 5; and b) the second binding region of the binding agent comprises a variable heavy chain (VH) region comprising the amino acid sequence presented in SEQ ID NO: 11 and a variable light chain (VL) region comprising the amino acid sequence presented in SEQ ID NO:

15.

5. A method according to any of the preceding claims, characterized in that the binding agent is a multispecific antibody, such as a bispecific antibody.

6. A method according to any of the preceding claims, characterized in that the binding agent is in the form of a full-length antibody or an antibody fragment.

7. A method according to any of the preceding claims, characterized in that the binding agent is an antibody comprising a first binding arm and a second binding arm, wherein the first binding arm comprises: i) a polypeptide comprising said first variable heavy chain (VH) region and a first constant heavy chain (CH) region, and ii) a polypeptide comprising said first variable light chain (VL) region and a first constant light chain (CL) region; and the second binding arm comprises: iii) a polypeptide comprising said second variable heavy chain (VH) region and a second constant heavy chain (CH) region, and Petition 870250088654, dated 09 / 30 / 2025, page 271 / 329 3 / 19 iv) a polypeptide comprising said second variable light chain (VL) region and a second constant light chain (CL) region.

8. A method according to any of the preceding claims, characterized in that said binding agent comprises: i) a first heavy chain and light chain comprising said antigen-binding region capable of binding to CD137, the first heavy chain comprising a first constant heavy chain region and the first light chain comprising a first constant light chain region; and ii) a second heavy chain and light chain comprising said antigen-binding region capable of binding to PD-L1, the second heavy chain comprising a second constant heavy chain region and the second light chain comprising a second constant light chain region.

9. Method according to claim 7 or 8, characterized in that (i) the amino acid at the position corresponding to F405 in a human IgG1 heavy chain according to EU numbering is L in said first heavy chain constant region (CH), and the amino acid at the position corresponding to K409 in a human IgG1 heavy chain according to EU numbering is R in said second heavy chain constant region (CH), or (ii) the amino acid at the position corresponding to K409 in a human IgG1 heavy chain according to EU numbering is R in said first heavy chain, and the amino acid at the position corresponding to F405 in a human IgG1 heavy chain according to EU numbering is L in said second heavy chain.

10. Method according to any one of claims 7 to 9, characterized in that the positions corresponding to the positions L234 and L235 in a human IgG1 heavy chain according to EU numbering are F and E, respectively, in said first and second heavy chains.

11. Method according to any one of claims 7 to 10, characterized in that the positions corresponding to positions L234, L235, and D265 in a human IgG1 heavy chain according to EU numbering are F, E, and A, respectively, in said first and second heavy chain constant regions (HCs).

12. Method according to any one of claims 7 to 11, characterized in that the positions corresponding to positions L234 and L235 in a human IgG1 heavy chain according to the EU numbering of both the first and second heavy chain constant regions are F and E, respectively, and wherein (i) the position corresponding to F405 in a human IgG1 heavy chain according to the EU numbering of the first heavy chain constant region is L, and the position corresponding to K409 in a human IgG1 heavy chain according to the EU numbering of the second heavy chain is R, or (ii) the position corresponding to K409 in a human IgG1 heavy chain according to the EU numbering of the first heavy chain constant region is R, and the position corresponding to F405 in a human IgG1 heavy chain according to the EU numbering of the second heavy chain is L.

13. Method according to any one of claims 7 to 12, characterized in that the positions corresponding to positions L234, L235, and D265 in a human IgG1 heavy chain according to the EU numbering of both the first and second constant regions of the heavy chain are F, E, and A, respectively, and wherein (i) the position corresponding to F405 in a human IgG1 heavy chain according to the EU numbering of the first constant region of the heavy chain is L, and the position corresponding to K409 in a human IgG1 heavy chain according to Petition 870250088654, dated 09 / 30 / 2025, p. 273 / 329 5 / 19 the EU numbering of the second heavy chain constant region is R, or (ii) the position corresponding to K409 in a human IgG1 heavy chain according to the EU numbering of the first heavy chain is R, and the position corresponding to F405 in a human IgG1 heavy chain according to the EU numbering of the second heavy chain is L.

14. A method according to any one of claims 7 to 13, characterized in that the constant region of said first and / or second heavy chain, such as the second heavy chain, comprises or consists essentially of, or consists of, an amino acid sequence selected from the group consisting of: a) the sequence presented in SEQ ID NO: 24 or 30 [IgG1Fc_FEAL]; b) a subsequence of the sequence in a), such as a subsequence in which 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 consecutive amino acids have been deleted, starting from the N-terminus or C-terminus of the sequence defined in a); and c) a sequence with at most 6 substitutions, such as at most 5 substitutions, at most 4 substitutions, at most 3, at most 2 substitutions or at most 1 substitution, compared to the amino acid sequence defined in a) or b).

15. Method according to any one of claims 7 to 14, characterized in that the constant region of said first and / or second heavy chain, such as the first heavy chain, comprises or consists essentially of, or consists of, an amino acid sequence selected from the group consisting of a) the sequence presented in SEQ ID NO: 23 or 29 [IgG1Fc_FEAR]; b) a subsequence of the sequence in a), such as a subsequence, in which 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 consecutive amino acids have been deleted, starting from the N-terminal or C-terminal of the sequence defined in a); (e) a sequence with at most 6 substitutions, such as at most 5 substitutions, at most 4, at most 3, at most 2 substitutions, or at most 1 substitution, compared to the amino acid sequence defined in (a) or (b).

16. Method according to any one of claims 7 to 15, characterized in that said linking agent comprises a constant region of kappa (κ) light chain.

17. Method according to any one of claims 7 to 16, characterized in that said linking agent comprises a constant region of lambda (λ) light chain.

18. Method according to any one of claims 7 to 17, characterized in that said first light chain constant region is a kappa (κ) light chain constant region or a lambda (λ) light chain constant region.

19. Method according to any one of claims 7 to 18, characterized in that said second light chain constant region is a lambda (λ) light chain constant region or a kappa (κ) light chain constant region.

20. Method according to any one of claims 7 to 19, characterized in that said first light chain constant region is a kappa (κ) light chain constant region and said second light chain constant region is a lambda (λ) light chain constant region, or said first light chain constant region is a lambda (λ) light chain constant region and said second light chain constant region is a kappa (κ) light chain constant region.

21. Method according to any one of claims 16 to 20, characterized in that the kappa (κ) light chain comprises Petition 870250088654, dated 09 / 30 / 2025, p. 275 / 329 7 / 19 an amino acid sequence selected from the group consisting of: a) the sequence presented in SEQ ID NO:35, b) a subsequence of the sequence in a), such as a subsequence, in which 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 consecutive amino acids have been deleted, starting from the N-terminal or C-terminal of the sequence defined in a); (e) a sequence with at most 10 substitutions, such as at most 9 substitutions, at most 8, at most 7, at most 6, at most 5, at most 4 substitutions, at most 3, at most 2 substitutions, or at most 1 substitution, compared to the amino acid sequence defined in (a) or (b).

22. A method according to any one of claims 17 to 21, characterized in that the lambda (λ) light chain comprises an amino acid sequence selected from the group consisting of a) the sequence presented in SEQ ID NO: 36, b) a subsequence of the sequence in a), such as a subsequence in which 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 consecutive amino acids have been deleted, starting from the N-terminus or C-terminus of the sequence defined in a); and c) a sequence with at most 10 substitutions, such as at most 9 substitutions, at most 8, at most 7, at most 6, at most 5, at most 4 substitutions, at most 3, at most 2 substitutions or at most 1 substitution, compared to the amino acid sequence defined in a) or b).

23. A method according to any of the preceding claims, characterized in that the binding agent is of an isotype selected from the group consisting of IgG1, IgG2, IgG3 and IgG4.

24. Method according to any of the preceding claims, characterized in that the binding agent is a full-length IgG1 antibody. Petition 870250088654, dated 09 / 30 / 2025, page 276 / 329 8 / 19 25. Method according to any of the preceding claims, characterized in that the binding agent is an antibody of the IgG1m(f) allotype.

26. Method according to any of the preceding claims, characterized in that the binding agent is a bispecific antibody that binds to CD137 and PD-L1, the bispecific antibody having i) a first heavy chain comprising the amino acid sequence presented in SEQ ID NO: 31 and a first light chain comprising the amino acid sequence presented in SEQ ID NO: 32, and ii) a second heavy chain comprising the amino acid sequence presented in SEQ ID NO: 33 and a second light chain comprising the amino acid sequence shown in SEQ ID NO:

34.

27. A method according to any of the preceding claims, characterized in that the binding agent is acasunlimab or a biosimilar thereof.

28. A method according to any of the preceding claims, characterized in that the binding agent is in a composition or formulation comprising histidine, sucrose and Polysorbate 80, and has a pH of 5 to 6.

29. A method according to any of the preceding claims, characterized in that the binding agent is in a composition or formulation comprising about 20 mM histidine, about 250 mM sucrose, about 0.02% Polysorbate-80, and having a pH of about 5.

5.

30. Method according to any of the preceding claims, characterized in that the binding agent is in a composition or formulation comprising 10 to 30 mg of binding agent / mL, such as 20 mg of binding agent / mL. Petition 870250088654, dated 09 / 30 / 2025, p. 277 / 329 9 / 19 31. A method according to any one of the preceding claims, characterized in that the binding agent is in a composition as defined in any one of claims 28 to 30 and is diluted in 0.9% NaCl (saline) before administration.

32. A method according to any of the preceding claims, characterized in that it comprises additionally administering a PD-1 inhibitor to said individual.

33. Method according to claim 32, characterized in that PD-1 is human PD-1, preferably PD-1 has or comprises the amino acid sequence as presented in SEQ ID NO: 113 or SEQ ID NO: 114, or the amino acid sequence of PD-1 has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% identity with the amino acid sequence as presented in SEQ ID NO: 113 or SEQ ID NO: 114, or is an immunogenic fragment thereof.

34. Method according to claim 32 or 33, characterized in that the PD-1 inhibitor is an antibody that binds to PD-1 or PD-L1, preferably an antibody that is an antagonist of the PD-1 / PD-L1 interaction and / or is a PD-1 or PD-L1 blocking antibody.

35. Method according to any one of claims 32 to 34, characterized in that the PD-1 inhibitor is an antibody of an isotype selected from the group consisting of IgG1, IgG2, IgG3 and IgG4, such as the IgG1 isotype.

36. Method according to any one of claims 32 to 35, characterized in that the PD-1 inhibitor is a full-length antibody or antibody fragment, such as a full-length IgG1 antibody.

37. Method according to any of the claims in Petition 870250088654, dated 09 / 30 / 2025, pp. 278 / 329 10 / 19 32 to 36, characterized in that the PD-1 inhibitor is a monospecific antibody.

38. Method according to any one of claims 32 to 37, characterized in that said PD-1 inhibitor is an antibody that binds to PD-1 comprising a variable heavy chain (VH) region comprising the CDR1, CDR2 and CDR3 sequences presented in SEQ ID NO: 59, 60 and 61, respectively, and a variable light chain (VL) region comprising the CDR1, CDR2 and CDR3 sequences presented in SEQ ID NO: 62, LAS and SEQ ID NO: 64, respectively.

39. Method according to any one of claims 32 to 38, characterized in that said PD-1 inhibitor is an antibody that binds to PD-1 comprising a VH region comprising the amino acid sequence of SEQ ID NO: 65 and a VL region comprising the amino acid sequence of SEQ ID NO:

66.

40. Method according to any one of claims 32 to 39, characterized in that said PD-1 inhibitor is an antibody that binds to PD-1 comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 67 and a light chain comprising the amino acid sequence of SEQ ID NO:

68.

41. A method according to any one of claims 32 to 40, characterized in that said PD-1 inhibitor is pembrolizumab or a biosimilar thereof.

42. Method according to any one of claims 32 to 41, characterized in that the binding agent is acasunlimab or a biosimilar thereof, and said PD-1 inhibitor is pembrolizumab or a biosimilar thereof.

43. Method according to any one of claims 32 to 37, characterized in that said PD-1 inhibitor is an antibody that binds to PD-1, or an antigen-binding fragment thereof, as per Petition 870250088654, dated 09 / 30 / 2025, page 279 / 329 11 / 19, wherein said PD-1-binding antibody comprises a VH CDR1, CDR2, and CDR3 region comprising the sequences as presented in SEQ ID Nos: 104, 101, and 100, respectively, and a VL CDR1, CDR2, and CDR3 region comprising the sequences as presented in SEQ ID NO: 107, QAS, and SEQ ID NO: 105, respectively.

44. Method according to claim 43, characterized in that the antibody binding to PD-1 comprises a variable heavy chain (VH) region comprising a sequence with at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% identity with the amino acid sequence of the VH sequence as presented in SEQ ID NO:

111.

45. Method according to claim 44, characterized in that the antibody binding to PD-1 comprises a variable heavy chain (VH) region, wherein the VH comprises the sequence in the manner presented in SEQ ID NO:

111.

46. ​​Method according to any one of claims 43 to 45, characterized in that the PD-1 binding antibody comprises a variable light chain (VL) region comprising a sequence with at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% identity with the amino acid sequence of the VL sequence as presented in SEQ ID NO:

112.

47. Method according to claim 46, characterized in that the antibody binding to PD-1 comprises a variable light chain (VL) region, wherein the VL comprises the sequence in the manner presented in SEQ ID NO:

112.

48. Method according to any one of claims 43 to 47, characterized in that the antibody that binds to PD-1 Petition 870250088654, dated 09 / 30 / 2025, page 280 / 329 12 / 19 comprises a variable heavy chain (VH) region and a variable light chain (VL) region, wherein the VH comprises or presents the sequence in the manner presented in SEQ ID NO: 111, and the VL comprises or presents the sequence in the manner presented in SEQ ID NO:

112.

49. Method according to any one of claims 43 to 48, characterized in that the PD-1 binding antibody comprises a constant heavy chain region, wherein the amino acid corresponding to position L234 in a human IgG1 heavy chain according to EU numbering is phenylalanine, the amino acid corresponding to position L235 in a human IgG1 heavy chain according to EU numbering is glutamate, and the amino acid corresponding to position G236 in a human IgG1 heavy chain according to EU numbering is arginine in said constant heavy chain region of the PD-1 binding antibody (L234F / L235E / G236R).

50. Method according to any one of claims 43 to 49, characterized in that the constant region of the antibody heavy chain that binds to PD-1 comprises a sequence with at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% identity with the amino acid sequence of the HC sequence in the manner shown in SEQ ID NO:

93.

51. Method according to any one of claims 43 to 50, characterized in that the constant region of the antibody heavy chain that binds to PD-1 comprises the sequence in the manner presented in SEQ ID NO:

93.

52. Method according to any one of claims 43 to 51, characterized in that the isotype of the antibody heavy chain constant region that binds to PD-1 is IgG1.

53. Method according to any of the claims in Petition 870250088654, dated 09 / 30 / 2025, pp. 281 / 329 13 / 19 43 to 52, characterized in that the antibody that binds to PD-1 is a monoclonal, chimeric or humanized antibody or a fragment of such an antibody.

54. A method according to any of the preceding claims, characterized in that the binding agent is acasunlimab or a biosimilar thereof, and said PD-1 inhibitor is an antibody that binds to PD-1 comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 152 and a light chain comprising the amino acid sequence of SEQ ID NO:

153.

55. Method according to any one of claims 32 to 36, characterized in that the PD-1 inhibitor is a multispecific antibody, just like a bispecific antibody.

56. A method according to any one of claims 32 to 37, characterized in that the PD-1 inhibitor is Pembrolizumab, Nivolumab, Cemiplimab, Dostarlimab, Espartalizumab, Camrelizumab, Sintilimab, Tislelizumab, Toripalimab, Retifanlimab, Pidilizumab, AMP-224, AMP-514, Atezolizumab, Avelumab, Durvalumab, Envafolimab, Cosibelimab, AUNP12, CA-170, BMS-986189, or a respective biosimilar thereof.

57. A method according to any one of claims 32 to 37, characterized in that the PD-1 inhibitor is Pembrolizumab, Nivolumab, Cemiplimab, Dostarlimab, Espartalizumab, Camrelizumab, Sintilimab, Tislelizumab, Toripalimab, Retifanlimab, Pidilizumab, AMP-514, Atezolizumab, Avelumab, Durvalumab, Envafolimab, Cosibelimab, or a respective biosimilar thereof.

58. Method according to any one of claims 32 to 37, characterized in that the PD-1 inhibitor is Pembrolizumab, Nivolumab, Cemiplimab, Dostarlimab, Espartalizumab, Camrelizumab, Sintilimab, Tislelizumab, Toripalimab, Retifanlimab, Pidilizumab, Petition 870250088654, dated 09 / 30 / 2025, page 282 / 329 14 / 19 AMP-514, or a respective biosimilar thereof.

59. A method according to any of the preceding claims, characterized in that the individual is a human individual.

60. A method according to any of the preceding claims, characterized in that the binding agent is administered in at least one treatment cycle, each treatment cycle being either three weeks (21 days) or six weeks (42 days).

61. Method according to any of the preceding claims, characterized in that a dose of the binding agent is administered every three weeks (1Q3W) or every six weeks (1Q6W).

62. A method according to any of the preceding claims, characterized in that a dose of the binding agent is administered on day 1 of each treatment cycle.

63. Method according to any of the preceding claims, characterized in that the amount of said binding agent administered in each dose and / or in each treatment cycle is 100 mg or 500 mg.

64. Method according to any of the preceding claims, characterized in that a dose of 100 mg of the binding agent is administered every three weeks (1Q3W).

65. A method according to any of the preceding claims, characterized in that a 100 mg dose of the binding agent is administered every three weeks (1Q3W) for two treatment cycles, followed by the administration of a 500 mg dose of the binding agent every six weeks (1Q6W) for one or more treatment cycles, preferably until complete tumor regression or disease progression.

66. Method according to any one of claims 32 to 64, characterized in that the PD-1 inhibitor is administered in at least one treatment cycle, each treatment cycle being three Petition 870250088654, dated 09 / 30 / 2025, p. 283 / 329 15 / 19 weeks (21 days) or six weeks (42 days).

67. Method according to any one of claims 32 to 64, 66, characterized in that a dose of the PD-1 inhibitor is administered every three weeks (1Q3W) or every six weeks (1Q6W).

68. Method according to any one of claims 32 to 64, 66, 67, characterized in that a dose of the PD-1 inhibitor is administered on day 1 of each treatment cycle.

69. Method according to any one of claims 32 to 64, 66 to 68, characterized in that the amount of said PD-1 inhibitor administered in each dose and / or in each treatment cycle is 200 mg or 400 mg.

70. Method according to any one of claims 32 to 64, 66 to 69, characterized in that a dose of 100 mg of the binding agent and a dose of 200 mg of the PD-1 inhibitor are administered every three weeks (1Q3W).

71. Method according to any one of claims 32 to 64, 66 to 70, characterized in that a dose of 100 mg of the binding agent, which is acasunlimab or a biosimilar thereof, and a dose of 200 mg of the PD-1 inhibitor, which is pembrolizumab or a biosimilar thereof, are administered every three weeks (1Q3W), such as on day one of each three-week treatment cycle.

72. Method according to any one of claims 32 to 64, 66 to 69, characterized in that a dose of 100 mg of the binding agent and a dose of 400 mg of the PD-1 inhibitor are administered every six weeks (1Q6W).

73. Method according to any one of claims 32 to 64, 66 to 69, 72, characterized in that a dose of 100 mg of the binding agent, which is acasunlimab or a biosimilar thereof, and a dose of 400 mg of the PD-1 inhibitor, which is pembrolizumab or a biosimilar thereof, are administered every six weeks (1Q6W), such as on day one of each six-week treatment cycle.

74. Method according to any one of claims 32 to 73, characterized in that the PD-1 inhibitor is administered first, followed by the binding agent, preferably the administration of the binding agent begins at least 30 minutes after the end of the administration of the PD-1 inhibitor.

75. A method according to any of the preceding claims, characterized in that said tumor or cancer is a solid tumor or leukemia, preferably a solid tumor.

76. A method according to any of the preceding claims, characterized in that said tumor or cancer is selected from the group consisting of colorectal cancer, gastric cancer, endometrial cancer, ovarian cancer, hepatobiliary tract cancer, pancreatic cancer, urinary tract cancer, bladder cancer, thyroid cancer, breast cancer, prostate cancer, ovarian cancer, central nervous system (CNS) cancer, and skin cancer such as melanoma, preferably selected from the group consisting of colorectal cancer, gastric cancer, and endometrial cancer.

77. Method according to claim 76, characterized in that said tumor or cancer is endometrial cancer.

78. Method according to claim 76, characterized in that said tumor or cancer is colorectal cancer.

79. Method according to claim 76, characterized in that said tumor or cancer is gastric cancer.

80. A method according to any of the preceding claims, characterized in that said tumor or cancer is unresectable, recurrent and / or metastatic.

81. Method according to any of the preceding claims Petition 870250088654, dated 09 / 30 / 2025, p. 285 / 329 17 / 19, characterized in that said individual experienced progression during or after at least 1 prior line of treatment regimen for said unresectable and / or metastatic tumor or cancer, preferably systemic chemotherapy such as platinum-based chemotherapy.

82. A method according to any of the preceding claims, characterized in that the individual has not received prior treatment with a checkpoint inhibitor, such as an anti-PD1 antibody, an anti-PD-L1 antibody, an anti-CTLA-4 antibody, an anti-LAG3 antibody, or an anti-TIGIT antibody.

83. A method according to any one of claims 1 to 81, characterized in that the individual has received prior treatment with a PD-1 inhibitor or a PD-L1 inhibitor, such as an anti-PD-1 antibody or an anti-PD-L1 antibody, the PD-1 inhibitor or PD-L1 inhibitor being administered as monotherapy or as part of a combination therapy.

84. Method according to claim 83, characterized in that the individual showed progression after treatment with a PD-1 inhibitor or a PD-L1 inhibitor, such as an anti-PD-1 antibody or an anti-PD-L1 antibody.

85. A binding agent, characterized by being for use in a method for treating a tumor or cancer in an individual, said method comprising administering to said individual the binding agent comprising a first binding region that binds to CD137 and a second binding region that binds to PD-L1, wherein said tumor or cancer exhibits high microsatellite instability (MSI-H) or deficiency in nonspecific binding repair (dMMR).

86. Bonding agent, characterized in that it is for use as defined in claim 85, wherein the method is as defined in Petition 870250088654, dated 09 / 30 / 2025, p. 286 / 329 18 / 19 any of claims 1 to 84, and / or the bonding agent is as defined in any of claims 1 to 84.

87. Pharmaceutical composition, characterized by being for use in a method for treating a tumor or cancer in an individual, comprising a binding agent comprising a first binding region that binds to CD137 and a second binding region that binds to PD-L1, and optionally a pharmaceutically acceptable carrier, wherein said tumor or cancer exhibits high microsatellite instability (MSI-H) or deficiency in nonspecific binding repair (dMMR).

88. Pharmaceutical composition, characterized in that it is for use as defined in claim 87, wherein the method is as defined in any one of claims 1 to 84, and / or the binding agent is as defined in any one of claims 1 to 84.

89. Use of a binding agent, characterized by being for the manufacture of a medicament to treat a tumor or cancer in an individual, wherein said tumor or cancer exhibits high microsatellite instability (MSI-H) or deficiency in nonspecific binding repair (dMMR), wherein the binding agent comprises a first binding region that binds to CD137 and a second binding region that binds to PD-L1.

90. Use of the linking agent according to claim 89, characterized in that the linking agent is as defined in any one of claims 1 to 84.

91. Kit, characterized by being for use in a method for treating a tumor or cancer in an individual, wherein said tumor or cancer exhibits high microsatellite instability (MSI-H) or deficiency in nonspecific binding repair (dMMR), comprising a) a binding agent comprising a first binding region that binds to CD137 and a second binding region that binds to PD-L1, and b) a PD-L1 inhibitor. Petition 870250088654, dated 09 / 30 / 2025, pp. 287 / 329 19 / 19 92. Kit, characterized in that it is for use as defined in claim 91, wherein the method is as defined in any one of claims 1 to 84, and / or the binding agent is as defined in any one of claims 1 to 84, and / or the PD-1 inhibitor is as defined in any one of claims 1 to 84.

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