Methods of treating cancer using bispecific EGFR x CD28 antibodies alone or in combination with anti-PD-1 antibodies

Abstract

The present disclosure provides methods of treating cancer using a multispecific antibody (EGFRxCD28), or an antigen-binding fragment thereof, that binds to EGFR and CD28. Such antibodies can be combined with another therapeutic agent, such as an anti-PD-1 antibody, such as a cemiplimab. Also provided are methods of treating cancer (e.g., EGFR-expressing cancer) by administering the antibodies (e.g., and combinations thereof with anti-PD-1).

Description

[0001] Related Applications

[0002] This application claims priority to U.S. Provisional Application No. 63 / 378,102, filed on October 3, 2022; U.S. Provisional Application No. 63 / 380,991, filed on October 26, 2022; and U.S. Provisional Application No. 63 / 495,189, filed on April 10, 2023. The entire content of each of the foregoing applications is hereby expressly incorporated by reference. Technical Field

[0003] The present disclosure relates to methods of treating or preventing cancer (e.g., EGFR-expressing cancer) in a subject (e.g., a human) in need thereof, comprising administering to the subject in combination an effective amount of a bispecific antibody or antigen-binding fragment thereof that binds epidermal growth factor (EGF) receptor and CD28, and an anti-PD-1 antibody or antigen-binding fragment thereof.

[0004] Sequence Listing

[0005] This application contains a Sequence Listing, which has been submitted electronically in XML format and is hereby incorporated by reference in its entirety. The XML copy, created on October 1, 2023, is named 118003-91020.XML and is 117,441 bytes in size. Background Art

[0006] The ability of T cells to recognize and kill their cellular targets (e.g., virus-infected cells or tumor cells) depends on a collection of co-interactions. The most important of these interactions is the recognition and binding of the target cell by the T cell receptor (TCR) complex (which includes associated CD3γ, δ, ε, and ζ chains), and this interaction is referred to as "signal 1", which is used for T cell activation. The TCR recognizes viral or tumor peptides presented in the groove of MHC proteins, which are expressed on the surface of the target cell. Since such binding generally has low affinity, successful triggering of "signal 1" requires the aggregation of many TCR complexes along the interface between the T cell and its target cell; this interface is called the "immunological synapse". T cell activation can be further promoted by other interactions. For example, T cells have a molecule called CD28 on their surface, which can provide a co-stimulatory "signal 2" to enhance activation via the TCR complex. T cell activation is enhanced when the T cell recognizes its target cell via its TCR complex and then also engages "signal 2" by binding to its cognate ligand on the target cell via CD28; like "signal 1", CD28-mediated "signal 2" is thought to occur through co-aggregation at the immunological synapse.

[0007] Agonistic anti-CD28 monoclonal antibodies can be applied to the continuous ex vivo expansion of cultured T cells; however, the use of antibodies against CD28 has been hampered because a series of acute and severe adverse events occurred in a phase I clinical trial of systemic testing of superagonist anti-CD28 monoclonal antibodies (Hünig, Nature Reviews Immunology. 2012; 12: 317-318). Local or targeted use of anti-CD28 monoclonal antibodies can promote anti-tumor immunity with lower risk. Jung et al., Int J Cancer. January 15, 2001; 91(2): 225-30.

[0008] It has been confirmed that families of different growth factors and growth factor receptors are associated with the spontaneous growth of cancer cells. Among them, the epidermal growth factor receptor (EGFR) and the EGF family of peptide growth factors play a central role in the pathogenic mechanisms and progression of different cancer types. The receptor family to which EGFR belongs encompasses three additional proteins, namely ErbB-2, ErbB-3, and ErbB-4. These proteins and the growth factors of the EGF family form an integrated system in which signals hitting individual receptor types are usually transmitted to other receptors of the same family.

[0009] Monoclonal antibodies (mAbs) aimed at enhancing T cell activation are being clinically developed as anti-tumor therapeutic agents. However, most current treatments are difficult to overcome the inhibitory nature of the tumor microenvironment, and thus cannot produce efficient tumor-specific T cell activation and subsequent tumor cell killing. Several blocking mAbs against checkpoint inhibitors (such as cytotoxic T lymphocyte-associated protein-4 (CTLA-4) and programmed cell death 1 (PD-1) / programmed cell death ligand 1 (PD-L1)) have been clinically approved for melanoma, renal cell carcinoma, non-small cell lung cancer, and advanced metastatic cutaneous squamous cell carcinoma. Blocking PD-1 eliminates the disruption of T cell activation, but its efficacy as a single agent is usually not sufficient to achieve tumor clearance and a durable anti-tumor response. SUMMARY OF THE INVENTION

[0010] The present disclosure provides a method for treating cancer in a subject, the method comprising administering to the subject a therapeutically effective amount of a combination of the following: a bispecific antibody or an antigen-binding fragment thereof, the bispecific antibody or antigen-binding fragment thereof comprising a first antigen-binding domain that binds to cluster of differentiation factor 28 (CD28) and a second antigen-binding domain that binds to epidermal growth factor receptor (EGFR); an antibody or an antigen-binding fragment thereof that specifically binds to programmed death receptor-1 (PD-1), thereby treating the cancer in the subject.

[0011] In some embodiments, the cancer is a solid tumor. In certain embodiments, the cancer is a cancer that expresses EGFR. In some embodiments, the cancer is selected from esophageal carcinoma, lung squamous cell carcinoma, lung adenocarcinoma, cervical cancer (including cervical squamous cell carcinoma), endometrial adenocarcinoma, bladder cancer, urothelial carcinoma, lung cancer, non-small cell lung cancer (NSCLC), colorectal cancer (such as microsatellite-stable colorectal cancer), sigmoid adenocarcinoma, rectal cancer, endometrial cancer, skin cancer, head and neck squamous cell carcinoma, brain cancer, glioblastoma multiforme, non-CNS tumors, cutaneous squamous cell carcinoma, breast cancer, gastric cancer, gastroesophageal cancer, gastroesophageal adenocarcinoma, pancreatic cancer, prostate cancer, ovarian cancer, melanoma, nasopharyngeal cancer, anal cancer, mesothelioma, renal cell carcinoma, gallbladder / bile duct cancer, pancreatic carcinoma, penile squamous cell carcinoma, or vulvovaginal cancer. In one embodiment, the cancer is cervical cancer. In one embodiment, the cervical cancer is cervical squamous cell carcinoma. In one embodiment, the cancer is microsatellite-stable colorectal cancer with liver metastasis. In one embodiment, the cancer is microsatellite-stable colorectal cancer without liver metastasis. In one embodiment, the cancer is microsatellite-stable colorectal cancer (MSS-CRC) with active liver and / or peritoneal metastasis. In one embodiment, the cancer is MSS-CRC with lung / lymph node metastasis. In one embodiment, the cancer is EGFR-mutated NSCLC after third-generation tyrosine kinase inhibitor (TKI). In one embodiment, the cancer is EGFR-mutated NSCLC after third-generation TKI and platinum doublet chemotherapy. In one embodiment, the cancer is cutaneous squamous cell carcinoma. In one embodiment, the cancer is triple-negative breast cancer.

[0012] In some embodiments, the method further comprises selecting a subject, wherein the subject has advanced solid tumors. In some embodiments, the subject meets at least one of the following criteria, or is selected based on at least one of the following criteria: (1) has metastatic disease or locally advanced disease and is not a candidate for curative surgery or curative radiotherapy; (2) is not a candidate for an approved anti-PD-1 or PD-L1 therapy indication, or such therapy is not available to the subject for other reasons (alone or in combination); (3) has exhausted all treatment options expected to provide meaningful clinical benefit due to disease recurrence, disease refractoriness, or intolerance, except for subjects with malignancies in which anti-PD-1 / PD-L1 therapy has shown clinical benefit; and / or (4) has any of the following cancer types: (a) colorectal cancer with local pathology documented as microsatellite stable; (b) gastric cancer or gastroesophageal junction cancer; (c) esophageal cancer; (d) breast cancer (ductal or lobular carcinoma, regardless of receptor status); (e) non-small cell lung cancer (NSCLC) (any PD-L1 expression); (f) head and neck squamous cell carcinoma (SCC); (g) nasopharyngeal carcinoma; (h) cervical cancer; (i) anal cancer; (j) mesothelioma; (k) prostatic adenocarcinoma; (l) renal cell carcinoma (chromophobe renal cell carcinoma, clear cell renal cell carcinoma, or papillary renal cell carcinoma); (m) gallbladder / bile duct cancer; (n) urothelial carcinoma; (o) pancreatic neuroendocrine tumor; (p) penile SCC; (q) vulvovaginal cancer; or (r) other non-CNS tumor types showing elevated EGFR expression in the tumor.

[0013] In some embodiments, the subject has been treated with a prior therapy selected from: radiotherapy, surgery, chemotherapy, PD-1 inhibitor, PD-L1 inhibitor, anti-VEGF therapy, CAR-T therapy, and / or anti-EGFR therapy. In some embodiments, the subject has not received a prior anti-PD-1 therapy or anti-PD-L1 therapy.

[0014] In some embodiments, the subject has microsatellite stable colorectal cancer (MSS CRC). In some embodiments, a subject with microsatellite stable colorectal cancer has at least one of the following attributes, or is selected based on at least one of the following attributes: (a) has metastatic CRC; (b) is not a candidate for curative surgery or curative radiotherapy; (c) may have active metastases in the liver and / or peritoneum at screening; (d) no active metastases are identified in the liver or peritoneum at screening and the disease site is present only in one or more lungs and / or pulmonary lymph nodes; (e) the pathology report is documented as microsatellite stable; (f) has received at least one line of therapy in the setting of recurrence / metastasis, wherein the therapy includes anti-EGFR therapy or anti-VEGF therapy; or (g) has never received anti-PD-1 / PD-L1, defined as never having been treated with a drug targeting PD-1.

[0015] In some embodiments, the subject has triple-negative breast cancer (TNBC). In some embodiments, the subject with TNBC has at least one of the following attributes, or is selected based on at least one of the following attributes: (a) has metastatic TNBC; (b) is not a candidate for radical surgery or radical radiotherapy; (c) is not a candidate for an indication of an approved anti-PD-1 or anti-PD-L1 therapy, or such therapy is not available to the subject for other reasons; (d) the pathology report records having triple-negative cancer (ER- / PR- / Her2-); or (e) has not received anti-PD-1 / PD-L1, defined as never having been treated with a drug targeting PD-1 previously.

[0016] In some embodiments, the subject has cutaneous squamous cell carcinoma (CSCC), where (i) the subject is not a candidate for radical surgery or radical radiotherapy or (ii) the subject has not received anti-PD-1 / PD-L1, defined as never having been treated with a drug targeting PD-1 previously.

[0017] In some embodiments, the subject has non-small cell lung cancer (NSCLC). In some embodiments, the subject has at least one of the following attributes, or is selected based on at least one of the following attributes: (a) the subject has locally advanced or metastatic EGFR-mutated non-squamous NSCLC disease previously histologically or cytologically recorded; (b) has advanced or metastatic NSCLC; (c) is not a candidate for radical surgery or radical radiotherapy; (d) has a previously recorded targetable EGFR mutation (EGFR exon 19 deletion, EGFR L858R mutation, EGFR exon 20 insertion, or exon 18 / 21 atypical mutation); (e) has not received chemotherapy; (f) has received treatment with platinum doublet chemotherapy; (e) has received treatment with a third-generation TKI; or (g) has not received anti-PD-1 / PD-L1, defined as never having been treated with a drug targeting PD-1 previously.

[0018] In some embodiments, the subject has head and neck squamous cell carcinoma (HNSCC). In some embodiments, the subject has at least one of the following attributes, or is selected based on at least one of the following attributes: (a) has advanced or metastatic disease; (b) is not a candidate for curative surgery or curative radiotherapy; (c) has a CPS of PD-L1 expression ≥ 1% by local IHC assay; (d) has not received prior systemic treatment for recurrent or metastatic HNSCC; and / or (e) has never received anti-PD-1 / PD-L1, defined as never having been treated with a drug targeting PD-1 previously.

[0019] In certain embodiments, the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof is administered at a dose of about 0.1 mg to about 3000 mg. In some embodiments, the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof is administered at a dose of about 0.01 mg, 0.03 mg, 0.05 mg, 0.1 mg, 0.3 mg, 0.5 mg, 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 8 mg, 10 mg, 15 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, 600 mg, 650 mg, 700 mg, 750 mg, 800 mg, 850 mg, 900 mg, 1000 mg, 1200 mg, 1500 mg, 1800 mg, 2000 mg, 2400 mg, 2700 mg, or 3000 mg.

[0020] In certain embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof is administered at a dose of about 50 mg to about 1500 mg. In some embodiments, the anti-PD-1 antibody is administered at a dose of 350 mg.

[0021] In some embodiments, the method comprises administering in combination one or more doses of the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof and one or more doses of the anti-PD-1 antibody or antigen-binding fragment thereof.

[0022] In some embodiments, each of one or more doses of the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof is from about 0.1 mg to about 3000 mg. In some embodiments, each of one or more doses is about 0.01 mg, 0.03 mg, 0.05 mg, 0.1 mg, 0.3 mg, 0.5 mg, 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 8 mg, 10 mg, 15 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, 600 mg, 650 mg, 700 mg, 750 mg, 800 mg, 850 mg, 900 mg, 1000 mg, 1200 mg, 1500 mg, 1800 mg, 2000 mg, 2400 mg, 2700 mg or 3000 mg.

[0023] In some embodiments, each of one or more doses of the anti-PD-1 antibody or antigen-binding fragment thereof is from about 50 mg to about 1500 mg. In some embodiments, each of one or more doses of the anti-PD-1 antibody is 350 mg.

[0024] In certain embodiments, each of one or more doses of the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof and / or each of one or more doses of the anti-PD-1 antibody or antigen-binding fragment thereof is administered 0.5 to 14 weeks after the previous dose.

[0025] In some embodiments, each of one or more doses of the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof and / or each of one or more doses of the anti-PD-1 antibody or antigen-binding fragment thereof is administered once a week, once every two weeks, once every three weeks, once every four weeks, once every five weeks or once every six weeks.

[0026] In some embodiments, each dose of the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof is administered once a week. In some embodiments, each of one or more doses of the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof is administered once every three weeks.

[0027] In some embodiments, each of one or more doses of the anti-PD-1 antibody or antigen-binding fragment thereof is administered once every three weeks.

[0028] In certain embodiments, the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof and / or the anti-PD-1 antibody or antigen-binding fragment thereof are administered intravenously. In some embodiments, the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof and / or the anti-PD-1 antibody or antigen-binding fragment thereof are administered subcutaneously.

[0029] In certain embodiments, the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof and the anti-PD-1 antibody or antigen-binding fragment thereof are administered on the same day. In some embodiments, the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof and the anti-PD-1 antibody or antigen-binding fragment thereof are administered on different days.

[0030] In certain embodiments, the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof is administered before or after the anti-PD-1 antibody or antigen-binding fragment thereof.

[0031] In certain aspects, the methods of the present disclosure comprise the steps of: (i) administering to a subject a dose of 0.1 mg to 3000 mg of the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof once a week or once every 3 weeks subcutaneously or intravenously for a period of monotherapy, wherein the period of monotherapy is at least 3 weeks; and (ii) administering to the subject a dose of 0.1 mg to 3000 mg of the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof once a week or once every 3 weeks subcutaneously or intravenously, and a dose of 150 mg to 500 mg of the anti-PD-1 antibody or antigen-binding fragment thereof once every 3 weeks intravenously or subcutaneously.

[0032] In some embodiments, the period of monotherapy is at least 3 weeks, at least 4 weeks, at least 5 weeks, or at least 6 weeks. In some embodiments, the period of monotherapy is less than one year, less than 9 months, less than 6 months, less than 3 months, less than 6 weeks, or less than 1 month.

[0033] In some embodiments, during step (ii), the anti-PD-1 antibody or antigen-binding fragment thereof and the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof are administered on different days. In some embodiments, during step (ii), the anti-PD-1 antibody or antigen-binding fragment thereof and the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof are administered on the same day.

[0034] In certain embodiments, the method further comprises administering to the subject one or more additional agents to treat one or more symptoms of an immune-related adverse event. In some embodiments, the one or more additional agents comprise an IL-6 receptor inhibitor (e.g., an anti-IL6R antibody), a corticosteroid, and / or a non-steroidal anti-inflammatory drug (NSAID).

[0035] In some embodiments, at least one week after co-administering a dose of from about 0.1 mg to about 3000 mg of the bispecific antibody or antigen-binding fragment thereof and an anti-PD-1 antibody or antigen-binding fragment thereof, the subject's disease is stable, with a partial response or a complete response.

[0036] In one embodiment, the subject has cervical cancer and has a partial response after administration. In one embodiment, the subject is a PD-1 negative patient, wherein the subject has cervical cancer and achieves a partial response after administration.

[0037] In one embodiment, the subject has microsatellite stable colorectal cancer (CRC) and the disease is stable or has a partial response after administration. In one embodiment, a bispecific antibody or antigen-binding fragment thereof and cemiplimab are co-administered to the subject, wherein the subject has CRC and the disease is stable or a partial response is achieved after administration.

[0038] In some embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof is cemiplimab, nivolumab, pembrolizumab, MEDI0608, BI 754091, spartalizumab (PDR001), camrelizumab (SHR-1210), JNJ-63723283, MCLA-134, toripalimab, sintilimab, tislelizumab, serplulimab, dostarlimab, retifanlimab, zimberelimab, penpulimab, pidilizumab, HX008, balstilimab, or ezabenlimab, or an antigen-binding fragment of any of the foregoing.

[0039] In certain embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof comprises: heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) of a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO:73, and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) of a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:74.

[0040] In some embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof comprises three HCDRs (HCDR1, HCDR2, and HCDR3) and three LCDRs (LCDR1, LCDR2, and LCDR3), wherein HCDR1 comprises the amino acid sequence of SEQ ID NO:75; HCDR2 comprises the amino acid sequence of SEQ ID NO:76; HCDR3 comprises the amino acid sequence of SEQ ID NO:77; LCDR1 comprises the amino acid sequence of SEQ ID NO:78; LCDR2 comprises the amino acid sequence of SEQ ID NO:79 (Ala Ala Ser, or AAS); and LCDR3 comprises the amino acid sequence of SEQ ID NO:80.

[0041] In some embodiments, the HCVR comprises the amino acid sequence of SEQ ID NO:73 and the LCVR comprises the amino acid sequence of SEQ ID NO:74.

[0042] In some embodiments, the anti-PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:81 and a light chain comprising the amino acid sequence of SEQ ID NO:82.

[0043] In some embodiments, the anti-PD-1 antibody is zimberelimab or an antigen-binding fragment thereof.

[0044] In some embodiments, the method comprises administering any one of the bispecific EGFRxCD28 antibodies disclosed herein.

[0045] In certain embodiments, the first antigen-binding domain that binds CD28 comprises: three heavy chain complementarity determining regions (CDR-H1, CDR-H2, and CDR-H3) contained within a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO:10; and three light chain complementarity determining regions (CDR-L1, CDR-L2, and CDR-L3) contained within a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:16.

[0046] In some embodiments, CDR-H1 of the first antigen-binding domain comprises the amino acid sequence: GGSISSYY (SEQ ID NO:12), CDR-H2 comprises the amino acid sequence: IYYSGIT (SEQ ID NO:6), and CDR-H3 comprises the amino acid sequence: ARWGVRRDYYYYGMDV (SEQ ID NO:14).

[0047] In some embodiments, CDR-L1 of the first antigen-binding domain comprises the amino acid sequence: QSVSSSY (SEQ ID NO:18), CDR-L2 comprises the amino acid sequence: GAS (SEQ ID NO:20), and CDR-L3 comprises the amino acid sequence: QQYGSSPWT (SEQ ID NO:22).

[0048] In some embodiments, the first antigen-binding domain comprises an HCVR comprising the amino acid sequence of SEQ ID NO:10 and an LCVR comprising the amino acid sequence of SEQ ID NO:16.

[0049] In certain embodiments, the second antigen-binding domain that binds to human EGFR comprises: three heavy-chain complementarity-determining regions (CDR-H1, CDR-H2, and CDR-H3) contained within a heavy-chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO:2; and three light-chain complementarity-determining regions (CDR-L1, CDR-L2, and CDR-L3) contained within a light-chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:16.

[0050] In some embodiments, CDR-H1 of the second antigen-binding domain comprises the amino acid sequence: GDSIITFY (SEQ ID NO:4), CDR-H2 comprises the amino acid sequence: IYYSGIT (SEQ ID NO:6), and CDR-H3 comprises the amino acid sequence: ARVSEDSYFHYGMDV (SEQ ID NO:8).

[0051] In some embodiments, CDR-L1 of the second antigen-binding domain comprises the amino acid sequence: QSVSSSY (SEQ ID NO:18); CDR-L2 comprises the amino acid sequence: GAS (SEQ ID NO:20); and CDR-L3 comprises the amino acid sequence: QQYGSSPWT (SEQ ID NO:22).

[0052] In some embodiments, the second antigen-binding domain comprises: an HCVR comprising the amino acid sequence of SEQ ID NO:2 and an LCVR comprising the amino acid sequence of SEQ ID NO:16.

[0053] In some aspects, the first antigen-binding domain that binds to human CD28 comprises a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:10, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:16; and the second antigen-binding domain that binds to human EGFR comprises a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:2; and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:16.

[0054] In some embodiments, the bispecific antibody comprises a first heavy chain comprising the amino acid sequence of SEQ ID NO:26.

[0055] In some embodiments, the bispecific antibody comprises a second heavy chain comprising the amino acid sequence of SEQ ID NO:24.

[0056] In some embodiments, the bispecific antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO:28.

[0057] In some embodiments, the bispecific antibody comprises: a first heavy chain comprising the amino acid sequence of SEQ ID NO:26, a second heavy chain comprising the amino acid sequence of SEQ ID NO:24, and a common light chain comprising the amino acid sequence of SEQ ID NO:28.

[0058] In some embodiments, the first antigen-binding domain comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:26 and a light chain comprising the amino acid sequence of SEQ ID NO:28.

[0059] In some embodiments, the second antigen-binding domain comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:24 and a light chain comprising the amino acid sequence of SEQ ID NO:28.

[0060] In some embodiments, the bispecific EGFRxCD28 antibody is REGN7075 or an antigen-binding fragment thereof.

[0061] In some embodiments, the method further comprises administering chemotherapy to the subject. In one embodiment, the chemotherapy is platinum-based chemotherapy. BRIEF DESCRIPTION OF THE DRAWINGS

[0062] Figure 1Schematic diagram showing the mechanism of the proposed combination therapy of REGN7075 and zimberelimab. REGN7075 (EGFRxCD28) binds to EGFR on tumor cells and CD28 on T cells, thereby bridging the two cell types. Zimberelimab (anti-PD-1) binds to PD-1 on T cells to prevent PD-1-mediated inhibitory signals. Tumor-associated antigens on MHC are presented to T cells.

[0063] Figures 2A - 2D Study flow chart showing weekly administration of REGN7075 during the REGN7075 monotherapy lead-in period ( Figure 2A ); study flow chart of the group receiving weekly administration of REGN7075 starting concurrently with zimberelimab ( Figure 2B ); study flow chart of the dose escalation group and continuous Q3W administration during the REGN7075 monotherapy lead-in period ( Figure 2C ); and study flow chart of dose escalation ( Figure 2D ). As disclosed elsewhere herein, REGN7075 can be administered in fractional doses or a titrated dosing schedule.

[0064] Figure 3 Schematic of a dose escalation study evaluating the safety, tolerability, pharmacokinetics, and initial anti-tumor activity of the combination of REGN7075 (EGFRxCD28) and zimberelimab (anti-PD-1) in subjects with advanced solid tumors. DL: Dose level; RP2D: Recommended Phase II dose.

[0065] Figure 4 Shows the mean serum concentration of REGN7075 after the first dose of REGN7075 administered IV. DL1, 0.03 mg; DL2, 0.1 mg; DL3, 0.3 mg; DL4, 1 mg; DL5, 3 mg; DL6, 10 mg; DL7, 30 mg.

[0066] Figures 5A - 5B Shows T cell activation-related cytokines in the sera of patients with detectable IL-2 ( Figure 5A ) and IFN-γ ( Figure 5B ) who received REGN7075 alone and in combination with zimberelimab. DL, Dose level; EOT, End of treatment; ET, Early termination; IFN, Interferon; IL, Interleukin; IV, Intravenous; LLOQ, Lower limit of quantification; Q3W, Every 3 weeks.

[0067] Figure 6Percentage change in target lesions relative to baseline in 18 patients receiving dose escalation therapy. Data are preliminary and the trial is ongoing. DL, dose level; MSS CRC, microsatellite stable colorectal cancer.

[0068] Figure 7 Percentage change in target lesions relative to baseline in 25 patients receiving dose escalation therapy based on data as of December 2022. Data are preliminary and the trial is ongoing. SD: stable disease; PD: progressive disease; CR / PR: complete response / partial response. Detailed implementation

[0069] Before describing the present disclosure, it is to be understood that the present disclosure is not limited to the specific methods and experimental conditions described, as such methods and conditions may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting, as the scope of the present disclosure is defined only by the appended claims.

[0070] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains. As used herein, when referring to a recited numerical value, the term "about" means that the value may vary from the recited value by no more than 1%. For example, as used herein, the expression "about 100" includes 99 and 101 and all values in between (e.g., 99.1, 99.2, 99.3, 99.4, etc.).

[0071] Although any methods and materials similar or equivalent to those described herein may be used in the practice or testing of the present disclosure, the preferred methods and materials are now described. All patents, applications, and non-patent disclosures mentioned in this specification are incorporated herein by reference in their entirety.

[0072] Definitions

[0073] Unless specified as from a non-human species (e.g., "mouse EGFR", "mouse EGFR fragment", "monkey EGFR", "monkey EGFR fragment", etc.), "EGFR" and "EGFR fragment" as used herein refer to the well-known human EGFR protein or fragments thereof. In one embodiment, human EGFR comprises the amino acid sequence set forth in NCBI accession number NP_005219.2. In one embodiment, human EGFR (amino acids L25 - A647 with accession number 005228.4) is displayed with a C-terminal CPGG.myc epitope (E1 - L10).GlyGly.myc epitope (E1 - L10).SerGly.6XHis.SSG tag (SEQ ID NO: 69).

[0074] Unless specified as being from a non-human species, as used herein, "CD28" refers to the well-known human CD28 protein that is expressed as a co-stimulatory receptor on T cells. In one embodiment, human CD28 comprises the amino acid sequence set forth in NCBI accession number NP_006130.1.

[0075] Unless specified as being from a non-human species, as used herein, "PD-1" or "programmed death receptor-1" refers to the well-known human PD-1 protein or a fragment thereof. In one embodiment, human PD-1 comprises the amino acid sequence set forth in NP_005009.

[0076] An "antibody" is an immunoglobulin molecule that comprises four polypeptide chains interconnected by disulfide bonds, two heavy chains (HC) and two light chains (LC). Each heavy chain (HC) comprises a heavy chain variable region (abbreviated herein as HCVR or V H ) and a heavy chain constant region (e.g., IgG, IgG1, or IgG4). The heavy chain constant region comprises three domains, namely C H 1, C H 2, and C H 3. Each light chain (LC) comprises a light chain variable region (abbreviated herein as LCVR or V L ) and a light chain constant region (e.g., λ or κ). The light chain constant region comprises one domain (C L 1). V H and V L regions can be further subdivided into hypervariable regions called complementarity determining regions (CDRs), interspersed with more conserved regions called framework regions (FRs). Each V H and V L comprises three CDRs and four FRs: arranged in the following order from the amino terminus to the carboxyl terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. Heavy chain CDRs can be referred to as HCDRs and light chain CDRs can be referred to as LCDRs. In various embodiments of the present disclosure, the FRs of the antibody (or its antigen-binding portion) can be identical to human germline sequences, or can be naturally or artificially modified.

[0077] For example, a bispecific antibody includes an arm that binds to a first antigen and another arm that binds to a second antigen. For example, an EGFRxCD28 bispecific antibody includes one arm that binds to EGFR and another arm that binds to CD28. A bispecific antigen-binding molecule (e.g., a bispecific antibody or an antigen-binding fragment or portion thereof) can have an effector arm that binds to a first antigen and a targeting arm that binds to a second antigen. The effector arm can be a first antigen-binding domain (e.g., anti-CD28) that binds to an antigen on an effector cell (e.g., a T cell). The targeting arm can be a second antigen-binding domain (e.g., an anti-EGFR antibody) that binds to an antigen on a target cell (e.g., a tumor cell). According to certain exemplary embodiments, the effector arm binds to CD28 and the targeting arm binds to EGFR. Throughout this specification, the terms "EGFRxCD28 bispecific antibody," "bispecific EGFRxCD28 antibody," and EGFRxCD28 can be used interchangeably.

[0078] The antigen-binding arm (e.g., the CD28 or EGFR-binding arm) of a Y-shaped IgG antibody refers to the structural portion of the antibody that confers antigen-binding specificity. For example, the antigen-binding arm of an IgG antibody has a heavy chain (HC) associated with a light chain (LC).

[0079] As used herein, the "antigen-binding portion" of an antibody, the "antigen-binding fragment" of an antibody, etc. include any naturally occurring, enzymatically obtainable, synthetic, or genetically engineered polypeptide or glycoprotein that specifically binds an antigen to form a complex. A multispecific antigen-binding fragment of an antibody binds to multiple antigens (e.g., two different antigens if the fragment is bispecific). Antigen-binding fragments of an antibody can be derived using any suitable standard techniques, such as from a whole antibody molecule, techniques such as proteolytic digestion or recombinant genetic engineering techniques that involve the manipulation and expression of DNA encoding the variable domain and optionally the constant domain of the antibody. Non-limiting examples of antigen-binding fragments include: (i) Fab fragments; (ii) F(ab')2 fragments; (iii) Fd fragments; (iv) Fv fragments; (v) single-chain Fv (scFv) molecules; and (vi) dAb fragments.

[0080] In one embodiment, the antigen-binding fragment of an antibody will comprise at least one variable domain. The variable domain can have any size or amino acid composition and will generally comprise at least one CDR adjacent to or in-frame with one or more framework sequences. In an antigen-binding fragment having a V L domain associated with a V H domain, the V H and V L domains can be positioned relative to each other in any suitable arrangement. For example, the variable region can be dimeric and contain V H -V H 、VH -V L or V L -V L dimer. Alternatively, an antigen-binding fragment of an antibody can comprise monomeric V H and V L domains.

[0081] In some embodiments, an antigen-binding fragment of an antibody can comprise at least one variable domain covalently linked to at least one constant domain. Non-limiting exemplary configurations of variable and constant domains found within antigen-binding fragments of antibodies disclosed herein include: (i) V H -C H 1; (ii) V H -C H 2; (iii) V H -C H 3; (iv) V H -C H 1-C H 2; (v) V H -C H 1-C H 2-C H 3; (vi) V H -C H 2-C H 3; (vii) V H -C L ; (viii) V L -C H 1; (ix) V L -C H 2; (x) V L -C H 3; (xi) V L -C H 1-C H 2; (xii) V L -C H 1-C H 2-C H 3; (xiii) V L -C H 2-C H 3; and (xiv) V L -C L。In any configuration of the variable and constant domains (including any of the exemplary configurations listed above), the variable and constant domains can be directly connected to each other or can be connected through a full or partial hinge region or linker region. The hinge region can consist of at least 2 (e.g., 5, 10, 15, 20, 40, 60 or more) amino acids that create a flexible or semi-flexible linkage between adjacent variable and / or constant domains in a single polypeptide molecule. Additionally, an antigen-binding fragment of an antibody of the present disclosure can comprise a homodimer or heterodimer (or other multimer) of any of the variable and constant domain configurations listed above, which are non-covalently associated with each other and / or non-covalently associated with one or more monomeric V H or V L domains (e.g., via disulfide bonds).

[0082] An "isolated" antigen-binding protein (e.g., an antibody or an antigen-binding fragment thereof), polypeptide, polynucleotide, and vector are at least partially free of other biomolecules from the cells or cell culture in which they are produced. Such biomolecules include nucleic acids, proteins, other antibodies or antigen-binding fragments, lipids, carbohydrates, or other substances such as cell debris and growth medium. An "isolated" antigen-binding protein can be further at least partially free of expression system components, such as biomolecules from the host cell or its growth medium. Generally, the term "isolated" is not intended to mean the complete absence of such biomolecules, or the absence of water, buffer, or salts, or the components of a pharmaceutical formulation that includes the antigen-binding protein (e.g., an antibody or an antigen-binding fragment).

[0083] The term "recombinant" antigen-binding protein, such as an antibody or an antigen-binding fragment thereof, refers to such molecules that are produced, expressed, isolated, or obtained by techniques or methods known in the art as recombinant DNA technology (including, for example, DNA splicing and transgenic expression). The term includes antibodies expressed in non-human mammals (including transgenic non-human mammals, such as transgenic mice) or host cells (e.g., Chinese hamster ovary (CHO) cells) or cell expression systems, or isolated from recombinant combinatorial human antibody libraries. The present disclosure includes recombinant antigen-binding proteins as described herein.

[0084] The term "specifically binds" and the like mean that an antibody or an antigen-binding fragment thereof forms a complex with an antigen that is relatively stable under physiological conditions. Methods for determining whether an antibody specifically binds an antigen are well known in the art and include, for example, equilibrium dialysis, surface plasmon resonance, etc. In some embodiments, the term "specifically binds" or "binds specifically" refers to having a binding affinity for an antigen (e.g., an EGFR, CD28, or PD-1 protein) of less than about 10-6 M (e.g. 10 -7 M, 10 -8 M, 10 -9 M, 10 -10 M, 10 -11 M or 10 -12 M) binding affinity (expressed as K D ) of these antigen-binding proteins (e.g., antibodies or antigen-binding fragments thereof), the binding affinity is determined by, for example, real-time, label-free biolayer interferometry (e.g., at 25° C. or 37° C., e.g. HTX biosensors), or by surface plasmon resonance (e.g., BIACORE TM ), or measured by solution affinity ELISA. "Anti-EGFR" refers to an antigen binding protein (or other molecules such as antigen binding arms) that specifically binds to EGFR, such as an antibody or an antigen binding fragment thereof. "Anti-CD28" refers to an antigen binding protein (or other molecules such as antigen binding arms) that specifically binds to CD28, such as an antibody or an antigen binding fragment thereof. "EGFRxCD28" refers to an antigen binding protein (or other molecules such as antigen binding arms) that specifically binds to EGFR and CD28 (and optionally binds to one or more other antigens), such as an antibody or an antigen binding fragment thereof. "Anti-PD-1" refers to an antigen binding protein (or other molecules such as antigen binding arms) that specifically binds to PD-1, such as an antibody or an antigen binding fragment thereof. However, an isolated antibody or antigen binding fragment that specifically binds to a human protein (e.g., PD-1) may have cross-reactivity with other antigens from other (non-human) species, such as proteins (e.g., PD-1 proteins).

[0085] The present disclosure includes methods of treating cancer that comprise administering an antigen-binding protein (e.g., an antibody or antigen-binding fragment) that binds the same EGFR and CD28 epitopes as an antigen-binding protein (e.g., REGN7075 (also referred to herein as bsAb7075); REGN6321 (also referred to herein as bsAb6321); REGN6322 (also referred to herein as bsAb6322); REGN6323 (also referred to herein as bsAb6323)). Other anti-EGFR X anti-CD28 antigen-binding proteins are shown in Tables 9A, 9B, and 9C. The amino acid sequences of the EGFR HCVR arms of the bispecific antibodies described herein are shown in Table 1, and the amino acid sequences of the CD28 HCVR arms of the bispecific antibodies described herein are shown in Table 3. Other EGFR parent antibodies for use in the present disclosure are described in WO2014 / 004427. The amino acid sequences of the EGFR HCVR arms and CD28 HCVR arms of REGN7075, REGN6321, REGN6322, and REGN6323 are shown in Table 6. The amino acid sequences of the common light chain variable regions described in the present disclosure are also shown in Table 6.

[0086] Generally, antibodies or antigen-binding fragments of the present disclosure that are modified in some way retain the ability to specifically bind EGFR and CD28, e.g., when activity is expressed in moles, retain at least 10% of their EGFR and CD28 binding activity (when compared to the parent antibody). Preferably, the antibodies or antigen-binding fragments of the present disclosure retain at least 20%, 50%, 70%, 80%, 90%, 95%, or 100% or more of the EGFR and CD28 binding affinity compared to the parent antibody. It is also meant that the antibody or antigen-binding fragment can include conservative or non-conservative amino acid substitutions that do not substantially alter its biological activity (referred to as "conservative variants" or "functionally conservative variants" of the antibody).

[0087] Polypeptides, such as immunoglobulin chains (e.g., REGN7075 (also referred to herein as bsAb7075); REGN6321 (also referred to herein as bsAb6321); REGN6322 (also referred to herein as bsAb6322); REGN6323 (also referred to herein as bsAb6323) V H 、V L, a "variant" of HC or LC or its CDR) refers to a polypeptide comprising an amino acid sequence that is at least about 70% to 99.9% (e.g., at least 70%, 72%, 74%, 75%, 76%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or 99.9%) identical or similar to a reference amino acid sequence set forth herein (e.g., SEQ ID NO: 6; 8; 10; 12; 14; 16; 18; 20; 22; 24; 26; 28; 30; 32; 34; 36; 38; 40; 42; 44; 46; 48; 50; 52; 54; 56; or any one of 58 - 68); when compared by the BLAST algorithm, wherein the parameters of the algorithm are selected to give the maximum match between the corresponding sequences over the entire length of the corresponding reference sequence (e.g., expect threshold: 10; word size: 3; maximum match in query range: 0; BLOSUM 62 matrix; gap costs: existence 11, extension 1; conditional compositional score matrix adjustment).

[0088] In addition, variants of the polypeptide may include, for example, immunoglobulin chains (e.g., REGN7075 (also referred to herein as bsAb7075); REGN6321 (also referred to herein as bsAb6321); REGN6322 (also referred to herein as bsAb6322); REGN6323 (also referred to herein as bsAb6323)V H , V L , a polypeptide of HC or LC or its CDR), which may include the amino acid sequence of a reference polypeptide, the amino acid sequence of the reference polypeptide being particularly set forth herein, and the variant of the polypeptide having one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) mutations, such as one or more missense mutations (e.g., conservative substitutions), nonsense mutations, deletions or insertions. By way of example, the present disclosure includes a CD28xEGFR antigen - binding protein, the antigen - binding protein including an EGFR - binding arm immunoglobulin light chain (or V L ) variant that comprises the amino acid sequence set forth in SEQ ID NO: 16 and has one or more of such mutations; and / or an immunoglobulin heavy chain (or V H) A variant that comprises the amino acid sequence set forth in SEQ ID NO:2 and has one or more of such mutations. In one embodiment, the CD28xEGFR antigen-binding protein comprises an immunoglobulin light chain variant comprising LCDR1, LCDR2, and LCDR3, wherein one or more (e.g., 1 or 2 or 3) of such CDRs have one or more of such mutations (e.g., conservative substitutions); and / or an immunoglobulin heavy chain variant comprising HCDR1, HCDR2, and HCDR3, wherein one or more (e.g., 1 or 2 or 3) of such CDRs have one or more of such mutations (e.g., conservative substitutions).

[0089] For example, a “conservatively modified variant” or “conservative substitution” of an immunoglobulin chain as described herein refers to a variant in which one or more amino acids in a polypeptide are substituted with other amino acids having similar characteristics (e.g., charge, side chain size, hydrophobicity / hydrophilicity, backbone conformation, and rigidity, etc.). Such changes can generally be made without significantly disrupting the biological activity of the antibody or fragment. Those skilled in the art recognize that, generally, single amino acid substitutions in non-essential regions of a polypeptide do not substantially alter biological activity (see, e.g., Watson et al. (1987) Molecular Biology of the Gene, The Benjamin / Cummings Pub. Co., page 224 (4th ed.)). Additionally, substitutions of amino acids that are structurally or functionally similar are not likely to significantly disrupt biological activity. The present disclosure includes bispecific EGFRxCD28 antibodies and antigen-binding fragments thereof that comprise such conservatively modified variant immunoglobulin chains.

[0090] Examples of groups of amino acids having side chains with similar chemical properties include 1) aliphatic side chains: glycine, alanine, valine, leucine, and isoleucine; 2) aliphatic hydroxyl side chains: serine and threonine; 3) amide-containing side chains: asparagine and glutamine; 4) aromatic side chains: phenylalanine, tyrosine, and tryptophan; 5) basic side chains: lysine, arginine, and histidine; 6) acidic side chains: aspartic acid and glutamic acid, and 7) sulfur-containing side chains: cysteine and methionine. Alternatively, a conservative substitution is any change that has a positive value in the PAM250 log-likelihood matrix disclosed in Gonnet et al. (1992) Science 256:1443-45.

[0091] The following references relate to the BLAST algorithms commonly used for sequence analysis: BLAST ALGORITHMS: Altschul et al. (2005) FEBS J. 272(20):5101-5109; Altschul, S.F. et al., (1990) J. Mol. Biol. 215:403-410; Gish, W. et al., (1993) Nature Genet. 3:266-272; Madden, T.L. et al., (1996) Meth. Enzymol. 266:131-141; Altschul, S.F. et al., (1997) Nucleic Acids Res. 25:3389-3402; Zhang, J. et al., (1997) Genome Res. 7:649-656; Wootton, J.C. et al., (1993) Comput. Chem. 17:149-163; Hancock, J.M. et al., (1994) Comput. Appl. Biosci. 10:67-70; ALIGNMENT SCORING SYSTEMS: Dayhoff, M.O. et al., “A model of evolutionary change in proteins.” in Atlas of Protein Sequence and Structure, (1978) Vol. 5, Suppl. 3. M.O. Dayhoff (ed.), pp. 345-352, Natl. Biomed. Res. Found., Washington, D.C.; Schwartz, R.M. et al., “Matrices for detecting distant relationships.” in Atlas of Protein Sequence and Structure, (1978) Vol. 5, Suppl. 3. M.O. Dayhoff (ed.), pp. 353-358, Natl. Biomed. Res. Found., Washington, D.C.; Altschul, S.F., (1991) J. Mol. Biol. 219:555-565; States, D.J. et al., (1991) Methods 3:66-70; Henikoff, S. et al., (1992) Proc. Natl. Acad. Sci. USA 89:10915-10919; Altschul, S.F. et al., (1993) J. Mol. Evol.36:290-300; Alignment Statistics: Karlin, S. et al., (1990) Proc. Natl. Acad. Sci. USA 87:2264-2268; Karlin, S. et al., (1993) Proc. Natl. Acad. Sci. USA 90:5873-5877; Dembo, A. et al., (1994) Ann. Prob. 22:2022-2039; and Altschul, S.F. “Evaluating the statistical significance of multiple distinct local alignments.” in Theoretical and Computational Methods in Genome Research (S. Suhai ed.), (1997) pp. 1-14, Plenum, N.Y.

[0092] As used herein, the term “subject” refers to a mammal (e.g., rat, mouse, cat, dog, cow, sheep, horse, goat, rabbit), preferably a human in need of prevention and / or treatment of cancer, such as cancer expressing EGFR. The subject may have cancer, such as cancer expressing EGFR, be predisposed to such a condition, and / or would benefit from inhibition or reduction of EGFR activity or depletion of EGFR+ cells. In one embodiment, the subject may have cancer or be at risk of developing cancer. In some embodiments, the term “subject” may be used interchangeably with the term “patient”.

[0093] As used herein, the expression “subject in need” means a human or non-human mammal exhibiting one or more symptoms or indications of cancer and / or diagnosed with cancer (including solid tumors) and in need of the same treatment.

[0094] As used herein, the terms “treat”, “treating”, etc. mean alleviating symptoms; temporarily or permanently eliminating the cause of symptoms; delaying or inhibiting tumor growth; reducing tumor cell burden or tumor burden; promoting tumor regression; causing the tumor to shrink, necrose, and / or disappear; preventing tumor recurrence and / or increasing the duration of the subject's survival.

[0095] As used herein, the term "solid tumor" refers to an abnormal mass of tissue that generally does not contain cysts or liquid areas. Solid tumors can be benign (non-cancerous) or malignant (cancerous). For the purposes of this disclosure, the term "solid tumor" means a malignant solid tumor. The term includes different types of solid tumors named for the cell types from which they form, namely sarcomas, carcinomas, and lymphomas. However, the term does not include leukemias. In various embodiments, the term "solid tumor" includes: cancers caused by connective or supportive tissues (such as bone or muscle) (referred to as sarcomas), cancers caused by body glandular cells and epithelial cells within body tissues (referred to as carcinomas), and cancers of lymphoid organs (such as lymph nodes, spleen, and thymus) (referred to as lymphomas). Lymphocytes are present in almost all tissues of the body and thus, lymphomas can occur in a wide variety of organs. In certain embodiments, the term "solid tumor" includes cancers that include (but are not limited to) colorectal cancer, ovarian cancer, prostate cancer, breast cancer, brain cancer, cervical cancer, bladder cancer, anal cancer, uterine cancer, colon cancer, liver cancer, pancreatic cancer, lung cancer, endometrial cancer, bone cancer, testicular cancer, skin cancer, kidney cancer, gastric cancer, esophageal cancer, head and neck cancer, salivary gland cancer, and myeloma. In certain embodiments, the term "solid tumor" includes cancers that include (but are not limited to) hepatocellular carcinoma, non-small cell lung cancer, squamous cell carcinoma of the head and neck, basal cell carcinoma, breast cancer, squamous cell carcinoma of the skin, chondrosarcoma, angiosarcoma, cholangiocarcinoma, soft tissue sarcoma, colorectal cancer, melanoma, Merkel cell carcinoma, and glioblastoma multiforme. In certain embodiments, the term "solid tumor" encompasses more than one solid tumor lesion that are distinct from each other in location, such as 2, more than 2, more than 5, more than 10, more than 15, more than 20, or more than 25 lesions in a subject in need of treatment. In certain embodiments, more than one lesion are distal to each other within the same organ. In certain other embodiments, the tumor lesions can be located in different organs.

[0096] As used herein, the phrase "in combination with" means that a first therapeutic agent (such as a bispecific EGFRxCD28 antibody or an antigen-binding fragment thereof) is administered before, after, or simultaneously with a second therapeutic agent (such as an anti-PD-1 antibody or an antigen-binding fragment thereof). The term "in combination with" also includes sequential or simultaneous administration of a first therapeutic agent (such as a bispecific EGFRxCD28 antibody or an antigen-binding fragment thereof) and a second therapeutic agent (such as an anti-PD-1 antibody or an antigen-binding fragment thereof).

[0097] Combination therapy methods for treating or inhibiting cancer growth

[0098] The present disclosure provides methods for treating, alleviating, or reducing the severity of at least one symptom or indication of cancer in a subject, or inhibiting the growth of said cancer, comprising administering in combination to the subject an effective dose of a bispecific EGFRxCD28 antibody (e.g., REGN7075; or any combination where the anti-EGFR HCVR is paired with the HCVR from any of the CD28 antibodies described herein) and an effective dose of a PD-1 inhibitor, such as an antibody (e.g., cemipimab) or an antigen-binding fragment thereof.

[0099] Combination therapies comprising the bispecific antigen-binding EGFRxCD28 molecule of the present disclosure and an anti-PD-1 antibody or antigen-binding portion thereof are particularly suitable for treating any cancer in which the stimulation, activation, and / or targeting of an immune response would be beneficial. Specifically, the bispecific EGFRxCD28 antibodies or antigen-binding molecules of the present disclosure can be used to treat, prevent, and / or alleviate cancers, such as cancers associated with or mediated by EGFR expression or activity or EGFR+ cell proliferation. The mechanisms of action by which the treatment methods of the present disclosure are achieved include killing EGFR-expressing cells in the presence of effector cells, such as T cells. EGFR-expressing cells that can be inhibited or killed using the antigen-binding molecules of the present disclosure include, for example, lung cancer cells.

[0100] For the purposes herein, cancer refers to a disease characterized by abnormal, excessive, and / or uncontrolled cell growth. Exemplary cancers include (but are not limited to) esophageal tumors or esophageal cancer, lung squamous cell carcinoma, lung adenocarcinoma, cervical squamous cell carcinoma or cervical cancer, glioma, thyroid cancer, lung cancer (e.g., non-small cell lung cancer), colorectal cancer, colon cancer, bladder cancer, rectal cancer, head and neck cancer, gastric cancer, liver cancer, pancreatic cancer, kidney cancer, urothelial cancer, prostate cancer or prostate adenocarcinoma, testicular cancer, breast cancer (e.g., ductal carcinoma or lobular carcinoma), cervical cancer or cervical tumor, endometrial cancer, ovarian cancer, gastroesophageal cancer (e.g., gastroesophageal adenocarcinoma), non-central nervous system (CNS) tumors, melanoma, nasopharyngeal cancer, anal cancer, mesothelioma, renal cell carcinoma (e.g., chromophobe renal cell carcinoma, clear cell renal cell carcinoma, or papillary renal cell carcinoma), gallbladder / bile duct cancer, pancreatic tumor, penile squamous cell carcinoma, or vulvovaginal cancer. In one embodiment, the cancer is an EGFR-expressing cancer. A wide range of cancers express EGFR. Accordingly, the methods of the present disclosure can be used to treat a wide range of cancers.

[0101] Cancer characterized by solid tumor cells or cancerous blood cells can be, for example, cancers that express EGFR, where EGFR expression in the cells of a particular subject being treated has been confirmed, and the cancers include esophageal tumors, lung squamous cell carcinoma, lung adenocarcinoma, cervical squamous cell carcinoma, endometrial adenocarcinoma, urothelial carcinoma of the bladder, lung cancer (e.g., non-small cell lung cancer), colorectal cancer, rectal cancer, endometrial cancer, skin cancer (e.g., head and neck squamous cell carcinoma), brain cancer (e.g., glioblastoma multiforme), breast cancer, gastroesophageal cancer (e.g., gastroesophageal adenocarcinoma), prostate cancer, and / or ovarian cancer.

[0102] The methods of the present disclosure can also be used to treat or alleviate primary and / or metastatic tumors that arise, for example, in colon cancer, lung cancer, breast cancer, kidney cancer, and bladder cancer (or from any of the cancers discussed herein).

[0103] The present disclosure also includes methods for treating or alleviating residual cancer in a subject. As used herein, the term "residual cancer" means that one or more cancer cells are present or remain in a subject after treatment with an anti-cancer therapy.

[0104] In certain embodiments, the methods of the present disclosure can be used to treat subjects exhibiting elevated levels of one or more cancer-related biomarkers [e.g., programmed death ligand 1 (PD-L1), CA125, CA19-9, prostate specific antigen (PSA), lactate dehydrogenase, KIT, carcinoembryonic antigen, epidermal growth factor receptor (EGFR), ALK gene rearrangement, or circulating tumor DNA]. For example, the methods of the present disclosure include administering, in combination, a therapeutically effective amount of an anti-PD-1 antibody and a bispecific EGFRxCD28 antibody to a subject having elevated levels of PD-L1 and / or EGFR. In one embodiment, the methods of the present disclosure are used in subjects with cancer, and the subjects are selected based on PD-L1 expression in cancer tissue. In certain embodiments, the methods of the present disclosure are used to treat subjects with cancer, wherein the subjects are selected based on at least 1%, at least 2%, at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, or at least 50% PD-L1 expression in cancer tissue and / or immune cells. Methods for determining PD-L1 expression in cancer tissue and / or immune cells are well known in the art. In certain embodiments, PD-L1 expression in tumor tissue is determined by any assay known in the art, such as by ELISA assay or by immunohistochemistry (IHC) assay (e.g., Ventana SP263 assay) or as described in PCT publication WO2016124558 or WO2016191751 or U.S. Patent Application Publication US20160305947. In certain embodiments, PD-L1 expression is determined by quantifying RNA expression, such as by in situ hybridization or by RT-PCR. In certain embodiments, PD-L1 expression is determined by imaging with a labeled anti-PD-L1 antibody, such as by immuno-positron emission tomography or iPET [see, e.g., The Oncologist, 12:1379 (2007); Journal of Nuclear Medicine, 52(8):1171 (2011) or U.S. Patent No. 10,736,976, the entire contents of which are hereby expressly incorporated by reference in their entirety].

[0105] In one embodiment, the method includes determining whether the cancer of the subject expresses EGFR. If such expression is observed, then a bispecific EGFRxCD28 antibody is administered in combination with an anti-PD-1 antibody or an antigen-binding fragment thereof. For example, in one embodiment, the method includes obtaining a biopsy of the cancer and determining whether the cells of the cancer express EGFR, and if EGFR expression is present, then administering to the subject a bispecific EGFRxCD28 antigen-binding protein and an anti-PD-1 antibody or an antigen-binding fragment thereof in combination. In one embodiment, expression is tested by immunohistochemistry (IHC) or by enzyme linked immunosorbent assay (ELISA).

[0106] According to certain aspects, the present disclosure provides methods for treating, alleviating, or reducing the severity of at least one symptom or indication of cancer, such as cancer associated with EGFR expression (e.g., lung cancer), or inhibiting the growth of the cancer, comprising, for example, administering to a subject in combination one or more of the bispecific EGFRxCD28 antibodies or antigen-binding molecules described herein (e.g., REGN7075), and an anti-PD-1 antibody or an antigen-binding fragment thereof (e.g., cemiplimab), for example, after it has been confirmed that the subject is unresponsive to other types of anti-cancer therapies.

[0107] For example, the present disclosure includes methods for treating cancer, such as lung cancer, comprising administering to a subject a bispecific EGFRxCD28 antibody or antigen-binding molecule, such as REGN7075, and an anti-PD-1 antibody or an antigen-binding fragment thereof 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, or 4 weeks, 2 months, 4 months, 6 months, 8 months, 1 year, or more after the subject has received standard of care for a subject with the cancer (e.g., lung cancer).

[0108] In certain embodiments, the combined administration of a bispecific EGFRxCD28 antibody or antigen-binding fragment thereof and an anti-PD-1 antibody or antigen-binding fragment thereof results in increased tumor regression, tumor shrinkage, and / or disappearance. In certain embodiments, the administration of an anti-PD-1 antibody and a bispecific EGFRxCD28 antibody retards tumor growth and development, e.g., tumor growth is retarded by about 3 days, more than 3 days, about 7 days, more than 7 days, more than 15 days, more than 1 month, more than 3 months, more than 6 months, more than 1 year, more than 2 years, or more than 3 years compared to an untreated subject or a subject treated with the antibody as monotherapy. In certain embodiments, the administration of an anti-PD-1 antibody and a bispecific EGFRxCD28 antibody prevents tumor recurrence and / or increases the duration of survival of the subject, e.g., the duration of survival is increased by more than 15 days, more than 1 month, more than 3 months, more than 6 months, more than 12 months, more than 18 months, more than 24 months, more than 36 months, or more than 48 months compared to an untreated subject or a subject administered the antibody as monotherapy. In certain embodiments, the administration of an anti-PD-1 antibody and a bispecific EGFRxCD28 antibody increases progression-free survival or overall survival. In certain embodiments, the administration of an anti-PD-1 antibody and a bispecific EGFRxCD28 antibody increases the response and duration of response of the subject, e.g., by more than 2%, more than 3%, more than 4%, more than 5%, more than 6%, more than 7%, more than 8%, more than 9%, more than 10%, more than 20%, more than 30%, more than 40%, or more than 50% relative to an untreated subject or a subject that has received the antibody as monotherapy.

[0109] In certain embodiments, the administration of an anti-PD-1 antibody and a bispecific EGFRxCD28 antibody to a subject with cancer results in at least a 30% or greater reduction in tumor cells or tumor size (“partial response”). In certain embodiments, the administration of an anti-PD-1 antibody and a bispecific EGFRxCD28 antibody to a subject with cancer results in the complete disappearance of all evidence of tumor cells (“complete response”). In certain embodiments, the administration of an anti-PD-1 antibody and a bispecific EGFRxCD28 antibody to a subject with cancer results in the complete or partial disappearance of tumor cells / lesions (including new measurable lesions). Tumor reduction can be measured by any of the methods known in the art, such as X-ray, positron emission tomography (PET), computed tomography (CT), magnetic resonance imaging (MRI), cytology, histology, or molecular genetic analysis.

[0110] In certain embodiments, the methods of the present disclosure include administering, in combination, a therapeutically effective amount of a bispecific EGFRxCD28 antibody and an anti-PD-1 antibody to a subject in need thereof, wherein the combination administration results in an increase in the overall survival (OS) or progression-free survival (PFS) of the subject as compared to a patient receiving a standard-of-care (SOC) therapy (e.g., chemotherapy, surgery, or radiation therapy). In certain embodiments, the PFS is increased by at least one month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 1 year, at least 2 years, or at least 3 years as compared to a patient receiving any one or more SOC therapies. In certain embodiments, the OS is increased by at least one month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 1 year, at least 2 years, or at least 3 years as compared to a patient receiving any one or more SOC therapies.

[0111] (I) Dosage and Timing

[0112] An "effective" or "therapeutically effective" dose of an anti-PD-1 antibody or an antigen-binding fragment thereof, or a bispecific EGFRxCD28 antibody or an antigen-binding fragment thereof (e.g., REGN7075) for treating or preventing cancer (e.g., cancer expressing EGFR) is an amount of the antibody or antigen-binding fragment sufficient to alleviate one or more signs and / or symptoms of the disease in the treated subject, whether by inducing regression or elimination of such signs and / or symptoms or by inhibiting the progression of such signs and / or symptoms.

[0113] The dosage of the antigen-binding molecule administered to a subject can vary depending on the age and size of the patient, the target disease, the condition, the route of administration, etc. Preferred dosages are typically calculated based on body weight or body surface area. The frequency and duration of treatment can be adjusted according to the severity of the condition.

[0114] In some embodiments of the present disclosure, the therapeutically effective dose of the bispecific EGFRxCD28 antibody, such as REGN7075, is from 0.1 to 3000 mg. The dose may vary depending on the age and body size of the subject to whom it is administered, the target disease, the condition, the route of administration, and the like. In some embodiments, the bispecific EGFRxCD28 antibody is administered at a dose of about 0.1 to 900 mg, 1 to 1500 mg, 100 to 1200 mg, 300 to 1000 mg, 500 to 1500 mg, 800 to 1000 mg, 800 to 1500 mg, 300 to 2000 mg, or 300 to 3000 mg. In some embodiments, the bispecific EGFRxCD28 antibody is administered at a dose of about 0.01 mg, 0.03 mg, 0.05 mg, 0.1 mg, 0.3 mg, 0.5 mg, 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 8 mg, 10 mg, 15 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, 600 mg, 650 mg, 700 mg, 750 mg, 800 mg, 850 mg, 900 mg, 1000 mg, 1200 mg, 1500 mg, 1800 mg, 2000 mg, 2400 mg, 2700 mg, or 3000 mg. In some embodiments, the bispecific EGFRxCD28 antibody is administered at a dose of about 0.1 to 50 mg / kg, 1 to 45 mg / kg, 5 to 10 mg / kg, 10 to 30 mg / kg, 15 to 25 mg / kg, 20 to 30 mg / kg, or 25 to 40 mg / kg of the subject's body weight. In some embodiments, the bispecific EGFRxCD28 antibody is administered at a dose of about 0.1 mg / kg, 0.5 mg / kg, 1 mg / kg, 2 mg / kg, 3 mg / kg, 4 mg / kg, 5 mg / kg, 6 mg / kg, 7 mg / kg, 8 mg / kg, 9 mg / kg, 10 mg / kg, 15 mg / kg, 20 mg / kg, 25 mg / kg, 30 mg / kg, 35 mg / kg, 45 mg / kg, or 50 mg / kg.

[0115] In some embodiments, the bispecific EGFRxCD28 antibody is administered weekly. In some embodiments, the bispecific EGFRxCD28 antibody is administered every two weeks. In some embodiments, the bispecific EGFRxCD28 antibody is administered every three weeks.

[0116] In some embodiments, the bispecific EGFRxCD28 antibody is administered intravenously (IV). In some embodiments, the IV infusion takes about 60 minutes. In some embodiments, the bispecific EGFRxCD28 antibody is administered subcutaneously.

[0117] In some embodiments of the present disclosure, a therapeutically effective dose of the bispecific EGFRxCD28 antibody, such as REGN7075, is 0.1 to 3000 mg IV or SC per week or every three weeks.

[0118] In some embodiments of the present disclosure, a therapeutically effective dose of the anti-PD-1 antibody or antigen-binding fragment thereof, such as cemipimab, is 50 to 1500 mg, such as 350 mg. The dose may vary depending on the age and body size of the subject to be administered, the target disease, the condition, the route of administration, etc. In some embodiments of the present disclosure, the therapeutically effective dose of the anti-PD-1 antibody or antigen-binding fragment thereof (such as cemipimab) is about 50 to 1500 mg, 100 to 1250 mg, 150 to 1000, 200 to 750 mg, 300 to 500 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, 600 mg, 650 mg, 700 mg, 750 mg, 800 mg, 850 mg, 900 mg, 950 mg, 1000 mg, 1100 mg, 1200 mg, 1300 mg, 1400 mg or 1500 mg. In some embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof is administered at a dose of about 0.1 to 20 mg / kg, 0.5 to 15 mg / kg, 1 to 12 mg / kg, 2 to 10 mg / kg, 5 to 10 mg / kg or 7.5 to 10 mg / kg of the subject body weight. In some embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof is administered at a dose of about 0.1 mg / kg, 0.2 mg / kg, 0.3 mg / kg, 0.4 mg / kg, 0.5 mg / kg, 1 mg / kg, 2 mg / kg, 3 mg / kg, 4 mg / kg, 5 mg / kg, 6 mg / kg, 7 mg / kg, 8 mg / kg, 9 mg / kg, 10 mg / kg, 15 mg / kg or 20 mg / kg.

[0119] In some embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof is administered weekly. In some embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof is administered every two weeks. In some embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof is administered every three weeks.

[0120] In some embodiments, the anti-PD-1 antibody or an antigen-binding fragment thereof is administered intravenously (IV). In some embodiments, the IV infusion takes about 30 minutes or about 60 minutes. In some embodiments, the anti-PD-1 antibody or an antigen-binding fragment thereof is administered subcutaneously.

[0121] In some embodiments, a therapeutically effective dose of the anti-PD-1 antibody or an antigen-binding fragment thereof (such as zimberelimab) is 50 to 1500 mg, such as 350 mg, intravenously (IV) or subcutaneously (SC), once a week (QW), every two weeks (Q2W), or every three weeks (Q3W).

[0122] In some embodiments, the bispecific EGFRxCD28 antibody or antigen-binding molecule is administered concomitantly with the anti-PD-1 antibody or an antigen-binding portion thereof. In some embodiments, the bispecific EGFRxCD28 antibody or antigen-binding molecule is administered on the same day as the anti-PD-1 antibody or an antigen-binding portion thereof. In some embodiments, the bispecific EGFRxCD28 antibody or antigen-binding molecule is administered before the anti-PD-1 antibody or an antigen-binding portion thereof, such as 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 12 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, or 21 days before. In some embodiments, the anti-PD-1 antibody or antigen-binding molecule is administered before the bispecific EGFRxCD28 antibody or an antigen-binding portion thereof, such as 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 12 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, or 21 days before.

[0123] In certain embodiments, a second or additional subsequent dose of the antigen-binding protein may be administered after the initial dose, in an amount that may be substantially the same as, less than, or more than the initial dose, wherein the subsequent doses are separated by, for example, about 1 week, 2 weeks, 3 weeks, 10 days, 20 days, or 30 days.

[0124] Multiple doses of an antigen-binding molecule (e.g., a bispecific antigen-binding molecule that specifically binds to EGFR and CD28, or an anti-PD-1 antibody or antigen-binding fragment thereof) can be administered to a subject within a limited time course. The methods according to this aspect of the present disclosure include sequentially administering multiple doses of the antigen-binding molecule of the present disclosure to a subject. As used herein, "sequentially administering" means administering each dose of the antigen-binding molecule to the subject at different time points, e.g., on the same day at a predetermined interval (e.g., hours) apart or on different days at a predetermined interval (e.g., hours, days, weeks, or months) apart. The present disclosure includes methods that include sequentially administering a single initial dose of an antigen-binding molecule to a subject, followed by administering one or more second doses of the same or different antigen-binding molecules, and optionally followed by administering one or more third doses of the antigen-binding molecule.

[0125] The terms "initial dose", "second dose", and "third dose" refer to the chronological order of administration of the antigen-binding molecule of the present disclosure. Thus, an "initial dose" is the dose administered at the start of a treatment regimen (also referred to as a "baseline dose"); a "second dose" is the dose administered after the initial dose; and a "third dose" is the dose administered after the second dose. The initial, second, and third doses can all contain the same amount of the antigen-binding molecule, but can differ from each other in terms of dosing frequency. However, in certain embodiments, the amounts of the antigen-binding molecule contained in the initial, second, and / or third doses differ from each other during the course of treatment (e.g., upregulated or downregulated as needed). In certain embodiments, two or more doses are administered at the start of a treatment regimen as a "loading dose", followed by subsequent doses administered less frequently (e.g., a "maintenance dose").

[0126] In one exemplary embodiment of the present disclosure, each second and / or third dose is administered one to several weeks after the previous dose. As used herein, the phrase "immediately preceding dose" means the dose of the antigen-binding molecule administered to a subject before administering the next dose in the order of multiple administrations, with no intervening dose.

[0127] The methods according to this aspect of the present disclosure can include administering any number of second and / or third doses of an antigen-binding molecule (e.g., a bispecific antigen-binding molecule that specifically binds to EGFR and CD28 and / or an anti-PD-1 antibody or antigen-binding fragment thereof) to a subject. For example, in certain embodiments, only a single second dose is administered to the subject. In other embodiments, two or more second doses are administered to the subject. Similarly, in certain embodiments, only a single third dose is administered to the subject. In other embodiments, two or more third doses are administered to the subject.

[0128] In embodiments involving multiple second doses, each second dose may be administered at the same frequency as the other second doses. Similarly, in embodiments involving multiple third doses, each third dose may be administered at the same frequency as the other third doses. Alternatively, the frequency of administration of the second and / or third doses to a subject may vary during the course of the treatment regimen. The dosing frequency may also be adjusted by a physician after a clinical examination during the course of treatment, depending on the needs of the individual subject.

[0129] In some embodiments, the bispecific EGFRxCD28 antibody (e.g., REGN7075) or an antigen-binding fragment thereof is administered weekly, every 2 weeks, every 3 weeks, every 10 days, every 20 days, every 30 days, monthly, every 2 months, or every 3 months during the course of treatment. In some embodiments, the anti-PD-1 antibody or an antigen-binding fragment thereof, such as cemipimab, is administered weekly, every 2 weeks, every 3 weeks, every 10 days, every 20 days, every 30 days, monthly, every 2 months, or every 3 months during the course of treatment.

[0130] In certain aspects, the methods of the present disclosure comprise the steps of: (i) administering subcutaneously or intravenously to a subject a dose of 0.1 mg to 3000 mg of a bispecific EGFRxCD28 antibody or an antigen-binding fragment thereof once a week or once every 3 weeks for a period of monotherapy, wherein the period of monotherapy is at least 3 weeks; and (ii) administering subcutaneously or intravenously to a subject a dose of 0.1 mg to 3000 mg of a bispecific EGFRxCD28 antibody or an antigen-binding fragment thereof once every 3 weeks, and administering intravenously or subcutaneously to a subject a dose of 150 mg to 500 mg of an anti-PD-1 antibody or an antigen-binding fragment thereof once every 3 weeks. In some embodiments, the period of monotherapy is at least 3 weeks, at least 4 weeks, at least 5 weeks, or at least 6 weeks. In some embodiments, the period of monotherapy is less than one year, less than 9 months, less than 6 months, less than 3 months, less than 6 weeks, or less than 1 month. In some embodiments, the period of monotherapy is at least 3 weeks but less than 1 year.

[0131] In some embodiments, during step (ii), the anti-PD-1 antibody or an antigen-binding fragment thereof is administered on a different day than the bispecific EGFRxCD28 antibody or an antigen-binding fragment thereof. In some embodiments, during step (ii), the anti-PD-1 antibody or an antigen-binding fragment thereof is administered on the same day as the bispecific EGFRxCD28 antibody or an antigen-binding fragment thereof.

[0132] In some embodiments, the bispecific EGFRxCD28 antibody is administered before, concomitantly with, or after administration of an anti-PD-1 antibody or an antigen-binding fragment thereof to a subject. In some embodiments, the anti-PD-1 antibody or an antigen-binding fragment thereof is administered before, concomitantly with, or after administration of the bispecific EGFRxCD28 antibody or an antigen-binding fragment thereof to a subject.

[0133] In some embodiments, the bispecific EGFRxCD28 antibody is administered in monotherapy form at least 2, 3, 4, or 5 times or more, followed by administration of an anti-PD-1 antibody or an antigen-binding fragment thereof. As a non-limiting example, the bispecific EGFRxCD28 antibody or fragment thereof can be administered by IV infusion or by subcutaneous injection at a dose of 0.1 mg to 3000 mg per week (QW) or every 3 weeks (Q3W) for at least one week, followed by administration of the anti-PD-1 antibody or an antigen-binding fragment thereof by IV infusion or by subcutaneous injection at a dose of 350 mg every 3 weeks (Q3W).

[0134] (II) Route of administration

[0135] The present disclosure provides methods for administering a bispecific EGFRxCD28 antibody (such as REGN7075; REGN6321; REGN6322; REGN6323; or any combination of an anti-EGFR HCVR paired with an HCVR from any of the CD28 antibodies described herein) or a pharmaceutical composition thereof, alone or in combination with an anti-PD-1 antibody (such as cemipimab) or an antigen-binding fragment thereof, to a subject (such as a human, who is, for example, suffering from cancer), the method comprising introducing the antigen-binding protein or pharmaceutical composition into the subject (such as a human) by, for example, intravenous or subcutaneous means. For example, the method comprises piercing the body of the subject with a syringe needle and injecting the antigen-binding protein or pharmaceutical composition into the subject, such as into a vein, artery, skin, tumor, muscle tissue, or subcutaneous tissue of the subject.

[0136] The mode of administration of the antibody or its pharmaceutical composition may vary. Routes of administration include parenteral, enteral, oral, rectal, mucosal, intestinal, parenteral; intramuscular, subcutaneous, intradermal, intramedullary, intrathecal, direct intraventricular, intravenous, intraperitoneal, intranasal, intraocular, inhaled, insufflated, topical, dermal, intraocular, intravitreal, transdermal, or intraarterial.

[0137] In some embodiments, the bispecific EGFRxCD28 antibody or antigen-binding portion thereof is administered intravenously, and the anti-PD-1 antibody or antigen-binding portion thereof is administered subcutaneously. In some embodiments, the bispecific EGFRxCD28 antibody or antigen-binding portion thereof is administered subcutaneously, and the anti-PD-1 antibody or antigen-binding portion thereof is administered intravenously. In some embodiments, the bispecific EGFRxCD28 antibody or antigen-binding portion thereof is administered intravenously, and the anti-PD-1 antibody or antigen-binding portion thereof is administered intravenously. In some embodiments, the bispecific EGFRxCD28 antibody or antigen-binding portion thereof is administered subcutaneously, and the anti-PD-1 antibody or antigen-binding portion thereof is administered subcutaneously.

[0138] In some other embodiments, the bispecific EGFRxCD28 antibody and / or the anti-PD-1 antibody are administered to a subject over about 10 to 120, 20 to 100, 30 to 90, or 45 to 75 minutes. In some embodiments, the bispecific EGFRxCD28 antibody and / or the anti-PD-1 antibody are administered to a subject over about 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, or 90 minutes.

[0139] The present disclosure provides a container (e.g., a plastic or glass vial or ampoule, which may have, for example, a cap or chromatography column, a hollow bore needle, or a syringe barrel) that contains the bispecific EGFRxCD28 antigen-binding protein of the present disclosure or a pharmaceutical composition thereof.

[0140] The present disclosure also provides an injection device that contains one or more antigen-binding proteins (e.g., antibodies or antigen-binding fragments) that specifically bind to EGFR and CD28 (EGFRxCD28) or a pharmaceutical formulation thereof. The injection device may be packaged in a kit. The injection device is a device for introducing a substance into a subject by a parenteral route (e.g., intramuscular, subcutaneous, or intravenous). For example, the injection device may be a syringe or an autoinjector (e.g., prefilled with a pharmaceutical formulation), which includes, for example, a barrel or cylinder for containing the injected fluid (e.g., containing an antibody or fragment or a pharmaceutical formulation thereof), a needle for piercing the skin, blood vessel, or other tissue to inject the fluid; and a plunger for pushing the fluid out of the barrel and through the needle into the subject.

[0141] A prefilled syringe is a syringe that has been filled with a composition (e.g., a pharmaceutical composition containing a multispecific antigen-binding protein and a pharmaceutically acceptable carrier) before being sold or transferred to an end user (e.g., a physician or caregiver) who will administer the composition to a subject.

[0142] The pharmaceutical composition is described in more detail below.

[0143] (III) Subject Selection

[0144] In some embodiments, the methods described herein further comprise one or more steps of selecting an object. A patient may be selected, for example, based on inclusion criteria, or may be excluded, for example, based on exclusion criteria. Inclusion and exclusion criteria are described in more detail in Example 2 below.

[0145] In some embodiments, the method comprises selecting an object having advanced solid tumors.

[0146] In some embodiments, the object meets at least one of the following criteria or is selected based on at least one of the following criteria: (1) has metastatic or locally advanced disease and is not a candidate for curative surgery or curative radiotherapy; (2) is not a candidate for an approved anti-PD-1 or PD-L1 therapy indication, or such therapy is not available for the object for other reasons (alone or in combination); (3) has exhausted all treatment options expected to provide meaningful clinical benefit due to disease recurrence, disease refractoriness, or intolerance, except for objects with malignancies in which anti-PD-1 / PD-L1 therapy has demonstrated clinical benefit; and / or (4) has any of the following cancer types: (a) colorectal cancer with local pathology documented as microsatellite stable; (b) gastric cancer or gastroesophageal junction cancer; (c) esophageal cancer; (d) breast cancer (ductal or lobular carcinoma, regardless of receptor status); (e) non-small cell lung cancer (NSCLC) (any PD-L1 expression); (f) head and neck squamous cell carcinoma (SCC); (g) nasopharyngeal carcinoma; (h) cervical cancer; (i) anal cancer; (j) mesothelioma; (k) prostate adenocarcinoma; (l) renal cell carcinoma (chromophobe renal cell carcinoma, clear cell renal cell carcinoma, or papillary renal cell carcinoma); (m) gallbladder / bile duct cancer; (n) urothelial carcinoma; (o) pancreatic neoplasm; (p) penile SCC; (q) vulvovaginal cancer; or (r) other non-CNS tumor types showing elevated EGFR expression in the tumor.

[0147] In some embodiments, the method comprises selecting an object who has been treated with a prior therapy. In one embodiment, the prior therapy is radiotherapy, surgery, chemotherapy, a PD-1 inhibitor, a PD-L1 inhibitor, an anti-VEGF therapy, a CAR-T therapy, and / or an anti-EGFR therapy. In some embodiments, the method comprises selecting an object who has not received a prior anti-PD-1 therapy or anti-PD-L1 therapy.

[0148] In some embodiments, the method comprises selecting a subject having microsatellite stable colorectal cancer (MSS CRC). In some embodiments, the subject having microsatellite stable colorectal cancer has at least one of the following attributes, or is selected based on at least one of the following attributes: (a) having metastatic CRC; (b) not being a candidate for curative surgery or curative radiotherapy; (c) likely having active metastases in the liver and / or peritoneum at screening; (d) the pathology report documenting microsatellite stability; (e) having received at least one line of therapy in the setting of recurrence / metastasis, where the therapy comprises anti-EGFR therapy or anti-VEGF therapy; and / or (f) having not received anti-PD-1 / PD-L1, defined as never having been treated with a drug targeting PD-1 previously.

[0149] In some embodiments, the method comprises selecting a subject having microsatellite stable colorectal cancer (MSS CRC). In some embodiments, the subject having microsatellite stable colorectal cancer has at least one of the following attributes, or is selected based on at least one of the following attributes: (a) having metastatic CRC; (b) not being a candidate for curative surgery or curative radiotherapy; (c) no active metastases identified in the liver or peritoneum at screening; (d) the disease site being present only in one or more lungs and / or pulmonary lymph nodes; (e) the pathology report documenting microsatellite stability; (f) having received at least one line of therapy in the setting of recurrence / metastasis, where the therapy comprises anti-EGFR therapy or anti-VEGF therapy; and / or (g) having not received anti-PD-1 / PD-L1, defined as never having been treated with a drug targeting PD-1 previously.

[0150] In some embodiments, the method comprises selecting a subject having triple-negative breast cancer (TNBC). In some embodiments, the subject having TNBC has at least one of the following attributes, or is selected based on at least one of the following attributes: (a) having metastatic TNBC; (b) not being a candidate for curative surgery or curative radiotherapy; (c) not being a candidate for an approved anti-PD-1 or anti-PD-L1 therapy indication, or such therapy not being available to the subject for other reasons; (d) the pathology report documenting triple-negative cancer (ER- / PR- / Her2-); and / or (e) having not received anti-PD-1 / PD-L1, defined as never having been treated with a drug targeting PD-1 previously.

[0151] In some embodiments, the method comprises selecting a subject having cutaneous squamous cell carcinoma (CSCC). In one embodiment, the subject is not a candidate for curative surgery or curative radiotherapy. In one embodiment, the subject has not received anti-PD-1 / PD-L1, defined as never having been treated with a drug targeting PD-1 previously.

[0152] In some embodiments, the method comprises selecting a subject having non-small cell lung cancer (NSCLC). In some embodiments, the subject has at least one of the following attributes, or is selected based on at least one of the following attributes: (a) having advanced or metastatic NSCLC; (b) not being a candidate for curative surgery or curative radiotherapy; (c) having no targetable molecular alterations (such as ALK, ROS1, EGFR, etc.); (d) not having received prior systemic therapy for recurrent or metastatic NSCLC; and / or (e) not having received anti-PD-1 / PD-L1, defined as never having been treated with a drug targeting PD-1 previously.

[0153] In some embodiments, the method comprises selecting a subject having histologically or cytologically documented locally advanced or metastatic EGFR-mutated non-squamous NSCLC disease. In some embodiments, the subject has at least one of the following attributes, or is selected based on at least one of the following attributes: (a) having advanced or metastatic NSCLC; (b) not being a candidate for curative surgery or curative radiotherapy; (c) having a previously documented targetable EGFR mutation (EGFR exon 19 deletion, EGFR L858R mutation, EGFR exon 20 insertion, or exon 18 / 21 atypical mutation); (d) not having received chemotherapy; (e) having received treatment with a third-generation TKI; and / or (e) not having received anti-PD-1 / PD-L1, defined as never having been treated with a drug targeting PD-1 previously.

[0154] In some embodiments, the method comprises selecting a subject having histologically or cytologically documented locally advanced or metastatic EGFR-mutated non-squamous NSCLC disease. In some embodiments, the subject has at least one of the following attributes, or is selected based on at least one of the following attributes: (a) having advanced or metastatic NSCLC; (b) not being a candidate for curative surgery or curative radiotherapy; (c) having a previously documented targetable EGFR mutation (EGFR exon 19 deletion, EGFR L858R mutation, EGFR exon 20 insertion, or exon 18 / 21 atypical mutation); (d) having received treatment with platinum doublet chemotherapy; (e) having received treatment with a third-generation TKI; and / or (f) not having received anti-PD-1 / PD-L1, defined as never having been treated with a drug targeting PD-1 previously.

[0155] In some embodiments, the method comprises selecting a subject having head and neck squamous cell carcinoma (HNSCC). In some embodiments, the subject has at least one of the following attributes, or is selected based on at least one of the following attributes: (a) having advanced or metastatic disease; (b) not being a candidate for curative surgery or curative radiotherapy; (c) having a CPS of PD-L1 expression ≥ 1% by local IHC assay; (d) not having received prior systemic therapy for recurrent or metastatic HNSCC; and / or (e) never having received anti-PD-1 / PD-L1, defined as never having been treated with a drug targeting PD-1 previously.

[0156] (IV) Adverse events

[0157] In some embodiments, the subject develops one or more mild symptoms of immune-related adverse events after administration of the bispecific EGFRxCD28 antibody alone or in combination with an anti-PD-1 antibody. In some embodiments, one or more symptoms of the immune-related adverse events are colitis, diarrhea, hypothyroidism, hyperthyroidism, hypophysitis, adrenal insufficiency, diabetes, hepatitis, neurotoxicity, pneumonia, renal events, uveitis, myocarditis, pericarditis, or a combination thereof.

[0158] In some embodiments, the subject receives one or more additional therapies to treat one or more symptoms of the mild immune-related adverse events.

[0159] In some embodiments, when the subject develops one or more mild symptoms of immune-related adverse events, treatment with the bispecific EGFRxCD28 antibody alone or in combination with an anti-PD-1 antibody is suspended and resumed when one or more symptoms resolve.

[0160] Multispecific EGFR x CD28 antigen-binding molecule

[0161] The present disclosure provides methods of using antigen-binding proteins that are multispecific (e.g., bispecific) and bind at least EGFR and CD28 for use in combination with an anti-PD-1 antibody or an antigen-binding portion thereof (e.g., cemipimab) in the treatment of cancer. As used herein, such multispecific antigen-binding proteins are referred to as format AxB, where A refers to the binding arm in the multispecific molecule that binds to EGFR, and B refers to the binding arm in the multispecific molecule that binds to CD28, or vice versa. EGFRxCD28 or CD28xEGFR refers to a multispecific antigen-binding protein that binds to EGFR and CD28. The specific EGFR- and CD28-binding arms in the multispecific antigen-binding protein may also be designated in the AxB format, where A refers to the specific arm and B refers to the other specific arm. For example, 085Nx14226P2 refers to a multispecific antigen-binding protein having an anti-EGFR binding arm of 085N as set forth herein and an anti-CD28 binding arm of 14426P2 as set forth herein. For example, 085N is a binding arm comprising the 085N immunoglobulin heavy and light chains or variable regions thereof or CDRs thereof, the sequences of which are specifically set forth herein or variants thereof.

[0162] Multispecific binding refers to the binding to two or more different antigenic determinants (EGFR and CD28 or more) that may be located on the same or different antigens. Multispecificity includes bispecificity, trispecificity, and tetraspecificity. An antibody or fragment thereof may be functionally linked (e.g., by chemical conjugation, gene fusion, non-covalent association, or other means) to one or more other molecular entities, such as another antibody or antibody fragment, to produce a bispecific or multispecific antibody having a second binding specificity.

[0163] In certain embodiments, the multispecific antigen-binding protein comprises a bispecific antigen-binding protein. As used herein, the term "bispecific antigen-binding protein" means a protein, polypeptide, or molecular complex (e.g., an antibody or an antigen-binding fragment thereof) comprising at least a first antigen-binding domain and a second antigen-binding domain. Each antigen-binding domain within the bispecific antigen-binding molecule comprises at least one CDR that specifically binds to a specific antigen, either alone or in combination with one or more other CDRs and / or FRs. In the context of the present disclosure, the first antigen-binding domain specifically binds to CD28, and the second antigen-binding domain specifically binds to EGFR.

[0164] The present disclosure includes: a method of administering any one of the following multispecific antigen-binding proteins (e.g., bispecific antibodies or antigen-binding fragments thereof): REGN7075, REGN6321, REGN6322, REGN6323, and bispecific antibodies prepared by combining any one of the EGFR HCVR arms of Tables 1 and 8 (e.g., the HCVR arms of parental monoclonal antibodies mAb12999P2, mAb13008P2, mAb35193P2, and mAb13006P2) with any one of the CD28R HCVR arms of Tables 3 and 8 (e.g., the HCVR arms of parental mAb14226, mAb14193, and mAb14216), and methods of using the same for treating cancer as set forth herein.

[0165] As used herein, the term "antigen-binding molecule" means a protein, polypeptide, or molecular complex that includes or consists of at least one complementarity determining region (CDR) that specifically binds to a specific antigen, either alone or in combination with one or more other CDRs and / or framework regions (FRs). In certain embodiments, the antigen-binding molecule is an antibody or antibody fragment, as defined elsewhere herein.

[0166] As used herein, the term "bispecific antigen-binding molecule" means a protein, polypeptide, or molecular complex (e.g., an antibody or antigen-binding fragment thereof) that includes at least a first antigen-binding domain and a second antigen-binding domain. Each antigen-binding domain within the bispecific antigen-binding molecule includes at least one CDR that specifically binds to a specific antigen, either alone or in combination with one or more other CDRs and / or FRs. In the context of the present disclosure, the first antigen-binding domain specifically binds to a first antigen (e.g., CD28), and the second antigen-binding domain specifically binds to a different second antigen (e.g., EGFR). In certain exemplary embodiments of the present disclosure, the bispecific antigen-binding molecule is a bispecific antibody. Each antigen-binding domain of the bispecific antibody includes a heavy chain variable region (HCVR) and a light chain variable region (LCVR).

[0167] The first antigen-binding domain and the second antigen-binding domain can be directly or indirectly linked to each other to form the bispecific antigen-binding molecule of the present disclosure. Alternatively, the first antigen-binding domain and the second antigen-binding domain can each be linked to a separate multimerization domain. The association of one multimerization domain with another multimerization domain facilitates the association between the two antigen-binding domains, thereby forming the bispecific antigen-binding molecule. As used herein, a "multimerization domain" is any macromolecule, protein, polypeptide, peptide, or amino acid capable of associating with a second multimerization domain having the same or similar structure or composition. For example, the multimerization domain can be an immunoglobulin C HA polypeptide of 3 domains. Non-limiting examples of the multimerization component are the Fc portion of an immunoglobulin (comprising C H 2-C H 3 domains), such as the Fc domain of IgG selected from isotypes IgG1, IgG2, IgG3, and IgG4 and any allotype within each isotype group.

[0168] The bispecific antigen-binding molecules of the present disclosure will generally comprise two multimerization domains, such as two Fc domains, each of which is separately part of a separate antibody heavy chain. The first and second multimerization domains may have the same IgG isotype, such as IgG1 / IgG1, IgG2 / IgG2, IgG4 / IgG4. Alternatively, the first and second multimerization domains may have different IgG isotypes, such as IgG1 / IgG2, IgG1 / IgG4, IgG2 / IgG4, etc.

[0169] In certain embodiments, the multimerization domain is an Fc fragment or amino acid sequence 1 to about 200 amino acids in length containing at least one cysteine residue. In other embodiments, the multimerization domain is a cysteine residue or a cysteine-containing short peptide. Other multimerization domains include peptides or polypeptides that comprise, consist of, or are composed of a leucine zipper, a helix-loop-helix motif, or a coiled-coil motif.

[0170] Any bispecific antibody format or technology can be used to prepare the bispecific antigen-binding molecules of the present disclosure. For example, an antibody or an antigen-binding fragment thereof having a first antigen-binding specificity can be functionally linked (e.g., by chemical conjugation, gene fusion, non-covalent association, or other means) to one or more other molecular entities, such as another antibody or antibody fragment having a second antigen-binding specificity, to produce a bispecific antigen-binding molecule. Specific exemplary bispecific formats that can be used in the context of the present disclosure include (but are not limited to) bispecific formats based on scFv or bispecific antibodies, IgG-scFv fusions, dual variable domain (DVD)-Ig, quadroma, knobs-into-holes, common light chain (e.g., common light chain with knobs-into-holes, etc.), CrossMab, CrossFab, (SEEO)body, leucine zipper, Ouobody, IgG1 / IgG2, dual action Fab (DAF)-IgG, and Mab 2 bispecific formats (for a review of the foregoing formats, see, e.g., Klein et al. 2012, mAbs 4:6, 1-11, and the references cited therein).

[0171] In the context of the bispecific antigen-binding molecules of the present disclosure, the multimerization domain (e.g., the Fc domain) can contain one or more amino acid changes (e.g., insertions, deletions, or substitutions) compared to the wild-type, naturally occurring Fc domain form. By way of example, the present disclosure includes bispecific antigen-binding molecules that contain one or more modifications in the Fc domain, where the one or more modifications result in a modified binding interaction (e.g., enhanced or diminished) between the modified Fc domain and FcRn. In one embodiment, the bispecific antigen-binding molecule contains a modification in the C H 2 or C H 3 region, where the modification increases the affinity of the Fc domain for FcRn in an acidic environment (e.g., an endosome where the pH value ranges from about 5.5 to about 6.0). Non-limiting examples of such Fc modifications include, for example, modifications at position 250 (e.g., E or Q); 250 and 428 (e.g., L or F); 252 (e.g., LN / FIW or T), 254 (e.g., S or T), and 256 (e.g., S / R / Q / EID or T); or modifications at position 428 and / or 433 (e.g., UR / S / P / Q or K) and / or 434 (e.g., H / F or V); or modifications at position 250 and / or 428; or modifications at position 307 or 308 (e.g., 308F, V308F) and 434. In one embodiment, the modification comprises 428L (e.g., M428L) and 434S (e.g., N434S) modifications; 428L, 2591 (e.g., V2591), and 308F (e.g., V308F) modifications; 433K (e.g., H433K) and 434 (e.g., 434Y) modifications; 252, 254, and 256 (e.g., 252Y, 254T, and 256E) modifications; 250Q and 428L modifications (e.g., T250Q and M428L); and 307 and / or 308 modifications (e.g., 308F or 308P).

[0172] The present disclosure also includes bispecific antigen-binding molecules that contain a first C H 3 domain and a second Ig C H 3 domain, where the first and second Ig C H 3 domains differ from each other by at least one amino acid, and where the at least one amino acid difference reduces the binding of the bispecific antibody to Protein A compared to a bispecific antibody that does not have the amino acid difference. In one embodiment, the first Ig C H 3 domain binds to Protein A and the second Ig C H 3 domain contains a mutation that reduces or eliminates Protein A binding, such as the H95R modification (according to IMGT exon numbering; H435R, according to EU numbering). The second C H3 may further comprise a Y96F modification (according to IMGT; Y436F, according to EU). Other modifications that may be found within the second C H include, in the case of an IgG1 antibody, D16E, L18M, N44S, K52N, V57M, and V821 (according to IMGT; D356E, L358M, N384S, K392N, V397M, and V4221, according to EU); in the case of an IgG2 antibody, N44S, K52N, and V821 (IMGT; N384S, K392N, and V4221, according to EU); and in the case of an IgG4 antibody, Q15R, N44S, K52N, V57M, R69K, E79Q, and V821 (according to IMGT; Q355R, N384S, K392N, V397M, R409K, E419Q, and V4221, according to EU).

[0173] In certain embodiments, the Fc domain may be a chimeric, combinatorial Fc sequence derived from more than one immunoglobulin isotype. For example, a chimeric Fc domain may comprise a portion or all of the C H 2 region of human IgG1, human IgG2, or human IgG4 C H 2 sequence, and a portion or all of the C H 3 sequence of human IgG1, human IgG2, or human IgG4. The chimeric Fc domain may also contain a chimeric hinge region. For example, the chimeric hinge may comprise a combination of an "upper hinge" sequence from the hinge region of human IgG1, human IgG2, or human IgG4, and a "lower hinge" sequence from the hinge region of human IgG1, human IgG2, or human IgG4. Specific examples of chimeric Fc domains that may be included in any of the antigen-binding molecules set forth herein include, from the N-terminus to the C-terminus: [IgG4 C H 1]-[IgG4 upper hinge]-[IgG2 lower hinge]-[IgG4 CH2]-[IgG4 C H 3]. Another example of a chimeric Fc domain that may be included in any of the antigen-binding molecules set forth herein includes, from the N-terminus to the C-terminus: [IgG1 C H 1]-[IgG1 upper hinge]-[IgG2 lower hinge]-[IgG4 CH2]-[IgG1 C H 3]. These and other examples of chimeric Fc domains that may be included in any of the antigen-binding molecules of the present disclosure are described in WO2014 / 022540A1, and chimeric Fc domains and variants thereof having these general structural arrangements may have altered Fc receptor binding, which in turn affects Fc effector functions.

[0174] The antibodies and antigen-binding fragments of the present disclosure comprise immunoglobulin chains that include the amino acid sequences (and variants thereof) specifically set forth herein, as well as cellular and in vitro post-translational modifications of the antibody or fragment. For example, the present disclosure includes antibodies and antigen-binding fragments thereof that specifically bind to EGFR and CD28 comprising the heavy chain and / or light chain amino acid sequences set forth herein, and antibodies and fragments in which one or more asparagine, serine, and / or threonine residues are glycosylated, one or more asparagine residues are deamidated, one or more residues (such as Met, Trp, and / or His) are oxidized, the N-terminal glutamine is pyroglutamic acid (pyroE), and / or the C-terminal lysine or other amino acid is lost.

[0175] The present disclosure also provides antigen-binding proteins that specifically bind to the CD28 protein or an antigenic fragment thereof (such as the extracellular domain of CD28), such as antibodies (such as human antibodies, monoclonal antibodies, and recombinant antibodies) and antigen-binding fragments thereof. Antigen-binding proteins that bind to the same antigenic determinant on CD28 or compete for binding to CD28 as any of the antigen-binding proteins set forth herein are also part of the present disclosure.

[0176] The multi-specific EGFRxCD28 antigen-binding proteins of the present disclosure bind to CD28 on the surface of T cells and co-stimulate CD28 signaling to enhance T cell activation and / or proliferation, which may be referred to herein as "co-stimulatory" or "costimulatory". T cell activation begins after the T cell receptor (TCR) / CD3 complex binds to the peptide-MHC complex ("signal 1"); subsequently, activation is enhanced by engaging a second "co-stimulatory" receptor, such as the CD28 receptor on the T cell that binds to its cognate ligand on the target cell ("signal 2"). For example, activation of T cells by CD28 bispecific antibodies can be caused by signal amplification in response to recognition of endogenous tumor antigens through the TCR / CD3 complex, or "signal 1" activation through CD3 bispecific antibodies.

[0177] (I) Antibodies comprising Fc variants

[0178] According to certain embodiments of the present disclosure, provided are anti-EGFR X anti-CD28 bispecific antigen-binding molecules comprising an Fc domain, the Fc domain comprising one or more mutations that enhance or reduce the binding of the antibody to the FcRn receptor, for example, at an acidic pH compared to neutral pH. For example, the present disclosure includes at C of the Fc domain H 2 or C HThe antibody and antigen-binding molecule in Region 3 contains a mutation that increases the affinity of the Fc domain for FcRn in an acidic environment (such as an endosome where the pH value is in the range of about 5.5 to about 6.0). Such mutations can increase the serum half-life of the antibody when administered to an animal. Non-limiting examples of such Fc modifications include, for example, modifications at position 250 (such as E or Q); 250 and 428 (such as L or F); 252 (such as L / Y / F / W or T); 254 (such as S or T) and / or 256 (such as S / R / Q / E / D or T); or modifications at position 428 and / or 433 (such as H / L / R / S / P / Q or K) and / or 434 (such as H / F or Y); or modifications at position 250 and / or 428; or modifications at position 307 or 308 (such as 308F, V308F) and / or 434.

[0179] In one embodiment, the modification comprises

[0180] · 428L (such as M428L) and 434S (such as N434S) modifications;

[0181] · 428L, 259I (such as V259I) and 308F (such as V308F) modifications;

[0182] · 433K (such as H433K) and 434 (such as 434Y) modifications;

[0183] · 252, 254 and 256 (such as 252Y, 254T and 256E) modifications;

[0184] · 250Q and 428L modifications (such as T250Q and M428L); and / or

[0185] · 307 and / or 308 modifications (such as 308F or 308P).

[0186] By way of example, the present disclosure includes an EGFRxCD28 bispecific antigen-binding molecule comprising an Fc domain, the Fc domain comprising one or more mutation pairs or groups selected from the group consisting of:

[0187] · 250Q and 248L (such as T250Q and M248L);

[0188] · 252Y, 254T and 256E (such as M252Y, S254T and T256E);

[0189] · 428L and 434S (such as M428L and N434S); and

[0190] · 433K and 434F (such as H433K and N434F).

[0191] All possible combinations of the foregoing Fc domain mutations and other mutations within the antibody variable domains disclosed herein are encompassed within the scope of the present disclosure.

[0192] Anti-PD-1 antibodies and antigen-binding fragments thereof

[0193] According to certain exemplary embodiments of the present disclosure, the method comprises administering in combination a therapeutically effective amount of an anti-PD-1 antibody or an antigen-binding fragment thereof and a bispecific EGFRxCD28 antigen-binding molecule or an antigen-binding fragment thereof. According to certain embodiments, the anti-PD-1 antibody or an antigen-binding fragment thereof used in the methods of the present disclosure specifically binds to PD-1. For example, as used in the context of the present disclosure, an antibody that "specifically binds" to PD-1 includes an antibody or an antigen-binding fragment thereof that binds to PD-1 or a portion thereof, wherein K D is less than about 500 nM, less than about 300 nM, less than about 200 nM, less than about 100 nM, less than about 90 nM, less than about 80 nM, less than about 70 nM, less than about 60 nM, less than about 50 nM, less than about 40 nM, less than about 30 nM, less than about 20 nM, less than about 10 nM, less than about 5 nM, less than about 4 nM, less than about 3 nM, less than about 2 nM, less than about 1 nM or less than about 0.5 nM, as measured in a surface plasmon resonance assay. However, an isolated antibody or antigen-binding fragment that specifically binds to human PD-1 may have cross-reactivity with other antigens, such as PD-1 molecules from other (non-human) species.

[0194] According to certain exemplary embodiments of the present disclosure, an anti-PD-1 antibody or an antigen-binding fragment thereof comprises a heavy-chain variable region (HCVR), a light-chain variable region (LCVR), and / or a complementarity-determining region (CDR) that comprises any of the amino acid sequences of the anti-PD-1 antibodies set forth in U.S. Patent No. 9,987,500. In certain exemplary embodiments, an anti-PD-1 antibody or an antigen-binding fragment thereof that can be used in the context of the methods of the present disclosure comprises: a heavy-chain complementarity-determining region (HCDR) of a heavy-chain variable region (HCVR) that comprises the amino acid sequence of SEQ ID NO:73 and a light-chain complementarity-determining region (LCDR) of a light-chain variable region (LCVR) that comprises the amino acid sequence of SEQ ID NO:74. According to certain embodiments, an anti-PD-1 antibody or an antigen-binding fragment thereof comprises three HCDRs (HCDR1, HCDR2, and HCDR3) and three LCDRs (LCDR1, LCDR2, and LCDR3), wherein HCDR1 comprises the amino acid sequence of SEQ ID NO:75; HCDR2 comprises the amino acid sequence of SEQ ID NO:76; HCDR3 comprises the amino acid sequence of SEQ ID NO:77; LCDR1 comprises the amino acid sequence of SEQ ID NO:78; LCDR2 comprises the amino acid sequence of SEQ ID NO:79 (AAS); and LCDR3 comprises the amino acid sequence of SEQ ID NO:80. In still other embodiments, an anti-PD-1 antibody or an antigen-binding fragment thereof comprises: an HCVR that comprises the amino acid sequence of SEQ ID NO:73 and an LCVR that comprises the amino acid sequence of SEQ ID NO:74. In certain embodiments, the methods of the present disclosure comprise using an anti-PD-1 antibody, wherein the antibody comprises a heavy chain that comprises the amino acid sequence of SEQ ID NO:81. In some embodiments, the anti-PD-1 antibody comprises a light chain that comprises the amino acid sequence of SEQ ID NO:82. An exemplary antibody that comprises an HCVR that comprises the amino acid sequence of SEQ ID NO:73 and an LCVR that comprises the amino acid sequence of SEQ ID NO:74 is a fully human anti-PD-1 antibody, called cemipimab (REGN2810; ). According to certain exemplary embodiments, the methods of the present disclosure comprise using cemipimab or a bioequivalent thereof. As used herein, the term "bioequivalent" refers to an anti-PD-1 antibody or a PD-1 binding protein or a fragment thereof that is a pharmaceutical equivalent or a pharmaceutical alternative whose rate and / or extent of absorption does not exhibit a significant difference from the rate and / or extent of absorption of cemipimab when administered in a single dose or multiple doses at the same molar dose under similar experimental conditions. In the context of the present disclosure, the term refers to an antigen-binding protein that binds to PD-1 and that has no clinically meaningful difference from cemipimab in terms of safety, purity, and / or performance.

[0195] Other anti-PD-1 antibodies that can be used in the context of the methods of the present disclosure include, for example, the following antibodies mentioned and known in the art: nivolumab (US 8008449), pembrolizumab (US 8354509), MEDI0608 (US8609089), BI 754091, spartalizumab (also known as PDR001), camrelizumab (also known as SHR-1210), JNJ-63723283, MCLA-134, toripalimab, sintilimab, tilsotolizumab, surufatinib, dostarlimab, rivoceranib, sepantronium bromide, penpulimab, pidilizumab, HX008, bintrafusp alfa, ebemlyzumab, or any of the anti-PD-1 antibodies described in U.S. Patent Nos. 6,808,710, 7,488,802, 8,008,449, 8,168,757, 8,354,509, 8,609,089, 8,686,119, 8,779,105, 8,900,587, and 9,987,500 and patent publications WO2006 / 121168, WO 2009 / 114335; or antigen-binding fragments of any of the foregoing.

[0196] Bioequivalent

[0197] The present disclosure encompasses methods comprising administering an antigen-binding molecule having an amino acid sequence that is different from the amino acid sequence of the described antibody but retains the ability to bind CD28 and EGFR or PD-1. When compared to the parental sequence, such variant molecules contain one or more amino acid additions, deletions, or substitutions, but exhibit biological activity that is substantially equivalent to the biological activity of the described antigen-binding molecule. Similarly, when compared to the disclosed sequences, the DNA sequences encoding the antigen-binding molecules of the present disclosure encompass sequences that contain one or more nucleotide additions, deletions, or substitutions but encode an antigen-binding molecule that is substantially bioequivalent to the antigen-binding molecules described in the present disclosure. Examples of such variant amino acid and DNA sequences are described above.

[0198] The present disclosure includes methods of administering an antigen-binding molecule that is bioequivalent to any of the exemplary antigen-binding molecules set forth herein. For example, two antigen-binding proteins or antibodies are considered bioequivalent if they are pharmaceutical equivalents or pharmaceutical alternatives such that when administered at the same molar dose in single or multiple doses under similar experimental conditions, neither the rate nor the extent of absorption is shown to be significantly different. If some antibodies are equivalent in their extent of absorption but not in their rate of absorption, the antibodies are considered equivalents or pharmaceutical alternatives and may also be considered bioequivalent because such differences in the rate of absorption are intentional and reflected in the labeling, are not necessary to achieve an effective concentration of the subject drug, for example, in chronic use, and are considered medically inconsequential for the particular drug product being studied.

[0199] In one embodiment, two antigen-binding proteins are bioequivalent if there are no clinically meaningful differences in safety, purity, and performance.

[0200] In one embodiment, two antigen-binding proteins are bioequivalent if a subject can switch between a reference product and a biological product one or more times and there is no expected increased risk of adverse reactions (including clinically significant changes in immunogenicity or waning of efficacy) compared to continuous therapy without such switching.

[0201] In one embodiment, two antigen-binding proteins are bioequivalent if both act via one or more common mechanisms of action under one or more conditions of use to an extent known for such mechanisms.

[0202] Bioequivalence can be demonstrated by in vivo and in vitro methods. Bioequivalence metrics include, for example, (a) in vivo tests in humans or other mammals in which the concentration of an antibody or its metabolite in blood, plasma, serum, or other biological fluid is measured over time; (b) in vitro tests that have been correlated with and reasonably predict human in vivo bioavailability data; (c) in vivo tests in humans or other mammals in which the appropriate acute pharmacological effect of an antibody (or its target) is measured over time; and (d) well-controlled clinical trials that establish the safety, efficacy, or bioavailability or bioequivalence of an antibody.

[0203] Bioequivalent variants of the exemplary bispecific antigen-binding molecules described herein can be constructed, for example, by making various substitutions of residues or sequences, or by deleting terminal or internal residues or sequences that are not essential for biological activity. For example, cysteine residues that are not essential for biological activity can be deleted or replaced with other amino acids to prevent the formation of unwanted or inappropriate intramolecular disulfide bridges upon refolding. In other cases, bioequivalent antibodies can include the exemplary bispecific antigen-binding molecules described herein that contain amino acid changes that modify the glycosylation profile of the antibody, such as mutations that eliminate or remove glycosylation.

[0204] Pharmaceutical Formulations and Kits

[0205] The present disclosure provides compositions comprising a bispecific EGFRxCD28 antibody (e.g., REGN7075) and / or an anti-PD-1 antibody or antigen-binding portion thereof, and one or more components; and methods of using the same.

[0206] The present disclosure provides pharmaceutical compositions comprising a bispecific EGFRxCD28 antibody or antigen-binding fragment thereof and / or an anti-PD-1 antibody or antigen-binding fragment thereof of the present disclosure. The pharmaceutical compositions of the present disclosure can be formulated with suitable carriers, excipients, and other agents that provide improved transfer, delivery, tolerability, etc. Numerous suitable formulations can be found in formularies known to all pharmaceutical chemists: Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, PA. These formulations include, for example, powders, pastes, ointments, gels, waxes, oils, lipids, vesicles containing lipids (cationic or anionic) (e.g., LIPOFECTIN TM , Life Technologies, Carlsbad, CA), DNA conjugates, anhydrous absorption pastes, water-in-oil and oil-in-water emulsions, emulsion carbowax (polyethylene glycol of various molecular weights), semi-solid gels, and semi-solid mixtures containing carbowax. See also Powell et al. “Compendium of excipients for parenteral formulations” PDA (1998) J Pharm Sci Technol 52:238-311.

[0207] For the preparation of pharmaceutical formulations of antigen-binding proteins, such as antibodies and antigen-binding fragments thereof (e.g., REGN7075; REGN6321; REGN6322; REGN6323; or cemipimab), the antigen-binding protein is admixed with a pharmaceutically acceptable carrier or excipient. See, e.g., Remington's Pharmaceutical Sciences and U.S. Pharmacopeia: National Formulary, Mack Publishing Company, Easton, Pa. (1984); Hardman et al. (2001) Goodman and Gilman's The Pharmacological Basis of Therapeutics, McGraw-Hill, New York, N.Y.; Gennaro (2000) Remington: The Science and Practice of Pharmacy, Lippincott, Williams and Wilkins, New York, N.Y.; Avis et al. (eds.) (1993) Pharmaceutical Dosage Forms: Parenteral Medications, Marcel Dekker, NY; Lieberman et al. (eds.) (1990) Pharmaceutical Dosage Forms: Tablets, Marcel Dekker, NY; Lieberman et al. (eds.) (1990) Pharmaceutical Dosage Forms: Disperse Systems, Marcel Dekker, NY; Weiner and Kotkoskie (2000) Excipient Toxicity and Safety, Marcel Dekker, Inc., New York, N.Y. In one embodiment, the pharmaceutical formulation is sterile. Such compositions are part of the present disclosure.

[0208] The pharmaceutical formulations of the present disclosure include bispecific EGFRxCD28 antigen-binding proteins and / or anti-PD-1 antibodies or antigen-binding fragments thereof, and a pharmaceutically acceptable carrier (including, for example, water, buffers, preservatives, and / or detergents).

[0209] The scope of the present disclosure includes dried, e.g., lyophilized, compositions that comprise a bispecific EGFRxCD28 antigen-binding protein, or an anti-PD-1 antibody or antigen-binding fragment thereof, or a pharmaceutical formulation thereof that includes a pharmaceutically acceptable carrier but is substantially free of water.

[0210] A variety of delivery systems are known and can be used to administer the pharmaceutical compositions of the present disclosure, such as encapsulated in liposomes, microparticles, microcapsules, recombinant cells capable of expressing mutant viruses, receptor-mediated endocytosis (see, e.g., Wu et al., 1987, J. Biol. Chem. 262:4429-4432). Introduction methods include (but are not limited to) intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, and oral routes. The compositions can be administered by any convenient route, such as by infusion or bolus injection, by absorption through epithelial or mucocutaneous linings (such as oral mucosa, rectal, and intestinal mucosa, etc.) and can be administered together with other bioactive agents. Administration can be systemic or local.

[0211] As discussed herein, the present disclosure provides a container (e.g., a plastic or glass vial) or an injection device (e.g., a syringe, a prefilled syringe, or an autoinjector) that contains any of the antigen-binding proteins herein (e.g., an antibody or an antigen-binding fragment thereof), or a pharmaceutical formulation that contains a pharmaceutically acceptable carrier or excipient thereof.

[0212] The pharmaceutical compositions of the present disclosure can be delivered subcutaneously or intravenously using standard needles and syringes. Additionally, with respect to subcutaneous delivery, pen-type delivery devices are readily applicable in delivering the pharmaceutical compositions of the present disclosure. Such pen-type delivery devices can be reusable or disposable. Reusable pen-type delivery devices generally utilize replaceable cartridges containing the pharmaceutical composition. After all of the pharmaceutical composition in the cartridge has been administered and the cartridge is emptied, the empty cartridge can be easily discarded and replaced with a new cartridge containing the pharmaceutical composition. Subsequently, the pen-type delivery device can be reused. In disposable pen-type delivery devices, there is no replaceable cartridge. In fact, disposable pen-type delivery devices are prefilled with the pharmaceutical composition stored in a reservoir within the device. After emptying the reservoir having the pharmaceutical composition, the entire device is discarded.

[0213] Many reusable and disposable pen and autoinjector delivery devices are used in subcutaneously delivering the pharmaceutical compositions of the present disclosure. See, e.g., AUTOPEN TM (Owen Mumford, Inc., Woodstock, UK) or HUMIRA TM Pen (Abbott Labs, Abbott Park, IL).

[0214] In certain instances, the pharmaceutical composition may be delivered in a controlled release system. In one embodiment, a pump may be used (see Langer, supra; Sefton, 1987, CRC Crit. Ref. Biomed. Eng. 14:201). In another embodiment, polymeric materials may be used; see Medical Applications of Controlled Release, Langer and Wise (eds.), 1974, CRC Pres., Boca Raton, Florida. In yet another embodiment, the controlled release system may be placed proximal to the target of the composition, such that only a fraction of the systemic dose is required (see, e.g., Goodson, 1984, in Medical Applications of Controlled Release, supra, vol. 2, pp. 115-138). Other controlled release systems are discussed in the review by Langer, 1990, Science 249:1527-1533.

[0215] Injectable preparations may include dosage forms for intravenous, subcutaneous, intradermal, and intramuscular injection, drip infusion, etc. These injectable preparations can be prepared by publicly known methods. For example, an injectable preparation can be prepared, for example, by dissolving, suspending, or emulsifying the antibody or its salt described above in a sterile aqueous medium or an oily medium conventionally used for injection. As the aqueous medium for injection, there are, for example, physiological saline and other isotonic solutions that can be used in combination with a suitable solubilizer. Injectable oily media are also part of the present disclosure. Such oily media can be combined with a solubilizer.

[0216] The pharmaceutical compositions for oral or parenteral use described above are preferably prepared in dosage unit forms suitable for fitting the dose of the active ingredient. Such dosage unit forms include, for example, tablets, pills, capsules, injections (ampoules), suppositories, etc. The amount of the aforementioned antibody contained in a unit dose is generally about 0.1 to about 2000 mg per dosage form; especially in the form of an injection.

[0217] The present disclosure also provides a kit comprising a bispecific EGFRxCD28 antibody and an anti-PD-1 antibody or antigen-binding portion thereof for therapeutic use. The kit generally includes a label and instructions for use indicating the intended use of the contents of the kit. The term label includes any written or recorded material on, provided with, or otherwise accompanying the kit. Accordingly, the present disclosure provides a kit for treating a subject suffering from cancer, the kit comprising: (a) a dose of the antibody or antigen-binding portion thereof; and (b) a dose of the bispecific EGFRxCD28 antibody or antigen-binding portion thereof; and (c) instructions for using the antibody in any of the treatment methods disclosed herein. In certain embodiments, the dose of the anti-PD-1 antibody or antigen-binding fragment thereof ranges from 150 to 550 mg, such as 350 mg. In certain embodiments, the dose of the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof ranges from 0.1 mg to 3000 mg.

[0218] Additional combination therapies

[0219] The present disclosure provides methods of treating and preventing diseases (such as cancer) by administering a bispecific EGFRxCD28 antibody (e.g., REGN7075) or antigen-binding fragment thereof alone in a monotherapy format, or in combination with an anti-PD-1 antibody or antigen-binding fragment thereof; and optionally in combination with one or more therapeutic agents (e.g., at least one third therapeutic agent or therapy).

[0220] In certain embodiments, the third therapeutic agent or therapy is selected from surgery, radiotherapy, chemotherapy (e.g., anti-cancer chemotherapy such as paclitaxel, docetaxel, vincristine, cisplatin, carboplatin, or oxaliplatin), CAR-T cell therapy, cancer vaccines, oncolytic viruses, cytokines, anti-VEGF therapy, anti-EGFR therapy, or anti-cancer drugs. As used herein, "anti-cancer drug" means any agent suitable for treating cancer, including (but not limited to) cytotoxins and agents such as: antimetabolites, alkylating agents, anthracyclines, antibiotics, anti-mitotic agents, procarbazine, hydroxyurea, asparaginase, corticosteroids, mitotane (O,P'-(DDD)), biological agents (such as antibodies and interferons), and radiopharmaceuticals. As used herein, "cytotoxin or cytotoxic agent" also refers to a chemotherapeutic agent and means any agent that is harmful to cells. Examples include (Paclitaxel), temozolamide, cytochalasin B, gramicidin D, ethidium bromide, emetine, cisplatin, mitomycin, etoposide, tenoposide, vincristine, vinbiastine, colchicine, doxorubicin, daunorubicin, dihydroxy anthracin dione, mitoxantrone, mithramycin, actinomycin D, 1-dehydrotestosterone, glucocorticoid, procaine, tetracaine, lidocaine, propranolol, and puromycin and their analogs or homologs. In certain embodiments, the method comprises administering an agent to reduce or alleviate or treat the symptoms of an immune-related adverse event. In one embodiment, the agent is selected from an IL-6 inhibitor (e.g., an anti-IL6 receptor antibody, e.g., tocilizumab or sarilumab), a corticosteroid, or a non-steroidal anti-inflammatory agent.

[0221] In another embodiment, according to the Physicians' Desk Reference 2003 (Thomson Healthcare; 57th Edition (November 1, 2002)) or the approved prescribing information typically provided with a particular agent, another therapeutic agent is administered to the subject, which is administered to the subject in combination with the bispecific EGFRxCD28 antibody or its antigen-binding fragment and the anti-PD-1 antibody or its antigen-binding fragment.

[0222] The term "in combination with" indicates that a component, a bispecific EGFRxCD28 antibody or an antigen-binding fragment thereof (e.g., REGN7075), an anti-PD-1 antibody or an antigen-binding fragment thereof (e.g., cemipimab) can be formulated into a single composition together with another therapeutic agent, e.g., for simultaneous delivery, or into two or more compositions separately (e.g., a kit comprising each component). Alternatively, components administered "in combination" with each other can be administered to a subject at different times than other components are administered; for example, as part of a treatment regimen, each administration can be provided at intervals within a given time period non-simultaneously (e.g., separately or sequentially). The individual components administered in combination with each other can also be administered sequentially (although substantially simultaneously) during the same administration phase. In addition, the individual components administered in combination with each other can be administered to the subject by the same or different routes.

[0223] In some embodiments, the third therapeutic agent is administered on the same day as the bispecific EGFRxCD28 antibody and / or the anti-PD-1 antibody. In some embodiments, the third therapeutic agent is administered on the same day as the bispecific EGFRxCD28 antibody and / or the anti-PD-1 antibody and after the bispecific EGFRxCD28 antibody and / or the anti-PD-1 antibody. In some embodiments, the third therapeutic agent is administered on the same day as the bispecific EGFRxCD28 antibody and / or the anti-PD-1 antibody and before the bispecific EGFRxCD28 antibody and / or the anti-PD-1 antibody. In some embodiments, the third therapeutic agent is administered on the same day as the bispecific EGFRxCD28 antibody and after the bispecific EGFRxCD28 antibody, but on a different day than the anti-PD-1 antibody. In some embodiments, the third therapeutic agent is administered on the same day as the bispecific EGFRxCD28 antibody and before the bispecific EGFRxCD28 antibody, but on a different day than the anti-PD-1 antibody. In some embodiments, the third therapeutic agent is administered on the same day as the anti-PD-1 antibody and after the anti-PD-1 antibody, but on a different day than the bispecific EGFRxCD28 antibody. In some embodiments, the third therapeutic agent is administered on the same day as the anti-PD-1 antibody and before the anti-PD-1 antibody, but on a different day than the bispecific EGFRxCD28 antibody.

[0224] In some embodiments, the third therapeutic agent is administered on different days from the bispecific EGFRxCD28 antibody and / or the anti-PD-1 antibody. In some embodiments, the third therapeutic agent is administered on different days from the bispecific EGFRxCD28 antibody and / or the anti-PD-1 antibody and is administered after the bispecific EGFRxCD28 antibody and / or the anti-PD-1 antibody. In some embodiments, the third therapeutic agent is administered on different days from the bispecific EGFRxCD28 antibody and / or the anti-PD-1 antibody and is administered before the bispecific EGFRxCD28 antibody and / or the anti-PD-1 antibody. In some embodiments, the third therapeutic agent is administered on different days from the bispecific EGFRxCD28 antibody and is administered after the bispecific EGFRxCD28 antibody, but is administered on different days from the anti-PD-1 antibody. In some embodiments, the third therapeutic agent is administered on different days from the bispecific EGFRxCD28 antibody and is administered before the bispecific EGFRxCD28 antibody, but is administered on different days from the anti-PD-1 antibody.

[0225] In some embodiments, a combination therapy of a bispecific EGFRxCD28 antibody (e.g., REGN7075) and an anti-PD-1 antibody (e.g., cemipimab) is administered alone during a lead-in period before initiating treatment with the third therapeutic agent.

[0226] In some embodiments, the combination therapy with the third therapeutic agent is administered at least 2, 3, 4, 5, 6, 7, or more times.

[0227] In certain embodiments, the subject does not exhibit side effects or does not experience any side effects due to administration of the bispecific EGFRxCD28 antibody alone or in combination with the anti-PD-1 antibody, and / or due to administration of the bispecific EGFRxCD28 antibody alone or in combination with the third therapeutic agent.

[0228] Examples

[0229] The following examples are provided to give a complete disclosure and description of how to make and use the methods and compositions of the present disclosure to those of ordinary skill in the art and are not intended to limit the scope of what the inventors regard as their disclosure.

[0230] Example 1: Bispecific EGFRxCD28 Antibody

[0231] Generation of Anti - EGFR and Anti - CD28 Antibodies

[0232] Anti-EGFR and anti-CD28 antibodies were obtained as described in WO2020 / 198009, the entire content of which is hereby incorporated by reference in its entirety.

[0233] Table 1 sets forth the amino acid sequence identifiers of the heavy and light chain variable regions and CDRs of the selected anti-EGFR antibodies of the present disclosure. The identifiers of the corresponding nucleic acid sequences are set forth in Table 2.

[0234] Table 1: Amino Acid Sequence Identifiers of Parental EGFR Monoclonal Antibodies (mAbs)

[0235]

[0236] SEQ ID NO:20: Gly Ala Ser (GAS)

[0237] Table 2: Nucleic Acid Sequence Identifiers of Parental EGFR Monoclonal Antibodies (mAbs)

[0238]

[0239] SEQ ID NO:19: ggggcaagt

[0240] Table 3 sets forth the amino acid sequence identifiers of the heavy and light chain variable regions and CDRs of the selected anti-CD28 antibodies of the present disclosure. The identifiers of the corresponding nucleic acid sequences are set forth in Table 4.

[0241] Table 3: Amino Acid Sequence Identifiers of Parental CD28 Monoclonal Antibodies (mAbs)

[0242]

[0243] Table 4: Nucleic Acid Sequence Identifiers of Parental CD28 Antibodies

[0244]

[0245] Generation of Bispecific Antibodies (bsAb) that Bind CD28 and EGFR

[0246] Bispecific antibodies comprising an anti-EGFR specific binding domain and an anti-CD28 specific binding domain are constructed using standard methods, wherein the anti-EGFR antigen binding domain and the anti-CD28 antigen binding domain each comprise different, distinct HCVRs paired with a common LCVR. In some cases, bispecific antibodies are constructed using the heavy chain from an anti-CD28 antibody, the heavy chain from an anti-EGFR antibody, and a common light chain comprising the components, amino acids, and nucleic acid sequences encoding the antibodies shown in Tables 5, 6, 7, and 8 below. Additional bispecific antibodies that bind EGFR and CD28 can be prepared using parental monoclonal antibodies having the names shown in Tables 9A, 9B, and 9C.

[0247] Table 5: Overview of Antibody Names for the HCVR Arms of Anti-EGFR x Anti-CD28 Bispecific Antibodies

[0248]

[0249] Table 6: Amino acid sequence of anti-EGFR x anti-CD28 bispecific antibody

[0250]

[0251] Table 7: Nucleic acid sequence encoding anti-EGFR x anti-CD28 bispecific antibody

[0252]

[0253] Table 8. Amino acid and nucleotide sequences of the full-length immunoglobulin chains of bispecific antibodies bsAb7075, bsAb6321, bsAb6322, and bsAb6323

[0254]

[0255] D = Nucleotide sequence of DNA encoding the designated sequence

[0256] P = Amino acids of the polypeptide of the designated sequence

[0257] Numbers refer to SEQ ID NO of the designated sequence

[0258] HC is the full-length heavy chain of the designated antibody

[0259] LC is the full-length light chain of the designated antibody

[0260] Additional bispecific antibodies comprising one HCVR from a parental EGFR antibody and the other HCVR arm from a parental CD28 antibody can be prepared using the techniques described herein. The parental EGFR antibodies used to generate these additional anti-EGFR X anti-CD28 bispecific antibodies have the HCVR sequences described in WO2014 / 004427. The parental CD28 antibodies used to generate these additional anti-EGFR X anti-CD28 bispecific antibodies have the amino acid sequences described in Table 3 above. These anti-EGFR and anti-CD28 binding domain (pairs) are shown in Tables 9A, 9B, and 9C below.

[0261] Table 9A: Overview of parental antibody names for the HCVR arms of additional anti-EGFR x anti-CD28 bispecific antibodies

[0262]

[0263]

[0264] Table 9B: Overview of parental antibody names for the HCVR arms of additional anti-EGFR x anti-CD28 bispecific antibodies

[0265]

[0266] Table 9C: Overview of parental antibody names for the HCVR arms of additional anti-EGFR x anti-CD28 bispecific antibodies

[0267]

[0268]

[0269] Exemplary bispecific antibodies were manufactured to have a modified (chimeric) IgG4 Fc domain as set forth in U.S. Patent Application Publication No. US20140243504A1, published August 28, 2014.

[0270] The bispecific antibodies described in this example comprise two separate antigen-binding domains (i.e., binding arms). The first antigen-binding domain comprises a heavy chain variable region derived from an anti-CD28 antibody ("CD28-VH"), and the second antigen-binding domain comprises a heavy chain variable region derived from an anti-EGFR antibody ("EGFR-VH"). Both anti-EGFR and anti-CD28 share a common light chain. The CD28-VH / EGFR-VH pairing results in an antigen-binding domain that specifically recognizes CD28 on T cells and EGFR on tumor cells.

[0271] Example 2: Evaluation of the treatment with REGN7075 alone and in combination with cemipimab

[0272] This example describes a Phase 1 clinical study of the combination of REGN7075 (an EGFRxCD28 costimulatory bispecific antibody) and cemipimab in subjects with advanced solid tumors.

[0273] Study Objectives

[0274] The primary objective of dose escalation was to evaluate the safety and tolerability of the induction phase of REGN7075 monotherapy and the combination of REGN7075 and cemipimab in subjects with advanced solid tumors.

[0275] The primary objective of dose expansion was to evaluate the initial efficacy of the combination of REGN7075 and cemipimab within selected advanced solid tumor-specific groups, as measured by the objective response rate (ORR) according to Response Evaluation Criteria in Solid Tumors version 1.1 (RECIST 1.1) and / or composite response criteria (depending on the subject's baseline evaluation criteria).

[0276] The secondary objectives of dose escalation were: (i) to characterize the pharmacokinetics (PK) of REGN7075 alone and in combination with cemipimab; (ii) to evaluate the initial efficacy of the combination of REGN7075 and cemipimab, measured by objective response rate (ORR), overall survival (OS), progression-free survival (PFS), duration of response (DOR), complete response (CR) rate, and disease control rate (DCR) according to RECIST 1.1 and / or combination response criteria (depending on the baseline assessment criteria of the subject); (iii) to evaluate the immunogenicity of REGN7075 and cemipimab.

[0277] The secondary objectives of dose expansion were: (i) to evaluate the initial efficacy of the combination of REGN7075 and cemipimab within selected groups of subjects with advanced solid tumors, measured by OS, PFS, DOR, CR rate, and DCR according to RECIST 1.1 and / or combination response criteria (depending on the baseline assessment criteria of the subject); (ii) to evaluate the safety and tolerability of the combination of REGN7075 and cemipimab with or without chemotherapy; (iii) to characterize the PK of REGN7075 alone and in combination with cemipimab with or without chemotherapy; (iv) to evaluate the immunogenicity of REGN7075 and cemipimab with or without chemotherapy; (v) to evaluate the effect of REGN7075 on subject-reported outcomes, including health-related quality of life (HRQoL), measured by the validated tools European Organisation for Research and Treatment of Cancer Quality of Life Questionnaire (EORTC QLQ-C30), EORTC-QLQ-BR23 (breast cancer patients only), EORTC QLQ-CR29 (CRC subjects only), EORTC QLQ-LC13 (NSCLC subjects only), and EORTC QLQ-HN35 (HNSCC subjects only) and EQ-5D-5L.

[0278] Exploratory objectives include: (i) measuring circulating tumor DNA content over time; (ii) exploring tumor gene alterations detected in circulating tumor DNA (ctDNA) at baseline and after dosing that are relevant to efficacy and safety; (iii) evaluating the efficacy of REGN7075 alone and in combination with cemipimab in patients with low and high tumor mutation burden (TMB) within selected advanced solid tumor cohorts, with or without prior chemotherapy; and (iv) assessing EGFR and programmed cell death ligand (PD-L1) expression in tumor and immune cell populations by evaluating proteins and RNA in tumor tissue.

[0279] Study Design

[0280] This is an open-label, Phase 1 / 2, first-in-human (FIH) study that evaluates the safety, tolerability, PK, and initial anti-tumor activity of REGN7075 alone and in combination with cemipimab in patients with advanced solid tumors.

[0281] This study has two parts:

[0282] Dose Escalation

[0283] During dose escalation, patients will receive a 3-week monotherapy lead-in of REGN7075 at the designated dose level (DL) QW IV, followed by combination therapy with REGN7075 at the designated DL and cemipimab 350 mg Q3W IV. Dose levels of REGN7075 and cemipimab 350 mg Q3W IV that can be studied with simultaneous start (i.e., no lead-in period) will be explored. Once the MTD / RP2D of REGN7075 IV is identified and after recruitment and DLT monitoring of the simultaneous start dose escalation cohort ( Figure 2B ), recruitment of the exploratory SC dosing cohort can be initiated. Combinations of SC administration of REGN7075 with IV or SC cemipimab will be explored.

[0284] Based on emerging PK data, recruitment of the lead-in and simultaneous start dose escalation cohorts can be conducted at REGN7075 QW ( Figure 2A and Figure 2B ). To address infusion-related reactions (IRR), the escalating dosing of REGN7075 will include escalating doses in Week 1 and target doses from Week 2 onwards. The initial escalating dose will be 30 mg. The dose and / or rate of this escalating dose can be adjusted. Additionally, the escalating dose and the first target dose can be administered separately over 2 days.

[0285] Dose Expansion

[0286] The RP2D will be determined based on all data. Dose escalation will be based on the RP2D. During dose escalation, patients will receive the combination therapy of REGN7075 at the specified DL (e.g., RP2D) and schedule. Dose escalation will consist of patients with various different mixed advanced solid tumor types. Dose expansion will consist of the following tumor-specific expansion cohorts:

[0287] Cohort A: Triple-negative breast cancer (TNBC)

[0288] Cohort B: Cutaneous squamous cell carcinoma (CSCC)

[0289] Cohort C: Non-small cell lung cancer (NSCLC)

[0290] Cohort D: Head and neck squamous cell carcinoma (HNSCC)

[0291] Cohort E: Microsatellite stable colorectal cancer (MSS-CRC) with active liver metastases and / or active peritoneal metastases

[0292] Cohort F: MSS-CRC with isolated lung / lymph node metastases (without active liver metastases and without active peritoneal metastases)

[0293] Cohort G: EGFR-mutated NSCLC after third-generation TKI

[0294] Cohort H: EGFR-mutated NSCLC after third-generation TKI and after platinum doublet chemotherapy

[0295] The dose expansion cohorts will be recruited after identifying the REGN7075 RP2D in combination with zimberelimab. Patients in Cohorts C and G will also receive 4 cycles of platinum-based chemotherapy. Before recruiting the expansion cohorts, it may be assumed that the RP2D recruits additional patients to further evaluate safety and bioactivity.

[0296] Patients will be treated with the study drug until they meet the criteria of definite disease progression, intolerable adverse events (AEs), withdrawal of consent, or other study discontinuation criteria.

[0297] Treatment will consist of a 3-week lead-in period of REGN7075 at the RP2D QW or start simultaneously with its combination with zimberelimab 350 mg Q3W IV. Patients in Cohorts C and G will also receive 4 cycles of platinum-based chemotherapy, which will be administered on the same day as zimberelimab (Q3W).

[0298] If the dose levels of simultaneous start schedules (i.e., starting REGN7075 and cemipimab without a REGN7075 lead-in period) are not studied, or if simultaneous start dose levels are tested and found to be intolerable (e.g., cytokine release syndrome (CRS) or other DLTs increase compared to the monotherapy lead-in schedule), then the expansion cohort will be opened at the previously tolerable dose level and will include a REGN7075 lead-in period. If the simultaneous start dose level is found to be tolerable, then the expansion cohort will be opened and the simultaneous schedule will be used.

[0299] Study duration

[0300] The total duration for each subject will vary based on the occurrence of one or more of the following: disease progression, intolerable AEs, withdrawal of consent, or study discontinuation criteria.

[0301] The study schedule consists of: screening (up to 28 days), treatment (variable duration and timing), and follow-up (approximately 90 days from the last dose).

[0302] The timing of the doses is outlined in Figure 2.

[0303] Treatment will continue until disease progression, intolerable adverse events, withdrawal of consent, or other treatment discontinuation criteria are met. Disease assessments will be performed after Cycle 1, after Cycle 2, and then every 12 weeks thereafter. In the absence of clinical deterioration, it may be optional to continue treating the subject on the same schedule until radiographic progression is confirmed (PD on 2 consecutive scans; the second scan must be performed at least 4 weeks after the first scan and must not be later than the subject's next scheduled scan). Further treatment beyond confirmed radiographic progression will be considered upon request, provided that the treatment remains well tolerated and is considered to be in the subject's interest. Responses will be evaluated according to RECIST 1.1 and / or composite response criteria (depending on the subject's baseline assessment criteria), but scan results will be collected and saved, and central review may be optional.

[0304] Dose Escalation

[0305] The study will initiate dose escalation of the combination of REGN7075 and a fixed dose of cemipimab in patients with advanced solid tumors who have never been treated with anti-PD-1 or anti-PD-L1 therapy previously ("treatment-naïve anti-PD-1 / PD-L1"). Treatment at each DL (Table 10) consists of a 3-week monotherapy lead-in period of REGN7075, followed by combination therapy of REGN7075 and cemipimab for a 6-week combined DLT period (see Figures 2A - 2D )

[0306] Table 10: Planned Dose Escalation Cohorts and Enrollment

[0307]

[0308]

[0309] At the specified DL, each patient will start with a lead-in period of REGN7075 monotherapy, during which the safety and PK of REGN7075 monotherapy will be evaluated. The lead-in period is planned to last 21 days and include 3 doses of REGN7075 QW. This will be followed by combination therapy of REGN7075 with cemipimab 350 mg Q3W on a QW or Q3W schedule (same nominal dose as QW). Combination therapy will only start when the patient 1) has received at least three doses of REGN7075 and 2) has not had a ≥ Grade 2 CRS, rash, or imAE related to the most recent REGN7075 monotherapy infusion. If a ≥ Grade 2 CRS, rash, or imAE is observed during the third dose of the lead-in period but resolves before the next scheduled dose of REGN7075, then REGN7075 monotherapy will continue for the fourth dose. If a ≥ Grade 2 CRS, rash, or imAE is observed during the fourth dose of monotherapy, then the patient will not start combination therapy and will be given the option to continue monotherapy. Monotherapy dosing will continue weekly at Week 3 (Day 15). These patients will be considered uninformative with respect to combination therapy dose selection and may be replaced if necessary, but will be allowed to continue in the study at the discretion of the investigator. In this case, the patient will complete all evaluations except for cemipimab treatment. The patient will only receive the combination of REGN7075 plus cemipimab when REGN7075 is administered and no ≥ Grade 2 CRS, rash, or imAE has occurred. One or more of the designated expansion cohorts may be opened and confirmed safe at the selected dose level while continuing dose escalation.

[0310] Simultaneously Starting Cohorts

[0311] Additional cohorts may be enrolled without a 3-week lead-in period of REGN7075 monotherapy (i.e., cemipimab and REGN7075 start together; considered the "start simultaneously" schedule in this document). These start simultaneously cohorts dosed with REGN7075 QW and / or cemipimab Q3W will only be opened after reviewing the safety and tolerability of the REGN7075 monotherapy lead-in period and the prospective dose of cemipimab.

[0312] In - Patient Dose Escalation

[0313] Patients with partial response (PR), stable disease (SD), or PD patients who were previously granted approval to treat beyond progression (and had at least 2 scans) at Day 1 of Cycle 3 or later and were tolerant to the therapy (observed maximum grade ≤2 treatment-related toxicity during previous treatment cycles), but did not show further improvement at their assigned dose level, may be considered for within-patient dose escalation with REGN7075 until the highest DL considered safe is reached (i.e., if dose escalation is ongoing, 1 level lower than the currently actively enrolled DL; if dose escalation is completed and the RP2D is determined, the current dose level). PR patients must not show further improvement in at least 3 consecutive scans. Patients tolerant to the therapy may be considered for further escalation, but not more frequently than once every 6 weeks.

[0314] Subcutaneous Dose Escalation Cohorts

[0315] To further inform safety, efficacy, and dosing administration, an SC dose escalation cohort may be initiated. Subcutaneous dose escalation will follow the DL with the starting dose based on preclinical, PK, and clinical data to date. Cemiplimab may continue to be administered IV (350 mg IV Q3W), or alternatively, SC administration may be done (the development results of SC cemiplimab in other studies are pending). The SC cohort will be denoted with the milligram dose as "SC" (e.g., if DL8 is the assumed RP2D, then DL_SC_100, where 100 represents the milligram dose of REGN7075).

[0316] Dose Expansion

[0317] Groups consisting of specific solid tumor types will be treated with REGN7075, with the associated doses and schedules demonstrated to be safe during dose escalation.

[0318] If the dose levels of the simultaneous start schedule (i.e., starting REGN7075 and cemiplimab without a REGN7075 lead-in period) were not studied, or the tested dose levels of the simultaneous start were found to be intolerable (e.g., increased CRS or other DLTs compared to the monotherapy lead-in schedule), then the expansion cohort will be opened at the previously tolerable dose level and will include a REGN7075 lead-in period. If the simultaneous start dose levels are found to be tolerable, then the expansion cohort will be opened and the simultaneous schedule will be used. The expansion cohorts are listed below:

[0319] Cohort A: Triple - Negative Breast Cancer

[0320] · Patients with previously documented metastatic TNBC or locally advanced TNBC who are not candidates for curative surgery or curative radiotherapy, and

[0321] · Have not received anti-PD-1 / PD-L1

[0322] Cohort B: Cutaneous Squamous Cell Carcinoma

[0323] · Patients with metastatic CSCC or locally advanced CSCC who are not candidates for curative surgery or curative radiotherapy, and

[0324] · Additionally not candidates for cemipimab monotherapy (or another anti-PD-1 / PD-L1 agent), and

[0325] · Have not received anti-PD-1 / PD-L1

[0326] Cohort C: Non - Small Cell Lung Cancer

[0327] · Patients with metastatic NSCLC or locally advanced NSCLC who are not candidates for curative surgery or curative radiotherapy, and

[0328] · Do not have a previously documented targetable molecular driver mutation (e.g., ALK, ROS1, EGFR, RET fusion, MET exon-14 skipping mutation), and

[0329] · Have not received anti-PD-1 / PD-L1

[0330] · Have not received prior systemic therapy for recurrent or metastatic NSCLC (note that adjuvant or neoadjuvant systemic therapy excluding anti-PD-1 / PD-L1 therapy will not be considered a prior line).

[0331] Cohort D: Head and Neck Squamous Cell Carcinoma

[0332] · Patients with metastatic HNSCC or locally advanced HNSCC who are not candidates for curative surgery or curative radiotherapy, and

[0333] · Patients with a previously documented PD-L1 expression CPS ≥ 1% by IHC. CPS is defined as the number of PD-L1 stained cells (tumor cells, lymphocytes, macrophages) divided by the total number of viable tumor cells multiplied by 100.

[0334] · Have not received anti-PD-1 / PD-L1

[0335] Cohort E: Microsatellite - Stable Colorectal Cancer with Active Liver Metastasis and / or Active Peritoneal Metastasis

[0336] · Patients with metastatic CRC or locally advanced CRC who are not candidates for curative surgery or curative radiotherapy, and

[0337] · Active metastatic disease in the liver or peritoneum at screening

[0338] - Ablated and resected liver diseases are not considered active.

[0339] Note: MSS-CRC patients with RECIST evaluable disease that is not independent of the lung / lymph nodes but not occurring in the liver / peritoneum may also be recruited into this group.

[0340] · The disease is MSS according to previously recorded results, and

[0341] · Has received at least one previous line of therapy

[0342] Patients with previously recorded RAS wild-type disease must have received anti-EGFR therapy or have a recorded reason why anti-EGFR therapy is not appropriate

[0343] Patients must have received anti-VEGF therapy or have a recorded reason why anti-VEGF therapy is not suitable

[0344] · Has not received anti-PD-1 / PD-L1

[0345] Cohort F: Microsatellite - Stable Colorectal Cancer with Inter - segmental Lung / Lymph Node Metastasis (without Active Liver Metastasis and without Active Peritoneal Metastasis)

[0346] · Patients with metastatic CRC or locally advanced CRC who are not candidates for curative surgery or curative radiotherapy, and

[0347] · No active metastases were identified in the liver or peritoneum at screening,

[0348] Ablated and resected liver diseases are not considered active.

[0349] · The metastatic sites are only the lung and / or hilar lymph nodes

[0350] · The disease is MSS according to previously recorded results, and

[0351] · Has received at least one previous line of therapy

[0352] Patients with previously recorded RAS wild-type disease must have received anti-EGFR therapy or have a recorded reason why anti-EGFR therapy is not appropriate

[0353] Patients must have received anti-VEGF therapy or have a recorded reason why anti-VEGF therapy is not suitable

[0354] · Has not received anti-PD-1 / PD-L1

[0355] Cohort G: EGFR - Mutant NSCLC after 3rd - Generation TKI

[0356] · Patients with histologically or cytologically documented locally advanced or metastatic non-squamous NSCLC who are not candidates for curative surgery or curative radiotherapy, and

[0357] · With a previously documented targetable EGFR mutation:

[0358] - NSCLC with EGFR exon 19 deletion

[0359] - NSCLC with EGFR L858R mutation

[0360] - NSCLC with activating EGFR exon 20 insertion

[0361] - NSCLC with exon 18 / 21 atypical mutations

[0362] Note: The EGFR deletion / mutation must have been previously documented by a certified test (allowing tissue from blood or ctDNA)

[0363] · Documentation of EGFR mutation status can be obtained at any time since the initial diagnosis of non-small cell lung cancer.

[0364] · Have not received anti-PD1 / PD-L1, and

[0365] · Have received third-generation TKI treatment

[0366] - For patients with tumors with a previously documented EFGR exon 19 deletion or L858R mutation, previous osimertinib or other third-generation TKI treatment is required

[0367] · Exclude small cell transformation

[0368] Cohort H: EGFR - Mutant NSCLC after 3rd - Generation TKI and Platinum Dual Chemotherapy

[0369] · Patients with histologically or cytologically documented locally advanced or metastatic non-squamous NSCLC who are not candidates for curative surgery or curative radiotherapy, and

[0370] · With a previously documented targetable EGFR mutation:

[0371] - NSCLC with EGFR exon 19 deletion

[0372] - NSCLC with EGFR L858R mutation

[0373] - NSCLC with activating EGFR exon 20 insertion

[0374] - NSCLC with exon 18 / 21 atypical mutations

[0375] Note: EGFR deletion / mutation must have been previously documented by a validated test (tissue or ctDNA from blood is allowed).

[0376] · Have not received anti-PD1 / PD-L1, and

[0377] · Have received third-generation TKI treatment

[0378] - For patients with tumors carrying a previously documented EFGR exon 19 deletion or L858R mutation, prior osimertinib or other third-generation TKI treatment is required

[0379] · Have received prior treatment with platinum doublet chemotherapy

[0380] · Small cell transformation is excluded

[0381] Study population

[0382] Dose escalation: The study population will consist of patients with selected advanced solid tumors who have exhausted treatment options expected to provide meaningful clinical benefit, except for patients with malignancies in which anti-PD-1 / PD-L1 therapy has shown clinical benefit.

[0383] Dose expansion: The study population will consist of a specified cohort of patients with selected advanced solid tumors who have not received anti-PD-1 / PD-L1.

[0384] Inclusion Criteria

[0385] Patients must meet the following criteria to be eligible for inclusion in the study:

[0386] 1. ≥ 18 years of age

[0387] 2. Have an Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1

[0388] 3. Have histologically or cytologically confirmed cancer that meets the following criteria:

[0389] a. Dose Escalation:

[0390] a. Have metastatic disease or locally advanced disease and are not candidates for curative surgery or curative radiotherapy.

[0391] b. Are not candidates for an approved anti-PD-1 or PD-L1 therapy indication, or such therapy is not available to the patient for other reasons (alone or in combination). The reason for ineligibility must be documented.

[0392] c. Due to disease recurrence, disease refractoriness, or intolerance, all treatment options that are expected to provide meaningful clinical benefits have been exhausted. Due to this requirement, patients with malignancies in whom anti-PD-1 / PD-L1 therapies have demonstrated clinical benefits are excluded.

[0393] d. Have any of the following cancer types:

[0394] a. Colorectal cancer with locally documented microsatellite stability

[0395] b. Gastric cancer or gastroesophageal junction cancer

[0396] c. Esophageal cancer

[0397] d. Breast cancer (ductal or lobular carcinoma, regardless of receptor status)

[0398] e. NSCLC (any PD-L1 expression)

[0399] f. Head and neck squamous cell carcinoma (SCC)

[0400] g. Nasopharyngeal carcinoma

[0401] h. Cervical cancer

[0402] i. Anal cancer

[0403] j. Mesothelioma

[0404] k. Prostatic adenocarcinoma

[0405] l. Renal cell carcinoma (e.g., chromophobe renal cell carcinoma, clear cell renal cell carcinoma, or papillary renal cell carcinoma)

[0406] m. Gallbladder / bile duct cancer

[0407] n. Urothelial carcinoma

[0408] o. Pancreatic carcinoma

[0409] p. Penile SCC

[0410] q. Vulvovaginal cancer

[0411] r. Other non-CNS tumor types in which, after discussion with the sponsor, the investigator is able to confirm elevated EGFR expression in the tumor may be eligible.

[0412] Note: Small cell / large cell / neuroendocrine, and sarcomatoid histologies are excluded.

[0413] b. Dose Expansion Cohorts:

[0414] a. Cohort A: Patients with metastatic TNBC who:

[0415] a. Not a candidate for curative surgery or curative radiotherapy

[0416] b. Not a candidate for an approved anti-PD-1 or anti-PD-L1 therapy indication, or such therapy is unavailable to the patient for other reasons (e.g., PD-L1 ICS did not achieve the score for the indicated use, the patient cannot obtain anti-PD-1 or anti-PD-L1 therapy through reimbursement, contraindication to protein-bound paclitaxel, etc.)

[0417] c. Has a previously documented triple-negative cancer (ER- / PR- / Her2-) by local pathology

[0418] b. Group B: Patients with metastatic CSCC or locally advanced CSCC who are not candidates for curative surgery or curative radiotherapy

[0419] c. Group C: Patients with histologically or cytologically documented locally advanced or metastatic NSCLC disease who:

[0420] a. Not a candidate for curative surgery or curative radiotherapy

[0421] b. Do not have a previously documented targetable molecular alteration (e.g., ALK, ROS1, EGFR, Met Ex14, etc.)

[0422] c. Have not received prior systemic therapy for recurrent or metastatic NSCLC (adjuvant or neoadjuvant systemic therapy will not be considered a prior line)

[0423] d. Group D: Patients with histologically or cytologically documented locally advanced or metastatic HNSCC disease who:

[0424] a. Not a candidate for curative surgery or curative radiotherapy

[0425] b. Have a previously documented CPS of PD-L1 expression ≥ 1% by IHC assay on specimens collected within the past 3 months

[0426] c. Have not received prior systemic therapy for recurrent or metastatic HNSCC (adjuvant or neoadjuvant systemic therapy will not be considered a prior line)

[0427] e. Group E: Patients with metastatic CRC who have a previously documented MSS disease and who:

[0428] a. Not a candidate for curative surgery or curative radiotherapy

[0429] b. May have active metastases in the liver and / or peritoneum at the time of screening

[0430] Note: Ablated and resected diseases are not considered active.

[0431] c. Based on the patient's medical history and the previously recorded results in the local pathology report, microsatellite stable

[0432] d. Have received at least one line of therapy in the setting of recurrence / metastasis

[0433] · Patients with previously recorded RAS wild-type disease must have received anti-EGFR therapy or have a recorded reason why anti-EGFR therapy is inappropriate.

[0434] · Patients must have received anti-VEGF therapy or have a recorded reason why anti-VEGF therapy is not suitable.

[0435] f. Group F: Patients with metastatic CRC who have previously recorded MSS disease and who:

[0436] a. Are not candidates for curative surgery or curative radiotherapy

[0437] b. No active metastases were identified in the liver or peritoneum at screening. Note: Ablated and resected diseases are not considered active.

[0438] c. The disease is present only in one or more lungs and / or pulmonary lymph nodes

[0439] d. Based on the patient's medical history and the previously recorded results in the local pathology report, microsatellite stable

[0440] e. Have received at least one line of therapy in the setting of recurrence / metastasis

[0441] · Patients with previously recorded RAS wild-type disease must have received anti-EGFR therapy or have a recorded reason why anti-EGFR therapy is inappropriate.

[0442] · Patients must have received anti-VEGF therapy or have a recorded reason why anti-VEGF therapy is not suitable.

[0443] g. Group G: Patients with previously histologically or cytologically recorded locally advanced or metastatic EGFR-mutant non-squamous NSCLC disease who:

[0444] a. Are not candidates for curative surgery or curative radiotherapy

[0445] b. Have a previously recorded targetable EGFR mutation:

[0446] - NSCLC carrying an EGFR exon 19 deletion.

[0447] - NSCLC patients with EGFR L858R mutation.

[0448] - NSCLC patients with activating EGFR exon 20 insertions

[0449] - NSCLC patients with exon 18 / 21 atypical mutations

[0450] Note: According to the patient's medical history, EGFR deletions / mutations must have been previously documented by an approved test (allowing tissue from blood or ctDNA).

[0451] b. Have not received prior chemotherapy

[0452] c. Have received treatment with a third-generation TKI

[0453] a. For patients with tumors harboring a previously documented EFGR exon 19 deletion or L858R mutation, prior osimertinib or other third-generation TKI treatment is required

[0454] Note:

[0455] - CNS disease stability is allowed. Stability is defined as no signs of progression for at least 6 weeks on imaging obtained during the screening period; no signs of new or enlarging brain metastases, and the patient does not require any immunosuppressive dose of systemic corticosteroids to control brain metastases within 4 weeks of the first dose of the study drug.

[0456] - Small cell transformation is excluded

[0457] h. Cohort H: Patients with histologically or cytologically documented locally advanced or metastatic non-squamous NSCLC disease who:

[0458] a. Are not candidates for curative surgery or curative radiotherapy

[0459] b. Have a previously documented targetable EGFR mutation:

[0460] - NSCLC patients with EGFR exon 19 deletions.

[0461] - NSCLC patients with EGFR L858R mutation.

[0462] - NSCLC patients with activating EGFR exon 20 insertions

[0463] - NSCLC patients with exon 18 / 21 atypical mutations

[0464] Note: According to the patient's medical history, EGFR deletions / mutations must have been previously documented by any approved test (allowing tissue from blood or ctDNA).

[0465] c. Have received treatment with a third-generation TKI

[0466] a. For patients with tumors carrying a previously documented EGFR exon 19 deletion or L858R mutation, prior osimertinib or other third-generation TKI treatment is required

[0467] d. Have received treatment with platinum doublet chemotherapy

[0468] Note:

[0469] - CNS disease stability is allowed. Stability is defined as no signs of progression for at least 6 weeks on imaging obtained during the screening period; no signs of new or enlarging brain metastases, and the patient does not require any immunosuppressive dose of systemic corticosteroids to control brain metastases within 4 weeks of the first dose of the study drug

[0470] - Small cell transformation is excluded

[0471] 4. Only Expansion Cohorts: Have never received anti-PD-1 / PD-L1, defined as having never been treated with a drug targeting PD-1

[0472] Note: In dose escalation, prior anti-PD-1 / PD-L1 therapy is allowed provided that ≥1 month has passed since the first dose of the study therapy, and the patient has recovered from immune-mediated adverse events or returned to baseline for at least 1 month prior to starting the study drug. Endocrinopathies and vitiligo that are well controlled with hormone replacement therapy are not excluded. Patients who have permanently discontinued prior anti-PD-1 / PD-L1 therapy due to drug-related toxicities are ineligible

[0473] 5. Have at least 1 lesion that meets the study criteria

[0474] · If progression has been demonstrated in such lesions after radiotherapy (confirmed by an increase in size or tumor viability on biopsy meeting the RECIST 1.1 criteria for PD), then tumor lesions in the previously irradiated area are considered measurable

[0475] Only applicable to patients in the CSCC expansion cohort:

[0476] a. If digital medical photography is performed, then there must be at least 1 measurable baseline lesion with a longest diameter (LD) and perpendicular diameter both ≥10 mm. Non-measurable disease is defined as a unidimensionally measurable lesion, a tumor with ill-defined margins, or a lesion with a maximum perpendicular diameter less than 10 mm. Patients with non-measurable disease at baseline are ineligible for the study

[0477] 6. Willingness to provide tumor tissue from newly obtained sections (at least core biopsies) from previously unirradiated tumor sites.

[0478] Note:

[0479] a. If the previously irradiated lesion is the only available lesion and it is documented that the lesion has progressed after radiotherapy, then the lesion may be used.

[0480] b. If the patient has only a single RECIST 1.1 measurable and biopsy - available lesion, then a biopsy is recommended prior to baseline imaging to ensure accurate post - biopsy measurements.

[0481] c. If the investigator believes that the biopsy would pose a life - threatening risk to the patient (e.g., if a post - cardiac mass is the only lesion), then, after discussion between the investigator and the sponsor, the biopsy requirement may be waived on a limited basis. In these cases, archival tissue from a biopsy obtained within the past 6 months is required.

[0482] 7. Normal organ and bone marrow function, recorded as:

[0483] a. Hemoglobin ≥ 8.0 g / dL

[0484] b. Absolute neutrophil count (ANC) ≥ 1.0×10 9 / L

[0485] Note: For African - American patients with a documented history of benign ethnic neutropenia, an ANC ≥ 0.5×10 9 / L

[0486] c. Platelet count ≥ 75×10 9 / L

[0487] 8. Serum creatinine ≤ 1.5×ULN or estimated glomerular filtration rate > 30 mL / min (using methods recognized by the investigator's institution, e.g., Modification of Diet in Renal Disease [MDRD], Chronic Kidney Disease Epidemiology Collaboration [CKD - EPI] formula). A 24 - hour urine creatinine collection may be substituted for the calculated creatinine clearance to meet eligibility criteria.

[0488] 9. Hypercalcemia < grade 3 or controlled to < grade 3 with bisphosphonate therapy

[0489] 10. Liver function meets the following criteria:

[0490] a. Total bilirubin ≤ 1.5 × upper limit of normal (ULN) (if the tumor involves the liver, then ≤ 3 × ULN)

[0491] b. Aspartate aminotransferase (AST) ≤ 2.5 × ULN (if the tumor involves the liver, then ≤ 5 × ULN)

[0492] c. Alanine aminotransferase (ALT) ≤ 2.5 × ULN (if the tumor involves the liver, then ≤ 5 × ULN)

[0493] d. Alkaline phosphatase (ALP) ≤ 2.5 × ULN (if the tumor involves the liver or bone, then ≤ 5 × ULN)

[0494] Note:

[0495] Patients with tumor involvement of the liver and AST content ≥ 3 × ULN or ALT ≥ 3 × ULN, and bilirubin content ≥ 2 × ULN will be excluded regardless of the above criteria.

[0496] Patients with Gilbert's syndrome do not need to meet the total bilirubin requirement, provided that their total bilirubin does not exceed 150% of their historical level and is less than 2 mg / dL (34.2 μmol / L). Gilbert's syndrome must be properly documented as a past medical history.

[0497] 11. Rapid and aggressive therapy is not required, so it is expected that there will be no clinically significant delay in the mavacamten component of the therapy during the 3-week lead-in period of REGN7075 monotherapy before the start of the combination of REGN7075 plus mavacamten.

[0498] Note: This criterion does not apply to patients recruited into the group that starts using REGN7075 and mavacamten simultaneously.

[0499] 12. Have a life expectancy of at least 3 months.

[0500] 13. Willing and able to comply with clinical interviews and study-related procedures.

[0501] 14. Able to provide an informed consent form signed by the study participant or a legally acceptable representative.

[0502] Exclusion Criteria

[0503] Patients who meet any of the following criteria will be excluded from the study:

[0504] 1. Currently participating in another study of a therapeutic agent.

[0505] 2. Have participated in any study of investigational agents or investigational devices within 4 weeks of the first administration of the study drug.

[0506] a. Exceptions: Patients who have received treatment with an investigational immuno-PET (iPET) reagent or who are enrolled in a study involving such treatment may be recruited in this study, provided that the iPET reagent does not target EGFR, CD28, PD-1, PD-L1 or PD-L2.

[0507] 3. Have received treatment with an approved systemic therapy or have not recovered from any acute toxicity (i.e., Grade 1 or baseline) within 4 weeks of the first administration of the study drug.

[0508] 4. Have received therapy with an anti-EGFR antibody therapy within the following drug-specific window (approximately 5 half-lives):

[0509] 5. Cetuximab: Within 4 weeks of the first administration of REGN7075

[0510] 6. Panitumumab: Within 6 weeks of the first administration of REGN7075

[0511] 7. Necitumumab: Within 10 weeks of the first administration of REGN7075

[0512] 8. Any other investigational agent that blocks EGFR by interacting with the extracellular binding domain: within 5 half-lives of the first administration of REGN7075, after sponsor approval.

[0513] a. Note: Kinase inhibitors targeting EGFR will be permitted within the scope of Exclusion Criterion 3.

[0514] 9. Have received radiotherapy or major surgery within 14 days of the first administration of the study drug, or have not recovered from an adverse event (i.e., Grade 1 or baseline).

[0515] 10. Have received any prior systemic non-immunomodulatory biotherapy within 4 weeks of the first administration of the study drug.

[0516] 11. Have undergone prior anti-cancer immunotherapy within 5 half-lives before the study drug. Examples of immunomodulators include blockers of CTLA-4, 4-1BB (CD137) or OX-40, therapeutic vaccines, PI3K-δ inhibitors or cytokine anti-cancer therapeutic agents.

[0517] a. Prior CAR-T therapy will be permitted regardless of cellular half-life / persistence.

[0518] 12. Have not recovered from immune-mediated adverse events prior to starting the investigational drug (i.e., at baseline). Endocrinopathies that are adequately controlled with hormone replacement therapy are not excluded.

[0519] 13. Have a contraindication to receiving anti-PD-1 therapy

[0520] 14. Have a known (previously documented) high microsatellite instability (MSI-H) or mismatch repair-deficient cancer.

[0521] 15. Have another malignancy that is progressing or requires active treatment, except for:

[0522] a. Non-melanoma skin cancer that has undergone potentially curative therapy

[0523] b. Any tumor considered treatable effectively with definitive local control

[0524] 16. Have any condition that requires continuous / consecutive corticosteroid therapy (>10 mg prednisone per day or anti-inflammatory equivalent) within 1 to 2 weeks prior to the first dose of the investigational drug. Physiological replacement doses are permitted, even if they are >10 mg prednisone per day or equivalent, provided they are not administered for immunosuppressive purposes. Inhaled or topical steroids are permitted provided they are not used to treat autoimmune or skin conditions.

[0525] a. Note: Patients who require short-term steroid courses (up to 2 days in the week prior to enrollment), for prophylaxis (e.g., contrast dye allergy) or for treatment of non-autoimmune conditions (e.g., delayed-type hypersensitivity reaction caused by contact with an allergen), or physiological replacement therapy, may be enrolled in the study.

[0526] 17. Have persistent or recent (within 5 years) signs of significant autoimmune disease or any other condition that requires treatment with systemic immunosuppressive agents. The following are not excluded: vitiligo, resolved childhood asthma, endocrinopathies that require only hormone replacement therapy (e.g., hypothyroidism or type 1 diabetes).

[0527] 18. Have an untreated or active primary brain tumor, CNS metastases, leptomeningeal disease, or spinal cord compression.

[0528] a. Note: Patients with pre-treated CNS metastases or spinal cord compression are not excluded if:

[0529] b. No signs of progression for at least 4 weeks prior to the first dose of the investigational drug, and any neurological symptoms have returned to baseline, and

[0530] c. No signs of new or enlarging central nervous system metastases, and

[0531] d. Systemic corticosteroids were not required within 4 weeks prior to the first dose of the investigational drug to control central nervous system metastases or spinal cord compression

[0532] 19. Having had encephalitis, meningitis, organic brain disease (such as Parkinson's disease) or uncontrolled epilepsy within 1 year prior to the first dose of the investigational drug

[0533] 20. The baseline QT / QTc interval is significantly prolonged or there are risk factors for QTc prolongation, such as:

[0534] a. Repeated demonstration of a QTc interval > 470 milliseconds at baseline

[0535] b. A history of other risk factors for polymorphic ventricular tachycardia (torsade de pointes; TdP), or a family history of long QT syndrome

[0536] 21. Having a history of myocardial infarction, cardiac event, congestive heart failure (NYHA class II or higher) or cardiac arrhythmia within the last 12 months.

[0537] a. Note: A history of arrhythmia that is transient and does not require any medical intervention will be permitted, provided it did not occur within the last 6 months prior to enrollment. In any case, premature atrial contractions (PAC) or premature ventricular contractions (PVC) that do not meet the specific arrhythmia criteria are permitted.

[0538] 22. Any patient with active inflammatory skin disease (such as psoriasis, eczema, etc.) or inflammatory skin disease (regardless of activity) that requires continuous medication (topical or systemic), or a history of inflammatory skin disease within the last 5 years.

[0539] 23. At baseline, persistently presenting any of the following skin examination findings, regardless of grade:

[0540] a. Bullous dermatitis

[0541] b. Exfoliative dermatitis

[0542] c. Erythroderma

[0543] d. Erythema multiforme

[0544] e. Acneiform rash

[0545] f. Generalized rash

[0546] g. Maculo-papular rash

[0547] h. Other unspecified skin toxicities (other than ≤ Grade 1) with any inflammatory or destructive features (e.g., infiltration, erythema, ulceration, blistering, exfoliation, etc.).

[0548] i. Note: Skin findings expected to be transient ≤ Grade 1 will not be excluded. Additionally, hypersensitivity reactions expected to be transient ≤ Grade 1 will not be excluded.

[0549] 24. A known history or any signs of interstitial lung disease or active, non-infectious pneumonia within 5 years prior to the first dose of the investigational drug. A history of radiation pneumonitis in the radiation field is permitted, provided that the pneumonia resolved ≥ 6 months prior to the first dose of the investigational therapy.

[0550] 25. Having any indwelling drainage tubes or stents (e.g., nephrostomy tubes, biliary stents, etc.) intended to preserve organ function or facilitate occlusive drainage.

[0551] a. Note

[0552] b. Catheters in the bladder or neobladder will not be excluded

[0553] c. Ports or indwelling intravenous lines are permitted

[0554] d. Coronary stents will not be excluded, provided that the patient meets the criteria of #17

[0555] 26. Having uncontrolled human immunodeficiency virus infection, hepatitis B or hepatitis C infection; or being diagnosed with immunodeficiency.

[0556] a. Note:

[0557] b. Patients will be tested for hepatitis C virus (HCV) and hepatitis B virus (HBV) at screening.

[0558] c. Patients known to have HIV infection with controlled infection (undetectable viral load (HIV RNA PCR) and CD4 count above 350, either spontaneously or based on a stable antiviral regimen) are permitted. For patients with controlled HIV infection, monitoring will be performed according to local standards.

[0559] d. Allow patients with hepatitis B (HepBsAg+) whose infection is under control (serum hepatitis B virus DNA PCR below the limit of detection and receiving antiviral therapy for hepatitis B). Patients with controlled infection must undergo regular monitoring of HBV DNA. After the last dose of the investigational study drug, patients must continue antiviral therapy for at least 6 months.

[0560] e. Patients with positive hepatitis C virus antibody (HCV Ab+) whose infection is under control (PCR-undetectable HCV RNA, spontaneously or in response to a successful previous course of anti-HCV therapy) can be recruited into the study.

[0561] 27. Require treatment with anti-infective agents within 4 weeks of the first dose of the study drug, or have any ongoing infection (except as otherwise specified in the exclusion criteria).

[0562] 28. Have received a live vaccine within 4 weeks of the planned start of the study drug.

[0563] 29. Have undergone a previous allogeneic stem cell transplant, autologous stem cell transplant, or solid organ transplant at any time.

[0564] 30. Known to have an allergic or hypersensitivity reaction to cemipimab or a component of the study drug.

[0565] 31. Have a mental or substance abuse disorder known to interfere with the requirements of participating in the study.

[0566] 32. Have any medical condition, comorbidity, physical examination finding, metabolic dysfunction, or clinical laboratory abnormality that makes the patient ineligible to participate in the clinical study due to high safety risks and / or potential interference with the interpretation of study results

[0567] 33. Members of the clinical site study team and / or their immediate family, unless prior approval has been granted.

[0568] 34. Patients with a positive serum hCG pregnancy test must be medically excluded from pregnancy to be eligible to participate in the study. Lactating women are also excluded.

[0569] 35. Women of childbearing potential (WOCBP)* or sexually active men who are unwilling to use highly effective contraceptive measures for at least 6 months before the initial dose / first treatment start, during the study, and after the last dose. Highly effective contraceptive measures include:

[0570] a. Stable use of combined (estrogen and progestogen-containing) hormonal contraception (oral, intravaginal, transdermal) or progestogen-only hormonal contraception (oral, injectable, implantable), which is associated with suppression of ovulation starting in 2 or more menstrual cycles prior to screening

[0571] b. Intrauterine device (IUD); Intrauterine hormone releasing system (IUS);

[0572] c. Bilateral tubal ligation or occlusion

[0573] d. Vasectomized partner (provided that the vasectomized male partner is the sole partner of the WOCBP study participant and the vasectomized partner has received a medical assessment of the success of the surgical procedure)

[0574] e. And / or abstinence

[0575] 36. WOCBP is defined as a fertile female after menarche until becoming postmenopausal, unless permanently infertile. Permanent sterilization methods include hysterectomy, bilateral salpingectomy, and bilateral oophorectomy.

[0576] a. Postmenopausal status is defined as 12 consecutive months without menses, without alternative medical causes. Elevated follicle-stimulating hormone (FSH) levels within the postmenopausal range can be used to confirm postmenopausal status in women not using hormonal contraception or hormone replacement therapy. However, in the absence of 12 months of amenorrhea, a single FSH measurement is insufficient to determine the occurrence of postmenopausal status. The above definitions are according to the Clinical Trial Facilitation Group (CTFG) guidelines. Postmenopausal or permanently infertile women do not require pregnancy testing and contraception. Abstinence is considered an effective method only when defined as avoiding heterosexual intercourse throughout the entire risk period associated with the study drug. The reliability of abstinence needs to be evaluated with reference to the duration of the clinical trial and the patient's preferred and common lifestyle.

[0577] b. Periodic abstinence (calendar method, symptothermal method, postovulatory method), withdrawal (coitus interruptus), spermicides only, and lactational amenorrhoea method (LAM) are unacceptable contraceptive methods. Female and male condoms should not be used together.

[0578] 37. Applies only to groups G and H: Small cell transformation.

[0579] Research treatment

[0580] REGN7075: Administered at 0.1 mg to 900 mg once weekly (QW) or 0.1 mg to 2700 mg QW or every three weeks (Q3W) by intravenous (IV) infusion or by subcutaneous (SC) injection at the target dose level. For doses of 100 mg and higher, REGN7075 will be administered in a step - up manner with or without a fractionated dosing schedule.

[0581] Cemiplimab: Will be administered at 350 mg Q3W by IV infusion over 30 minutes or by SC injection.

[0582] When both drugs are administered on the same day, REGN7075 will be administered before cemiplimab.

[0583] Q3W IV administration of platinum - based doublet chemotherapy for 4 cycles (subsequently pemetrexed maintenance therapy will be administered to those patients initially assigned to receive a regimen containing pemetrexed).

[0584] The choice of chemotherapy will be one of the regimens shown in Table 11.

[0585] Table 11: Guidelines for platinum - based chemotherapy regimens

[0586]

[0587]

[0588] Abbreviations: AUC = area under the curve; IV = intravenous; N / A = not applicable, Q3W = every three weeks

[0589] Chemotherapy will be administered after the completion of cemiplimab administration. When chemotherapy is administered on the same day as REGN7075 and cemiplimab, the full dose of REGN7075 will be administered first, followed by cemiplimab, and finally chemotherapy.

[0590] Study endpoints

[0591] The primary endpoint of treatment will be determined based on the following assessments.

[0592] Dose escalation:

[0593] · Incidence of dose - limiting toxicity (DLT) during the DLT period

[0594] · Incidence and severity of treatment emergent adverse events (TEAE), adverse events of special interest (AESI), serious adverse events (SAE), and laboratory abnormalities of grade ≥3

[0595] Dose escalation:

[0596] · Objective response rate (ORR) according to Response Evaluation Criteria in Solid Tumors version 1.1 (RECIST 1.1) and / or Composite Response Criteria (depending on the subject's baseline evaluation criteria).

[0597] Secondary endpoints of treatment were determined based on the following assessments:

[0598] Dose escalation

[0599] · Drug concentration of REGN7075 in serum and drug concentration of zimberelimab in serum

[0600] · ORR, progression-free survival (PFS), duration of response (DOR), complete response (CR) rate, and disease control rate (DCR) according to RECIST 1.1 and / or Composite Response Criteria (depending on the subject's baseline evaluation criteria)

[0601] · Overall survival (OS)

[0602] · Incidence of anti-drug antibodies (ADA) against REGN7075 and zimberelimab

[0603] Dose escalation

[0604] · Incidence and severity of TEAE / AESI / SAE and laboratory abnormalities of grade ≥3

[0605] · Drug concentration of REGN7075 in serum and drug concentration of zimberelimab in serum

[0606] · Incidence of ADA against REGN7075 and zimberelimab

[0607] · PFS, DOR, DCR, and CR rate according to RECIST 1.1 and / or Composite Response Criteria (depending on the subject's baseline evaluation criteria)

[0608] · OS

[0609] · Patient-reported QoL, symptoms, function, and general health status (CRC patients according to EORTC QLQ-C30, EORTC QLQ-CR29, breast cancer patients according to EORTC QLQ-BR23, NSCLC patients according to EORTC QLQ-LC13, HNSCC patients according to EORTC QLQ-HN35, and EQ-5D-5L).

[0610] Procedures and Assessments

[0611] The following procedures and assessments will be used to evaluate the treatment:

[0612] · Only at screening / baseline: height, brain imaging, coagulation assessment, and assessment of archived or fresh tumor.

[0613] · Efficacy: radiographic disease assessment (computed tomography [CT], magnetic resonance imaging [MRI], and / or digital radiography). For subjects with diseases that can be measured radiographically, the disease will be evaluated radiographically according to RECIST 1.1. For subjects with CSCC lesions assessable on the skin, the combined response criteria should be used in combination with radiographic imaging as appropriate.

[0614] · Safety: vital signs (including body temperature, sitting and standing blood pressure, pulse, and respiration), physical examination, Eastern Cooperative Oncology Group (ECOG) performance status, weight, electrocardiogram, adverse events (AE), hematology, blood chemistry, C-reactive protein (CRP), thyroid-stimulating hormone (TSH), pregnancy test, urine analysis

[0615] · Pharmacokinetic and immunogenicity sampling

[0616] · Biomarkers and exploratory studies: serum cytokines, exploratory tumor biopsies (or archived tissue), whole blood for immune monitoring, plasma for ctDNA, peripheral blood mononuclear cells (PBMC) for immune expression profiling, DNA / RNA whole blood for T cell repertoire analysis, serum and plasma for exploratory biomarkers.

[0617] Results

[0618] Initial efficacy and safety results are described in the following examples. Results to date include partial and complete responses in many cohorts.

[0619] Example 3: Initial dose-escalation results of a phase 1 / 2 study of the combination of REGN7075 and cemipimab (anti-PD-1) in subjects with advanced solid tumors

[0620] Conduct an open-label, phase 1 / 2 dose escalation and expansion study in humans for the first time to evaluate the safety, tolerability, pharmacokinetics, and initial anti-tumor activity of the combination of REGN7075 (EGFRxCD28) and cemipimab (anti-PD-1) in subjects with advanced solid tumors ( Figure 3 ). The study protocol is described in detail in Example 2.

[0621] During the dose escalation period (Bayesian optimal interval design; Part 1), heavily pre-treated subjects with advanced solid tumors received a 3-week lead-in period of REGN7075 monotherapy weekly, followed by combination therapy with 350 mg of cemipimab every 3 weeks. The planned dose levels (DL) of REGN7075 were 0.03, 0.1, 0.3, 1, 3, 10, 30, 100, 300, and 900 mg. The primary objective was to evaluate the safety and tolerability of the combination of REGN7075 and cemipimab.

[0622] As of the data cutoff date, 18 subjects (median age, 53.5 years, 56% female) were treated with REGN7075 at up to 30 mg DL and cemipimab during the dose escalation period (Table 12). Most subjects (67%) were treated for microsatellite stable colorectal cancer. No subject experienced dose-limiting toxicity; the maximum tolerated dose was not reached.

[0623] Table 13 summarizes treatment-emergent adverse events (TEAE) and treatment-related adverse events (TRAE). The most frequent TEAE (any grade) were increased aspartate aminotransferase (AST), constipation, and fatigue (each 33% [n = 6]). The most frequent TRAE (any grade) were fatigue (17% [n = 3]), increased AST, diarrhea, hypothyroidism, fever, and rash (each 11% [n = 2]). One subject developed cytokine release syndrome, characterized by a single grade 1 fever without hypotension or hypoxia. There were five recorded deaths that did not occur during study treatment and were not attributed to the study drug.

[0624] Serum concentrations of REGN7075 were measured after the first dose of REGN7075 was administered intravenously. The maximum concentration (Cmax) and area under the curve (AUC) values were greater than dose-proportional at the 30 mg dose, while the minimum concentration (Cmin) values were dose-proportional at all doses ( Figure 4 ). Sustained PK evaluations indicated that there may be a target-mediated effect.

[0625] T cell activation-related cytokines were detected in monotherapy induction and combination dosing. One patient induced IL-2 ( Figure 5A ) after the first dose of REGN7075 (1 mg). Multiple patients induced IFN-γ ( Figure 5B ) after receiving REGN7075 alone or in combination with cemipimab.

[0626] Treatment and evaluation in the dose escalation phase are ongoing. Nevertheless, initial data showed that among all 18 subjects treated in the dose escalation, 1 PD-1 negative subject receiving 1 mg REGN7075 and cemipimab to treat cervical cancer unexpectedly achieved a durable partial response (DL4[c], Figure 6 ).

[0627] In this dose escalation study, REGN7075 was safely combined with cemipimab at dose levels up to 30 mg without dose-limiting toxicity. Early data suggest that the novel agent REGN7075 is generally well tolerated and has initial anti-tumor activity. No signs of large-scale immune activation or cytokine release syndrome were observed with the CD28 superagonist TGN1412.

[0628] Table 12. Demographic data and baseline characteristics

[0629]

[0630] Percentage of tumor cells stained with PD-L1 with any intensity above background in the tumor cell membrane.

[0631] Except for <1, data were collected as whole integers.

[0632] ECOG PS, Eastern Cooperative Oncology Group Performance Status

[0633] PD-L1, programmed cell death-ligand 1; Q, quartile.

[0634] Table 13. Overview of treatment-emergent adverse events (TEAE) and treatment-related adverse events (TRAE)

[0635]

[0636] Example 4: Updated results of a clinical trial using REGN7075 (EGFRxCD28) alone and in combination with an anti-PD-1 antibody (e.g., cemipimab)

[0637] An open-label, phase 1 / 2, first-in-human study was conducted in subjects with advanced solid tumors to evaluate the safety, tolerability, pharmacokinetics, and initial anti-tumor activity of REGN7075 (EGFRxCD28) alone and in combination with zimberelimab (anti-programmed cell death [PD]-1) (see Example 2). Subjects must have advanced solid tumors as defined by the protocol, an Eastern Cooperative Oncology Group performance status of 0 or 1, and have not received anti-PD-1 / anti-PD-ligand (L)1 therapy.

[0638] This study included a dose-escalation (Bayesian optimal interval design; Part 1) and a dose-expansion phase (Part 2). In Part 1, heavily pretreated subjects with advanced solid tumors received a 3-week induction period of REGN7075 monotherapy weekly, followed by combination therapy with zimberelimab 350 mg every 3 weeks. The planned dose levels (DL) of REGN7075 were 0.03, 0.1, 0.3, 1, 3, 10, 30, 100, 300, and 900 mg.

[0639] After determining the recommended phase 2 dose in Part 1, five tumor-specific expansion cohorts will be opened in Part 2: colorectal cancer (microsatellite stable [MSS]), non-small cell lung cancer (NSCLC, PD-L1 ≥ 50%), triple-negative breast cancer, cutaneous squamous cell carcinoma, and head and neck squamous cell carcinoma (select PD-L1, combined positive score ≥ 1). Subjects with MSS-CRC with RAS or BRAF wild-type mutations must have received anti-EGFR therapy or anti-vascular endothelial growth factor (VEGF) therapy. The primary endpoint of Part 1 was the safety and tolerability of REGN7075 alone and in combination with zimberelimab, and the primary endpoint of Part 2 was the objective response rate according to Response Evaluation Criteria in Solid Tumors version 1.1. For Part 2, secondary objectives were to evaluate the effect of REGN7075 on subject-reported outcomes, including health-related quality of life, which was measured by several validated instruments including the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire (EORTC QLQ-C30) and EORTC QLQ-CR29 (only CRC subjects).

[0640] As of September 13, 2022, 30 subjects were treated with REGN7075 at up to 300 mg DL and zimberelimab in the dose-escalation period.

[0641] Updated results as of December 2022 showed that two patients with MSS-CRC (one each in DL1 and DL9) had initial partial responses, and the cervical cancer patient in DL4 had a complete response ( Figure 7 ).

[0642] Example 5: Updated Clinical Results

[0643] This example describes updated results of an open-label, phase 1 / 2, first-in-human global study to evaluate the safety, tolerability, PK, and anti-tumor activity of REGN7075 ± zimberelimab in patients with advanced solid tumors (NCT04626635) (see Example 2).

[0644] The study included a dose-escalation (Part 1) and a dose-expansion phase (Part 2). Patients must have had advanced solid tumor types that may express EGFR, an ECOG performance status of 0 / 1, and have not received anti-PD(L)-1 therapy (Part 2 only). In Part 1, patients received a 3-week lead-in period of REGN7075 monotherapy QW (except for cohorts C and G) before receiving combination therapy of REGN7075 QW or Q3W + zimberelimab Q3W. Q3W dosing with zimberelimab could be initiated at a dose level tolerated during the lead-in period. After determining the tolerable dose and drug administration schedule in Part 1, Part 2 could include 8 tumor-specific expansion cohorts: triple-negative breast cancer (A); cutaneous squamous cell carcinoma (B); non-small cell lung cancer (NSCLC; C); head and neck squamous cell carcinoma (D); microsatellite stable colorectal cancer with active liver and / or peritoneal metastases (MSS-CRC) (E); MSS-CRC with lung / lymph node metastases (F); EGFR-mutated NSCLC after third-generation TKI (G); EGFR-mutated NSCLC after third-generation TKI and platinum doublet chemotherapy (H). Cohorts C and G will also receive 4 cycles of platinum-based chemotherapy and zimberelimab. Primary endpoint: In Part 1, safety and tolerability of REGN7075 ± zimberelimab; in Part 2, ORR (REGN7075 + zimberelimab ± chemotherapy; RECIST 1.1). Secondary objectives for Parts 1 and 2 included: OS, PFS, DoR, CR rate, and DCR; immunogenicity of REGN7075 and zimberelimab; and characterization of PK.

[0645] The study was expected to enroll approximately 769 patients: approximately 221 patients in Part 1 and approximately 548 patients in Part 2.

[0646] As of July 19, 2023, it has been confirmed that REGN7075 was well tolerated in more than 60 patients with advanced solid tumors treated with up to 900 mg of REGN7075 administered IV alone and in combination with cemipimab. No patient experienced a treatment-related adverse event leading to death. No patient experienced a treatment-related AE leading to permanent study drug discontinuation. During the dose-limiting toxicity period, no patient developed a dose-limiting toxicity as defined by the protocol. Most IRR events were grade 2 or 1 and IRR symptoms remained clinically manageable.

[0647] As of September 5, 2023, 93 patients have been enrolled in Part 1. Several partial and complete responses have been reported in the patients in the study. The study is ongoing and open for enrollment.

[0648] The scope of the present disclosure is not limited to the specific embodiments described herein. Indeed, various modifications of the present disclosure will become apparent to those skilled in the art in addition to those described herein, in light of the foregoing description and the accompanying drawings. Such modifications are intended to fall within the scope of the appended claims.

Claims

1. A method for treating cancer in a subject, the method comprising administering to the subject a therapeutically effective amount of a combination of the following: a bispecific antibody or an antigen-binding fragment thereof, which comprises a first antigen-binding domain that binds to cluster of differentiation factor 28 (CD28) and a second antigen-binding domain that binds to epidermal growth factor receptor (EGFR); and an antibody or an antigen-binding fragment thereof that specifically binds to programmed death receptor-1 (PD-1), thereby treating the cancer in the subject.

2. The method according to claim 1, wherein the cancer is a solid tumor.

3. The method according to claim 1 or 2, wherein the cancer is a cancer that expresses EGFR.

4. The method according to any one of claims 1 to 3, wherein the cancer is selected from esophageal tumor, lung squamous cell carcinoma, lung adenocarcinoma, cervical cancer, endometrial adenocarcinoma, bladder cancer, urothelial carcinoma, lung cancer, non-small cell lung cancer, colorectal cancer, sigmoid adenocarcinoma, rectal cancer, endometrial cancer, skin cancer, head and neck squamous cell carcinoma, brain cancer, glioblastoma multiforme, non-CNS tumor, cutaneous squamous cell carcinoma, breast cancer, gastric cancer, gastroesophageal cancer, gastroesophageal adenocarcinoma, pancreatic cancer, prostate cancer, ovarian cancer, melanoma, nasopharyngeal cancer, anal cancer, mesothelioma, renal cell carcinoma, gallbladder / bile duct cancer, pancreatic tumor, penile squamous cell carcinoma or vulvovaginal cancer.

5. The method according to any one of claims 1 to 4, further comprising selecting a subject, wherein the subject has advanced solid tumors.

6. The method according to any one of claims 1 to 5, wherein the subject meets at least one of the following criteria, or is selected based on at least one of the following criteria: a. Having metastatic disease or locally advanced disease and not being a candidate for curative surgery or curative radiotherapy; b. Not being a candidate for an approved anti-PD-1 or PD-L1 therapy indication, or such therapy being unavailable for the subject for other reasons (alone or in combination); c. Having exhausted all treatment options expected to provide meaningful clinical benefit due to disease recurrence, disease refractoriness or intolerance, except for subjects with malignancies in whom anti-PD-1 / PD-L1 therapy has shown clinical benefit; and / or d. Having any one of the following cancer types: (a) Colorectal cancer with local pathology recorded as microsatellite stable; (b) Gastric cancer or gastroesophageal junction cancer; (c) Esophageal cancer; (d) Breast cancer (ductal or lobular carcinoma, regardless of receptor status); (e) NSCLC (any PD-L1 expression); (f) Head and neck squamous cell carcinoma (SCC); (g) Nasopharyngeal cancer; (h) Cervical cancer; (i) Anal cancer; (j) Mesothelioma; (k) Prostatic adenocarcinoma; (l) Renal cell carcinoma (chromophobe renal cell carcinoma, clear cell renal cell carcinoma or papillary renal cell carcinoma); (m) Gallbladder / bile duct cancer; (n) Urothelial carcinoma; (o) Pancreatic tumor; (p) Penile SCC; (q) Vulvovaginal cancer; or (r) Other non-CNS tumor types showing elevated EGFR expression in the tumor.

7. The method according to any one of claims 1 to 6, wherein the subject has been treated with a previous therapy selected from: radiotherapy, surgery, chemotherapy, PD-1 inhibitor, PD-L1 inhibitor, anti-VEGF therapy, CAR-T therapy, and / or anti-EGFR therapy.

8. The method according to any one of claims 1 to 7, wherein the subject has not received a previous anti-PD-1 therapy or anti-PD-L1 therapy.

9. The method according to any one of claims 1 to 8, wherein the subject has microsatellite-stable colorectal cancer (MSS CRC).

10. The method according to claim 9, wherein the subject with microsatellite-stable colorectal cancer has at least one of the following attributes, or is selected based on at least one of the following attributes: (a) having metastatic CRC; (b) not being a candidate for curative surgery or curative radiotherapy; (c) having possible active metastases in the liver and / or peritoneum at the time of screening; (d) having no active metastases identified in the liver or peritoneum at the time of screening, and the disease site being present only in one or more lungs and / or pulmonary lymph nodes; (e) the pathology report being recorded as microsatellite stable; (f) having received at least one line of therapy in the setting of recurrence / metastasis, wherein the therapy comprises anti-EGFR therapy or anti-VEGF therapy; or (g) having never received anti-PD-1 / PD-L1, defined as never having been treated with a drug targeting PD-1 previously.

11. The method according to any one of claims 1 to 8, wherein the subject has triple-negative breast cancer (TNBC).

12. The method according to claim 11, wherein the subject with TNBC has at least one of the following attributes, or is selected based on at least one of the following attributes: (a) having metastatic TNBC; (b) not being a candidate for curative surgery or curative radiotherapy; (c) not being a candidate for an approved anti-PD-1 or anti-PD-L1 therapy indication, or such therapy being unavailable to the subject for other reasons; (d) the pathology report being recorded as having triple-negative cancer (ER- / PR- / Her2-); or (e) having never received anti-PD-1 / PD-L1, defined as never having been treated with a drug targeting PD-1 previously.

13. The method according to any one of claims 1 to 8, wherein the subject has cutaneous squamous cell carcinoma (CSCC), wherein: (i) The subject is not a candidate for curative surgery or curative radiotherapy; or (ii) The subject has never received anti-PD-1 / PD-L1, defined as never having been treated with a drug targeting PD-1 previously.

14. The method according to any one of claims 1 to 8, wherein the subject has non-small cell lung cancer (NSCLC).

15. The method according to claim 14, wherein the subject has at least one of the following attributes, or is selected based on at least one of the following attributes: (a) the subject has a histologically or cytologically recorded locally advanced or metastatic EGFR-mutated non-squamous NSCLC disease; (b) has advanced or metastatic NSCLC; (c) is not a candidate for radical surgery or radical radiotherapy; (d) has a previously recorded targetable EGFR mutation (EGFR exon 19 deletion, EGFR L858R mutation, EGFR exon 20 insertion, or exon 18 / 21 atypical mutation); (e) has not received chemotherapy; (f) has received treatment with platinum doublet chemotherapy; (e) has received treatment with a third-generation TKI; or (g) has not received anti-PD-1 / PD-L1, defined as never having been treated with a drug targeting PD-1 previously.

16. The method according to any one of claims 1 to 8, wherein the subject has head and neck squamous cell carcinoma (HNSCC).

17. The method according to claim 16, wherein the subject has at least one of the following attributes, or is selected based on at least one of the following attributes: (a) has advanced or metastatic disease; (b) is not a candidate for radical surgery or radical radiotherapy; (c) has a CPS of PD-L1 expression ≥ 1% by local IHC assay; (d) has not received a previous systemic treatment for recurrent or metastatic HNSCC; or (e) has not received anti-PD-1 / PD-L1, defined as never having been treated with a drug targeting PD-1 previously.

18. The method according to any one of claims 1 to 17, wherein the bispecific EGFRxCD28 antibody or an antigen-binding fragment thereof is administered at a dose of about 0.1 mg to about 3000 mg.

19. The method according to any one of claims 1 to 18, wherein the bispecific EGFRxCD28 antibody or an antigen-binding fragment thereof is administered at a dose of about 0.01 mg, 0.03 mg, 0.05 mg, 0.1 mg, 0.3 mg, 0.5 mg, 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 8 mg, 10 mg, 15 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, 600 mg, 650 mg, 700 mg, 750 mg, 800 mg, 850 mg, 900 mg, 1000 mg, 1200 mg, 1500 mg, 1800 mg, 2000 mg, 2400 mg, 2700 mg or 3000 mg.

20. The method according to any one of claims 1 to 19, wherein the anti-PD-1 antibody or antigen-binding fragment thereof is administered at a dose of about 50 mg to about 1500 mg.

21. The method according to any one of claims 1 to 20, wherein the anti-PD-1 antibody is administered at a dose of 350 mg.

22. The method according to any one of claims 1 to 17, wherein the method comprises co-administering one or more doses of the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof and one or more doses of the anti-PD-1 antibody or antigen-binding fragment thereof.

23. The method according to claim 22, wherein each of the one or more doses of the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof is about 0.1 mg to about 3000 mg.

24. The method according to claim 23, wherein each of the one or more doses is about 0.01 mg, 0.03 mg, 0.05 mg, 0.1 mg, 0.3 mg, 0.5 mg, 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 8 mg, 10 mg, 15 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, 600 mg, 650 mg, 700 mg, 750 mg, 800 mg, 850 mg, 900 mg, 1000 mg, 1200 mg, 1500 mg, 1800 mg, 2000 mg, 2400 mg, 2700 mg or 3000 mg.

25. The method according to any one of claims 22 to 24, wherein each of the one or more doses of the anti-PD-1 antibody or antigen-binding fragment thereof is about 50 mg to about 1500 mg.

26. The method according to any one of claims 22 to 25, wherein each of the one or more doses of the anti-PD-1 antibody is 350 mg.

27. The method according to any one of claims 22 to 26, wherein each of the one or more doses of the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof and / or each of the one or more doses of the anti-PD-1 antibody or antigen-binding fragment thereof is administered 0.5 to 14 weeks after the previous dose.

28. The method according to any one of claims 22 to 27, wherein each of the one or more doses of the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof and / or each of the one or more doses of the anti-PD-1 antibody or antigen-binding fragment thereof is administered once a week, once every two weeks, once every three weeks, once every four weeks, once every five weeks or once every six weeks.

29. The method according to any one of claims 22 to 28, wherein each dose of the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof is administered once a week.

30. The method according to any one of claims 22 to 29, wherein each of the one or more doses of the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof is administered once every three weeks.

31. The method according to any one of claims 22 to 30, wherein each of the one or more doses of the anti-PD-1 antibody or antigen-binding fragment thereof is administered once every three weeks.

32. The method according to any one of claims 1 to 31, wherein the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof and / or the anti-PD-1 antibody or antigen-binding fragment thereof are administered intravenously.

33. The method according to any one of claims 1 to 31, wherein the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof and / or the anti-PD-1 antibody or antigen-binding fragment thereof are administered subcutaneously.

34. The method according to any one of claims 1 to 33, wherein the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof and the anti-PD-1 antibody or antigen-binding fragment thereof are administered on the same day.

35. The method according to any one of claims 1 to 33, wherein the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof and the anti-PD-1 antibody or antigen-binding fragment thereof are administered on different days.

36. The method according to claim 35, wherein the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof is administered before or after the anti-PD-1 antibody or antigen-binding fragment thereof.

37. The method according to any one of claims 1 to 36, comprising the steps of: (i) administering to the subject a dose of 0.1 mg to 3000 mg of the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof once a week or once every three weeks, subcutaneously or intravenously, for a period of monotherapy, wherein the period of monotherapy is at least 3 weeks; and (ii) administering to the subject a dose of 0.1 mg to 3000 mg of the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof once a week or once every three weeks, subcutaneously or intravenously, and a dose of 150 mg to 500 mg of the anti-PD-1 antibody or antigen-binding fragment thereof once every three weeks, intravenously or subcutaneously.

38. The method according to claim 37, wherein the period of monotherapy is at least 3 weeks, at least 4 weeks, at least 5 weeks or at least 6 weeks.

39. The method according to claim 37 or 38, wherein during step (ii), the anti-PD-1 antibody or antigen-binding fragment thereof and the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof are administered on different days.

40. The method according to claim 37 or 38, wherein during step (ii), the anti-PD-1 antibody or antigen-binding fragment thereof and the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof are administered on the same day.

41. The method according to any one of claims 1 to 40, further comprising administering to the subject one or more additional agents to treat one or more symptoms of immune-related adverse events.

42. The method according to claim 41, wherein the one or more additional agents comprise an IL-6 receptor inhibitor, a corticosteroid, and / or a non-steroidal anti-inflammatory drug (NSAID).

43. The method according to any one of claims 1 to 42, wherein the disease of the subject is stable, with a partial response or a complete response, at least one week after the combined administration of a dose of from about 0.1 mg to about 3000 mg of the bispecific antibody or antigen-binding fragment thereof and the anti-PD-1 antibody or antigen-binding fragment thereof.

44. The method according to any one of claims 1 to 43, wherein the anti-PD-1 antibody or antigen-binding fragment thereof is cemiplimab, nivolumab, pembrolizumab, MEDI0608, BI 754091, spartalizumab (PDR001), camrelizumab (SHR-1210), JNJ-63723283, MCLA-134, toripalimab, sintilimab, tislelizumab, serplulimab, dostarlimab, retifanlimab, zimberelimab, penpulimab, pidilizumab, HX008, balstilimab, or ezabenlimab, or an antigen-binding fragment of any of the foregoing.

45. The method according to any one of claims 1 to 43, wherein the anti-PD-1 antibody or antigen-binding fragment thereof comprises: heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) of a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 73, and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) of a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:

74.

46. The method according to any one of claims 1 to 43, wherein the anti-PD-1 antibody or antigen-binding fragment thereof comprises three HCDRs (HCDR1, HCDR2, and HCDR3) and three LCDRs (LCDR1, LCDR2, and LCDR3), wherein HCDR1 comprises the amino acid sequence of SEQ ID NO: 75; HCDR2 comprises the amino acid sequence of SEQ ID NO: 76; HCDR3 comprises the amino acid sequence of SEQ ID NO: 77; LCDR1 comprises the amino acid sequence of SEQ ID NO: 78; LCDR2 comprises the amino acid sequence of SEQ ID NO: 79; and LCDR3 comprises the amino acid sequence of SEQ ID NO:

80.

47. The method according to claim 46, wherein the HCVR comprises the amino acid sequence of SEQ ID NO: 73 and the LCVR comprises the amino acid sequence of SEQ ID NO:

74.

48. The method according to any one of claims 45 to 47, wherein the anti-PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 81 and a light chain comprising the amino acid sequence of SEQ ID NO:

82.

49. The method according to any one of claims 1 to 48, wherein the anti-PD-1 antibody is zimberelimab or an antigen-binding fragment thereof.

50. The method according to any one of claims 1 to 49, wherein the first antigen-binding domain that binds CD28 comprises three heavy chain complementarity-determining regions (CDR-H1, CDR-H2, and CDR-H3) contained within a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 10; and three light chain complementarity-determining regions (CDR-L1, CDR-L2, and CDR-L3) contained within a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:

16.

51. The method according to claim 50, wherein CDR-H1 comprises the amino acid sequence: GGSISSYY (SEQ ID NO: 12), CDR-H2 comprises the amino acid sequence: IYYSGIT (SEQ ID NO: 6), and CDR-H3 comprises the amino acid sequence: ARWGVRRDYYYYGMDV (SEQ ID NO: 14).

52. The method according to claim 50, wherein CDR-L1 comprises the amino acid sequence: QSVSSSY (SEQ ID NO: 18), CDR-L2 comprises the amino acid sequence: GAS (SEQ ID NO: 20), and CDR-L3 comprises the amino acid sequence: QQYGS SPWT (SEQ ID NO: 22).

53. The method according to any one of claims 50 to 52, wherein the first antigen-binding domain comprises a HCVR comprising the amino acid sequence of SEQ ID NO: 10 and a LCVR comprising the amino acid sequence of SEQ ID NO:

16.

54. The method according to any one of claims 1 to 53, wherein the second antigen-binding domain that binds to human EGFR comprises: three heavy-chain complementarity-determining regions (CDR-H1, CDR-H2, and CDR-H3), which are contained within a heavy-chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO:2; and three light-chain complementarity-determining regions (CDR-L1, CDR-L2, and CDR-L3), which are contained within a light-chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:

16.

55. The method according to claim 54, wherein CDR-H1 comprises the amino acid sequence: GDSIITFY (SEQ ID NO:4), CDR-H2 comprises the amino acid sequence: IYYSGIT (SEQ ID NO:6), and CDR-H3 comprises the amino acid sequence: ARVSEDSYFHYGMDV (SEQ ID NO:8).

56. The method according to claim 54, wherein CDR-L1 comprises the amino acid sequence: QSVSSSY (SEQ ID NO:18); CDR-L2 comprises the amino acid sequence: GAS (SEQ ID NO:20); and CDR-L3 comprises the amino acid sequence: QQYGSSPWT (SEQ ID NO:22).

57. The method according to any one of claims 54 to 56, wherein the second antigen-binding domain comprises: an HCVR comprising the amino acid sequence of SEQ ID NO:2 and an LCVR comprising the amino acid sequence of SEQ ID NO:

16.

58. The method according to any one of claims 1 to 57, wherein: (a) the first antigen-binding domain that binds to human CD28 comprises: a heavy-chain variable region comprising the amino acid sequence set forth in SEQ ID NO:10, and a light-chain variable region comprising the amino acid sequence set forth in SEQ ID NO:16; and (b) the second antigen-binding domain that binds to human EGFR comprises: a heavy-chain variable region comprising the amino acid sequence set forth in SEQ ID NO:2, and a light-chain variable region comprising the amino acid sequence set forth in SEQ ID NO:

16.

59. The method according to any one of claims 50 to 58, wherein the bispecific antibody comprises a first heavy chain comprising the amino acid sequence of SEQ ID NO:

26.

60. The method according to any one of claims 50 to 58, wherein the bispecific antibody comprises a second heavy chain comprising the amino acid sequence of SEQ ID NO:

24.

61. The method according to any one of claims 50 to 58, wherein the bispecific antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO:

28.

62. The method according to any one of claims 50 to 58, wherein the bispecific antibody comprises: a first heavy chain comprising the amino acid sequence of SEQ ID NO: 26, a second heavy chain comprising the amino acid sequence of SEQ ID NO: 24, and a common light chain comprising the amino acid sequence of SEQ ID NO:

28.

63. The method according to any one of claims 50 to 58, wherein the first antigen-binding domain comprises: a heavy chain comprising the amino acid sequence of SEQ ID NO: 26 and a light chain comprising the amino acid sequence of SEQ ID NO:

28.

64. The method according to any one of claims 50 to 58, wherein the second antigen-binding domain comprises: a heavy chain comprising the amino acid sequence of SEQ ID NO: 24 and a light chain comprising the amino acid sequence of SEQ ID NO:

28.

65. The method according to any one of claims 1 to 64, wherein the bispecific EGFRxCD28 antibody is REGN7075 or an antigen-binding fragment thereof.

66. The method according to any one of claims 1 to 65, further comprising administering chemotherapy to the subject, optionally platinum-based chemotherapy.

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