Cancer treatment method using bispecific EGFRxCD28 antibody alone or in combination with anti-PD-1 antibody

Abstract

The present disclosure provides a method of treating cancer in a subject, the method comprising administering to the subject a therapeutically effective amount of a 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), in combination with an antibody or antigen-binding fragment thereof that specifically binds to programmed death receptor-1 (PD-1), thereby treating the cancer in the subject. The present disclosure provides a method of treating cancer using a multispecific antibody or antigen-binding fragment thereof that binds to EGFR and CD28 (EGFRxCD28). Such an antibody may be combined with an additional therapeutic agent, such as an anti-PD-1 antibody, e.g., cemiplimab. Methods of treating cancer (e.g., an EGFR-expressing cancer) by administering an antibody (e.g., and a combination thereof with an anti-PD-1 antibody) are also provided.

Description

[Technical Field]

[0001] Related Applications This application is related to and claims priority to U.S. Provisional Application No. 63 / 378,102, filed October 3, 2022, U.S. Provisional Application No. 63 / 380,991, filed October 26, 2022, and U.S. Provisional Application No. 63 / 495,189, filed April 10, 2023, the entire contents of each of the foregoing applications being expressly incorporated herein by reference.

[0002] The present disclosure relates to methods for treating or preventing cancer (e.g., an EGFR-expressing cancer) in a subject (e.g., a human) in need thereof, comprising administering to the subject an effective amount of a bispecific antibody, or antigen-binding fragment thereof, that binds to epidermal growth factor (EGF) receptor and CD28, in combination with an anti-PD-1 antibody or antigen-binding fragment thereof.

[0003] Sequence Listing This application contains a Sequence Listing that has been submitted electronically in XML format, and is incorporated herein 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 technology]

[0004] The ability of T cells to recognize and kill cellular targets (such as virus-infected or tumor cells) depends on a series of coordinated interactions. The most important of these is the recognition and binding of target cells by the T cell receptor (TCR) complex (containing the associated CD3γ, δ, ε, and ζ chains), an interaction referred to as "signal 1" of T cell activation. The TCR recognizes viral or tumor peptides presented in the groove of the MHC protein expressed on the surface of target cells. Because such binding is generally low affinity, successful triggering of "signal 1" requires the clustering of many TCR complexes along the interface between the T cell and its target cell, a region referred to as the "immune synapse." T cell activation can be further promoted by additional interactions. For example, T cells carry a molecule called CD28 on their surface, which can provide a costimulatory "signal 2" to enhance activation via the TCR complex. T cell activation is enhanced when a T cell recognizes its target cell via its TCR complex and also engages "signal 2" via CD28 binding to its cognate ligand(s) on the target cell; like "signal 1," CD28-mediated "signal 2" is thought to occur via co-clustering at the immune synapse.

[0005] Agonist anti-CD28 monoclonal antibodies can be applied to sustained ex vivo expansion of cultured T cells; however, a series of acute and severe adverse events in a phase I clinical trial testing a superagonist anti-CD28 monoclonal antibody systemically resulted in the discontinuation of the use of antibodies against CD28 (Huenig, Nature Reviews Immunology. 2012;12:317-318). Local or targeted use of anti-CD28 monoclonal antibodies can be used to promote antitumor immunity with reduced risk. Jung et al., Int J Cancer. 2001 Jan 15;91(2):225-30.

[0006] Various families of growth factors and growth factor receptors have been shown to be involved in the autonomous growth of cancer cells. Among these, the epidermal growth factor receptor (EGFR) and the EGF family of peptide growth factors play a central role in the development and progression of different cancer types. EGFR belongs to a family of receptors that includes three additional proteins: ErbB-2, ErbB-3, and ErbB-4. These proteins and the EGF family of growth factors form an integrated system in which signals impinging on individual receptor types are always transmitted to other receptors in the same family. Monoclonal antibodies (mAbs) aimed at enhancing T cell activation are currently in clinical development as antitumor therapeutics. However, most current treatments have difficulty overcoming the inhibitory properties of the tumor microenvironment and therefore fail to generate efficient tumor-specific T cell activation and subsequent tumor cell killing. Several mAbs targeting checkpoint inhibitors, such as CTLA-4 (cytotoxic T lymphocyte-associated protein) 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. While PD-1 inhibition relieves the inhibition on T cell activation, its efficacy as a single agent is not always sufficient to result in tumor elimination and sustained antitumor responses. [Prior art documents] [Non-patent literature]

[0007] [Non-Patent Document 1] Huenig,Nature Reviews Immunology.2012;12:317-318 [Non-patent document 2] Jung et al., Int J Cancer. January 15, 2001;91(2):225-30 Summary of the Invention [Means for solving the problem]

[0008] The present disclosure provides a method of treating cancer in a subject, the method comprising administering to the subject a therapeutically effective amount of a 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), in combination with an antibody, or antigen-binding fragment thereof, that specifically binds to programmed death receptor-1 (PD-1), thereby treating the cancer in the subject.

[0009] In some embodiments, the cancer is a solid tumor. In certain embodiments, the cancer is an EGFR-expressing cancer. In some embodiments, the cancer is selected from esophageal cancer, 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 colon adenocarcinoma, rectal cancer, endometrial cancer, skin cancer, head and neck squamous cell carcinoma, brain cancer, glioblastoma multiforme, non-central nervous system tumors, cutaneous squamous cell carcinoma, breast cancer, gastric cancer, gastroesophageal cancer, gastroesophageal adenocarcinoma, pancreatic cancer, prostate cancer, ovarian cancer, melanoma, nasopharyngeal carcinoma, anal cancer, mesothelioma, renal cell carcinoma, gallbladder / bile duct cancer, pancreatic cancer, 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 with active liver and / or peritoneal metastasis (MSS-CRC). 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.

[0010] In some embodiments, the method further comprises selecting a subject with an advanced solid tumor. In some embodiments, the subject (1) has metastatic or locally advanced disease that is not a candidate for definitive surgery or definitive radiation, (2) is not a candidate for an approved indication for anti-PD-1 or PD-L1 therapy, or such therapy is not otherwise available to the subject (alone or in combination), (3) has exhausted all treatment options expected to provide significant clinical benefit through disease recurrence, treatment-resistant disease, or intolerance, except for subjects with malignancies in which anti-PD-1 or PD-L1 therapy has shown clinical benefit, and / or (4) has (a) microsatellite-stable colorectal cancer documented by local pathology, (b) gastric cancer or gastric cancer. (c) esophageal junction 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, clear cell, or papillary), (m) gallbladder / bile duct carcinoma, (n) urothelial carcinoma, (o) pancreatic cancer, (p) penile SCC, (q) vulvovaginal cancer, or (r) additional non-CNS tumor types in which elevated EGFR expression has been demonstrated in the tumor.

[0011] In some embodiments, the subject has been treated with a prior therapy selected from radiation therapy, 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 subject has not received prior anti-PD-1 or anti-PD-L1 therapy.

[0012] In some embodiments, the subject has microsatellite-stable colorectal cancer (MSS CRC). In some embodiments, a subject with microsatellite-stable colorectal cancer has, or is selected based on, at least one of the following attributes: (a) has metastatic CRC; (b) is not a candidate for definitive surgery or definitive radiation; (c) may have active liver and / or peritoneal metastases at the time of screening; (d) has no identified active liver or peritoneal metastases at the time of screening, with disease sites limited to the lung(s) and / or lymph nodes; (e) has microsatellite stability documented by a pathology report; (f) has received at least one line of therapy in the recurrent / metastatic setting, where the therapy includes anti-EGFR therapy or anti-VEGF therapy; or (g) is anti-PD-1 / PD-L1 naive, defined as having never received treatment with an agent that targets PD-1.

[0013] In some embodiments, the subject has triple-negative breast cancer (TNBC). In some embodiments, the subject with TNBC has or is selected based on at least one of the following attributes: (a) has metastatic TNBC, (b) is not a candidate for definitive surgery or definitive radiation, (c) is not a candidate for anti-PD-1 or anti-PD-L1 therapy in an approved indication, or such therapy is not otherwise available to the subject, (d) has triple-negative cancer (ER- / PR- / Her2-) documented by a pathology report, or (e) is anti-PD-1 / PD-L1 naive, defined as never having been treated with an agent that targets PD-1.

[0014] In some embodiments, the subject has cutaneous squamous cell carcinoma (CSCC) and (i) the subject is not a candidate for definitive surgery or definitive radiation, or (ii) the subject is anti-PD-1 / PD-L1 naive, defined as having never been treated with an agent that targets PD-1.

[0015] In some embodiments, the subject has non-small cell lung cancer (NSCLC). In some embodiments, the subject has, or is selected based on, at least one of the following attributes: (a) the subject has previously histologically documented or cytologically documented locally advanced or metastatic EGFR-mutated non-squamous NSCLC disease, (b) has advanced or metastatic NSCLC, (c) is not a candidate for definitive surgery or definitive radiation, (d) has a previously documented targetable EGFR mutation (EGFR exon 19 deletion, EGFR L858R mutation, EGFR exon 20 insertion, or exon 18 / 21 atypical mutation), (e) is chemotherapy-naive, (f) has been treated with platinum-doublet chemotherapy, (e) has been treated with a third-generation TKI, or (g) is anti-PD-1 / PD-L1 naive, defined as having never been treated with an agent that targets PD-1.

[0016] In some embodiments, the subject has head and neck squamous cell carcinoma (HNSCC). In some embodiments, the subject has or is selected based on at least one of the following attributes: (a) has advanced or metastatic disease, (b) is not a candidate for definitive surgery or definitive radiation, (c) has PD-L1 expression of CPS ≥ 1% by local IHC assay, (d) has not received prior systemic treatment for recurrent or metastatic HNSCC, and / or (e) is anti-PD-1 / PD-L1 naive, defined as not having received treatment with an agent that targets PD-1.

[0017] 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 , 150mg, 200mg, 250mg, 300mg, 350mg, 400mg, 450mg, 500mg, 550mg, 600mg, 650mg, 700mg, 750mg, 800mg, 850mg, 900mg, 1000mg, 1200mg, 1500mg, 1800mg, 2000mg, 2400mg, 2700mg, or 3000mg.

[0018] 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, hi some embodiments, the anti-PD-1 antibody is administered at a dose of 350 mg.

[0019] In some embodiments, the methods comprise administering one or more doses of a bispecific EGFRxCD28 antibody, or antigen-binding fragment thereof, in combination with one or more doses of an anti-PD-1 antibody, or antigen-binding fragment thereof.

[0020] In some embodiments, each of the 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 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.

[0021] In some embodiments, 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, hi some embodiments, each of the one or more doses of the anti-PD-1 antibody is 350 mg.

[0022] In certain embodiments, each of the one or more doses of the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof and / or 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 immediately preceding dose.

[0023] In some embodiments, each of the one or more doses of the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof and / or the one or more doses of the anti-PD-1 antibody or antigen-binding fragment thereof is administered once every week, once every two weeks, once every three weeks, once every four weeks, once every five weeks, or once every six weeks.

[0024] In some embodiments, each of the doses of the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof is administered once every week, hi some embodiments, each of the one or more doses of the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof is administered once every three weeks.

[0025] In some embodiments, each of the one or more doses of the anti-PD-1 antibody or antigen-binding fragment thereof is administered once every three weeks.

[0026] In certain embodiments, the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof and / or the anti-PD-1 antibody or antigen-binding fragment thereof is 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 is administered subcutaneously.

[0027] 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.

[0028] 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.

[0029] In certain aspects, the methods of the disclosure comprise the steps of: (i) administering to a subject a bispecific EGFRxCD28 antibody or antigen-binding fragment thereof subcutaneously or intravenously at a dose of 0.1 mg to 3000 mg once every week or once every three weeks for a monotherapy period, wherein the monotherapy period is at least three weeks; and (ii) administering to the subject a bispecific EGFRxCD28 antibody or antigen-binding fragment thereof subcutaneously or intravenously at a dose of 0.1 mg to 3000 mg once every week or once every three weeks, and administering to the subject an anti-PD-1 antibody or antigen-binding fragment thereof intravenously or subcutaneously at a dose of 150 mg to 500 mg once every three weeks.

[0030] In some embodiments, the monotherapy period is at least 3 weeks, at least 4 weeks, at least 5 weeks, or at least 6 weeks, hi some embodiments, the monotherapy period is less than 1 year, less than 9 months, less than 6 months, less than 3 months, less than 6 weeks, or less than 1 month.

[0031] In some embodiments, during step (ii), the anti-PD-1 antibody or antigen-binding fragment thereof is administered on a different day from the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof. In some embodiments, during step (ii), the anti-PD-1 antibody or antigen-binding fragment thereof is administered on the same day as the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof.

[0032] In certain embodiments, the method further comprises administering to the subject one or more additional agents to treat one or more symptoms of the 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 nonsteroidal anti-inflammatory drug (NSAID).

[0033] In some embodiments, the subject experiences stable disease, a partial response, or a complete response when administered a dose of about 0.1 mg to about 3000 mg of the bispecific antibody or antigen-binding fragment thereof in combination with an anti-PD-1 antibody or antigen-binding fragment thereof for at least one week.

[0034] In one embodiment, the subject has cervical cancer and experiences a partial response after administration. In one embodiment, the subject is a PD-1 negative patient, and the subject has cervical cancer and achieves a partial response after administration.

[0035] In one embodiment, the subject has microsatellite-stable colorectal cancer (CRC) and experiences stable disease or a partial response following the administration. In one embodiment, the subject is administered the bispecific antibody or antigen-binding fragment thereof in combination with cemiplimab, and the subject has CRC and experiences stable disease or a partial response following the administration.

[0036] In some embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof is cemiplimab, nivolumab, pembrolizumab, MEDI0608, BI754091, spartalizumab (PDR001), camrelizumab (SHR-1210), JNJ-63723283, MCLA-134, toripalimab, sintilimab, tislelizumab, selplulimab, dostallimab, retifanlimab, zimberelimab, penprimab, pidilizumab, HX008, balstilimab, or ezabenlimab, or an antigen-binding fragment of any of the foregoing.

[0037] In certain embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof comprises a heavy chain complementarity-determining region (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.

[0038] 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.

[0039] 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.

[0040] 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.

[0041] In some embodiments, the anti-PD-1 antibody is cemiplimab or an antigen-binding fragment thereof.

[0042] In some embodiments, the method comprises administering any one of the bispecific EGFRxCD28 antibodies disclosed herein.

[0043] In a specific embodiment, the first antigen-binding domain that binds to 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.

[0044] 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).

[0045] 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).

[0046] 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.

[0047] In a specific embodiment, 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.

[0048] 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).

[0049] 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).

[0050] In some embodiments, the first 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.

[0051] In a specific embodiment, 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.

[0052] In some embodiments, the bispecific antibody comprises a first heavy chain comprising the amino acid sequence of SEQ ID NO:26.

[0053] In some embodiments, the bispecific antibody comprises a second heavy chain comprising the amino acid sequence of SEQ ID NO:24.

[0054] In some embodiments, the bispecific antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO:28.

[0055] 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.

[0056] 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.

[0057] 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.

[0058] In some embodiments, the bispecific EGFRxCD28 antibody is REGN7075 or an antigen-binding fragment thereof.

[0059] In some embodiments, the method further comprises administering chemotherapy to the subject. In one embodiment, the chemotherapy is a platinum-based chemotherapy. [Brief explanation of the drawings]

[0060] [Figure 1] A schematic diagram of the proposed mechanism of combination therapy of REGN7075 and cemiplimab is shown. REGN7075 (EGFRxCD28) binds to EGFR on tumor cells and CD28 on T cells, thereby bridging the two cell types. Cemiplimab (anti-PD-1) binds to PD-1 on T cells and blocks PD-1-mediated inhibitory signals. Tumor-associated antigens on MHC are presented to T cells. [Figure 2A]

[0023] Figure 1 shows a study flow diagram of weekly dosing of REGN7075 with a lead-in period for REGN7075 monotherapy. REGN7075 may be administered as a split dose or step-up dosing schedule, as disclosed elsewhere herein. [Figure 2B]

[0023] Figure 1 shows a study flow diagram for weekly REGN7075 administration, co-initiation cohort with cemiplimab. REGN7075 may be administered as a split dose or step-up dosing schedule, as disclosed elsewhere herein. [Figure 2C]

[0023] Figure 1 shows a study flow diagram of dose escalation cohorts with a lead-in period for REGN7075 monotherapy followed by Q3W dosing. REGN7075 may be administered as a split dose or step-up dosing schedule, as disclosed elsewhere herein. [Figure 2D]

[0033] Figure 1 shows a dose escalation study flow diagram. REGN7075 may be administered as a split dose or step-up dosing schedule, as disclosed elsewhere herein. [Figure 3] Schematic diagram of a dose-escalation study to evaluate the safety, tolerability, pharmacokinetics, and preliminary antitumor activity of REGN7075 (EGFRxCD28) in combination with cemiplimab (anti-PD-1) in subjects with advanced solid tumors. DL: dose level; RP2D: recommended phase II dose. [Figure 4] The mean serum concentrations of REGN7075 after the first IV dose of REGN7075 are shown: DL1, 0.03 mg; DL2, 0.1 mg; DL3, 0.3 mg; DL4, 1 mg; DL5, 3 mg; DL6, 10 mg; DL7, 30 mg. [Figure 5A] IL-2 is a T cell activation-related cytokine detected in the serum of patients receiving REGN7075 alone and in combination with cemiplimab. DL, dose level; EOT, end of treatment; ET, early discontinuation; IFN, interferon; IL, interleukin; IV, intravenous; LLOQ, lower limit of quantitation; Q3W, every 3 weeks. [Figure 5B]IFN-γ, a T-cell activation-related cytokine, was detected in the serum of patients receiving REGN7075 alone and in combination with cemiplimab. DL, dose level; EOT, end of treatment; ET, early discontinuation; IFN, interferon; IL, interleukin; IV, intravenous; LLOQ, lower limit of quantitation; Q3W, every 3 weeks. [Figure 6] Percent change from baseline in target lesions in 18 patients treated with dose escalation is shown. Data are preliminary and the study is still ongoing. DL, dose level; MSS CRC, microsatellite stable colorectal cancer. [Figure 7] Percent change from baseline in target lesions in 25 patients treated with dose escalation is shown, based on data as of December 2022. Data are preliminary and the study is still ongoing. SD: stable disease; PD: progressive disease; CR / PR: complete response / partial response. DETAILED DESCRIPTION OF THE INVENTION

[0061] Before the present disclosure is described, it is to be understood that this disclosure is not limited to the particular 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 limit the scope of the present disclosure, which will be limited only by the appended claims.

[0062] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. As used herein, the term "about," when used in connection with a specific recited numerical value, means that the value may vary by 1% or less from the recited value. For example, as used herein, the expression "about 100" includes 99 and 101, and all values ​​therebetween (e.g., 99.1, 99.2, 99.3, 99.4, etc.).

[0063] Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of this disclosure, the preferred methods and materials are now described. All patents, applications, and non-patent publications mentioned herein are incorporated by reference in their entirety.

[0064] definition As used herein, "EGFR" and "EGFR fragment" refer to the well-known human EGFR protein or fragments thereof (e.g., "mouse EGFR," "mouse EGFR fragment," "simian EGFR," "simian EGFR fragment," etc.), unless specified to be derived from a non-human species. In one embodiment, the human EGFR comprises the amino acid sequence set forth in NCBI Accession No. NP_005219.2. In one embodiment, the human EGFR (amino acids L25-A647 of Accession No. 005228.4) is depicted with a C-terminal CPGG.myc epitope (E1-L10).GlyGly.myc epitope (E1-L10).SerGly.6XHis.SSG tag (SEQ ID NO: 69).

[0065] As used herein, "CD28" refers to the well-known human CD28 protein expressed on T cells as a costimulatory receptor, unless specified to be from a non-human species. In one embodiment, human CD28 comprises the amino acid sequence set forth in NCBI Accession No. NP_006130.1.

[0066] As used herein, "PD-1" or "programmed death receptor-1" refers to the well-known human PD-1 protein or a fragment thereof, unless specified to be from a non-human species. In one embodiment, human PD-1 comprises the amino acid sequence set forth in NP_005009.

[0067] An "antibody" is an immunoglobulin molecule comprising four polypeptide chains, two heavy chains (HC) and two light chains (LC), interconnected by disulfide bonds. Each heavy chain (HC) contains a heavy chain variable region (HCVR or VLC) Hand a heavy chain constant region (e.g., IgG, IgG1, or IgG4). The heavy chain constant region is H 1. C H 2 and C H Each light chain (LC) contains three domains: a light chain variable region (LCVR or VVR) and a VV ... L and a light chain constant region (e.g., lambda or kappa). The light chain constant region comprises one domain (C L 1) V H Area and V L The regions can be further subdivided into regions of hypervariability, called complementarity-determining regions (CDRs), interspersed with more conserved regions, called framework regions (FRs). H and V L comprises three CDRs and four FRs arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The heavy chain CDRs may be referred to as HCDRs, and the light chain CDRs may be referred to as LCDRs. In different embodiments of the present disclosure, the FRs of the antibody (or antigen-binding portion thereof) may be identical to the human germline sequence or may be modified naturally or artificially.

[0068] 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 antigen-binding fragment or portion thereof) may have an effector arm that binds to a first antigen and a targeting arm that binds to a second antigen. The effector arm may 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 may 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. The terms "EGFRxCD28 bispecific antibody," "bispecific EGFRxCD28 antibody," and EGFRxCD28 are used interchangeably throughout this specification.

[0069] The antigen-binding arm of a Y-shaped IgG antibody (e.g., a CD28- or EGFR-binding arm) refers to the structural portion of the antibody that confers binding specificity to an antigen. For example, the antigen-binding arm of an IgG antibody comprises a heavy chain (HC) associated with a light chain (LC).

[0070] As used herein, an "antigen-binding portion" of an antibody, an "antigen-binding fragment" of an antibody, etc., includes naturally occurring, enzymatically obtainable, synthetic, or genetically engineered polypeptides or glycoproteins that specifically bind to an antigen to form a complex. Multispecific antigen-binding fragments of antibodies bind to multiple antigens (e.g., two different antigens if the fragment is bispecific). Antigen-binding fragments of antibodies can be derived from intact antibody molecules using any suitable standard technique, such as, for example, proteolytic digestion or recombinant genetic engineering techniques involving the manipulation and expression of DNA encoding antibody variable domains, and optionally, constant domains. 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.

[0071] An antigen-binding fragment of an antibody, in one embodiment, comprises at least one variable domain. A variable domain may be of any size or amino acid composition and generally comprises at least one CDR adjacent to or in-frame with one or more framework sequences. L V associated with the domain H For antigen-binding fragments containing domains, V H Domain and V L The domains can be positioned relative to each other in any suitable configuration. For example, the variable region is a dimer, with the V H -V H , V H -V L or V L -V L Alternatively, the antigen-binding fragment of an antibody may comprise a monomeric V dimer. H or V L It may include a domain.

[0072] In certain embodiments, an antigen-binding fragment of an antibody may comprise at least one variable domain covalently linked to at least one constant domain. Non-limiting exemplary configurations of variable and constant domains that may be found in an antigen-binding fragment of an antibody of the present disclosure include: (i) a 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 H2. (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 arrangement of variable and constant domains, including any of the exemplary arrangements listed above, the variable and constant domains may be directly linked to each other or may be linked by a complete or partial hinge or linker region. A hinge region may consist of at least two (e.g., 5, 10, 15, 20, 40, 60, or more) amino acids that provide a flexible or semi-flexible connection between adjacent variable and / or constant domains in a single polypeptide molecule. Furthermore, antigen-binding fragments of antibodies of the present disclosure may be linked to each other and / or to one or more monomeric V H Domain or V L It may comprise homodimers or heterodimers (or other multimers) of any of the variable and constant domain arrangements listed above in non-covalent association with the domains (e.g., by disulfide bond(s)).

[0073] "Isolated" antigen binding proteins (e.g., antibodies or antigen-binding fragments thereof), polypeptides, polynucleotides, and vectors are at least partially free from other biological molecules derived from the cell or cell culture in which they are produced. Such biological molecules include nucleic acids, proteins, other antibodies or antigen-binding fragments, lipids, carbohydrates, or other substances such as cell debris and growth medium. Isolated antigen binding proteins may also be at least partially free from expression system components, such as biological molecules derived from the host cell or its growth medium. In general, the term "isolated" does not imply the complete absence of such biological molecules, or the absence of water, buffers, or salts, or components of a pharmaceutical formulation that comprises the antigen binding protein (e.g., antibody or antigen-binding fragment).

[0074] The term "recombinant" antigen binding protein (such as an antibody or antigen-binding fragment thereof) refers to a molecule made, 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 a non-human mammal (including a transgenic non-human mammal, e.g., a transgenic mouse), or in a host cell (e.g., Chinese hamster ovary (CHO) cell) or cellular expression system, or antibodies isolated from a recombinant combinatorial human antibody library. The present disclosure includes recombinant antigen binding proteins as described herein.

[0075] Terms such as "specifically bind" mean that an antibody or 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 to an antigen are well known in the art and include, for example, equilibrium dialysis, surface plasmon resonance, and the like. In some embodiments, the terms "specifically binds" or "binds specifically" refer to the formation of a K complex to an antigen, such as an EGFR, CD28, or PD-1 protein. DThe binding affinity, expressed as a function of the binding affinity of the antibody, is about 10 when measured, for example, at 25° C. or 37° C. by, for example, a real-time label-free biolayer interferometry assay, such as, for example, an Octet® HTX biosensor, or by surface plasmon resonance, such as, for example, a BIACORE™, or by solution affinity ELISA. -6 M (e.g., 10 -7 M, 10 -8 M, 10 -9 M, 10 -10 M, 10 -11 M or 10 -12 "Anti-EGFR" refers to an antigen-binding protein (e.g., an antibody or antigen-binding fragment thereof) that specifically binds to EGFR. "Anti-CD28" refers to an antigen-binding protein (or other molecule, such as an antigen-binding arm), e.g., an antibody or antigen-binding fragment thereof, that specifically binds to CD28. "EGFRxCD28" refers to an antigen-binding protein (or other molecule, such as an antigen-binding arm), e.g., an antibody or antigen-binding fragment thereof, that specifically binds to EGFR and CD28 (and, optionally, one or more other antigens). "Anti-PD-1" refers to an antigen-binding protein (or other molecule, such as an antigen-binding arm), e.g., an antibody or antigen-binding fragment thereof, that specifically binds to PD-1. However, an isolated antibody or antigen-binding fragment that specifically binds to a human protein, e.g., PD-1, may have cross-reactivity to other antigens, such as proteins of other (non-human) species, e.g., PD-1 protein.

[0076] The present disclosure includes methods of treating cancer comprising administering an antigen binding protein, e.g., an antibody or antigen-binding fragment that binds to the same EGFR and CD28 epitopes as the 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 can be found in Tables 9A, 9B, and 9C. The amino acid sequences of the EGFR HCVR arms of the bispecific antibodies described herein can be found in Table 1, while the amino acid sequences of the CD28 HCVR arms of the bispecific antibodies described herein can be found 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 found in Table 6. The amino acid sequences of the consensus light chain variable regions described in this disclosure are also found in Table 6.

[0077] Typically, antibodies or antigen-binding fragments of the present disclosure, modified in some way, retain the ability to specifically bind to EGFR and CD28, e.g., retain at least 10% of their EGFR and CD28 binding activity (compared to the parent antibody), when that activity is expressed on a molar basis. Preferably, 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 of the parent antibody. It is also intended that antibodies or antigen-binding fragments include conservative or non-conservative amino acid substitutions (referred to as "conservative variants" or "function-conservative variants" of antibodies) that do not substantially alter their biological activity.

[0078] "Variant" of a polypeptide, e.g., an immunoglobulin chain comprising an amino acid sequence specifically set forth herein (e.g., the V of 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)) H , V L , HC or LC, or CDRs thereof) may have 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, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 118, 119, 120, 122, 124, 126, 128, 129, 130, 131, 132, 133, 134, 135, 136, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, "A" refers to a polypeptide containing an amino acid sequence that is identical or similar to a polypeptide containing an amino acid sequence that is 95, 96, 97, 98, 99, 99.5 or 99.9% of the total amino acid sequence. When the comparison is performed by the BLAST algorithm, the algorithm parameters are selected to give the largest match between the respective sequences over the entire length of the respective reference sequences (e.g., expectation threshold: 10; word size: 3; maximum match within query: 0; BLOSUM62 matrix; gap cost: 11 for presence, 1 for extension; conditional composition score matrix adjustment).

[0079] Additionally, variants of a polypeptide may include 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, which may include the amino acid sequence of a reference polypeptide having an amino acid sequence specifically set forth herein. H , V L, HC, or LC, or CDRs thereof), but may contain 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. For example, the present disclosure provides an EGFR-binding arm immunoglobulin light chain (or VH chain) comprising the amino acid sequence set forth in SEQ ID NO: 16, but with one or more such mutations. L ) variants, and / or immunoglobulin heavy chains (or V) comprising the amino acid sequence set forth in SEQ ID NO:2 but with one or more such mutations. H 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., one or two or three) such CDRs have one or more such mutations (e.g., conservative substitutions)), and / or an immunoglobulin heavy chain variant comprising HCDR1, HCDR2 and HCDR3 (wherein one or more (e.g., one or two or three) such CDRs have one or more such mutations (e.g., conservative substitutions)).

[0080] "Conservatively modified variants" or "conservative substitutions," e.g., of immunoglobulin chains described herein, refer to variants in which one or more amino acids in a polypeptide are substituted with other amino acids having similar properties (e.g., charge, side chain size, hydrophobicity / hydrophilicity, backbone conformation and rigidity, etc.). Such changes can frequently be made without significantly impairing the biological activity of the antibody or fragment. Those skilled in the art recognize that, in general, 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., p. 224(4 th(See, e.g., EGFRxCD28, Vol. 1, No. 1, pp. 111-114, 2003). Furthermore, substitution of structurally or functionally similar amino acids is unlikely to significantly impair biological activity. The present disclosure includes bispecific EGFRxCD28 antibodies and antigen-binding fragments thereof comprising such conservatively modified variant immunoglobulin chains.

[0081] Examples of groups of amino acids with side chains that have 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 replacement is any change that has a positive value in the PAM250 log-likelihood matrix disclosed in Gonnet et al. (1992) Science 256:1443-45.

[0082] The following references relate to the BLAST algorithm, which is 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. al.,(1993)Comput.Chem.17:149-163;Hancock,JMet al.,(1994)Comput.Appl.Biosci.10:67-70;ALIGNMENT SCORING SYSTEMS:Dayhoff,MO,et al.,“A model of evolutionary change in proteins.”in Atlas of Protein Sequence and Structure,(1978)vol.5,suppl.3.MODayhoff(ed.),pp.345-352,Natl.Biomed.Res.Found.,Washington,DC;Schwartz,RM,et al.,“Matrices for detecting distant relationships.”in Atlas of Protein Sequence and Structure,(1978)vol.5,suppl.3.''MODayhoff(ed.),pp.353-358,Natl.Biomed.Res.Found.,Washington,DC;Altschul,SF,(1991)J.Mol.Biol.219:555-565;States,DJ,et al.,(1991)Methods 3:66-70;Henikoff,S.,et al.,(1992)Proc.Natl.Acad.Sci.USA 89:10915-10919;Altschul,SF,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, SF “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, NY

[0083] 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, e.g., an EGFR-expressing cancer. The subject may have cancer, e.g., an EGFR-expressing cancer, be predisposed to developing such a condition, and / or 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 many embodiments, the term "subject" may be used interchangeably with the term "patient."

[0084] As used herein, the phrase "subject in need thereof" refers to a human or non-human mammal that exhibits one or more symptoms or signs of cancer and / or has been diagnosed with cancer, including solid tumors, and is in need of treatment.

[0085] As used herein, the terms "treatment", "treating" and the like mean alleviating symptoms, temporarily or permanently eliminating the cause of symptoms, slowing or inhibiting tumor growth, reducing tumor cell mass or tumor burden, promoting tumor regression, causing tumor shrinkage, necrosis and / or disappearance, preventing tumor recurrence, and / or prolonging the survival of a subject.

[0086] As used herein, the term "solid tumor" refers to an abnormal mass of tissue that usually does not contain cysts or areas of fluid. Solid tumors may be benign (not cancerous) or malignant (cancer). For purposes of this disclosure, the term "solid tumor" refers to a malignant solid tumor. This term includes different types of solid tumors named for the cell type that forms them: sarcomas, carcinomas, and lymphomas. However, this term does not include leukemias. In various embodiments, the term "solid tumor" includes cancers that begin in connective or supportive tissue (e.g., bone or muscle) (called sarcomas), cancers that begin in the epithelial cells lining the body's glandular cells and body tissues (called carcinomas), and cancers of lymphatic organs, such as the lymph nodes, spleen, and thymus (called lymphomas). Because lymphocytes occur in almost every tissue in the body, lymphomas can arise in a variety of organs. In certain embodiments, the term "solid tumor" includes cancers including, but 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, stomach cancer, esophageal cancer, head and neck cancer, salivary gland cancer, and myeloma. In certain embodiments, the term "solid tumor" includes cancers including, but 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" includes multiple solid tumor lesions located apart from one another in a subject in need of treatment, e.g., 2, more than 2, more than 5, more than 10, more than 15, more than 20, or more than 25 lesions. In certain embodiments, the multiple lesions are located distally from one another within the same organ, while in other certain embodiments, the tumor lesions may be located in different organs.

[0087] As used herein, the phrase "in combination with" means that a first therapeutic agent, e.g., a bispecific EGFRxCD28 antibody or antigen-binding fragment thereof, is administered before, after, or simultaneously with a second therapeutic agent, e.g., an anti-PD-1 antibody or antigen-binding fragment thereof. The term "in combination with" also includes sequential or simultaneous administration of a first therapeutic agent, e.g., a bispecific EGFRxCD28 antibody or antigen-binding fragment thereof, and a second therapeutic agent, e.g., an anti-PD-1 antibody or antigen-binding fragment thereof.

[0088] Combination Therapy for Treating or Inhibiting Cancer Growth - Patent application The present disclosure provides methods for treating, ameliorating, or reducing the severity of at least one symptom or sign, or inhibiting the growth of cancer in a subject, comprising administering to the subject an effective amount of a bispecific EGFRxCD28 antibody (e.g., REGN7075, or any combination of an anti-EGFR HCVR paired with a HCVR from any of the CD28 antibodies described herein), in combination with an effective amount of a PD-1 inhibitor, such as an antibody (e.g., cemiplimab) or antigen-binding fragment thereof.

[0089] 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 useful, inter alia, for the treatment of any cancer in which stimulating, activating, and / or targeting an immune response is beneficial. In particular, the bispecific EGFRxCD28 antibody or antigen-binding molecule of the present disclosure can be used to treat, prevent, and / or ameliorate cancer, for example, cancers associated with or mediated by EGFR expression or activity or the proliferation of EGFR+ cells. The mechanism of action achieved by the therapeutic methods of the present disclosure involves killing EGFR-expressing cells in the presence of effector cells, e.g., 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.

[0090] For purposes of this specification, cancer refers to a disease characterized by abnormal, excessive and / or uncontrolled cell growth. Exemplary cancers include, but are not limited to, esophageal carcinoma 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, stomach cancer, liver cancer, pancreatic cancer, kidney cancer, urothelial carcinoma, prostate cancer or prostate adenocarcinoma, testicular cancer, breast cancer (e.g., ductal carcinoma or lobular carcinoma), cervical cancer or cervical carcinoma, endometrial cancer, ovarian cancer, gastroesophageal cancer (e.g., gastroesophageal adenocarcinoma), tumors outside the central nervous system (CNS), melanoma, nasopharyngeal carcinoma, anal cancer, mesothelioma, renal cell carcinoma (e.g., chromophobe, clear cell, or papillary), gallbladder / bile duct carcinoma, penile squamous cell carcinoma, or vulvovaginal cancer. In one embodiment, the cancer is an EGFR-expressing cancer. A wide range of cancers express EGFR. Therefore, the methods of the present disclosure can be used to treat a wide range of cancers.

[0091] Cancers characterized by solid tumor cells or cancerous blood cells (which may be, for example, EGFR-expressing cancers in which EGFR expression is confirmed in the cells of a particular subject to be treated) include esophageal cancer, lung squamous cell carcinoma, lung adenocarcinoma, cervical squamous cell carcinoma, endometrial adenocarcinoma, bladder urothelial carcinoma, lung cancer (e.g., non-small cell lung cancer), colon 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.

[0092] The methods of the present disclosure may also be used to treat or ameliorate primary and / or metastatic tumors occurring, for example, in colon, lung, breast, kidney, and bladder cancer (or any cancer discussed herein).

[0093] The present disclosure also includes methods for treating or ameliorating residual cancer in a subject. As used herein, the term "residual cancer" refers to the presence or persistence of one or more cancerous cells in a subject after treatment with an anti-cancer therapy.

[0094] In certain embodiments, the methods of the present disclosure can be used to treat subjects who exhibit elevated levels of one or more cancer-associated 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 rearrangements, or circulating tumor DNA). For example, the methods of the present disclosure include administering a therapeutically effective amount of an anti-PD-1 antibody in combination with a bispecific EGFRxCD28 antibody to a subject with elevated levels of PD-L1 and / or EGFR. In one embodiment, the methods of the present disclosure are used in subjects with cancers selected based on PD-L1 expression in cancer tissue. In certain embodiments, the current methods are used to treat subjects with cancers selected based on PD-L1 expression in cancer tissues and / or immune cells of 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%. Methods for determining PD-L1 expression in cancer tissues 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 an ELISA assay, or by an immunohistochemistry (IHC) assay (e.g., the 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, for example, by in situ hybridization or RT-PCR. In certain embodiments, PD-L1 expression is determined by imaging with a labeled anti-PD-L1 antibody, for example by immunopositron 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 expressly incorporated herein by reference in their entireties).

[0095] In one embodiment, the method includes determining whether the subject's cancer expresses EGFR. If such expression is observed, a bispecific EGFRxCD28 antibody is administered in combination with an anti-PD-1 antibody or antigen-binding fragment thereof. For example, in one embodiment, the method includes obtaining a biopsy of the cancer and determining whether cells of the cancer express EGFR, and if EGFR expression is present, administering to the subject a bispecific EGFRxCD28 antigen-binding protein in combination with an anti-PD-1 antibody or antigen-binding fragment thereof. In one embodiment, EGFR expression is examined immunohistochemically (IHC) or by ELISA (enzyme-linked immunosorbent assay).

[0096] According to certain aspects, the present disclosure provides methods for treating, ameliorating, or reducing the severity of at least one symptom or sign of, or inhibiting the growth of cancer, e.g., a cancer associated with EGFR expression (e.g., lung cancer), comprising administering to a subject one or more bispecific EGFRxCD28 antibodies or antigen-binding molecules described herein, e.g., REGN7075, in combination with an anti-PD-1 antibody or antigen-binding fragment thereof (e.g., cemiplimab), e.g., after the subject has been shown to be non-responsive to other types of anti-cancer therapy.

[0097] For example, the disclosure includes a method of treating cancer, such as lung cancer, comprising administering to a subject a bispecific EGFRxCD28 antibody or antigen-binding molecule, e.g., REGN7075, and an anti-PD-1 antibody or 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 a standard of care treatment for a subject with cancer, e.g., lung cancer.

[0098] In certain embodiments, administration of a bispecific EGFRxCD28 antibody or antigen-binding fragment thereof in combination with an anti-PD-1 antibody or antigen-binding fragment thereof results in increased tumor regression, tumor shrinkage and / or elimination. In certain embodiments, administration of an anti-PD-1 antibody and a bispecific EGFRxCD28 antibody slows tumor growth and progression, for example, may slow tumor growth 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 untreated subjects or subjects treated with either antibody as monotherapy. In certain embodiments, administration of the anti-PD-1 antibody and the bispecific EGFRxCD28 antibody prevents tumor recurrence and / or increases subject survival, e.g., by at least 15 days, at least 1 month, at least 3 months, at least 6 months, at least 12 months, at least 18 months, at least 24 months, at least 36 months, or at least 48 months relative to untreated subjects or subjects receiving either antibody as monotherapy. In certain embodiments, administration of the anti-PD-1 antibody and the bispecific EGFRxCD28 antibody increases progression-free survival or overall survival. In certain embodiments, administration of the anti-PD-1 antibody and the bispecific EGFRxCD28 antibody increases response and duration of response in a subject, e.g., by at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, at least 7%, at least 8%, at least 9%, at least 10%, at least 20%, at least 30%, at least 40%, or at least 50% relative to untreated subjects or subjects receiving either antibody as monotherapy.

[0099] In certain embodiments, 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 (a "partial response"). In certain embodiments, 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 (a "complete response"). In certain embodiments, 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 shrinkage can be measured by any method known in the art, for example, by x-ray, positron emission tomography (PET), computed tomography (CT), magnetic resonance imaging (MRI), cytology, histology, or molecular genetic analysis.

[0100] In certain embodiments, methods of the disclosure comprise administering a therapeutically effective amount of a bispecific EGFRxCD28 antibody in combination with an anti-PD-1 antibody to a subject in need thereof, wherein administration of the combination increases the patient's overall survival (OS) or progression-free survival (PFS) compared to patients administered a standard of care (SOC) therapy (e.g., chemotherapy, surgery, or radiation). In certain embodiments, PFS is increased by at least 1 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 compared to patients administered any one or more SOC therapies. In certain embodiments, OS is increased by at least 1 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 compared to patients administered any one or more SOC therapies.

[0101] (I) Dosage and timing An "effective" or "therapeutically effective" amount of an anti-PD-1 antibody or antigen-binding fragment thereof, or a bispecific EGFRxCD28 antibody or antigen-binding fragment thereof, such as REGN7075, for treating or preventing cancer, such as an EGFR-expressing cancer, is the amount of the antibody or antigen-binding fragment sufficient to alleviate one or more signs and / or symptoms of the disease in a treated subject, by inducing regression or elimination of such signs and / or symptoms, or inhibiting the progression of such signs and / or symptoms.

[0102] The dose of an antigen-binding molecule administered to a subject may vary depending on the age and size of the subject, the target disease, condition, route of administration, etc. A preferred dose is typically calculated based on body weight or body surface area. The frequency and duration of treatment can be adjusted depending on the severity of the condition.

[0103] In some embodiments of the present disclosure, the therapeutically effective amount of a bispecific EGFRxCD28 antibody, e.g., REGN7075, is 0.1-3000 mg. The dose may vary depending on the age and size of the subject, the target disease, condition, route of administration, etc. In some embodiments, the bispecific EGFRxCD28 antibody is administered at a dose of approximately 0.1-900 mg, 1-1500 mg, 100-1200 mg, 300-1000 mg, 500-1500 mg, 800-1000 mg, 800-1500 mg, 300-2000 mg, or 300-3000 mg. In some embodiments, the bispecific EGFRxCD28 antibody is administered at a concentration 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 In some embodiments, the bispecific EGFRxCD28 antibody is administered at a dose of about 0.1-50 mg / kg, 1-45 mg / kg, 5-10 mg / kg, 10-30 mg / kg, 15-25 mg / kg, 20-30 mg / kg, or 25-40 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.

[0104] 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.

[0105] In some embodiments, the bispecific EGFRxCD28 antibody is administered intravenously (IV). In some embodiments, the IV infusion is administered over about 60 minutes. In some embodiments, the bispecific EGFRxCD28 antibody is administered subcutaneously.

[0106] In some embodiments of the disclosure, the therapeutically effective dose of a bispecific EGFRxCD28 antibody, e.g., REGN7075, is 0.1 to 3000 mg IV or SC every week or every three weeks.

[0107] In some embodiments of the present disclosure, the therapeutically effective amount of an anti-PD-1 antibody or antigen-binding fragment thereof, e.g., cemiplimab, is 50 to 1500 mg, e.g., 350 mg. The dose may vary depending on the age and size of the subject, the target disease, condition, route of administration, etc. In some embodiments of the present disclosure, the therapeutically effective amount of an anti-PD-1 antibody or antigen-binding fragment thereof, such as cemiplimab, is about 50-1500 mg, 100-1250 mg, 150-1000, 200-750 mg, 300-500 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 0.1-20 mg / kg, 0.5-15 mg / kg, 1-12 mg / kg, 2-10 mg / kg, 5-10 mg / kg, or 7.5-10 mg / kg of the subject's 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 of the subject's body weight.

[0108] 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.

[0109] In some embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof is administered intravenously (IV). In some embodiments, the IV infusion is administered over about 30 minutes or about 60 minutes. In some embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof is administered subcutaneously.

[0110] In some embodiments, the therapeutically effective amount of an anti-PD-1 antibody or antigen-binding fragment thereof, e.g., cemiplimab, is 50-1500 mg, e.g., 350 mg, administered intravenously (IV) or subcutaneously (SC) every week (QW), Q2W (every 2 weeks), or Q3W (every 3 weeks).

[0111] In some embodiments, the bispecific EGFRxCD28 antibody or antigen-binding molecule is administered simultaneously with the anti-PD-1 antibody or 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 antigen-binding portion thereof. In some embodiments, the bispecific EGFRxCD28 antibody or antigen-binding molecule is administered before, for example, 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, the anti-PD-1 antibody or antigen-binding portion thereof. In some embodiments, the anti-PD-1 antibody or antigen-binding molecule is administered prior to administration of the bispecific EGFRxCD28 antibody, or antigen-binding portion thereof, e.g., 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.

[0112] In certain embodiments, the initial dose is followed by a second or multiple subsequent doses of the antigen binding protein(s) in an amount that is about the same as, less than, or greater than the initial dose, with the subsequent doses being spaced apart, for example, by about 1 week, 2 weeks, 3 weeks, 10 days, 20 days, or 30 days.

[0113] 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) may be administered to a subject over a defined time course. The method according to this aspect of the present disclosure comprises sequentially administering multiple doses of the antigen-binding molecule of the present disclosure to the subject. As used herein, "sequentially administering" means that each dose of the antigen-binding molecule is administered to the subject at different times, for example, on the same day with a predetermined interval (e.g., several hours) between them, or on different days with a predetermined interval (e.g., several hours, days, weeks, or months) between them. The present disclosure includes methods comprising sequentially administering to a subject a single initial dose of an antigen-binding molecule, followed by one or more secondary doses of the same or different antigen-binding molecule, and then optionally one or more tertiary doses of the antigen-binding molecule.

[0114] The terms "initial dose," "secondary dose," and "tertiary dose" refer to the time sequence of administration of an antigen-binding molecule of the present disclosure. Thus, an "initial dose" is a dose administered at the start of a treatment regimen (also referred to as a "baseline dose"), a "secondary dose" is a dose administered after the initial dose, and a "tertiary dose" is a dose administered after the second dose. The initial dose, secondary dose, and tertiary dose may all contain the same amount of antigen-binding molecule but may differ from one another in terms of administration frequency. However, in certain embodiments, the amount of antigen-binding molecule contained in the initial dose, secondary dose, and / or tertiary dose differs from one another during the course of treatment (e.g., adjusted up or down as needed). In certain embodiments, two or more doses are administered as "loading doses" at the start of a treatment regimen, followed by subsequent doses (e.g., "maintenance doses") that are administered on a less frequent basis.

[0115] In one exemplary embodiment of the present disclosure, each secondary and / or tertiary dose is administered one to several weeks after the immediately preceding dose. As used herein, the phrase "immediately preceding dose" refers to a dose of an antigen-binding molecule administered to a subject prior to the administration of a subsequent dose in a series of multiple doses, with no intervening doses.

[0116] The method according to this aspect of the disclosure may include administering any number of secondary and / or tertiary 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 secondary dose is administered to a subject. In other embodiments, two or more secondary doses are administered to a subject. Similarly, in certain embodiments, only a single tertiary dose is administered to a subject. In other embodiments, two or more tertiary doses are administered to a subject.

[0117] In embodiments involving multiple secondary doses, each secondary dose can be administered at the same frequency as the other secondary doses. Similarly, in embodiments involving multiple tertiary doses, each tertiary dose can be administered at the same frequency as the other tertiary doses. Alternatively, the frequency with which the secondary doses and / or tertiary doses are administered to a subject can vary over the course of a treatment regimen. The administration frequency can also be adjusted by a physician during the course of treatment, depending on the needs of each individual subject after clinical examination.

[0118] In some embodiments, the bispecific EGFRxCD28 antibody, e.g., REGN7075, or 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 a course of treatment. In some embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof, e.g., cemiplimab, 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 a course of treatment.

[0119] In certain aspects, the methods of the disclosure comprise the steps of: (i) administering to a subject a bispecific EGFRxCD28 antibody or antigen-binding fragment thereof subcutaneously or intravenously at a dose of 0.1 mg to 3000 mg once every week or once every three weeks for a monotherapy period, wherein the monotherapy period is at least three weeks; and (ii) administering to the subject a bispecific EGFRxCD28 antibody or antigen-binding fragment thereof subcutaneously or intravenously at a dose of 0.1 mg to 3000 mg once every three weeks, and administering to the subject an anti-PD-1 antibody or antigen-binding fragment thereof intravenously or subcutaneously at a dose of 150 mg to 500 mg once every three weeks. In some embodiments, the monotherapy period is at least three weeks, at least four weeks, at least five weeks, or at least six weeks. In some embodiments, the monotherapy period is less than one year, less than nine months, less than six months, less than three months, less than six weeks, or less than one month. In some embodiments, the monotherapy period is at least 3 weeks but less than 1 year.

[0120] In some embodiments, during step (ii), the anti-PD-1 antibody or antigen-binding fragment thereof is administered on a different day from the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof. In some embodiments, during step (ii), the anti-PD-1 antibody or antigen-binding fragment thereof is administered on the same day as the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof.

[0121] In some embodiments, the bispecific EGFRxCD28 antibody is administered to the subject before, concurrently with, or after administration of the anti-PD-1 antibody or antigen-binding fragment thereof. In some embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof is administered to the subject before, concurrently with, or after administration of the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof.

[0122] In some embodiments, the bispecific EGFRxCD28 antibody is initially administered at least two, three, four, or five or more times as monotherapy, followed by administration of the anti-PD-1 antibody or antigen-binding fragment thereof. As a non-limiting example, the bispecific EGFRxCD28 antibody or fragment thereof may be administered by IV infusion or subcutaneous injection at a dose of 0.1 mg to 3000 mg every week (QW) or every three weeks (Q3W) at least one week before administration of the anti-PD-1 antibody or antigen-binding fragment thereof by IV infusion or subcutaneous injection at a dose of 350 mg every three weeks (Q3W).

[0123] (II) Administration route The present disclosure provides methods for administering a bispecific EGFRxCD28 antibody, e.g., 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, e.g., cemiplimab, or an antigen-binding fragment thereof, to a subject (e.g., a human, e.g., a human suffering from cancer), comprising introducing the antigen-binding protein or pharmaceutical composition into the body of the subject (e.g., a human), e.g., intravenously or subcutaneously. For example, the method comprises puncturing the body of the subject with the needle of a syringe and injecting the antigen-binding protein or pharmaceutical composition into the body of the subject, e.g., into a vein, artery, skin, tumor, muscle tissue, or subcutaneous tissue of the subject.

[0124] The antibody or pharmaceutical composition thereof can be administered by a variety of methods, including parenteral, non-parenteral, oral, rectal, transmucosal, intestinal, parenteral, intramuscular, subcutaneous, intradermal, intramedullary, intrathecal, direct intraventricular, intravenous, intraperitoneal, intranasal, intraocular, inhalation, insufflation, topical, dermal, intraocular, intravitreal, transdermal, or intra-arterial.

[0125] 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.

[0126] In some further embodiments, the bispecific EGFRxCD28 antibody and / or anti-PD-1 antibody is administered to the 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 anti-PD-1 antibody is administered to the subject over about 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, or 90 minutes.

[0127] The present disclosure also provides a container (e.g., a plastic or glass vial or ampoule with a cap or chromatography column, hollow needle or syringe cylinder) comprising a bispecific EGFRxCD28 antigen binding protein of the disclosure or a pharmaceutical composition thereof.

[0128] The present disclosure also provides an injection device comprising 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. An injection device is a device that introduces a substance into a subject's body via a parenteral route, for example, intramuscularly, subcutaneously, or intravenously. For example, the injection device may be a syringe or auto-injector (e.g., pre-filled with a pharmaceutical formulation) that includes a cylinder or barrel for holding the fluid to be injected (e.g., comprising an antibody or fragment thereof or a pharmaceutical formulation thereof), a needle for suturing the skin, blood vessel, or other tissue to inject the fluid, and a plunger that forces the fluid from the cylinder through the needle bore and into the subject's body.

[0129] A pre-filled syringe is a syringe that is filled with a composition (e.g., a pharmaceutical composition comprising 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.

[0130] Pharmaceutical compositions are described in more detail below.

[0131] (III) Selection of Subjects In some embodiments, the methods described herein further comprise one or more steps of selecting subjects. Patients may be selected, for example, based on inclusion criteria, and excluded, for example, based on exclusion criteria. The inclusion and exclusion criteria are described in detail in Example 2 below.

[0132] In some embodiments, the method comprises selecting a subject with an advanced solid tumor.

[0133] In some embodiments, the subject (1) has metastatic or locally advanced disease that is not a candidate for curative surgery or curative radiation; (2) is not a candidate for an approved indication for anti-PD-1 or PD-L1 therapy, or such therapy is not otherwise available to the subject (alone or in combination); (3) has exhausted all treatment options expected to provide significant clinical benefit through disease recurrence, treatment-resistant disease, or intolerance, except for subjects with malignancies in which anti-PD-1 or PD-L1 therapy has shown clinical benefit; and / or (4) has (a) microsatellite-stable colorectal cancer documented by local pathology, (b) gastric cancer or gastric cancer. (c) esophageal junction 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, clear cell, or papillary), (m) gallbladder / bile duct carcinoma, (n) urothelial carcinoma, (o) pancreatic cancer, (p) penile SCC, (q) vulvovaginal cancer, or (r) additional non-CNS tumor types in which elevated EGFR expression has been demonstrated in the tumor.

[0134] In some embodiments, the method includes selecting a subject who has been treated with a previous therapy. In one embodiment, the previous therapy is radiation, surgery, chemotherapy, a PD-1 inhibitor, a PD-L1 inhibitor, anti-VEGF therapy, CAR-T therapy, and / or anti-EGFR therapy. In some embodiments, the method includes selecting a subject who has not received a previous anti-PD-1 or anti-PD-L1 therapy.

[0135] In some embodiments, the method includes selecting a subject with microsatellite-stable colorectal cancer (MSS CRC). In some embodiments, the subject with microsatellite-stable colorectal cancer has, or is selected based on, at least one of the following attributes: (a) has metastatic CRC; (b) is not a candidate for definitive surgery or definitive radiation; (c) may have active liver and / or peritoneal metastases at the time of screening; (d) has microsatellite stability documented by a pathology report; (e) has received at least one line of therapy in the recurrent / metastatic setting, where the therapy includes anti-EGFR or anti-VEGF therapy; and / or (f) is anti-PD-1 / PD-L1 naive, defined as not having received treatment with an agent that targets PD-1.

[0136] In some embodiments, the method includes selecting a subject with microsatellite-stable colorectal cancer (MSS CRC). In some embodiments, the subject with microsatellite-stable colorectal cancer has, or is selected based on, at least one of the following attributes: (a) has metastatic CRC; (b) is not a candidate for definitive surgery or definitive radiation; (c) has no active metastases identified in the liver or peritoneum at screening; (d) has disease sites limited to the lung(s) and / or lung lymph nodes; (e) has microsatellite stability documented by a pathology report; (f) has received at least one line of therapy in the setting of recurrence / metastasis, where the therapy includes anti-EGFR therapy or anti-VEGF therapy; and / or (g) is anti-PD-1 / PD-L1 naive, defined as having never received treatment with an agent that targets PD-1.

[0137] In some embodiments, the method includes selecting a subject with triple-negative breast cancer (TNBC). In some embodiments, the subject with TNBC has, or is selected based on, at least one of the following attributes: (a) has metastatic TNBC, (b) is not a candidate for definitive surgery or definitive radiation, (c) is not a candidate for anti-PD-1 or anti-PD-L1 therapy in an approved indication, or such therapy is not otherwise available to the subject, (d) has triple-negative cancer (ER- / PR- / Her2-) documented by a pathology report, and / or (e) is anti-PD-1 / PD-L1 naive, defined as never having been treated with an agent that targets PD-1.

[0138] In some embodiments, the method includes selecting a subject with cutaneous squamous cell carcinoma (CSCC). In one embodiment, the subject is not a candidate for definitive surgery or definitive radiation. In one embodiment, the subject is anti-PD-1 / PD-L1 naive, defined as having never been treated with an agent that targets PD-1.

[0139] In some embodiments, the method includes selecting a subject with non-small cell lung cancer (NSCLC). In some embodiments, the subject has, or is selected based on, at least one of the following attributes: (a) has advanced or metastatic NSCLC, (b) is not a candidate for definitive surgery or definitive radiation, (c) does not have a targetable molecular alteration (e.g., ALK, ROS1, EGFR, etc.), (d) has not received systemic treatment for recurrent or metastatic NSCLC, and / or (e) is anti-PD-1 / PD-L1 naive, defined as not having received treatment with an agent that targets PD-1.

[0140] In some embodiments, the method comprises selecting a subject with previously histologically documented or cytologically documented locally advanced or metastatic EGFR-mutated non-squamous NSCLC disease. In some embodiments, the subject has, or is selected based on, at least one of the following attributes: (a) advanced or metastatic NSCLC, (b) is not a candidate for definitive surgery or definitive radiation, (c) has a previously documented targetable EGFR mutation (EGFR exon 19 deletion, EGFR L858R mutation, EGFR exon 20 insertion, or exon 18 / 21 atypical mutation), (d) is chemotherapy-naive, (e) has been treated with a third-generation TKI, and / or (e) is anti-PD-1 / PD-L1 naive, defined as having never been treated with an agent that targets PD-1.

[0141] In some embodiments, the method comprises selecting a subject with previously histologically documented or cytologically documented locally advanced or metastatic EGFR-mutated non-squamous NSCLC disease. In some embodiments, the subject has, or is selected based on, at least one of the following attributes: (a) advanced or metastatic NSCLC, (b) is not a candidate for definitive surgery or definitive radiation, (c) has a previously documented targetable EGFR mutation (EGFR exon 19 deletion, EGFR L858R mutation, EGFR exon 20 insertion, or exon 18 / 21 atypical mutation), (d) has been treated with platinum-doublet chemotherapy, (e) has been treated with a third-generation TKI, and / or (f) is anti-PD-1 / PD-L1 naive, defined as not having been treated with an agent that targets PD-1.

[0142] In some embodiments, the method includes selecting a patient with head and neck squamous cell carcinoma (HNSCC). In some embodiments, the subject has or is selected based on at least one of the following attributes: (a) has advanced or metastatic disease, (b) is not a candidate for definitive surgery or definitive radiation, (c) has PD-L1 expression of CPS ≥ 1% by local IHC assay, (d) has not received prior systemic treatment for recurrent or metastatic HNSCC, and / or (e) is anti-PD-1 / PD-L1 naive, defined as not having received treatment with an agent that targets PD-1.

[0143] (IV) Adverse events In some embodiments, the subject develops one or more mild symptoms of an immune-related adverse event after administration of the bispecific EGFRxCD28 antibody alone or in combination with an anti-PD-1 antibody. In some embodiments, the one or more symptoms of the immune-related adverse event are colitis, diarrhea, hypothyroidism, hyperthyroidism, hypophysitis, adrenal insufficiency, diabetes, hepatitis, neurotoxicity, pneumonitis, renal events, uveitis, myocarditis, pericarditis, or a combination thereof.

[0144] In some embodiments, the subject receives one or more additional therapies to treat one or more symptoms of the mild immune-related adverse event.

[0145] In some embodiments, treatment with the bispecific EGFRxCD28 antibody alone or in combination with an anti-PD-1 antibody is paused when the subject develops one or more mild symptoms of an immune-related adverse event and resumed when one or more symptoms have resolved.

[0146] Multispecific EGFRxCD28 antigen binding molecule The present disclosure provides methods of using antigen binding proteins that are multispecific (e.g., bispecific) and bind to at least EGFR and CD28 in combination with an anti-PD-1 antibody or antigen-binding portion thereof, such as cemiplimab, to treat cancer. As used herein, such multispecific antigen binding proteins are referred to in the format AxB, where A refers to the binding arm of the multispecific molecule that binds EGFR and B refers to the binding arm of the multispecific molecule that binds CD28, or vice versa. EGFRxCD28 or CD28xEGFR refers to a multispecific antigen binding protein that binds EGFR and CD28. Particular EGFR and CD28 binding arms of a multispecific antigen binding protein may also be designated in the format AxB, where A refers to a particular arm and B refers to another particular arm. For example, 085Nx14226P2 refers to a multispecific antigen binding protein having an anti-EGFR binding arm of 085N as shown herein and an anti-CD28 binding arm of 14426P2 as shown herein. For example, 085N is a binding arm comprising the 085N immunoglobulin heavy and light chains or variable regions thereof, or CDRs whose sequences are specifically set forth herein or are variants thereof.

[0147] Multispecific binding refers to binding to two or more different epitopes (EGFR and CD28 or more), which can be on the same antigen or different antigens. Multispecificity includes bispecificity, trispecificity, and tetraspecificity. An antibody or fragment thereof can be operatively linked (e.g., by chemical bonding, genetic fusion, noncovalent bonding, or other methods) 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.

[0148] In certain embodiments, a multispecific antigen-binding protein comprises a bispecific antigen-binding protein. As used herein, the term "bispecific antigen-binding protein" refers to a protein, polypeptide, or molecular complex (e.g., an antibody or antigen-binding fragment thereof) comprising at least a first antigen-binding domain and a second antigen-binding domain. Each antigen-binding domain in a bispecific antigen-binding molecule comprises at least one CDR that specifically binds to a particular antigen, either alone or in combination with one or more additional 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.

[0149] The present disclosure includes methods comprising administering any 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 of the EGFR HCVR arms in Tables 1 and 8 (e.g., the HCVR arms of parental monoclonal antibodies mAb12999P2, mAb13008P2, mAb35193P2, and mAb13006P2) with any of the CD28 HCVR arms in Tables 3 and 8 (e.g., the HCVR arms of parental mAb14226, mAb14193, and mAb14216), and methods of use thereof to treat cancer as described herein.

[0150] As used herein, the term "antigen-binding molecule" refers to a protein, polypeptide, or molecular complex comprising or consisting of at least one complementarity-determining region (CDR), alone or in combination with one or more additional CDRs and / or framework regions (FRs), that specifically binds to a particular antigen. In certain embodiments, the antigen-binding molecule is an antibody or an antibody fragment, as defined elsewhere herein.

[0151] As used herein, the term "bispecific antigen-binding molecule" refers to a protein, polypeptide, or molecular complex (e.g., an antibody or antigen-binding fragment thereof) comprising at least a first antigen-binding domain and a second antigen-binding domain. Each antigen-binding domain in a bispecific antigen-binding molecule comprises at least one CDR that specifically binds to a particular antigen, either alone or in combination with one or more additional 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 second, different 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 a bispecific antibody comprises a heavy chain variable domain (HCVR) and a light chain variable domain (LCVR).

[0152] The first antigen-binding domain and the second antigen-binding domain can be directly or indirectly connected to each other to form a bispecific antigen-binding molecule of the present disclosure. Alternatively, the first antigen-binding domain and the second antigen-binding domain can each be connected to a separate multimerization domain. The association of one multimerization domain with another multimerization domain promotes the association between the two antigen-binding domains, thereby forming a bispecific antigen-binding molecule. As used herein, a "multimerization domain" is any macromolecule, protein, polypeptide, peptide, or amino acid that has the ability to bind to a second multimerization domain of the same or similar structure or composition. For example, a multimerization domain can be connected to a second multimerization domain of the same or similar structure or composition. H A non-limiting example of a multimerizing component is a polypeptide comprising the Fc portion of an immunoglobulin (C H 2~C H 3 domains), for example, the Fc domain of IgG selected from the isotypes IgG1, IgG2, IgG3, and IgG4, as well as any allotype within each isotype group.

[0153] A bispecific antigen-binding molecule of the present disclosure typically comprises two multimerization domains, e.g., two Fc domains, each part of a separate antibody heavy chain. The first and second multimerization domains may be of the same IgG isotype, e.g., IgG1 / IgG1, IgG2 / IgG2, or IgG4 / IgG4. Alternatively, the first and second multimerization domains may be of different IgG isotypes, e.g., IgG1 / IgG2, IgG1 / IgG4, or IgG2 / IgG4.

[0154] In certain embodiments, the multimerization domain is an Fc fragment or an amino acid sequence of 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 short cysteine-containing peptide. Other multimerization domains include peptides or polypeptides comprising or consisting of a leucine zipper, a helix loop motif, or a coiled-coil motif.

[0155] Any bispecific antibody format or technology can be used to generate the bispecific antigen-binding molecules of the present disclosure. For example, an antibody or antigen-binding fragment thereof having a first antigen-binding specificity can be operably linked (e.g., by chemical conjugation, genetic fusion, non-covalent bonding, or other methods) to one or more other molecular entities, such as another antibody or antibody fragment having a second antigen-binding specificity, to generate the 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, scFv-based or diabody bispecific formats, IgG-scFv fusions, dual variable domain (OVO)-Ig, quadroma, knobs-into-holes, common light chains (e.g., common light chains with knobs-into-holes), CrossMab, CrossFab, (SEEO)body, leucine zipper, Ouobody, IgG1 / IgG2, dual acting Fab (OAF)-IgG, and Mab. 2Bispecific formats are included (for a review of the aforementioned formats, see, e.g., Klein et al. 2012, mAbs 4:6, 1-11 and references cited therein).

[0156] In the context of the bispecific antigen-binding molecules of the present disclosure, the multimerization domain, e.g., the Fc domain, may comprise one or more amino acid changes (e.g., insertions, deletions, or substitutions) compared to a wild-type, naturally occurring Fc domain. For example, the present disclosure includes bispecific antigen-binding molecules comprising one or more modifications in the Fc domain that result in a modification of the Fc domain that alters (e.g., enhances or decreases) the binding interaction between Fc and FcRn. In one embodiment, the bispecific antigen-binding molecule comprises a C H 2 or C H The FcRn-binding domain contains modifications in three regions that increase the affinity of the Fc domain for FcRn in an acidic environment (e.g., within an endosome at a pH of about 5.5 to about 6.0). Non-limiting examples of such Fc modifications include, for example, modifications at positions 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 positions 428 and / or 433 (e.g., UR / S / P / Q or K) and / or 434 (e.g., H / F or V), or modifications at positions 250 and / or 428, or modifications at positions 307 or 308 (e.g., 308F, V308F), and 434. In one embodiment, the modifications include the 428L (e.g., M428L) and 434S (e.g., N434S) modifications, the 428L, 2591 (e.g., V2591), and 308F (e.g., V308F) modifications, the 433K (e.g., H433K) and 434 (e.g., 434Y) modifications, the 252, 254, and 256 (e.g., 252Y, 254T, and 256E) modifications, the 250Q and 428L (e.g., T250Q and M428L) modifications, and the 307 and / or 308 (e.g., 308F or 308P) modifications.

[0157] The present disclosure also provides a first CH 3 domain and second Ig C H a bispecific antigen-binding molecule comprising three domains, a first and a second Ig C H The three domains differ from each other by at least one amino acid, and the at least one amino acid difference reduces binding of the bispecific antibody to Protein A compared to a bispecific antibody lacking the amino acid difference. H The 3 domain binds to protein A and the second Ig C H The 3 domain contains a mutation that reduces or abolishes binding to Protein A, such as the H95R modification (according to IMGT exon numbering, H435R according to EU numbering). H 3 may further comprise a Y96F modification (by IMGT, Y436F by EU). Additional modifications that may be found within the second CH3 include D16E, L18M, N44S, K52N, V57M, and V821 for IgG1 antibodies (D356E, L358M, N384S, K392N, V397M and V4221 by EU by IMGT), N44S, K52N and V821 for IgG2 antibodies (N384S, K392N and V4221 by IMGT, EU), and Q15R, N44S, K52N, V57M, R69K, E79Q and V821 for IgG4 antibodies (Q355R, N384S, K392N, V397M, R409K, E419Q and V4221 by EU by IMGT).

[0158] In certain embodiments, the Fc domain can be a chimera that combines Fc sequences from two or more immunoglobulin isotypes. For example, a chimeric Fc domain can be a chimeric Fc domain that combines the Fc sequences of human IgG1, human IgG2, or human IgG4. H C derived from 2 regions H Part or all of the 2 sequences, and C derived from human IgG1, human IgG2, or human IgG4 HThe chimeric Fc domain may comprise some or all of the three sequences. The chimeric Fc domain may also comprise a chimeric hinge region. For example, the chimeric hinge may comprise an "upper hinge" sequence derived from the hinge region of human IgG1, human IgG2, or human IgG4, combined with a "lower hinge" sequence derived from the hinge region of human IgG1, human IgG2, or human IgG4. A particular example of a chimeric Fc domain that can be included in any of the antigen-binding molecules described herein is one that comprises, from the N-terminus to the C-terminus, [IgG4 C H 1]-[IgG4 upper hinge]-[IgG2 lower hinge]-[IgG4 CH2]-[IgG4 C H Another example of a chimeric Fc domain that can be included in any of the antigen-binding molecules described herein includes, from the N-terminus to the C-terminus, [IgG1 C H 1]-[IgG1 upper hinge]-[IgG2 lower hinge]-[IgG4 C H 2]-[IgG1 C H These and other examples of chimeric Fc domains that can be included in any of the antigen-binding molecules of the present disclosure are described in WO2014 / 022540A1, and chimeric Fc domains having these general structural arrangements and variants thereof can alter Fc receptor binding and, consequently, affect Fc effector function.

[0159] The antibodies and antigen-binding fragments of the present disclosure include immunoglobulin chains comprising the amino acid sequences (and variants thereof) specifically set forth herein, as well as cellular and in vitro post-translational modifications to the antibodies or fragments. For example, the present disclosure includes antibodies and antigen-binding fragments thereof that specifically bind to EGFR and CD28, comprising the heavy and / or light chain amino acid sequences set forth herein, as well as 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 (e.g., 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 deleted.

[0160] The present disclosure also provides antigen binding proteins, such as antibodies (e.g., human antibodies, monoclonal antibodies, and recombinant antibodies) and antigen-binding fragments thereof, that specifically bind to the CD28 protein or antigenic fragments thereof (e.g., the extracellular domain of CD28). Antigen binding proteins that bind to the same epitope on CD28 as any of the antigen binding proteins provided herein or that compete with any of the antigen binding proteins described herein for binding to CD28 are also part of the present disclosure.

[0161] The multispecific EGFRxCD28 antigen binding proteins of the present disclosure bind to CD28 on the surface of T cells and promote CD28 signaling to promote T cell activation and / or proliferation, sometimes referred to herein as "costimulatory" or "costimulatory." T cell activation is initiated when the T cell receptor (TCR) / CD3 complex binds to a peptide-MHC complex ("signal 1"); activation is then enhanced by the engagement of a second "costimulatory" receptor, such as the CD28 receptor on the T cell, which binds to its cognate ligand(s) on the target cell ("signal 2"). For example, T cell activation by a CD28 bispecific antibody can be triggered by an amplified signal in response to recognition of an endogenous tumor antigen by the TCR / CD3 complex, or in response to activation of "signal 1" via CD3 bispecificity.

[0162] (I) Antibodies containing Fc variants According to certain embodiments of the present disclosure, there are provided anti-EGFR X anti-CD28 bispecific antigen binding molecules comprising an Fc domain comprising one or more mutations that enhance or decrease binding of the antibody to the FcRn receptor at, for example, acidic pH compared to neutral pH. For example, the present disclosure includes an Fc domain comprising a C H 2 or C HThe antibodies and antigen-binding molecules include those containing mutations in the three regions, which mutation(s) increase the affinity of the Fc domain for FcRn in acidic environments (e.g., within endosomes where the pH ranges from about 5.5 to about 6.0). Such mutations may result in an increase in the serum half-life of the antibody when administered to an animal. Non-limiting examples of such Fc modifications include, for example, modifications at positions 250 (e.g., E or Q), 250 and 428 (e.g., L or F), 252 (e.g., L / Y / F / W or T), 254 (e.g., S or T), and / or 256 (e.g., S / R / Q / E / D or T), or modifications at positions 428 and / or 433 (e.g., H / L / R / S / P / Q or K) and / or 434 (e.g., H / F or Y), or modifications at positions 250 and / or 428, or modifications at positions 307 or 308 (e.g., 308F, V308F), and / or 434.

[0163] In one embodiment, the modification is 428L (e.g., M428L) and 434S (e.g., N434S) modifications, 428L, 259I (e.g., V259I), and 308F (e.g., V308F) modifications, 433K (e.g., H433K) and 434 (e.g., 434Y) modifications, modifications of 252, 254 and 256 (e.g., 252Y, 254T and 256E), 250Q and 428L (e.g., T250Q and M428L) modifications, and / or - containing modifications of 307 and / or 308 (e.g., 308F or 308P).

[0164] For example, the disclosure includes an EGFRxCD28 bispecific antigen binding molecule comprising an Fc domain comprising one or more pairs or groups of mutations selected from the group consisting of: 250Q and 248L (e.g., T250Q and M248L), 252Y, 254T and 256E (e.g., M252Y, S254T and T256E), 428L and 434S (e.g., M428L and N434S), and 433K and 434F (e.g., H433K and N434F).

[0165] All possible combinations of the aforementioned Fc domain mutations, and other mutations in the antibody variable domains disclosed herein, are considered within the scope of this disclosure.

[0166] Anti-PD-1 antibodies and their antigen-binding fragments According to certain exemplary embodiments of the present disclosure, the method comprises administering a therapeutically effective amount of an anti-PD-1 antibody or antigen-binding fragment thereof in combination with a bispecific EGFRxCD28 antigen-binding molecule or antigen-binding fragment thereof. According to certain embodiments, the anti-PD-1 antibody or 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 has a K of 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 by surface plasmon resonance assay. D However, an isolated antibody or antigen-binding fragment that specifically binds human PD-1 may have cross-reactivity to other antigens, such as PD-1 molecules from other (non-human) species.

[0167] According to certain exemplary embodiments of the present disclosure, an anti-PD-1 antibody, or antigen-binding fragment thereof, comprises a heavy chain variable region (HCVR), a light chain variable region (LCVR), and / or a complementarity-determining region (CDR) comprising 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 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 the heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO:73, and a light chain complementarity-determining region (LCDR) of the light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:74. According to certain 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 (AAS), and LCDR3 comprises the amino acid sequence of SEQ ID NO: 80. In yet other embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof comprises a HCVR comprising SEQ ID NO: 73 and a LCVR comprising SEQ ID NO: 74. In still other embodiments, the methods of the disclosure comprise the use of an anti-PD-1 antibody, wherein the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 81. In some embodiments, the anti-PD-1 antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 82. An exemplary antibody comprising an HCVR comprising the amino acid sequence of SEQ ID NO:73 and an LCVR comprising the amino acid sequence of SEQ ID NO:74 is the fully human anti-PD-1 antibody known as cemiplimab (REGN2810, LIBTAYO®). According to certain exemplary embodiments, the methods of the disclosure comprise the use of cemiplimab or a bioequivalent thereof. As used herein, the term "bioequivalent" refers to an anti-PD-1 antibody or PD-1 binding protein or fragment thereof that is a pharmaceutical equivalent or pharmaceutical substitute whose rate and / or extent of absorption is not significantly different from that of cemiplimab when administered at the same molar dose under similar experimental conditions, either in a single dose or multiple doses.In the context of the present disclosure, this term refers to an antigen-binding protein that binds to PD-1 that has no clinically meaningful differences in safety, purity, and / or efficacy from cemiplimab.

[0168] Other anti-PD-1 antibodies that can be used in the context of the disclosed methods include, for example, nivolumab (US8008449), pembrolizumab (US8354509), MEDI0608 (US8609089), BI754091, spartalizumab (also known as PDR001), camrelizumab (also known as SHR-1210), JNJ-63723283, MCLA-134, toripalimab, sintilimab, tislelizumab, selplulimab, dostallimab, retifanlimab, zimberelimab, penprimimab, pidelimab, and pembrolizumab. Antibodies that can be used in the treatment of PD-1 include antibodies referred to and known in the art as tibiofibrillab, HX008, balstilimab, ezabenlimab, 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 and WO2009 / 114335, or antigen-binding fragments of any of the foregoing.

[0169] bioequivalence The present disclosure encompasses methods comprising administering antigen-binding molecules that have amino acid sequences different from those of the described antibodies but retain the ability to bind to CD28 and EGFR or PD-1. Such variant molecules contain one or more additions, deletions, or substitutions of amino acids compared to the parent sequence, but exhibit essentially the same biological activity as the described antigen-binding molecules. Similarly, DNA sequences encoding the antigen-binding molecules of the present disclosure encompass sequences that contain one or more additions, deletions, or substitutions of nucleotides compared to the disclosed sequences, but encode antigen-binding molecules that are essentially biologically equivalent to the described antigen-binding molecules of the present disclosure. Examples of such variant amino acid sequences and DNA sequences are discussed above.

[0170] The present disclosure includes methods comprising administering an antigen-binding molecule that is bioequivalent to any of the exemplary antigen-binding molecules described herein. Two antigen-binding proteins or antibodies are considered to be bioequivalent if, for example, they are pharmaceutical equivalents or pharmaceutical substitutes that do not exhibit significant differences in absorption rate and extent when administered at the same molar dose, either in single or multiple doses, under similar experimental conditions. Some antibodies are considered equivalents or pharmaceutical substitutes if their extent of absorption is comparable but their absorption rate is not; however, such differences in absorption rate may be considered bioequivalent because they are intentional, reflected in the labeling, and are not considered to be essential, for example, for achieving effective body drug concentrations during chronic use and are not considered medically significant for the particular pharmaceutical agent being studied.

[0171] In one embodiment, two antigen binding proteins are bioequivalent if there are no clinically significant differences in their safety, purity, and efficacy.

[0172] In one embodiment, two antigen binding proteins are bioequivalent if a subject can be switched between the reference product and the biological product one or more times without an expected increased risk of adverse effects, including clinically significant changes in immunogenicity or reduced efficacy, compared to continued treatment without switching.

[0173] In one embodiment, two antigen binding proteins are bioequivalent if they both operate by a common mechanism or mechanism of action for the condition(s) of use, to the extent that such mechanism is known.

[0174] Bioequivalence may be demonstrated by in vivo and / or in vitro methods. Measures of bioequivalence include, for example, (a) in vivo tests in humans or other mammals that measure the concentration of an antibody or its metabolites as a function of time in blood, plasma, serum, or other biological fluids, (b) in vitro tests that correlate with and are reasonably predictive of in vivo bioavailability data in humans, (c) in vivo tests in humans or other mammals that measure the relevant acute pharmacological effects of the antibody (or its target) as a function of time, and (d) well-controlled clinical trials that establish the safety, efficacy, or bioavailability or bioequivalence of the antibody.

[0175] Biologically equivalent 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 required 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 unnecessary or incorrect intramolecular disulfide bridge formation during renaturation. In other contexts, biologically equivalent antibodies can include exemplary bispecific antigen-binding molecules described herein that contain amino acid changes that modify the glycosylation characteristics of the antibody, for example, mutations that eliminate or remove glycosylation.

[0176] Pharmaceutical preparations and kits The present invention provides compositions comprising a bispecific EGFRxCD28 antibody, such as REGN7075, and / or an anti-PD-1 antibody or antigen-binding portion thereof, and one or more components, and methods of use thereof.

[0177] The present disclosure provides pharmaceutical compositions comprising the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof and / or 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 effect transportation, delivery, tolerance, etc. Many suitable formulations can be found in formularies known to all pharmaceutical chemists (e.g., Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, PA). These formulations include, for example, powders, pastes, ointments, jellies, waxes, oils, lipids, lipid (cationic or anionic)-containing vesicles (e.g., 11POFECTIN™, Life Technologies, Carlsbad, CA), DNA conjugates, anhydrous absorbent pastes, oil-in-water and water-in-oil 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.

[0178] To prepare pharmaceutical formulations of antigen binding proteins, such as antibodies and antigen-binding fragments thereof (e.g., REGN7075, REGN6321, REGN6322, REGN6323, or cemiplimab), the antigen binding protein is mixed with a pharmaceutically acceptable carrier or excipient. See, e.g., Remington's Pharmaceutical Sciences and US 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, NY; Gennaro (2000) Remington: The Science and Practice of Pharmacy, Lippincott, Williams, and Wilkins, New York, NY; Avis, et al. (eds.) (1993) Pharmaceutical Dosage Forms: Parenteral Medications, Marcel See 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, NY. In one embodiment, the pharmaceutical formulation is sterile. Such compositions are part of the present disclosure.

[0179] Pharmaceutical formulations of the disclosure include a bispecific EGFRxCD28 antigen-binding protein, and / or an anti-PD-1 antibody or antigen-binding fragment thereof, and a pharmaceutically acceptable carrier comprising, for example, water, a buffer, a preservative, and / or a detergent.

[0180] The scope of the present disclosure includes dried, e.g., lyophilized, compositions comprising 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.

[0181] Various delivery systems are well known and can be used to administer the pharmaceutical compositions of the present disclosure, such as encapsulation in liposomes, microparticles, microcapsules, recombinant cells capable of expressing mutant viruses, and receptor-mediated endocytosis (see, e.g., Wu et al., 1987, J. Biol. Chem. 262:4429-4432). Methods of introduction 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, for example, by infusion or bolus injection, or by absorption through epithelial or mucocutaneous linings (e.g., oral, rectal, and intestinal mucosa), and can be administered together with other biologically active agents. Administration can be systemic or local.

[0182] As discussed herein, the present disclosure provides a container (e.g., a plastic or glass vial) or injection device (e.g., a syringe, pre-filled syringe, or auto-injector) containing any of the antigen-binding proteins herein, e.g., antibodies or antigen-binding fragments thereof, or a pharmaceutical formulation comprising a pharmaceutically acceptable carrier or excipient thereof.

[0183] The pharmaceutical composition of the present disclosure can be delivered subcutaneously or intravenously using a standard needle and syringe. In addition, for subcutaneous delivery, a pen delivery device is easily used to deliver the pharmaceutical composition of the present disclosure. Such a pen delivery device can be reusable or disposable. Reusable pen delivery devices generally utilize a replaceable cartridge containing the pharmaceutical composition. Once all of the pharmaceutical composition in the cartridge has been administered and the cartridge is empty, the empty cartridge can be easily discarded and replaced with a new cartridge containing the pharmaceutical composition. The pen delivery device can then be reused. In disposable pen delivery devices, there is no replaceable cartridge. Rather, disposable pen delivery devices are pre-filled with the pharmaceutical composition held in a reservoir within the device. Once the reservoir is emptied of the pharmaceutical composition, the entire device is discarded.

[0184] Numerous reusable and disposable pen and autoinjector delivery devices have application in the subcutaneous delivery of the pharmaceutical compositions of the present disclosure, see, for example, AUTOPEN™ (Owen Mumford, Inc., Woodstock, UK) or HUMIRA™ Pen (Abbott Labs, Abbott Park, IL).

[0185] In certain circumstances, pharmaceutical compositions can be delivered in a controlled release system. In one embodiment, a pump can be used (see Langer, supra; Sefton, 1987, CRC Crit. Ref. Biomed. Eng. 14:201). In another embodiment, a polymeric material can be used; see Medical Applications of Controlled Release, Langer and Wise (eds.), 1974, CRC Pres., Boca Raton, Florida. In yet another embodiment, a controlled release system can be placed near the target of the composition, thus requiring only a fraction of the systemic dose (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.

[0186] Injectable preparations may include dosage forms for intravenous injection, subcutaneous injection, intradermal injection, intramuscular injection, drip infusion, etc. These injectable preparations may be prepared by publicly known methods. For example, injectable preparations may be prepared by dissolving, suspending, or emulsifying the above-mentioned antibody or a salt thereof in a sterile aqueous or oily medium conventionally used for injections. Aqueous media for injection include, for example, physiological saline and other isotonic solutions, which may be used in combination with an appropriate solubilizer. Injectable oily media are also part of the present disclosure. Such oily media may be combined with a solubilizer.

[0187] Advantageously, the above-described pharmaceutical compositions for oral or parenteral use are prepared into a suitable unit dosage form to fit the dose of the active ingredient. Such unit dosage forms include, for example, tablets, pills, capsules, injections (ampoules), suppositories, etc. The amount of the antibody contained therein is generally about 0.1 to about 2000 mg per unit dosage form, particularly in the form of an injection.

[0188] The present disclosure also provides kits containing a bispecific EGFRxCD28 antibody and an anti-PD-1 antibody, or antigen-binding portion thereof, for therapeutic use. The kit typically includes a label indicating the intended use and instructions for use of the kit contents. The term label includes any written or recorded material supplied on or with the kit, or that otherwise accompanies the kit. Thus, the present disclosure provides kits for treating a subject suffering from cancer, the kit including (a) a dosage of an antibody or antigen-binding portion thereof, (b) a dosage of a bispecific EGFRxCD28 antibody or antigen-binding portion thereof, and (c) instructions for using the antibody in any of the therapeutic methods disclosed herein. In certain embodiments, the dosage of the anti-PD-1 antibody or antigen-binding fragment thereof is in the range of 150 to 550 mg, e.g., 350 mg. In certain embodiments, the dosage of the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof is in the range of 0.1 mg to 3000 mg.

[0189] Additional combination therapy The present disclosure provides methods for treating and preventing diseases, e.g., cancer, by administering a bispecific EGFRxCD28 antibody, e.g., REGN7075 or an antigen-binding fragment thereof, alone as monotherapy, or together with an anti-PD-1 antibody or antigen-binding fragment thereof as combination therapy, optionally with one or more therapeutic agent(s), e.g., at least a third therapeutic agent or therapy.

[0190] In certain embodiments, the third therapeutic agent or therapy is selected from surgery, radiation, chemotherapy (e.g., an anti-cancer chemotherapy, e.g., paclitaxel, docetaxel, vincristine, cisplatin, carboplatin, or oxaliplatin), CAR-T cell therapy, a cancer vaccine, an oncolytic virus, a cytokine, an anti-VEGF therapy, an anti-EGFR therapy, or an anti-cancer agent. As used herein, "anti-cancer agent" means any agent that is useful for treating cancer, and includes, but is not limited to, cytotoxins and agents such as antimetabolites, alkylating agents, anthracyclines, antibiotics, antimitotic agents, procarbazine, hydroxyurea, asparaginase, corticosteroids, mitotane (O,P'-(DDD)), biologics (e.g., antibodies and interferons), and radioactive agents. As used herein, "cytotoxin or cytotoxic agent," which also refers to chemotherapeutic agents, means any agent that is detrimental to cells. Examples include Taxol® (paclitaxel), temozolamide, cytochalasin B, gramicidin D, ethidium bromide, emetine, cisplatin, mitomycin, etoposide, tenoposide, vincristine, vinbuastine, colchicine, doxorubicin, daunorubicin, dihydroxyanthracin dione, mitoxantrone, mithramycin, actinomycin D, 1-dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine, propranolol, and puromycin, as well as analogs or homologs thereof. In certain embodiments, the method comprises administering an agent to reduce, ameliorate, or treat a symptom of an immune-related adverse event. In one embodiment, the agent is selected from an IL-6 inhibitor (e.g., an anti-IL-6 receptor antibody such as tocilizumab or sarilumab), a corticosteroid, or a nonsteroidal anti-inflammatory agent.

[0191] In a further embodiment, the additional therapeutic agent administered to the subject in combination with the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof and the anti-PD-1 antibody or antigen-binding fragment thereof is selected from the group consisting of PD-1 antibodies, anti- ... thedition (November 1, 2002)), or is usually administered to a subject according to the approved prescribing information provided with the particular drug.

[0192] The term "in combination with" indicates that a component, such as a bispecific EGFRxCD28 antibody or antigen-binding fragment thereof, e.g., REGN7075, an anti-PD-1 antibody or antigen-binding fragment thereof, e.g., cemiplimab, along with an additional therapeutic agent, can be formulated in a single composition, e.g., for simultaneous delivery, or can be formulated separately in two or more compositions (e.g., a kit containing each component). Alternatively, components administered "in combination with" each other can be administered to a subject at a different time than the other components are administered; e.g., each administration may be spaced apart (e.g., separately or sequentially) over a predetermined period of time as part of a treatment regimen, rather than simultaneously. Separate components administered in combination with each other may be administered sequentially, albeit essentially simultaneously, during the same administration session. Furthermore, separate components administered in combination with each other may be administered to a subject by the same route or by different routes.

[0193] 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, or subsequent to, 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, or prior to, 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, or subsequent to, the bispecific EGFRxCD28 antibody, but on a separate day from the anti-PD-1 antibody. In some embodiments, the third therapeutic agent is administered on the same day as, or prior to, the bispecific EGFRxCD28 antibody, but on a separate day from the anti-PD-1 antibody. In some embodiments, the third therapeutic agent is administered on the same day as, or subsequent to, the anti-PD-1 antibody, but on a separate day from the bispecific EGFRxCD28 antibody. In some embodiments, the third therapeutic agent is administered on the same day as, or before, the anti-PD-1 antibody, but on a different day than, the bispecific EGFRxCD28 antibody.

[0194] In some embodiments, the third therapeutic agent is administered on a different day from the bispecific EGFRxCD28 antibody and / or the anti-PD-1 antibody. In some embodiments, the third therapeutic agent is administered on a different day after the bispecific EGFRxCD28 antibody and / or the anti-PD-1 antibody. In some embodiments, the third therapeutic agent is administered on a different day before the bispecific EGFRxCD28 antibody and / or the anti-PD-1 antibody. In some embodiments, the third therapeutic agent is administered on a different day after the bispecific EGFRxCD28 antibody, but on a different day from the anti-PD-1 antibody. In some embodiments, the third therapeutic agent is administered on a different day before the bispecific EGFRxCD28 antibody, but on a different day from the anti-PD-1 antibody.

[0195] In some embodiments, the combination therapy with a bispecific EGFRxCD28 antibody, e.g., REGN7075, and an anti-PD-1 antibody, e.g., cemiplimab, is administered alone for a lead-in period before initiating treatment with a third therapeutic agent.

[0196] In some embodiments, the combination therapy with the third therapeutic agent is administered at least two, three, four, five, six, seven, or more times.

[0197] In certain embodiments, the subject does not experience or suffers any side effects from the administration of the bispecific EGFRxCD28 antibody alone or in combination with a third therapeutic agent and / or the administration of the anti-PD-1 antibody alone or in combination with a third therapeutic agent. [Example]

[0198] The following examples are put forward so as to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the disclosed methods and compositions, and are not intended to limit the scope of what the inventors regard as the present disclosure.

[0199] Example 1: Bispecific EGFRxCD28 antibodies Generation of anti-EGFR and anti-CD28 antibodies The anti-EGFR antibody and anti-CD28 antibody can be obtained as described in WO2020 / 198009, the entire contents of which are expressly incorporated herein by reference.

[0200] Table 1 shows the amino acid sequence identifiers for the heavy and light chain variable regions and CDRs of selected anti-EGFR antibodies of the disclosure. The corresponding nucleic acid sequence identifiers are shown in Table 2. [Table 1] SEQ ID NO: 20: Gly Ala Ser (GAS) [Table 2] SEQ ID NO: 19: ggggcaagt

[0201] Table 3 shows the amino acid sequence identifiers for the heavy and light chain variable regions and CDRs of selected anti-CD28 antibodies of the disclosure. The corresponding nucleic acid sequence identifiers are shown in Table 4. [Table 3] [Table 4]

[0202] Generation of bispecific antibodies (bsAbs) that bind to CD28 and EGFR Bispecific antibodies comprising an anti-EGFR-specific binding domain and an anti-CD28-specific binding domain were constructed using standard methodologies, with the anti-EGFR antigen-binding domain and the anti-CD28 antigen-binding domain each comprising a distinct and separate HCVR paired with a common LCVR. In some examples, bispecific antibodies were constructed utilizing an anti-CD28 antibody heavy chain, an anti-EGFR antibody heavy chain, and a common light chain comprising the components, amino acid sequences, and nucleic acid sequences encoding the antibodies, as shown in Tables 5, 6, 7, and 8 below. Additional bispecific antibodies that bind to EGFR and CD28 can be prepared using the parent monoclonal antibodies with the designations shown in Tables 9A, 9B, and 9C. [Table 5] [Table 6] [Table 7] [Table 8]

[0203] Additional bispecific antibodies comprising one HCVR arm from a parent EGFR antibody and another HCVR arm from a parent CD28 antibody can be generated using the techniques described herein. The parent EGFR antibody used to generate these additional anti-EGFR x anti-CD28 bispecific antibodies has the HCVR sequence described in WO 2014 / 004427. The CD28 parent antibody used to generate these additional anti-EGFR x anti-CD28 bispecific antibodies has the amino acid sequence described in Table 3 above. These anti-EGFR and anti-CD28 binding domains (pairings) are shown below in Tables 9A, 9B, and 9C. [Table 9A] [Table 9B] [Table 9C]

[0204] Exemplary bispecific antibodies were produced with modified (chimeric) IgG4 Fc domains as described in U.S. Patent Application Publication No. US20140243504A1, published August 28, 2014.

[0205] The bispecific antibody described in this example comprises 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 pairing of CD28-VH / EGFR-VH creates an antigen-binding domain that specifically recognizes CD28 on T cells and EGFR on tumor cells.

[0206] Example 2: Evaluation of treatment with REGN7075 alone and in combination with cemiplimab This example describes a Phase 1 clinical trial of REGN7075 (an EGFRxCD28 costimulatory bispecific antibody) in combination with cemipimab in subjects with advanced solid tumors.

[0207] Research purpose The primary objective of dose escalation is to evaluate the safety and tolerability of REGN7075 monotherapy induction and the combination of REGN7075 with cemiplimab in subjects with advanced solid tumors.

[0208] The primary objective of the dose expansion is to evaluate the preliminary efficacy of REGN7075 in combination with cemiplimab, as measured by 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), within selected advanced solid tumor-specific cohorts.

[0209] Secondary objectives of the dose escalation study are: (i) to characterize the pharmacokinetics (PK) of REGN7075 alone and in combination with cemiplimab; (ii) to evaluate the preliminary efficacy of the combination of REGN7075 and cemiplimab as measured by 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 composite response criteria (depending on the subject's baseline evaluation criteria); and (iii) to evaluate the immunogenicity of REGN7075 and cemiplimab.

[0210] Secondary objectives of the dose expansion are (i) to evaluate the preliminary efficacy of the combination of REGN7075 and cemiplimab as measured by OS, PFS, DOR, CR rate, and DCR according to RECIST 1.1 and / or composite response criteria (according to the subject's baseline evaluation criteria) within selected advanced solid tumor-specific cohorts of subjects; (ii) to evaluate the safety and tolerability of the combination of REGN7075 and cemiplimab with or without chemotherapy; (iii) to characterize the PK of REGN7075 alone and in combination with cemiplimab with or without chemotherapy; (iv) to evaluate the immunogenicity of REGN7075 and cemiplimab with or without chemotherapy; and (v) to evaluate the efficacy and safety of the combination of REGN7075 and cemiplimab with or without chemotherapy using validated instruments: the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire (EORTC QLQ-C30), EORTC-QLQ-BR23 (among breast cancer patients only), EORTC-QLQ-CR29 (among CRC subjects only), and EORTC To assess the effects of REGN7075 on subject-reported outcomes, including health-related quality of life (HRQoL) measured by QLQ-LC13 (among NSCLC subjects only) and EORTC QLQ-HN35 (among HNSCC subjects only), and EQ-5D-5L.

[0211] Exploratory objectives include: (i) measuring circulating tumor DNA levels over time; (ii) investigating efficacy- and safety-related tumor genetic alterations detected in circulating tumor DNA (ctDNA) at baseline and post-dose; (iii) evaluating the efficacy of REGN7075 alone and in combination with cemiplimab (with or without chemotherapy) in patients with low and high tumor mutational burden (TMB) within a selected cohort of advanced solid tumors; and (iv) assessing EGFR and programmed cell death ligand (PD-L1) expression in tumor and immune cell populations by assessing protein and RNA in tumor tissue.

[0212] research design This is an open-label, Phase 1 / 2, first-in-human (FIH) study evaluating the safety, tolerability, PK, and preliminary antitumor activity of REGN7075 alone and in combination with cemiplimab in patients with advanced solid tumors.

[0213] There are two parts to this study:

[0214] Dose escalation During dose escalation, patients will receive a 3-week monotherapy induction of REGN7075 at their assigned dose level (DL) QW IV, followed by a combination of REGN7075 at their assigned DL and cemiplimab 350 mg Q3W IV. Concurrently initiated (i.e., no induction) dose levels of REGN7075 and cemiplimab 350 mg Q3W IV can be explored. Once the MTD / RP2D(s) for REGN7075 IV are identified and enrollment and DLT monitoring for the concurrently initiated dose escalation cohorts are complete (Figure 2B), enrollment into the exploratory SC cohorts can begin. SC REGN7075 in combination with either IV or SC cemiplimab can be explored.

[0215] Based on the new PK data, both induction and simultaneous dose-escalation cohorts can be enrolled with REGN7075 QW (Figures 2A and 2B). To manage infusion-related reactions (IRR), step-up dosing of REGN7075 will include a step-up dose in week 1 and a target dose in weeks 2 and beyond. The first step-up dose will be 30 mg. The dose and / or rate of this step-up dose may be adjusted. Additionally, there is an option to split the step-up dose and first target dose over 2 days.

[0216] Dose expansion The RP2D(s) will be determined based on the entire data. Dose expansion will be based on the RP2D(s). During dose expansion, patients will receive REGN7075 combination therapy at their assigned DL (e.g., RP2D(s)) and schedule. Dose escalation will consist of patients with a variety of mixed advanced solid tumors. Dose expansion will consist of the following tumor-specific expansion cohorts: Cohort A: Triple-negative breast cancer (TNBC) Cohort B: Cutaneous squamous cell carcinoma (CSCC) Cohort C: Non-small cell lung cancer (NSCLC) Cohort D: Head and neck squamous cell carcinoma (HNSCC) Cohort E: Microsatellite stable colorectal cancer (MSS-CRC) with active liver and / or peritoneal metastases Cohort F: MSS-CRC with isolated lung / lymph node metastasis (no active liver or peritoneal metastasis) Cohort G: EGFR-mutated NSCLC after third-generation TKIs Cohort H: EGFR-mutated NSCLC after third-generation TKI and platinum-doublet chemotherapy

[0217] Dose expansion cohorts will be enrolled after identifying the REGN7075 RP2D(s) in combination with cemiplimab. Patients in cohorts C and G will also receive four cycles of platinum-based chemotherapy. Additional patients may be enrolled at their estimated RP2D(s) to further evaluate safety and biological activity prior to enrollment in the expansion cohorts.

[0218] Patients will be treated with study drug until overt disease progression, intolerable adverse events (AEs), withdrawal of consent, or other study discontinuation criteria are met.

[0219] Treatment consists of a 3-week induction of REGN7075 at RP2D(s) QW or co-initiation in combination with cemiplimab 350mg Q3W IV. Patients in cohorts C and G will also receive 4 cycles of platinum-based chemotherapy administered Q3W on the same days as cemiplimab.

[0220] If a dose level for the concurrent initiation schedule (i.e., initiating REGN7075 and cemiplimab without a REGN7075 lead-in period) is not explored, or if a concurrent initiation dose level is tested and deemed unacceptable (e.g., increased occurrence of cytokine release syndrome (CRS) or other DLTs compared to the monotherapy induction schedule), expansion cohorts will be initiated at dose levels that were previously tolerated, including the REGN7075 induction schedule. If the concurrent initiation dose level is deemed acceptable, expansion cohorts will be initiated and will use the concurrent schedule.

[0221] Research period The total duration for each subject will vary based on disease progression, intolerable AE, withdrawal of consent, or occurrence of one or more of the following study discontinuation criteria.

[0222] The study timeline consists of screening (up to 28 days), treatment (variable in duration and timing), and follow-up (approximately 90 days after the last dose).

[0223] The timing of administration is summarized in Figure 2.

[0224] 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, Cycle 2, and every 12 weeks thereafter. If there is no clinical progression, patients have the option to continue treatment on the same event schedule until radiological progression is confirmed (PD on two consecutive scans, the second scan must be performed at least 4 weeks after the first scan and not later than the subject's next scheduled scan). If treatment remains well tolerated and is deemed to benefit the subject, further treatment beyond confirmed radiological progression will be considered upon request. Response according to RECIST 1.1 and / or composite response criteria (depending on the subject's baseline evaluation criteria) will be assessed while scans are collected and retained with the option of central review.

[0225] Dose escalation The study will begin with dose escalation of REGN7075 in combination with fixed-dose cemiplimab in patients with advanced solid tumors who have not previously been treated with anti-PD-1 or anti-PD-L1 therapy ("anti-PD-1 / PD-L1 naive"). Treatment at each DL (Table 10) will consist of a 3-week monotherapy induction of REGN7075, followed by a 6-week combination DLT period of REGN7075 and cemiplimab (see Figures 2A-2D). [Table 10]

[0226] In the assigned DL, each patient will begin induction of REGN7075 monotherapy, during which the safety and PK of REGN7075 monotherapy will be evaluated. Induction is planned to last 21 days and include three doses of REGN7075 QW. This will be followed by combination therapy with cemiplimab 350 mg Q3W, administered on a QW or Q3W schedule (same nominal dose as QW). Combination therapy will begin only after the patient 1) has received at least three doses of REGN7075 and 2) has not developed Grade ≥2 CRS, rash, or imAEs associated with their most recent REGN7075 monotherapy infusion. If Grade ≥2 CRS, rash, or imAEs are observed during the third dose of induction but resolve before the next scheduled dose of REGN7075, REGN7075 monotherapy will be continued for the fourth dose. If Grade ≥2 CRS, rash, or imAEs are observed during the fourth dose of monotherapy, patients will not begin combination therapy but will have the option to continue monotherapy. Monotherapy administration will continue weekly, as at Week 3 (Day 15). These patients will be considered uninformative for combination therapy dose selection and may be replaced as needed, but will be allowed to continue the study with monotherapy at the investigator's discretion. In this case, patients will complete all assessments except for cemiplimab treatment. Only if REGN7075 is administered and Grade ≥2 CRS, rash, or imAEs do patients receive the REGN7075 and cemiplimab combination. One or more designated expansion cohorts may be initiated at selected dose levels shown to be safe while simultaneously continuing dose escalation.

[0227] Simultaneous start cohorts Additional cohorts may be enrolled without the 3-week lead-in of REGN7075 monotherapy (i.e., cemiplimab and REGN7075 will be initiated together, considered herein as a "concurrent start" schedule). These concurrent start cohorts, with REGN7075 administered QW and / or Q3W with cemiplimab, will commence only after the lead-in of REGN7075 monotherapy and the combination with cemiplimab at the projected dose have been evaluated for safety and tolerability.

[0228] Intra-patient dose escalation Patients with a partial response (PR), stable disease (SD), or previously approved post-progression therapy after Day 1 of Cycle 3 (and at least two scans) who tolerate treatment (maximum grade of treatment-related toxicity observed during the previous treatment cycle ≤2) but do not demonstrate further improvement at their assigned dose level may be considered for intrapatient dose escalation (IPDE) with REGN7075 to the highest DL deemed safe (i.e., one level below the currently actively enrolled DL if dose escalation is ongoing, or the current dose level if dose escalation is complete and RP2D(s) have been determined). Patients with a PR must demonstrate further improvement on at least three consecutive scans. Patients who continue to tolerate treatment may be considered for further escalation, but no more frequently than once every 6 weeks.

[0229] Subcutaneous dose escalation cohort SC dose-escalation cohorts may be initiated to further inform safety, efficacy, and dosing management. Subcutaneous dose escalation will begin at a dose based on prior preclinical, PK, and clinical data and follow DL(s). Cemiplimab may continue to be administered IV (350 mg IV Q3W) or may instead be administered SC (pending SC development results for cemiplimab in other studies). SC cohort(s) will be designated "SC" along with the milligram dose (e.g., if DL8 is the estimated RP2D(s), then DL_SC_100, where 100 indicates the milligram dose of REGN7075).

[0230] Dose expansion Cohorts of specific solid tumor types will be treated with REGN7075 at doses and schedules of interest that have been demonstrated to be safe during dose escalation.

[0231] If a dose level for the concurrent start schedule (i.e., REGN7075 and cemiplimab initiated without a REGN7075 lead-in period) is not explored, or if a concurrent start dose level is tested and deemed unacceptable (e.g., increased incidence of CRS or other DLTs compared to the monotherapy lead-in schedule), expansion cohorts will be initiated at previously tolerated dose levels, including the REGN7075 lead-in schedule. If the concurrent start dose level is deemed acceptable, expansion cohorts will begin and use the concurrent schedule. The expansion cohorts are listed below: Cohort A: Triple-negative breast cancer Patients with previously documented metastatic or locally advanced TNBC who are not candidates for definitive surgery or definitive radiation, and Anti-PD-1 / PD-L1 naive Cohort B: cutaneous squamous cell carcinoma Patients with metastatic or locally advanced CSCC who are not candidates for definitive surgery or definitive radiation, Patients who would otherwise be candidates for cemiplimab monotherapy (or another anti-PD-1 / PD-L1 agent), and Anti-PD-1 / PD-L1 naive Cohort C: Non-small cell lung cancer Patients with metastatic or locally advanced NSCLC who are not candidates for definitive surgery or definitive radiation, No previously documented targetable molecular driver mutations (e.g., ALK, ROS1, EGFR, RET fusion, MET exon 14 skipping), and Anti-PD-1 / PD-L1 naive No prior systemic treatment for recurrent or metastatic NSCLC (note that adjuvant or neoadjuvant systemic treatment, excluding anti-PD-1 / PD-L1 treatment, does not count as prior line). Cohort D: Head and neck squamous cell carcinoma Patients with metastatic or locally advanced HNSCC who are not candidates for definitive surgery or definitive radiation, and Patients with previously documented PD-L1 expression by IHC with a CPS of ≥ 1%, 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. Anti-PD-1 / PD-L1 naive Cohort E: Microsatellite-stable colorectal cancer with active liver and / or peritoneal metastases Patients with metastatic or locally advanced CRC who are not candidates for definitive surgery or definitive radiation, and Active metastatic disease in the liver or peritoneum at the time of screening Resected and resected liver disease is not considered active. Note: MSS-CRC patients with RECIST-evaluable disease not confined to the lung / lymph nodes but not the liver / peritoneum may also be enrolled in this cohort The disease is MSS with previously documented results, Have received at least one prior line of treatment - Patients with previously documented RAS wild-type disease must have received anti-EGFR therapy or have a documented reason why anti-EGFR therapy was not appropriate - Patients must be receiving anti-VEGF therapy or have a documented reason why anti-VEGF therapy was not appropriate Anti-PD-1 / PD-L1 naive Cohort F: Microsatellite-stable colorectal cancer with isolated lung / lymph node metastasis (no active liver or peritoneal metastasis) Patients with metastatic or locally advanced CRC who are not candidates for definitive surgery or definitive radiation, and No active liver or peritoneal metastases identified at screening; Resected and resected liver disease is not considered active. -Metastasis is limited to the lungs and / or lung lymph nodes The disease is MSS with previously documented results, Have received at least one prior line of treatment - Patients with previously documented RAS wild-type disease must have received anti-EGFR therapy or have a documented reason why anti-EGFR therapy was not appropriate - Patients must be receiving anti-VEGF therapy or have a documented reason why anti-VEGF therapy was not appropriate Anti-PD-1 / PD-L1 naive Cohort G: EGFR-mutated NSCLC after third-generation TKIs Patients with histologically or cytologically documented locally advanced or metastatic non-squamous NSCLC who are not candidates for definitive surgery or definitive radiation, and With previously documented targetable EGFR mutations: -NSCLC with EGFR exon 19 deletion -NSCLC with EGFR L858R mutation -NSCLC with activating EGFR exon 20 insertions -NSCLC with exon 18 / 21 atypical mutations Note: EGFR deletions / mutations must be previously documented by a certified test (either from tissue or blood ctDNA is acceptable) Documentation of EGFR mutation status can be obtained any time after the initial diagnosis of non-small cell lung cancer. Anti-PD1 / PD-L1 naive, and -Previously received third-generation TKI treatment - For patients whose tumors have a previously documented EFGR exon 19 deletion or L858R mutation, prior osimertinib or other third-generation TKI therapy is required Small cell transformation is excluded Cohort H: EGFR-mutated NSCLC after third-generation TKI and platinum-doublet chemotherapy Patients with histologically or cytologically documented locally advanced or metastatic non-squamous NSCLC who are not candidates for definitive surgery or definitive radiation, and With previously documented targetable EGFR mutations: -NSCLC with EGFR exon 19 deletion -NSCLC with EGFR L858R mutation -NSCLC with activating EGFR exon 20 insertions -NSCLC with exon 18 / 21 atypical mutations Note: EGFR deletions / mutations must be previously documented by a certified test (either from tissue or blood ctDNA is acceptable) Anti-PD1 / PD-L1 naive, and -Previously received third-generation TKI treatment - For patients whose tumors have a previously documented EFGR exon 19 deletion or L858R mutation, prior osimertinib or other third-generation TKI therapy is required -Has received prior treatment with platinum-doublet chemotherapy Small cell transformation is excluded

[0232] Study population Dose Escalation: The study population consists of patients with selected advanced solid tumors who have exhausted treatment options expected to provide significant clinical benefit, excluding patients with malignancies for which anti-PD-1 or PD-L1 therapy has demonstrated clinical benefit.

[0233] Dose Expansion: The study population consists of a specific cohort of patients with selected advanced solid tumors who are anti-PD-1 / PD-L1 naive.

[0234] Selection Criteria Patients must meet the following criteria to be eligible for inclusion in the study: 1. ≥ 18 years old 2. Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1 3.Have histologically or cytologically confirmed cancer that meets the following criteria: a. Dose escalation: a. Have metastatic or locally advanced disease that is not a candidate for definitive surgery or definitive radiation. b. Not a candidate for an approved indication for anti-PD-1 or PD-L1 therapy, or such therapy is not otherwise available to the patient (alone or in combination). Reasons for ineligibility must be documented. c. Have exhausted all treatment options expected to provide significant clinical benefit, either through disease recurrence, refractory disease, or intolerance. Patients with malignancies for which anti-PD-1 or PD-L1 therapy has demonstrated clinical benefit are exempt from this requirement. d. Have any of the following cancer types: a. Microsatellite-stable colorectal cancer documented by local pathology b. Gastric cancer or gastroesophageal junction cancer c. Food tract 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. prostate adenocarcinoma l. Renal cell carcinoma (chromophobe, clear cell, or papillary) m. Gallbladder / bile duct cancer n.Urothelial carcinoma Pancreatic cancer p. Penile SCC Q. Vulvovaginal cancer r. Additional non-CNS tumor types where the investigator can demonstrate elevated EGFR expression in the tumor may be eligible after discussion with the sponsor Note: Small cell / large cell / neuroendocrine and sarcomatoid histology are excluded b. Dose expansion cohort: Cohort A: Patients with metastatic TNBC a. Not a candidate for definitive surgery or definitive radiation b. Not a candidate for anti-PD-1 or anti-PD-L1 therapy in the approved indication, or such therapy is not otherwise available to the patient (e.g., PD-L1 ICS does not meet the indicated use score, anti-PD-1 or anti-PD-L1 therapy is not accessible to the patient through reimbursement, contraindication to protein-bound paclitaxel, etc.) c. Previously documented triple-negative cancer (ER- / PR- / Her2-) by local pathology b. Cohort B: Patients with metastatic or locally advanced CSCC who are not candidates for definitive surgery or definitive radiation c. Cohort C: Patients with histologically or cytologically documented locally advanced or metastatic NSCLC disease a. Not a candidate for definitive surgery or definitive radiation b. No previously documented targetable molecular alterations (e.g., ALK, ROS1, EGFR, MetEx14, etc.) c. No prior systemic treatment for recurrent or metastatic NSCLC (adjuvant or neoadjuvant systemic treatment does not count as prior line) d. Cohort D: Patients with histologically or cytologically documented locally advanced or metastatic HNSCC disease a. Not a candidate for definitive surgery or definitive radiation b. Have PD-L1 expression of CPS ≥ 1% by previously documented IHC assay performed on a specimen collected within the past 3 months c. No prior systemic treatment for recurrent or metastatic HNSCC (adjuvant or neoadjuvant systemic treatment does not count as prior line) e. Cohort E: Patients with metastatic CRC and previously documented MSS disease a. Not a candidate for definitive surgery or definitive radiation b. Possible active liver and / or peritoneal metastases at the time of screening Note: Resected and resected disease is not considered active c. Microsatellite stable according to previously documented results by local pathology report from the patient's medical history d. Received at least one line of therapy in the recurrent / metastatic setting Patients with previously documented RAS wild-type disease must have received anti-EGFR therapy or have a documented reason why anti-EGFR therapy was not appropriate Patients must be receiving anti-VEGF therapy or have a documented reason why anti-VEGF therapy was not appropriate f. Cohort F: Patients with metastatic CRC and previously documented MSS disease a. Not a candidate for definitive surgery or definitive radiation b. No active liver or peritoneal metastases identified at screening. Note: Resected and resected disease is not considered active. c. The disease site is limited to the lung(s) and / or pulmonary lymph nodes d. Microsatellite stable according to previously documented results based on clinical history and documented by local pathology report e. Received at least one line of therapy in the recurrent / metastatic setting Patients with previously documented RAS wild-type disease must have received anti-EGFR therapy or have a documented reason why anti-EGFR therapy was not appropriate. Patients must be receiving anti-VEGF therapy or have a documented reason why anti-VEGF therapy was not appropriate g. Cohort G: Patients with previously histologically documented or cytologically documented locally advanced or metastatic EGFR-mutated non-squamous NSCLC disease a. Not a candidate for definitive surgery or definitive radiation b. Have a previously documented targetable EGFR mutation: - NSCLC with EGFR exon 19 deletion. - NSCLC with EGFR L858R mutation. -NSCLC with activating EGFR exon 20 insertions -NSCLC with exon 18 / 21 atypical mutations Note: EGFR deletions / mutations must be previously documented by certified testing from the patient's medical history (either from tissue or blood ctDNA is acceptable). b. Chemotherapy naive c. Received treatment with third-generation TKI a. For patients whose tumors have a previously documented EFGR exon 19 deletion or L858R mutation, prior osimertinib or other third-generation TKI treatment is required Note: - Stable CNS disease is permitted, defined as no evidence of progression for at least 6 weeks on imaging obtained during the screening period, no evidence of new or expanding brain metastases, and the patient not requiring any immunosuppressive doses of systemic corticosteroids for the management of brain metastases within 4 weeks of the first dose of study drug. - Small cell transformation is excluded h. Cohort H: Patients with histologically or cytologically documented locally advanced or metastatic non-squamous NSCLC disease a. Not a candidate for definitive surgery or definitive radiation b. Have a previously documented targetable EGFR mutation: - NSCLC with EGFR exon 19 deletion. - NSCLC with EGFR L858R mutation. -NSCLC with activating EGFR exon 20 insertions -NSCLC with exon 18 / 21 atypical mutations NOTE: EGFR deletions / mutations must be previously documented by any recognized test from the patient's medical history (either from tissue or blood ctDNA is acceptable) c. Received treatment with third-generation TKI a. For patients whose tumors have a previously documented EFGR exon 19 deletion or L858R mutation, prior osimertinib or other third-generation TKI treatment is required d. Have been treated with platinum doublet chemotherapy Note: - Stable CNS disease is permitted, defined as no evidence of progression for at least 6 weeks on imaging obtained during the screening period, no evidence of new or expanding brain metastases, and the patient not requiring any immunosuppressive doses of systemic corticosteroids for the management of brain metastases within 4 weeks of the first dose of study drug. - Small cell transformation is excluded 4. Expansion cohort only: Anti-PD-1 / PD-L1 naive, defined as those who have not received prior treatment with PD-1-targeted agents Note: For dose escalation, prior anti-PD-1 or PD-L1 therapy is permitted only if ≥1 month has elapsed before the first dose of study treatment and the patient has recovered from or returned to baseline from immune-mediated adverse events at least 1 month before initiating study drug. Endocrinopathy and vitiligo adequately controlled with hormone replacement therapy are not exclusionary. Patients who have permanently discontinued prior anti-PD-1 or PD-L1 therapy due to drug-related toxicity are not eligible. 5. Have at least one lesion meeting study criteria. Tumor lesions at previously irradiated sites are considered measurable if such lesions have demonstrated progression after radiation (either by an increase in size meeting RECIST 1.1 criteria for PD or by biopsy-proven viable tumor). For patients in the CSCC expansion cohort only: Following digital medical photography, there must be at least one measurable baseline lesion with both the longest diameter (LD) and perpendicular diameter ≥ 10 mm. Non-measurable disease is defined as either a unidimensionally measurable lesion, a tumor with poorly defined margins, or a lesion with a maximum perpendicular diameter < 10 mm. Patients without measurable disease at baseline are not eligible for the study. 6. Willingness to donate tumor tissue from a freshly obtained biopsy (minimum core biopsy) from a previously unirradiated tumor site. Note: a. Previously irradiated lesions may be used if they are the only accessible lesions and it is documented that the lesion has progressed since radiation. b. If the patient has only a single RECIST 1.1 measurable and biopsy-accessible lesion, a biopsy is recommended before baseline imaging to ensure post-biopsy measurements are accurate. c. This biopsy requirement may be waived on a limited basis after consultation between the investigator and sponsor if, in the investigator's opinion, the biopsy poses a life-threatening risk to the patient (e.g., if a retrocardiac mass is the only lesion). In these cases, archival tissue from a biopsy obtained within the past six months is required. 7. Have adequate organ and bone marrow function as documented by: Hemoglobin ≥ 8.0 g / dL b. Absolute neutrophil count (ANC) ≥ 1.0 × 10 9 / L Note: In African-American patients with a documented history of benign ethnic neutropenia, an ANC ≥ 0.5 × 10 9 / L is acceptable c. Platelet count ≧75×10 9 / L 8. Serum creatinine ≤ 1.5 x ULN or estimated glomerular filtration rate > 30 mL / min (per methodology accepted by the investigator's institution, e.g., Dietary Modification for Renal Response [MDRD], Chronic Kidney Disease Epidemiology Collaboration [CKD-EPI] formula). 24-hour urine creatinine collection can be substituted for calculated creatinine clearance to meet eligibility criteria. 9. Hypercalcemia of grade <3 or controlled to grade <3 with bisphosphonate therapy 10. Liver function meeting the following criteria: Total bilirubin ≤ 1.5 x upper limit of normal (ULN) (≤ 3 x ULN if tumor involves the liver) b. Aspartate aminotransferase (AST) ≤ 2.5 × ULN (≤ 5 × ULN if tumor invades the liver) c. Alanine aminotransferase (ALT) ≤ 2.5 × ULN (≤ 5 × ULN if tumor invades the liver) d. Alkaline phosphatase (ALP) ≤ 2.5 × ULN (≤ 5 × ULN if tumor invades liver or bone) Note: Patients with tumor invading the liver and AST levels ≥ 3 x ULN or ALT ≥ 3 x ULN, and bilirubin levels ≥ 2 x ULN will be excluded regardless of the above criteria. Patients with Gilbert syndrome do not need to meet the total bilirubin requirement as long as the total bilirubin does not exceed 150% of past levels and is less than 2 mg / dL (34.2 μmol / L). Gilbert syndrome should be properly documented in the past medical history. 11. Do not require rapidly effective therapy such that a 3-week REGN7075 monotherapy induction period prior to initiating the REGN7075 and cemiplimab combination is not expected to be a clinically meaningful delay in receiving the cemiplimab component of treatment. NOTE: This criterion does not apply to patients enrolling in cohorts with concurrent initiation of REGN7075 and cemiplimab. 12. Life expectancy is at least 3 months. 13. Willing and able to comply with clinic visit and study-related procedures. 14. Able to provide informed consent signed by the research participant or their legally authorized representative.

[0235] Exclusion criteria Patients who meet any of the following criteria will be excluded from the study: 1.Currently participating in another study of a therapeutic drug. 2. Have participated in any study of an investigational drug or device within 4 weeks of first dose of study drug. a. Exception: Patients undergoing or enrolled in a study involving treatment with an investigational immunoPET (iPET) reagent may enroll in the study as long as the iPET reagent does not target EGFR, CD28, PD-1, PD-L1, or PD-L2. 3. Have received approved systemic therapy within 4 weeks of first dose of study drug or have not yet recovered from acute toxicity (Grade 1 or baseline). 4. Received treatment with an anti-EGFR antibody therapy within the drug-specific window (approximately 5 half-lives) of: 5. Cetuximab: Within 4 weeks of the first dose of REGN7075 6. Panitumumab: Within 6 weeks of the first dose of REGN7075 7. Necitumumab: Within 10 weeks of the first dose of REGN7075 8. Any other investigational drug that inhibits EGFR via interaction with the extracellular binding domain: After sponsor approval, within 5 half-lives of the first dose of REGN7075. a. Note: Kinase inhibitors targeting EGFR are allowed within the guidelines of exclusion criterion 3. 9. Have undergone radiation therapy or major surgery within 14 days of first dose of study drug or have not recovered from an adverse event (Grade 1 or baseline). 10. Have received any previous systemic non-immunomodulatory biologic therapy within 4 weeks of first dose of study drug. 11. Have received prior anti-cancer immunotherapy within 5 half-lives prior to study drug. Examples of immunomodulatory agents include blockers of CTLA-4, 4-1BB (CD137), or OX-40, therapeutic vaccines, PI3K-delta inhibitors, or cytokine anti-cancer treatments. a. Prior CAR-T therapy is allowed regardless of cell half-life / persistence. 12. Not recovered from an immune-mediated adverse event prior to initiation of study drug (i.e., baseline). Endocrine disorders adequately controlled with hormone replacement therapy are not exclusionary. 13. Contraindication to anti-PD-1 therapy 14. Have a known (previously documented) microsatellite instability-high (MSI-high) or mismatch repair-deficient cancer. 15. Have a secondary malignancy that is progressing or requiring aggressive treatment, except for the following: Non-melanoma skin cancer that has been treated with potentially curative therapy b. Tumors that have been determined to be effectively treated with reliable local control 16. Have a condition requiring ongoing / continuous corticosteroid therapy (≥10 mg prednisone daily or equivalent anti-inflammatory) within 1-2 weeks prior to the first dose of study drug. Physiologic replacement therapy is permitted, even at ≥10 mg prednisone daily or equivalent, as long as it is not administered for immunosuppressive purposes. Inhaled or topical steroids are permitted only if not intended to treat an autoimmune or skin disorder. a. Note: Patients requiring short-term steroid administration (maximum 2 days in the week prior to enrollment), short-term corticosteroid administration for the prevention (e.g., contrast allergy) or treatment of non-autoimmune diseases (e.g., delayed hypersensitivity reactions to contact allergens), or physiological replacement may be enrolled in the study. 17. Current or recent (within the past 5 years) evidence of a significant autoimmune disease or any other condition requiring treatment with systemic immunosuppressive therapy. Vitiligo, resolved childhood asthma, and endocrine disorders requiring hormone replacement only (e.g., hypothyroidism or type 1 diabetes) are not excluded. 18. Untreated or active primary brain tumor, CNS metastasis, leptomeningeal disease, or spinal cord compression. a. Note: Patients previously treated for central nervous system metastases or spinal cord compression are not excluded if: b. No evidence of progression for at least 4 weeks prior to the first dose of study medication, and any neurological symptoms have returned to baseline; c. No evidence of new or spreading central nervous system metastases; and d. No systemic corticosteroids required for management of central nervous system metastases or spinal cord compression within 4 weeks prior to the first dose of study drug. 19. Encephalitis, meningitis, organic brain disease (e.g., Parkinson's disease), or uncontrolled seizures within 1 year prior to the first dose of study drug 20. Have significant baseline prolongation of the QT / QTc interval or risk factors for prolonged QTc, e.g. a. Repeated demonstration of a QTc interval of >470 ms at baseline b. Personal history of additional risk factors for torsade de pointes (TdP) or family history of long QT syndrome 21. History of myocardial infarction, cardiac events, congestive heart failure (NYHA class II or higher), or arrhythmia within the past 12 months. a. Note: A history of arrhythmias that were transient and did not require any medical intervention is acceptable as long as the episode did not occur within the past 6 months prior to enrollment. Premature atrial contractions (PACs) or premature ventricular contractions (PVCs) that do not meet the criteria for a specific arrhythmia are acceptable anyway. 22. Patients with active inflammatory skin disease (e.g., psoriasis, eczema) or inflammatory skin disease (regardless of activity) requiring ongoing medication (topical or systemic), or a history of inflammatory skin disease within the past 5 years. 23. Any of the following skin findings persist at baseline, regardless of grade: a. Bullous dermatitis b.Exfoliative dermatitis c. Erythroderma d. Erythema multiforme e. Acne-like rash f. Generalized rash g.Maculopapular rash h. Other unspecified skin toxicity (other than Grade ≤ 1) with any inflammatory or destructive features such as infiltration, erythema, ulceration, blistering, or desquamation. i. Note: Skin findings of Grade ≤ 1, which are expected to be transient, are not excluded. Additionally, hypersensitivity reactions of Grade ≤ 1, which are expected to be transient, are not excluded. 24. History or any evidence of interstitial lung disease or active non-infectious pneumonitis within 5 years prior to the first dose of study medication. A history of radiation pneumonitis in the radiation field is acceptable as long as the pneumonitis resolves at least 6 months prior to the first dose of study medication. 25. Any indwelling drain or stent intended to maintain organ function or facilitate drainage of the obstruction (e.g., nephrostomy tube, biliary stent, etc.). a.Note b. Urinary catheterization of the bladder or neobladder is not excluded c. Ports or indwelling intravenous lines are acceptable. d. Coronary stents are not excluded as long as the patient meets criterion #17 26. Uncontrolled infection with human immunodeficiency virus, hepatitis B, or hepatitis C infection, or diagnosed immunodeficiency. a.Note: b. Patients will be tested for Hepatitis C virus (HCV) and Hepatitis B virus (HBV) at screening. c. Patients with known HIV infection and whose infection is controlled (spontaneous or on a stable antiviral regimen, with undetectable viral load (HIV RNA PCR) and CD4 count >350) are permitted. For patients with controlled HIV infection, monitoring will be performed according to local standards. d. Patients with hepatitis B (HepBsAg+) whose infection is controlled (serum hepatitis B virus DNA PCR below the limit of detection and receiving hepatitis B antiviral therapy) are allowed. Patients with controlled infection should undergo regular HBV DNA monitoring. Patients should continue antiviral therapy for at least 6 months after the last dose of study drug. Patients who are eHepatitis C virus antibody positive (HCV Ab+) and whose infection is controlled (no detectable HCV RNA by PCR due to spontaneous or previous successful anti-HCV therapy) can be enrolled in the study. 27. Requiring treatment with anti-infectives within 4 weeks of the first dose of study drug or any ongoing infection (unless otherwise specified in the exclusion criteria). 28. Have received a live vaccine within 4 weeks of the planned start of study drug. 29. Have previously undergone allogeneic stem cell transplantation, autologous stem cell transplantation, or solid organ transplantation at any time. 30. Known allergy or hypersensitivity to cemiplimab or any component of the study drug. 31. Have a known psychiatric or substance abuse disorder that may prevent you from participating in the study requirements. 32. Have any medical condition, comorbidity, physical examination findings, metabolic dysfunction, or laboratory abnormality that would make them unsuitable for clinical trial participation because they pose a high safety risk and / or may affect the interpretation of study results. 33. Members of the research team at the clinical site and / or their immediate family members, unless approved in advance. 34. Patients with a positive serum hCG pregnancy test must have a medically excluded pregnancy to be eligible for the study. Lactating women will also be excluded. 35. Continuous sexual activity by women of childbearing potential (WOCBP)* or sexually active men who are unwilling to use highly effective contraception before the first dose / initiation of initial treatment, during the study, and for at least 6 months after the last dose. Highly effective contraception includes: a. Stable use of combined hormonal contraceptives (oral, vaginal, transdermal) (containing estrogen and progestogen) or progestogen-only hormonal contraceptives (oral, injectable, implantable) associated with ovulation suppression, with at least two menstrual cycles initiated prior to screening b. Intrauterine device (IUD), intrauterine hormone-releasing system (IUS), c. Bilateral tubal ligation or occlusion d. Vasectomized partner (provided the male partner who underwent vasectomy was the WOCBP study participant's only sexual partner and the vasectomized partner was medically evaluated for the success of the surgery) e. and / or sexual abstinence (*1, *2), 36. WOCBP are defined as women of childbearing potential from menarche until menopause, unless permanently infertile. Permanent methods of contraception include hysterectomy, bilateral salpingectomy, and bilateral oophorectomy. a. Postmenopausal status is defined as the absence of menstruation for 12 months without an alternative medical cause. High follicle-stimulating hormone (FSH) levels in the postmenopausal range may be used to confirm postmenopausal status in women not using hormonal contraception or hormone replacement therapy. However, a single FSH measurement is insufficient to determine the occurrence of postmenopausal status in the absence of 12 months of amenorrhea. The above definition follows guidance from the Clinical Trials Facilitation Group (CTFG). Pregnancy testing and contraception are not required for postmenopausal or permanently infertile women. (*1) Sexual abstinence is considered highly effective only when defined as abstinence from heterosexual intercourse for the entire duration of the risk associated with the study drug. The reliability of sexual abstinence needs to be evaluated in relation to the duration of the clinical trial and the patient's preferences and usual lifestyle. b. (*2) Periodic abstinence (calendar, symptom-thermal, or postovulatory methods), abortion (interrupted coitus), spermicide alone, and lactational amenorrhea (LAM) are not acceptable methods of contraception. Female condoms and male condoms should not be used together. 37. Cohorts G and H only: Small cell transformation.

[0236] Test treatment REGN7075: 0.1 mg up to 900 mg administered weekly (QW) or 0.1 mg up to 2700 mg administered QW or every 3 weeks (Q3W) by IV infusion or subcutaneous (SC) injection at the target dose level. For doses above 100 mg, REGN7075 may be administered using a step-up schedule with or without a split-dose schedule.

[0237] Cemiplimab: 350 mg administered concurrently by IV infusion over 30 minutes or SC injection Q3W.

[0238] If both drugs are administered on the same day, REGN7075 will be administered before cemiplimab.

[0239] Platinum-based doublet chemotherapy administered IV Q3W for four cycles (followed by pemetrexed maintenance for patients initially assigned to receive a pemetrexed-containing regimen).

[0240] The choice of chemotherapy is one of the regimens shown in Table 11. [Table 11]

[0241] Chemotherapy is administered after completion of semipimab administration. If chemotherapy is administered on the same day as REGN7075 and semipimab, the full dose of REGN7075 is administered first, followed by semipimab, and finally chemotherapy.

[0242] Study endpoints The primary endpoint of treatment will be determined based on the following assessments: Dose escalation: Incidence of dose-limiting toxicities (DLTs) during the DLT period Incidence and severity of treatment-emergent adverse events (TEAEs), adverse events of special interest (AESIs), serious adverse events (SAEs), and grade ≥ 3 laboratory abnormalities Dose expansion: Objective response rate (ORR) according to Response Evaluation Criteria in Solid Tumors version 1.1 (RECIST 1.1) and / or composite response criteria (according to the subject's baseline evaluation criteria).

[0243] Secondary treatment endpoints will be determined based on the following assessments: Dose escalation Serum concentrations of REGN7075 and cemiplimab Objective response rate (ORR) according to Response Evaluation Criteria in Solid Tumors version 1.1 (RECIST 1.1) and / or composite response criteria (according to the subject's baseline criteria), progression-free survival (PFS), duration of response (DOR), complete response (CR) rate, and disease control rate (DCR) ·Overall survival rate (OS) Incidence of anti-drug antibodies (ADA) to REGN7075 and cemiplimab Dose expansion Incidence and severity of TEAEs / AESIs / SAEs and grade ≥3 laboratory abnormalities. Serum concentrations of REGN7075 and cemiplimab Incidence of AD with REGN7075 and cemiplimab PFS, DOR, DCR, and CR rates according to RECIST 1.1 and / or composite response criteria (based on subject's baseline evaluation criteria) Operating System Patient-reported quality of life, symptoms, function, and general health status (according to EORTC QLQ-C30, EORTC QLQ-CR29 for CRC patients, EORTC QLQ-BR23 for breast cancer patients, EORTC QLQ-LC13 for NSCLC patients, EORTC QLQ-HN35 for HNSCC patients, and EQ-5D-5L).

[0244] Procedures and Evaluation Treatment will be evaluated using the following procedures and assessments: Screening / Baseline only: Height, brain imaging, coagulation assessment, and archival or fresh tumor evaluation. Efficacy: Radiographic disease assessment (computed tomography [CT], magnetic resonance imaging [MRI], and / or digital photography). For subjects with radiologically measurable disease, disease will be assessed radiologically according to RECIST 1.1. For subjects with CSCC lesions evaluable on the skin, composite response criteria should be used in combination with radiological imaging, when appropriate. Safety: Vital signs (including temperature, sitting blood pressure, pulse, and respiration), physical examination, Eastern Cooperative Oncology Group (ECOG) performance status, weight, electrocardiogram, adverse events (AEs), hematology, blood chemistry, C-reactive protein (CRP), thyroid-stimulating hormone (TSH), pregnancy test, and urinalysis Pharmacokinetic and immunogenicity sampling Biomarker and exploratory studies: serum cytokines, exploratory tumor biopsy (or archival tissue), whole blood for immune monitoring, plasma for ctDNA, peripheral blood mononuclear cells (PBMC) for immunophenotyping, whole blood for DNA / RNA for T cell repertoire analysis, serum and plasma for exploratory biomarkers.

[0245] result Preliminary efficacy and safety results are described in the Examples below. Outcomes to date include partial and complete responses in multiple cohorts.

[0246] Example 3: Initial Dose-Escalation Results of a Phase 1 / 2 Study of REGN7075 in Combination with Cemiplimab (Anti-PD-1) in Patients with Advanced Solid Tumors A first-in-human, open-label, Phase 1 / 2 dose-escalation and expansion study was conducted to evaluate the safety, tolerability, pharmacokinetics, and preliminary antitumor activity of REGN7075 (EGFRxCD28) in combination with cemiplimab (anti-PD-1) in subjects with advanced solid tumors (Figure 3). The study protocol is described in detail in Example 2.

[0247] In the dose-escalation phase (Bayesian optimal interval design), Part 1, heavily pretreated subjects with advanced solid tumors received induction of REGN7075 monotherapy weekly for 3 weeks, followed by combination therapy with cemipimab 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. The primary objective was to evaluate the safety and tolerability of the combination of REGN7075 and cemipimab.

[0248] As of the data cutoff date, 18 subjects (median age 53.5 years, 56% female) were treated with REGN7075 in combination with cemipimab up to a DL of 30 mg in the dose-escalation phase (Table 12). Most subjects (67%) were treated for microsatellite-stable colorectal cancer. No subjects experienced dose-limiting toxicities and the maximum tolerated dose was not reached.

[0249] Table 13 summarizes treatment-emergent adverse events (TEAEs) and treatment-related adverse events (TRAEs). The most frequent TEAEs (any grade) were aspartate aminotransferase (AST) elevation, constipation, and fatigue (33% [n=6] each). The most frequent TRAEs (any grade) were fatigue (17% [n=3]), AST elevation, diarrhea, hypothyroidism, fever, and rash (11% [n=2] each). One subject developed cytokine release syndrome, characterized by isolated grade 1 fever without hypotension or hypoxia. Five deaths were recorded that were not during study treatment or attributable to the study drug(s).

[0250] Serum concentrations of REGN7075 were measured after intravenous administration of the first dose of REGN7075. Maximum concentration (Cmax) and area under the curve (AUC) values ​​were greater than dose-proportional at the 30 mg dose, whereas minimum concentration (Cmin) values ​​were dose-proportional at all doses (Figure 4). Ongoing PK evaluation suggests a possible target-mediated effect.

[0251] T cell activation-related cytokines were detected during monotherapy and combination therapy. In one patient, IL-2 was induced after the first dose of REGN7075 (1 mg) (Figure 5A). In multiple patients, IFN-γ was induced after receiving REGN7075 alone or in combination with cemiplimab (Figure 5B).

[0252] Treatment and evaluation of the dose-escalation phase is ongoing. Nevertheless, preliminary data indicate that of all 18 subjects treated in dose-escalation, one PD-1-negative subject receiving 1 mg of REGN7075 plus cemipimab for cervical cancer achieved a striking ongoing partial response (DL4[c], Figure 6).

[0253] In this dose-escalation study, REGN7075 was safely administered up to the 30 mg dose level in combination with cemiplimab without dose-limiting toxicities. Early data indicate that the novel agent REGN7075 was generally well tolerated and has preliminary antitumor activity. There was no evidence of widespread immune activation or cytokine release syndrome, as observed with the CD28 superagonist TGN1412. [Table 12] [Table 13]

[0254] Example 4: Updated Results of Clinical Trials of REGN7075 (EGFRxCD28) Alone and in Combination with Anti-PD-1 Antibodies, e.g., Cemiplimab An open-label, Phase 1 / 2, first-in-human study was conducted to evaluate the safety, tolerability, pharmacokinetics, and preliminary antitumor activity of REGN7075 (EGFRxCD28) alone and in combination with cemiplimab (anti-programmed cell death [PD]-1) in subjects with advanced solid tumors (see Example 2). Subjects had to have protocol-defined advanced solid tumors, Eastern Cooperative Oncology Group performance status of 0 or 1, and be naïve to anti-PD-1 / anti-PD-ligand (L)1 therapy.

[0255] The 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 induction of REGN7075 monotherapy weekly for 3 weeks, followed by combination therapy with cemipimab 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.

[0256] Once the recommended dose for Phase 2 is determined in Part 1, five tumor-specific expansion cohorts will be initiated 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 (PD-L1 selected, combined positive score ≥ 1). Subjects with MSS-CRC harboring RAS or BRAF wild-type mutations must have received prior anti-EGFR or anti-vascular endothelial growth factor (VEGF) therapy. The primary endpoints for Part 1 are safety and tolerability of REGN7075 alone or in combination with cemiplimab, and for Part 2, objective response rate (ORR) based on Response Evaluation Criteria in Solid Tumors version 1.1. For Part 2, a secondary objective is to evaluate the effect of REGN7075 on subject-reported outcomes, including health-related quality of life, 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 (CRC subjects only).

[0257] As of September 13, 2022, 30 subjects have been treated with REGN7075 in combination with cemiplimab in the dose-escalation phase at a DL of up to 300 mg.

[0258] Updated results as of December 2022 show preliminary partial responses in two MSS-CRC patients (one each from DL1 and DL9) and a complete response in a patient with cervical cancer in DL4 (Figure 7).

[0259] Example 5: Updated Clinical Results This example describes updated results from an open-label, Phase 1 / 2, first-in-human, global study (NCT04626635) to evaluate the safety, tolerability, PK, and antitumor activity of REGN7075 ± cemiplimab in patients with advanced solid tumors (see Example 2).

[0260] This study includes a dose-escalation phase (Part 1) and a dose-expansion phase (Part 2). Patients must have advanced solid tumors likely to express EGFR, have an ECOG performance status of 0 / 1, and be naïve to anti-PD(L)-1 therapy (Part 2 only). In Part 1, patients will receive a 3-week induction of REGN7075 monotherapy QW before receiving REGN7075 QW or Q3W in combination with cemiplimab Q3W (excluding Cohorts C and G). Concurrent Q3W administration with cemiplimab may be administered at dose levels where induction is tolerated. After an acceptable drug dose and schedule are identified in Part 1, Part 2 will include eight 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 (MSS-CRC) with active liver and / or peritoneal metastases (E), MSS-CRC with lung / lymph node metastases (F), EGFR-mutant NSCLC after third-generation TKIs (G), and EGFR-mutant NSCLC after third-generation TKIs and platinum-doublet chemotherapy (H). Cohorts C and G will also receive four cycles of platinum-based chemotherapy concomitantly with cemiplimab. Primary endpoints: Part 1, safety and tolerability of REGN7075 with or without cemiplimab; Part 2, ORR (REGN7075 plus cemiplimab with or without chemotherapy; RECIST 1.1). Secondary objectives for Parts 1 and 2 include OS, PFS, DoR, CR rate and DCR, immunogenicity of REGN7075 and cemiplimab, and PK characterization.

[0261] The study is expected to enroll approximately 769 patients, with approximately 221 patients in part 1 and approximately 548 patients in part 2.

[0262] As of July 19, 2023, REGN7075 has been well tolerated in more than 60 patients with advanced solid tumors who received up to 900 mg of REGN7075 IV, either alone or in combination with cemiplimab. No patients experienced treatment-related adverse events leading to death. No patients experienced treatment-related AEs leading to permanent discontinuation of study drug. No patients had protocol-defined dose-limiting toxicities during the dose-limiting toxicity period. Most IRR events were grade 2 or grade 1, and IRR symptoms remained clinically manageable.

[0263] As of September 5, 2023, 93 patients have been enrolled in Part 1. Several partial and complete responses have been reported in patients in this study. This study is ongoing and continues to recruit participants.

[0264] The present disclosure is not limited in scope by the specific embodiments described herein. Indeed, various modifications of the present disclosure in addition to those described herein will become apparent to those skilled in the art from the foregoing description and accompanying figures. Such modifications are intended to be within the scope of the appended claims.

Claims

1. A method of treating cancer in a subject, comprising administering to the subject a therapeutically effective amount of a 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), in combination with an antibody or antigen-binding fragment thereof that specifically binds to programmed death receptor-1 (PD-1), thereby treating the cancer in the subject.

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

3. 3. The method of claim 1 or 2, wherein the cancer is an EGFR-expressing cancer.

4. 4. The method of claim 1, wherein the cancer is selected from esophageal cancer, lung squamous cell carcinoma, lung adenocarcinoma, cervical cancer, endometrial adenocarcinoma, bladder cancer, urothelial carcinoma, lung cancer, non-small cell lung cancer, colon cancer, sigmoid colon adenocarcinoma, rectal cancer, endometrial cancer, skin cancer, head and neck squamous cell carcinoma, brain cancer, glioblastoma multiforme, non-central nervous system tumors, cutaneous squamous cell carcinoma, breast cancer, gastric cancer, gastroesophageal cancer, gastroesophageal adenocarcinoma, pancreatic cancer, prostate cancer, ovarian cancer, melanoma, nasopharyngeal carcinoma, anal cancer, mesothelioma, renal cell carcinoma, gallbladder / bile duct cancer, pancreatic cancer, penile squamous cell carcinoma, or vulvovaginal cancer.

5. The method of any one of claims 1 to 4, further comprising selecting a subject with an advanced solid tumor.

6. The subject is a. have metastatic disease or locally advanced disease that is not a candidate for definitive surgery or definitive radiation; b. is not a candidate for an approved indication for anti-PD-1 or PD-L1 therapy, or such therapy is not otherwise available to said subject (alone or in combination); c. Exhaustion of all treatment options expected to provide significant clinical benefit through disease recurrence, refractory disease, or intolerance, except for subjects with malignancies in which anti-PD-1 or PD-L1 therapy has shown clinical benefit, and / or d.

6. The method of any one of claims 1-5, wherein the patient has at least one of the following criteria: (a) microsatellite-stable colorectal cancer documented by local pathology; (b) gastric 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 carcinoma; (h) cervical cancer; (i) anal cancer; (j) mesothelioma; (k) prostate adenocarcinoma; (l) renal cell carcinoma (chromophobe, clear cell, or papillary); (m) gallbladder / bile duct carcinoma; (n) urothelial carcinoma; (o) pancreatic cancer; (p) penile SCC; (q) vulvovaginal cancer; or (r) additional non-CNS tumor types in which elevated EGFR expression has been demonstrated in the tumor.

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

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

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

10. 10. The method of any one of claims 9, wherein the subject with microsatellite-stable colorectal cancer has 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 radiation; (c) may have active metastases in the liver and / or peritoneum at the time of screening; (d) has no active metastases identified in the liver or peritoneum at the time of screening, with disease sites limited to the lung(s) and / or lymph nodes; (e) has microsatellite stability documented by a pathology report; (f) has received at least one line of therapy in the setting of recurrence / metastasis, where the therapy includes anti-EGFR therapy or anti-VEGF therapy; or (g) is anti-PD-1 / PD-L1 naive, defined as having never received treatment with an agent targeting PD-1.

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

12. 12. The method of claim 11, wherein the subject with TNBC has or is selected based on at least one of the following attributes: (a) has metastatic TNBC; (b) is not a candidate for definitive surgery or definitive radiation; (c) is not a candidate for anti-PD-1 or anti-PD-L1 therapy in an approved indication, or such therapy is not otherwise available to the subject; (d) has triple-negative cancer (ER- / PR- / Her2-) documented by a pathology report; or (e) is anti-PD-1 / PD-L1 naive, defined as never having been treated with an agent that targets PD-1.

13. 9. The method of any one of claims 1-8, wherein the subject has cutaneous squamous cell carcinoma (CSCC) and (i) the subject is not a candidate for definitive surgery or definitive radiation, or (ii) the subject is anti-PD-1 / PD-L1 naive, defined as having never been treated with an agent that targets PD-1.

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

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

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

17. 17. The method of claim 16, wherein the subject has or is selected based on at least one of the following attributes: (a) has advanced or metastatic disease; (b) is not a candidate for definitive surgery or definitive radiation; (c) has PD-L1 expression of CPS > 1% by local IHC assay; (d) has not received prior systemic treatment for recurrent or metastatic HNSCC; or (e) is anti-PD-1 / PD-L1 naive, defined as not having received treatment with an agent that targets PD-1.

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

19. The bispecific EGFRxCD28 antibody or antigen-binding fragment thereof may be administered in 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, 350 mg, 400 mg, 550 mg, 600 mg, 700 mg, 800 mg, 90 ...

19. The method of any one of claims 1 to 18, wherein the compound is administered at a dose of 1 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. 20. The method of any one of claims 1-19, 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. 21. The method of any one of claims 1 to 20, wherein the anti-PD-1 antibody is administered at a dose of 350 mg.

22. 18. The method of any one of claims 1-17, wherein the method comprises administering one or more doses of the bispecific EGFRxCD28 antibody, or antigen-binding fragment thereof, in combination with one or more doses of the anti-PD-1 antibody, or antigen-binding fragment thereof.

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

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

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

27. 27. The method of any one of claims 22-26, wherein each of the one or more doses of the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof and / or 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 immediately preceding dose.

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

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

30. 30. The method of 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. 31. The method of any one of claims 22-30, 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. 32. The method of 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 is administered intravenously.

33. 32. The method of 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 is administered subcutaneously.

34. 34. The method of 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. 34. The method of 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. 36. The method of 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. (i) administering to the subject the bispecific EGFRxCD28 antibody, or antigen-binding fragment thereof, at a dose of 0.1 mg to 3000 mg subcutaneously or intravenously once every week or once every three weeks for a monotherapy period, wherein the monotherapy period is at least three weeks; (ii) administering to the subject the bispecific EGFRxCD28 antibody or antigen-binding fragment thereof at a dose of 0.1 mg to 3000 mg subcutaneously or intravenously once every week or once every three weeks, and administering to the subject an anti-PD-1 antibody or antigen-binding fragment thereof at a dose of 150 mg to 500 mg intravenously or subcutaneously once every three weeks.

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

39. 39. The method of claim 37 or 38, wherein during step (ii), the anti-PD-1 antibody, or antigen-binding fragment thereof, is administered on a different day than the bispecific EGFRxCD28 antibody, or antigen-binding fragment thereof.

40. 39. The method of claim 37 or 38, wherein during step (ii), the anti-PD-1 antibody, or antigen-binding fragment thereof, is administered on the same day as the bispecific EGFRxCD28 antibody, or antigen-binding fragment thereof.

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

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

43. 43. The method of any one of claims 1-42, wherein the subject exhibits stable disease, a partial response, or a complete response when administered for at least one week at a dose of about 0.1 mg to about 3000 mg of the bispecific antibody or antigen-binding fragment thereof in combination with the anti-PD-1 antibody or antigen-binding fragment thereof.

44. The method of 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, selplulimab, dostallimab, retifanlimab, zimberelimab, penprimab, pidilizumab, HX008, balstilimab, or ezabenlimab, or an antigen-binding fragment of any of the foregoing.

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

74.

46. The method of 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. 47. The method of 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 of 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 of any one of claims 1 to 48, wherein the anti-PD-1 antibody is cemiplimab or an antigen-binding fragment thereof.

50. The method of any one of claims 1 to 49, wherein the first antigen-binding domain that binds to 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. 51. The method of 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. 51. The method of 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: QQYGSSPWT (SEQ ID NO: 22).

53. The method of any one of claims 50 to 52, wherein 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.

54. The method of 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) 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.

55. 55. The method of 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. 55. The method of 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 of 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. (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; (b) the second antigen-binding domain that binds to human EGFR comprises a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 2 and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO:

16. The method of any one of claims 1 to 57.

59. 59. The method of 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. 59. The method of 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. 59. The method of 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. 59. The method of 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. 59. The method of 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. 59. The method of 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. 65. The method of any one of claims 1 to 64, wherein the bispecific EGFRxCD28 antibody is REGN7075 or an antigen-binding fragment thereof.

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

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