Treating cancers that lack EGFR activating mutations

Abstract

The present invention relates to the treatment of subjects with cancer whose tumors lack at least one activating EGFR mutation.

Description

[Technical field]

[0001] (CROSS REFERENCE TO RELATED APPLICATIONS) This application claims priority to U.S. Provisional Patent Application No. 63 / 158,552, filed March 9, 2021, the entire contents of which are incorporated herein by reference.

[0002] (Reference to electronically submitted sequence listing) This application contains a Sequence Listing, having a size of 29 KB, that has been submitted electronically via EFS-Web as an ASCII formatted Sequence Listing with the filename "JBI6507WOPCT1SEQLIST.txt" created on March 1, 2022. The Sequence Listing submitted via EFS-Web is a part of the present specification and is incorporated herein by reference in its entirety.

[0003] FIELD OF THEINVENTION The present invention relates to the treatment of subjects with cancer whose tumors lack at least one activating EGFR mutation. [Background technology]

[0004] The individual roles of both epidermal growth factor receptor (EGFR) and the receptor tyrosine kinase mesenchymal-epithelial transition factor (c-Met) in cancer are well established, making these targets attractive for combination therapy. Both receptors signal through the same survival and anti-apoptotic pathways (ERK and AKT), and thus joint inhibition of this pair may limit the potential activation of compensatory pathways, thereby improving overall efficacy.

[0005] Molecular segmentation of advanced non-small cell lung cancer (NSCLC) based on oncogenic driver mutations has improved overall survival and quality of life for patients with treatable driver mutations.

[0006] Amivantamab is a bispecific antibody targeting EGFR and c-MET. Its clinical activity has been explored across a range of EGFR activating mutations in clinical trials, but has not been evaluated for the treatment of lung cancers that are EGFR positive but lack EGFR activating mutations. Summary of the Invention [Means for solving the problem]

[0007] Improved therapeutic agents or combinations of therapeutic agents are needed to develop more effective treatments for cancers whose tumors contain EGFR that lack activating mutations.

[0008] The present disclosure provides a method of treating a subject having a cancer that is positive for EGFR and lacks at least one EGFR activating mutation, comprising administering to the subject having a cancer that is positive for EGFR and lacks at least one EGFR activating mutation a therapeutically effective amount of an isolated bispecific anti-epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) antibody.

[0009] The present disclosure also provides a method of treating a subject with cancer with a bispecific anti-EGFR / c-Met antibody, comprising: a) providing a biological sample from a subject; b) determining the presence or absence of EGFR activating mutations in the sample; and c) administering or providing for the administration of a bispecific anti-EGFR / c-Met antibody to a subject determined to lack an EGFR activating mutation.

[0010] In one embodiment, the at least one activating mutation is a mutation that increases at least one biological activity of EGFR.

[0011] In one embodiment, the at least one biological activity of EGFR is selected from the group consisting of tyrosine kinase activity, ligand-independent signaling, increased cell proliferation, signaling to the MAPK / ERK pathway, gene transcription, dimerization (EGFR:EGFR), and heterodimerization (EGFR:HER2 or EGFR:HER3).

[0012] In one embodiment, the at least one activating mutation that increases at least one biological activity of EGFR is a substitution of L718Q, G719A, G719X (X is any amino acid), L861X (X is any amino acid), L858R, E746K, L747S, E749Q, A750P, A755V, V765M, C797S, L858P or T790M, a deletion of E746 to A750, a deletion of R748 to P753, an insertion of Ala (A) between M766 and A767, an insertion of Ser, Val, and Ala (SVA) between S768 and V769, an insertion of H772 and H773, an insertion of H774 and H775, an insertion of H776 and H7 ... an insertion of Asn and Ser (NS) between D761 and E762, A763 and Y764, Y764 and Y765, M766 and A767, A767 and V768, S768 and V769, V769 and D770, D770 and N771, N771 and P772, P772 and H773, H773 and V774, V774 and C775, one or more deletions in EGFR exon 20, one or more insertions in EGFR exon 20, and at least one mutation selected from the group consisting of S768I, L861Q, and G719X (X is any amino acid).

[0013] In one embodiment, the method further comprises determining the presence or absence of at least one mutation in any one gene selected from the group consisting of KRAS, PIK3CA, and PTEN, and administering or providing for administration of a bispecific anti-EGFR / c-Met antibody to the subject determined to have EGFR lacking an activating mutation and determined to lack at least one mutation in any one gene selected from the group consisting of KRAS, PIK3CA, and PTEN.

[0014] In one embodiment, the at least one mutation in KRAS is selected from the group consisting of G12V, G12C, G12A and G12D.

[0015] In one embodiment, the at least one mutation in KRAS is G12C.

[0016] In one embodiment, the at least one mutation in PI3K is selected from the group consisting of E545K, H1047L, and PI3K amplification.

[0017] In one embodiment, the at least one mutation in PTEN is a PTEN deletion.

[0018] In one embodiment, the bispecific anti-EGFR / c-Met antibody comprises a first domain that specifically binds EGFR and a second domain that specifically binds c-Met, wherein the first domain comprises heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 1, HCDR2 of SEQ ID NO: 2, HCDR3 of SEQ ID NO: 3, light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 4, LCDR2 of SEQ ID NO: 5, and LCDR3 of SEQ ID NO: 6, and the second domain that binds c-Met comprises HCDR1 of SEQ ID NO: 7, HCDR2 of SEQ ID NO: 8, HCDR3 of SEQ ID NO: 9, LCDR1 of SEQ ID NO: 10, LCDR2 of SEQ ID NO: 11, and LCDR3 of SEQ ID NO: 12.

[0019] In one embodiment, a first domain that specifically binds to EGFR comprises a heavy chain variable region (VH) of SEQ ID NO: 13 and a light chain variable region (VL) of SEQ ID NO: 14, and a second domain that specifically binds to c-Met comprises a VH of SEQ ID NO: 15 and a VL of SEQ ID NO: 16.

[0020] In one embodiment, the bispecific anti-EGFR / c-Met antibody is of the IgG1 isotype.

[0021] In one embodiment, the bispecific anti-EGFR / c-Met antibody comprises a first heavy chain (HC1) of SEQ ID NO: 17, a first light chain (LC1) of SEQ ID NO: 18, a second heavy chain (HC2) of SEQ ID NO: 19, and a second light chain (LC2) of SEQ ID NO: 20.

[0022] In one embodiment, the bispecific anti-EGFR / c-Met antibody comprises one or more Fc silencing mutations.

[0023] In one embodiment, the one or more Fc silencing mutations reduce affinity for an Fcγ receptor.

[0024] In one embodiment, the one or more Fc silencing mutations include V234A / G237A / P238S / H268A / V309L / A330S / P331S.

[0025] In one embodiment, the bispecific anti-EGFR / c-Met antibody comprises a biantennary glycan structure having a fucose content of between about 1% and about 15%.

[0026] In one embodiment, the subject is relapsed or refractory to treatment with one or more previous anti-cancer therapies.

[0027] In one embodiment, the one or more previous anti-cancer therapies include one or more chemotherapeutic agents, checkpoint inhibitors, targeted anti-cancer therapies, or kinase inhibitors, or any combination thereof.

[0028] In one embodiment, the one or more prior anticancer therapies comprise carboplatin, paclitaxel, gemcitabine, cisplatin, vinorelbine, docetaxel, palbociclib, crizotinib, PD-(L)1 axis inhibitors, inhibitors of EGFR, inhibitors of c-Met, inhibitors of HER2, inhibitors of HER3, inhibitors of HER4, inhibitors of VEGFR, inhibitors of AXL, erlotinib, gefitinib, lapatinib, vandetanib, afatinib, osimertinib, lazertinib, poziotinib, cliotinib, cabozantinib, capmatinib, axitinib, lenvatinib, nintedanib, regorafenib, pazopanib, sorafenib, or sunitinib, or any combination thereof.

[0029] In one embodiment, the subject is treatment naive.

[0030] In one embodiment, the cancer positive for EGFR lacking activating mutations is positive for at least one mutation in a gene selected from the group consisting of ALK, APC, BRAF, BRCA1, BRCA2, CDKN2A, CDKN2B, CTNNB1, ERBB2, ERBB3, FGFR3, KIT, LRP1B, MET, MLH1, MSH3, NOTCH1, NTRK1, RET, ROS1, STK11, TP53, and VEGFA.

[0031] In one embodiment, the cancer is lung cancer, gastric cancer, colorectal cancer, brain cancer, cancer originating from epithelial cells, breast cancer, ovarian cancer, colorectal cancer, anal cancer, prostate cancer, kidney cancer, bladder cancer, head and neck cancer, pharyngeal cancer, nasal cancer, pancreatic cancer, skin cancer, oral cancer, tongue cancer, esophageal cancer, vaginal cancer, cervical cancer, spleen cancer, testicular cancer, stomach cancer, thymic cancer, colon cancer, thyroid cancer, liver cancer, hepatocellular carcinoma (HCC), or sporadic or hereditary papillary renal cell carcinoma (PRCC), or any combination thereof.

[0032] In one embodiment, the lung cancer is non-small cell lung cancer (NSCLC), small cell lung cancer (SCLC) or lung adenocarcinoma, lung sarcomatoid carcinoma, or any combination thereof.

[0033] In one embodiment, the method includes further administering to the subject one or more anti-cancer therapies.

[0034] In one embodiment, the one or more anti-cancer therapies include chemotherapy, radiation therapy, surgery, targeted anti-cancer therapy, kinase inhibitors, or any combination thereof.

[0035] In one embodiment, the kinase inhibitor is an inhibitor of EGFR, an inhibitor of c-Met, an inhibitor of HER2, an inhibitor of HER3, an inhibitor of HER4, an inhibitor of VEGFR, or an inhibitor of AXL.

[0036] In one embodiment, the kinase inhibitor is erlotinib, gefitinib, lapatinib, vandetanib, afatinib, osimertinib, lazertinib, poziotinib, cliotinib, cabozantinib, capmatinib, axitinib, lenvatinib, nintedanib, regorafenib, pazopanib, sorafenib, or sunitinib.

[0037] In one embodiment, the bispecific anti-EGFR / c-Met antibody is administered at a dose of between about 140 mg and about 2240 mg.

[0038] In one embodiment, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 700 mg, about 750 mg, about 800 mg, about 850 mg, about 900 mg, about 950 mg, about 1000 mg, about 1050 mg, about 1100 mg, about 1150 mg, about 1200 mg, about 1250 mg, about 1300 mg, about 1350 mg, about 1400 mg, about 1450 mg, about 1500 mg, about 1550 mg, about 1575 mg, about 1600 mg, about 1650 mg, about 1700 mg, about 1750 mg, about 1800 mg, about 1850 mg, about 1900 mg, about 1950 mg, about 2000 mg, about 2050 mg, about 2100 mg, about 2150 mg, about 2200, or about 2240 mg.

[0039] In one embodiment, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 1050 mg.

[0040] In one embodiment, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 1400 mg.

[0041] In one embodiment, the bispecific anti-EGFR / c-Met antibody is administered twice a week, once a week, once every two weeks, once every three weeks, or once every four weeks.

[0042] In one embodiment, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 1575 mg.

[0043] In one embodiment, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 1600 mg.

[0044] In one embodiment, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 2100 mg.

[0045] In one embodiment, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 2240 mg.

[0046] In one embodiment, the bispecific anti-EGFR / c-Met antibody is administered twice a week, once a week, once every two weeks, once every three weeks, or once every four weeks.

[0047] In one embodiment, the bispecific anti-EGFR / c-Met antibody is administered intravenously.

[0048] In one embodiment, the bispecific anti-EGFR / c-Met antibody is administered subcutaneously. [Brief description of the drawings]

[0049] [Figure 1A] Receptor expression as IHC scores plotted against signaling as PLA scores for EGFR (FIG. 1A) and MET (FIG. 1B). [Figure 1B] Receptor expression as IHC scores plotted against signaling as PLA scores for EGFR (FIG. 1A) and MET (FIG. 1B). [Figure 2A] Representative in vivo efficacy plots of amivantamab, Fc-silenced EGFR / MET, or isotype control in mouse xenograft tumors for LXFA677 (Figure 2A), LXFA1584 (Figure 2B), and LXFA2158 (Figure 2C). [Figure 2B] Representative in vivo efficacy plots of amivantamab, Fc-silenced EGFR / MET, or isotype control in mouse xenograft tumors for LXFA677 (Figure 2A), LXFA1584 (Figure 2B), and LXFA2158 (Figure 2C). [Figure 2C] Representative in vivo efficacy plots of amivantamab, Fc-silenced EGFR / MET, or isotype control in mouse xenograft tumors for LXFA677 (Figure 2A), LXFA1584 (Figure 2B), and LXFA2158 (Figure 2C). [Figure 3A]Amivantamab efficacy is shown as % tumor growth inhibition (%TGI) in selected PDX models harboring EGFR lacking activating mutations plotted with respect to EGFR IHC H score (Fig. 3A) and PLA score (Fig. 3B), and MET IHC H score (Fig. 3C) and PLA score (Fig. 3D). [Figure 3B] Amivantamab efficacy is shown as % tumor growth inhibition (%TGI) in selected PDX models harboring EGFR lacking activating mutations plotted with respect to EGFR IHC H score (Fig. 3A) and PLA score (Fig. 3B), and MET IHC H score (Fig. 3C) and PLA score (Fig. 3D). [Figure 3C] Amivantamab efficacy is shown as % tumor growth inhibition (%TGI) in selected PDX models harboring EGFR lacking activating mutations plotted with respect to EGFR IHC H score (Fig. 3A) and PLA score (Fig. 3B), and MET IHC H score (Fig. 3C) and PLA score (Fig. 3D). [Figure 3D] Amivantamab efficacy is shown as % tumor growth inhibition (%TGI) in selected PDX models harboring EGFR lacking activating mutations plotted with respect to EGFR IHC H score (Fig. 3A) and PLA score (Fig. 3B), and MET IHC H score (Fig. 3C) and PLA score (Fig. 3D). [Figure 4A] Expression (Figure 4A) and mutation status (Figure 4B) of common oncogenes in PDX models with EGFR lacking activating mutations are shown, and efficacy was tested and shown as % tumor growth inhibition (%TGI) (Figure 4C). Arrows indicate models with mutations in the KRAS or PI3K pathways. [Figure 4B] Expression (Figure 4A) and mutation status (Figure 4B) of common oncogenes in PDX models with EGFR lacking activating mutations are shown, and efficacy was tested and shown as % tumor growth inhibition (%TGI) (Figure 4C). Arrows indicate models with mutations in the KRAS or PI3K pathways. [Figure 4C]Expression (Figure 4A) and mutation status (Figure 4B) of common oncogenes in PDX models with EGFR lacking activating mutations are shown, and efficacy was tested and shown as % tumor growth inhibition (%TGI) (Figure 4C). Arrows indicate models with mutations in the KRAS or PI3K pathways. [Figure 5A] Correlation plots of amivantamab efficacy, shown as % tumor growth inhibition (%TGI) against combined IHC H-score and PLA score (IHC+PLA), for EGFR (Figure 5A) and MET (Figure 5B) in select models are shown. [Figure 5B] Correlation plots of amivantamab efficacy, shown as % tumor growth inhibition (%TGI) against combined IHC H-score and PLA score (IHC+PLA), for EGFR (Figure 5A) and MET (Figure 5B) in select models are shown. [Figure 6] FIG. 1 shows a correlation plot of amivantamab efficacy, shown as % tumor growth inhibition (%TGI) against expression of the EGFR ligand amphiregulin (AREG), as measured by RNA-Seq using Transcripts Per Kilobase Million (TPM). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0050] definition All publications, including but not limited to patents and patent applications, cited in this specification are herein incorporated by reference as if fully set forth.

[0051] It should be understood that the terms used herein are used only to describe particular embodiments and are not intended to be limiting. Unless otherwise specified, 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 invention belongs.

[0052] Although any methods and materials similar or equivalent to those described herein can be used in the practice for testing of the present invention, exemplary materials and methods are described herein. In describing and claiming the present invention, the following terminology is used.

[0053] When lists are presented, it is to be understood that each individual member of that list and every combination of that list is a separate embodiment, unless otherwise specified. For example, a list of embodiments presented as "A, B, or C" should be interpreted to include the embodiments "A," "B," "C," "A or B," "A or C," "B or C," or "A, B, or C."

[0054] As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the content clearly dictates otherwise. Thus, for example, reference to "a cell" includes a combination of two or more cells, and the like.

[0055] The conjunction phrase "and / or" between multiple listed elements is understood to encompass both individual and combined options. For example, when two elements are connected by "and / or," the first option refers to the first element being applicable without the second element. The second option refers to the second element being applicable without the first element. The third option refers to the first and second elements being applicable together. Any one of these options is understood to be within the meaning and thus meet the requirements of the term "and / or" as used herein. The simultaneous applicability of two or more of the options is also understood to be within the meaning and thus meet the requirements of the term "and / or."

[0056] The transitional phrases "comprising," "consisting essentially of," and "consisting of" are intended to connote their generally accepted meanings in patent language, i.e., (i) "comprising" is synonymous with "including," "containing," or "characterized by" and is inclusive or open-ended and does not exclude other unrecited elements or method steps, (ii) "consisting of" excludes any element, step, or ingredient not specified in the claim, and (iii) "consisting essentially of" limits the scope of the claim to the materials or steps specified, and those that do not materially affect the "basic and novel characteristics" of the claimed invention. Embodiments described with the phrase "comprising" (or its equivalents) are also provided as embodiments described independently with "consisting of" and "consisting essentially of."

[0057] "Co-administration," "administration with," "administration in combination," "in combination with," and the like are intended to encompass administration of selected therapeutic agents or drugs to a single patient, and include therapeutic regimens in which the therapeutic agents or drugs are administered by the same or different routes of administration or at the same or different times.

[0058] "Isolated" refers to a homogenous population of molecules (e.g., synthetic polynucleotides, polypeptides, vectors, or viruses) that have been substantially separated and / or purified away from other components of the system in which they are produced, such as recombinant cells, as well as to proteins that have been subjected to at least one purification or isolation step. "Isolated" refers to molecules that are substantially free of other cellular material and / or chemicals, and includes molecules that have been isolated to greater degrees of purity, e.g., 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% purity.

[0059] "Treating," "treating," or "treatment" of a disease or disorder, such as cancer, refers to achieving one or more of the following: reducing the severity and / or duration of the disorder, inhibiting the worsening of symptoms characteristic of the disorder being treated, limiting or preventing the recurrence of the disorder in a subject who previously had the disorder, or limiting or preventing the recurrence of symptoms in a subject who was previously symptomatic for the disorder.

[0060] "Prevent," "preventing," "prevention," or "prophylaxis" of a disease or disorder means barring the occurrence of the disorder in a subject.

[0061] "Diagnosing" or "diagnosis" refers to a method of determining whether a subject is afflicted with a given disease or condition, or may develop a given disease or condition in the future, or is likely to respond to treatment for a previously diagnosed disease or condition, i.e., stratifying a patient population for the likelihood of responding to treatment. Diagnosis is typically made by a physician based on a general guide to the disease being diagnosed, or other criteria that indicate that the subject is likely to respond to a particular treatment.

[0062] "Responsive," "responsive," or "likely to respond" refers to any type of improvement or positive response, whether detectable or undetectable, such as reduction or amelioration of one or more symptoms, reduction in the extent of disease, stabilized (i.e., not worsening) disease state, prevention of the spread of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, and remission (whether partial or total).

[0063] "Newly diagnosed" refers to a subject who has been diagnosed with an EGFR or c-Met expressing cancer but has not yet received treatment for multiple myeloma.

[0064] "Therapeutically effective amount" refers to an amount effective to achieve the desired therapeutic result at the dosage and duration required. Therapeutically effective amounts may vary depending on factors such as the individual's condition, age, sex, and weight, as well as the ability of the therapeutic agent or combination of therapeutic agents to elicit a desired response in the individual. Exemplary indicators of an effective therapeutic agent or combination of therapeutic agents include, for example, improved health of the patient.

[0065] "Refractory" refers to a disease that does not respond to treatment. A refractory disease can be resistant to treatment before or at the start of treatment, or a refractory disease can become resistant during treatment.

[0066] "Relapse" refers to the return of a disease or signs and symptoms of a disease after a period of improvement following prior treatment with a therapeutic agent.

[0067] A "subject" includes any human or non-human animal. "Non-human animals" include all vertebrates, e.g., mammals and non-mammals, such as non-human primates, sheep, dogs, cats, horses, cows, chickens, amphibians, reptiles, etc. The terms "subject" and "patient" are used interchangeably herein.

[0068] "About" means within an acceptable range of error for a particular value as determined by one of ordinary skill in the art, which will depend in part on the limitations of the method by which the value is measured or determined, i.e., the measurement system. In the context of a particular assay, result, or embodiment, unless expressly stated otherwise in the examples or elsewhere in the specification, "about" means within one standard deviation per the practice of the art, or within a range of up to 5%, whichever is greater.

[0069] "Cancer" refers to an abnormal growth of cells that tend to proliferate uncontrolled and, in some cases, metastasize (spread) to other areas of the patient's body.

[0070] "EGFR or c-Met expressing cancer" refers to a cancer that has detectable expression of EGFR or c-Met, or has mutations or amplifications of EGFR or c-Met. EGFR or c-Met expression, amplification, and mutation status can be detected using known methods such as sequencing, fluorescent in situ hybridization, immunohistochemistry, flow cytometry, or western blotting.

[0071] "Epidermal growth factor receptor" or "EGFR" refers to human EGFR (also known as HER1 or ErbB1 (Ullrich et al., Nature 309:418-425, 1984) having the amino acid sequence set forth in UniProt identifier: P00533-1 (SEQ ID NO:21), as well as naturally occurring variants or mutants thereof.

[0072] SEQ ID NO:21:

[0073] As used herein, "hepatocyte growth factor receptor" or "c-Met" refers to human c-Met having the amino acid sequence set forth in GenBank Accession No. NP_001120972 and naturally occurring variants thereof.

[0074] A "bispecific anti-EGFR / c-Met antibody" or "bispecific EGFR / c-Met antibody" refers to a bispecific antibody having a first domain that specifically binds EGFR and a second domain that specifically binds c-Met. The domains that specifically bind EGFR and c-Met are typically VH / VL paired, and the bispecific anti-EGFR / c-Met antibody is monovalent with respect to binding to EGFR and c-Met.

[0075] "Specific binding" or "specifically binds" or "specific binding" or "binds" refers to an antibody binding to an antigen or an epitope within an antigen with higher affinity than other antigens. Typically, antibodies bind to an antigen with an equilibrium dissociation constant (K D ) is the K for binding to non-specific antigens (e.g., BSA, casein) D At least 100 times smaller than about 5 × 10 -8 M or less, for example, about 1×10 -9 M or less, approximately 1×10 -10 M or less, approximately 1×10 -11 M or less, approximately 1×10 -12 K below M D The dissociation constant can be measured using known protocols. However, an antibody that binds to an antigen or an epitope within an antigen may have cross-reactivity to other related antigens, e.g., the same antigen (homolog) from other species, such as humans or monkeys, e.g., Macaca fascicularis (cynomolgus monkey, cyno) or Pan troglodytes (chimp). A monospecific antibody binds to one antigen or one epitope, whereas a bispecific antibody binds to two different antigens or two different epitopes.

[0076] "Antibody" has a broad meaning and includes monoclonal antibodies, including murine, human, humanized, and chimeric monoclonal antibodies, antigen-binding fragments, multispecific antibodies, such as bispecific, trispecific, tetraspecific, dimeric, tetrameric, or multimeric antibodies, single-chain antibodies, domain antibodies, and immunoglobulin molecules, including any other modified form of an immunoglobulin molecule that contains an antigen-binding site of the required specificity. A "full-length antibody" is composed of two heavy chains (HC) and two light chains (LC), and multimers thereof (e.g., IgM), interconnected by disulfide bonds. Each heavy chain is composed of a heavy chain variable region (VH) and a heavy chain constant region (consisting of domains CH1, hinge, CH2, and CH3). Each light chain is composed of a light chain variable region (VL) and a light chain constant region (CL). The VH and VL regions are further divided into regions of hypervariability called complementarity determining regions (CDRs) that are interspersed with framework regions (FRs). Each VH and VL is composed of three CDR and four FR segments, arranged from the amino terminus to the carboxy terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4.

[0077] "Complementarity determining region (CDR)" is the region of an antibody that binds to an antigen. CDRs can be defined using various descriptions, such as Kabat (Wu et al. (1970) J Exp Med 132:211-50) (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md., 1991), Chothia (Chothia et al. (1987) J Mol Biol 196:901-17), IMGT (Lefranc et al. (2003) Dev Comp Immunol 27:55-77), and AbM (Martin and Thornton (1996) J Bmol Biol 263:800-15). Correspondence between the various descriptions and numbering of the variable regions has been described (see, for example, Lefranc et al. (2003) Dev Comp Immunol 27:55-77; Honegger and Pluckthun, (2001) J Mol Biol 309:657-70; and the International ImMunoGeneTics (IMGT) database; Web resources, http: / / www_imgt_org). Available programs such as abYsis by UCL Business PLC can be used to delineate CDRs. As used herein, the terms "CDR", "HCDR1", "HCDR2", "HCDR3", "LCDR1", "LCDR2" and "LCDR3" include CDRs defined by any of the methods of Kabat, Chothia, IMGT or AbM as described above, unless otherwise expressly stated herein.

[0078] Immunoglobulins can be assigned to five major classes, IgA, IgD, IgE, IgG, and IgM, depending on the amino acid sequence of the heavy chain constant domain. IgA and IgG are further subdivided into isotypes IgA1, IgA2, IgG1, IgG2, IgG3, and IgG4. The antibody light chain of any vertebrate species can be assigned to one of two clearly distinct types, kappa (κ) and lambda (λ), based on the amino acid sequence of its constant domain.

[0079] "Antigen-binding fragment" refers to a portion of an immunoglobulin molecule that binds to an antigen. Antigen-binding fragments may be synthetic, enzymatically obtainable, or recombinant polypeptides, and include VH, VL, VH and VL, Fab, F(ab')2, Fd, and Fv fragments, domain antibodies (dAbs) consisting of one VH domain or one VL domain, shark variable IgNAR domains, camelized VH domains, and the minimum recognition unit consisting of amino acid residues reproducing the CDRs of an antibody, such as the FR3-CDR3-FR4 portion, HCDR1, HCDR2, and / or HCDR3, and LCDR1, LCDR2, and / or LCDR3. The VH and VL domains can be linked to each other via synthetic linkers to form various types of single chain antibody designs; when the VH and VL domains are expressed as separate single chain antibody constructs, the VH / VL domains can pair intramolecularly or intermolecularly to form monovalent antigen binding sites, such as single chain Fvs (scFvs) or diabodies, as described, for example, in WO 1998 / 44001, WO 1988 / 01649, WO 1994 / 13804, and WO 1992 / 01047.

[0080] "Monoclonal antibody" refers to an antibody obtained from a population of substantially homogeneous antibody molecules, i.e., the individual antibodies constituting the population are identical except for possible known modifications, such as removal of C-terminal lysine from the antibody heavy chain, or post-translational modifications such as isomerization or deamidation of amino acids, oxidation of methionine, or deamidation of asparagine or glutamine. A monoclonal antibody typically binds to one antigenic epitope. A bispecific monoclonal antibody binds to two different antigenic epitopes. A monoclonal antibody may have heterogeneous glycosylation within the antibody population. A monoclonal antibody may be monospecific or multispecific, such as bispecific, monovalent, bivalent, or multivalent.

[0081] "Recombinant" refers to DNA, antibodies, and other proteins that are prepared, expressed, produced, or isolated by recombinant means when segments from different sources are joined to produce recombinant DNA, antibody, or protein.

[0082] "Bispecific" refers to an antibody that specifically binds to two different antigens or to two different epitopes within the same antigen. Bispecific antibodies may have cross-reactivity to other related antigens, e.g., the same antigen (homolog) from other species such as humans or monkeys, e.g., Macaca cynomolgus (cyno) or chimpanzees, or may bind to an epitope that is shared between two or more different antigens.

[0083] "Antagonist" or "inhibitor" refers to a molecule that, when bound to a cellular protein, inhibits at least one response or activity induced by the protein's natural ligand. A molecule is an antagonist when at least one response or activity is inhibited by at least about 20%, 30%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% more than the at least one response or activity inhibited in the absence of the antagonist (e.g., a negative control), or when the inhibition is statistically significant compared to inhibition in the absence of the antagonist.

[0084] A "PD-(L)1 axis inhibitor" refers to a molecule that inhibits PD-1 downstream signaling. A PD-(L)1 axis inhibitor may be a molecule that binds to PD-1, PD-L1, or PD-L2.

[0085] A "biological sample" refers to a collection of fluids, cells, or tissues present within a subject, in addition to similar fluids, cells, or tissues isolated from a subject. Exemplary samples are biological fluids, such as blood, serum and serous fluid, plasma, lymphatic fluid, urine, saliva, cyst fluid, tears, feces, sputum, mucosal secretions of secretory tissues and organs, vaginal secretions, ascites, fluids of the pleural, pericardial, peritoneal, abdominal cavities, and other body cavities, fluids collected by bronchial lavage, synovial fluid, liquid solutions in contact with a subject or biological source, such as cell and organ culture media, including conditioned media of cells or organs, lavage fluids, and the like, tissue biopsies, tumor tissue biopsies, tumor tissue samples, fine needle aspirations, surgically resected tissues, organ cultures, or cell cultures.

[0086] As used herein, "low fucose" or "low fucose content" refers to an antibody having a fucose content of between about 1% and 15%.

[0087] As used herein, "normal fucose" or "normal fucose content" refers to an antibody having a fucose content of greater than about 50%, typically greater than 80% or greater than 85%.

[0088] As used herein, "silent Fc" refers to an Fc domain that has been modified to have decreased binding to Fcγ receptors (FcγR) or decreased effector function such as ADCC, ADCP and / or CDC compared to an unmodified Fc. The Fc modifications can be mutations at positions 214, 233, 234, 235, 236, 237, 238, 265, 267, 268, 270, 295, 297, 309, 327, 328, 329, 330, 331 or 365. Exemplary mutations that can be made alone or in combination are the mutations K214T, E233P, L234V, L234A, deletion of G236, V234A, F234A, L235A, G237A, P238A, P238S, D265A, S267E, H268A, H268Q, Q268A, N297A, A327Q, P329A, D270A, Q295A, V309L, A327S, L328F, A330S, and P331S in IgG1, IgG2, IgG3, or IgG4. Exemplary mutation combinations that result in antibodies with reduced ADCC include the mutations L234A / L235A in IgG1, V234A / G237A / P238S / H268A / V309L / A330S / P331S in IgG2, F234A / L235A in IgG4, S228P / F234A / L235A in IgG4, N297A in all Ig isotypes, V234A / G237A in IgG2, K214T / E233P / L234V / L235A / G236 deletion / A327G / in IgG1, P331A / D365E / L358M, H268Q / V309L / A330S / P331S in IgG2, S267E / L328F in IgG1, L234F / L235E / D265A in IgG1, L234A / L235A / G237A / P238S / H268A / A330S / P331S in IgG1, S228P / F234A / L235A / G237A / P238S in IgG4, and S228P / F234A / L235A / G236 deletion / G237A / P238S in IgG4. An exemplary mutation resulting in an antibody with reduced CDC is the K322A mutation.Residue numbering follows EU numbering (see, for example, IMGT® Web resources; IMGT® Repertoire (IG and TR); Proteins and alleles; allotypes).

[0089] Methods of the Disclosure Amivantamab or JNJ-61186372 (JNJ-372) is an IgG1 anti-EGFR / c-Met bispecific antibody described in U.S. Pat. No. 9,593,164.

[0090] The present disclosure is based, at least in part, on the discovery that amivantamab is effective in treating tumors that lack EGFR activating mutations.

[0091] EGFR activating mutations that may be associated with cancer include point mutations, deletion mutations, insertion mutations, inversions, or gene amplifications that result in an increase in at least one biological activity of EGFR (such as increased tyrosine kinase activity), enhanced ligand binding, ligand-independent signaling, increased cell proliferation, signaling to the MAPK / ERK pathway, gene transcription, receptor homodimer and heterodimer formation, dimerization (EGFR:EGFR), heterodimerization (EGFR:HER2 or EGFR:HER3). The mutations may be located in any part of the EGFR gene or the regulatory region associated with the EGFR gene, including mutations in exons 18, 19, 20, or 21, or mutations in the kinase domain. Other examples of EGFR activating mutations are known in the art (see, for example, U.S. Patent Application Publication No. 2005 / 0272083). Information regarding EGFR and other ErbB receptors, including receptor homo- and heterodimers, receptor ligands, autophosphorylation sites, and signaling molecules involved in ErbB-mediated signal transduction, is known in the art (see, e.g., Hynes and Lane, Nature Reviews Cancer 5:341-354, 2005).

[0092] In some embodiments, the EGFR activating mutation is a substitution of L718Q, G719A, G719X (where X is any amino acid), L861X (where X is any amino acid), L858R, E746K, L747S, E749Q, A750P, A755V, V765M, C797S, L858P, or T790M, a deletion of E746 to A750, a deletion of R748 to P753, an insertion of Ala (A) between M766 and A767, an insertion of Ser, Val, and Ala (SVA) between S768 and V769, a deletion of P746 to P753, an insertion of Ala (A) between S768 and V769 ... EGFR exon 20 mutations include Asn and Ser (NS) insertions between EGFR exon 20 and H773, D761 and E762, A763 and Y764, Y764 and Y765, M766 and A767, A767 and V768, S768 and V769, V769 and D770, D770 and N771, N771 and P772, P772 and H773, H773 and V774, V774 and C775, one or more amino acid insertions, one or more deletions in EGFR exon 20, or one or more insertions in EGFR exon 20, or any combination thereof. Subjects with EGFR exon 20 mutations (insertion of one or more amino acids) are generally resistant to EGFR tyrosine kinase inhibitors (TKIs) (see, e.g., WO 2018 / 094225).

[0093] In some embodiments, the EGFR activating mutations include one or more rare EGFR activating mutations, such as S768I, L861Q, and G719X.

[0094] EGFR mutation status can be detected using methods known in the art, such as Sanger sequencing, next-generation sequencing (NGS), whole exome sequencing (WES), RNA-Seq, fluorescent in situ hybridization, or immunohistochemistry.

[0095] The present disclosure provides a method of treating a subject having a cancer that lacks an activating EGFR mutation, the method comprising administering to a subject having a cancer that is positive for EGFR that lacks an activating mutation a therapeutically effective amount of an isolated bispecific anti-epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) antibody.

[0096] The disclosure also provides a method of treating a subject having a lung cancer that lacks an EGFR activating mutation, the method comprising administering to the subject having a lung cancer that lacks an EGFR activating mutation a therapeutically effective amount of an isolated bispecific anti-EGFR / c-Met antibody.

[0097] The present disclosure also provides a method of treating a subject having non-small cell lung cancer (NSCLC) that lacks an EGFR activating mutation, comprising administering to the subject having NSCLC that lacks an EGFR activating mutation a therapeutically effective amount of an isolated bispecific anti-EGFR / c-Met antibody.

[0098] The disclosure also provides a method of treating a subject having small cell lung cancer (SCLC) that lacks an EGFR activating mutation, comprising administering to a subject having SCLC that lacks an EGFR activating mutation a therapeutically effective amount of an isolated bispecific anti-EGFR / c-Met antibody.

[0099] The disclosure also provides a method of treating a subject having a lung adenocarcinoma positive for EGFR lacking activating mutations, comprising administering to a subject having a lung adenocarcinoma positive for EGFR lacking activating mutations a therapeutically effective amount of an isolated bispecific anti-EGFR / c-Met antibody.

[0100] The present disclosure also provides a method of treating a subject with cancer with a bispecific anti-EGFR / c-Met antibody, comprising: Providing a biological sample from a subject; determining the presence or absence of EGFR lacking an activating mutation in the sample; administering or providing for administration of a bispecific anti-EGFR / c-Met antibody to a subject determined to have an EGFR that lacks an activating mutation.

[0101] In some embodiments, the biological sample is a blood sample.

[0102] In some embodiments, the biological sample is a tumor tissue biopsy.

[0103] In some embodiments, the bispecific anti-EGFR / c-Met antibody is The antibody comprises a first domain that specifically binds to EGFR and a second domain that specifically binds to c-Met, wherein the first domain comprises heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 1, HCDR2 of SEQ ID NO: 2, HCDR3 of SEQ ID NO: 3, light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 4, LCDR2 of SEQ ID NO: 5, and LCDR3 of SEQ ID NO: 6, and the second domain comprises HCDR1 of SEQ ID NO: 7, HCDR2 of SEQ ID NO: 8, HCDR3 of SEQ ID NO: 9, LCDR1 of SEQ ID NO: 10, LCDR2 of SEQ ID NO: 11, and LCDR3 of SEQ ID NO: 12.

[0104] In some embodiments, a first domain that specifically binds EGFR comprises a heavy chain variable region (VH) of SEQ ID NO: 13 and a light chain variable region (VL) of SEQ ID NO: 14, and a second domain that specifically binds c-Met comprises a VH of SEQ ID NO: 15 and a VL of SEQ ID NO: 16.

[0105] In some embodiments, the bispecific anti-EGFR / c-Met antibody is of the IgG1 isotype.

[0106] In some embodiments, the bispecific anti-EGFR / c-Met antibody comprises a first heavy chain (HC1) of SEQ ID NO: 17, a first light chain (LC1) of SEQ ID NO: 18, a second heavy chain (HC2) of SEQ ID NO: 19, and a second light chain (LC2) of SEQ ID NO: 20.

[0107] In some embodiments, the bispecific anti-EGFR / c-Met antibody comprises a biantennary glycan structure having a fucose content of between approximately 1% and about 15%.

[0108] Antibodies with low fucose content can be produced using various methods that have been reported to successfully express relatively highly defucosylated antibodies with biantennary complex type Fc oligosaccharides, such as: control of the osmolarity of the culture (Konno et al., Cytotechnology 64(:249-65, 2012), application of the variant CHO line Lec13 as a host cell line (Shields et al., J Biol Chem 277:26733-26740, 2002), application of the variant CHO line EB66 as a host cell line (Olivier et al., MAbs;2(4),2010;Epub ahead of print;PMID:20562582), application of the rat hybridoma cell line YB2 / 0 as a host cell line (Shinkawa et al., J Biol Chem 278:3466-3473, 2003), introduction of small interfering RNA specific for the α1,6-fucosyltransferase (FUT8) gene (Morie et al., Biotechnol Bioeng 88:901-908, 2004), or co-expression of β-1,4-N-acetylglucosaminyltransferase III and Golgi α-mannosidase II or kifunensine, a potent α-mannosidase I inhibitor (Ferrara et al., J Biol Chem 281:5032-5036, 2006; Ferrara et al., Biotechnol Bioeng 93:851-861, 2006; Xhou et al., Biotechnol Bioeng 99:652-65, 2008). In general, reducing the fucose content in the glycans of antibodies enhances antibody-mediated cellular cytotoxicity (ADCC).

[0109] The disclosure also provides a method of treating a subject having a cancer that lacks an EGFR activating mutation, comprising administering to a subject having a cancer that is positive for EGFR that lacks an activating mutation a therapeutically effective amount of an isolated bispecific anti-EGFR / c-Met antibody, wherein the bispecific anti-EGFR / c-Met antibody comprises a first domain that specifically binds EGFR and a second domain that specifically binds c-Met, wherein the first domain comprises an HCDR1 of SEQ ID NO: 1, an HCDR2 of SEQ ID NO: 2, an HCDR3 of SEQ ID NO: 3, an LCDR1 of SEQ ID NO: 4, an LCDR2 of SEQ ID NO: 5, and an LCDR3 of SEQ ID NO: 6, and the second domain comprises an HCDR1 of SEQ ID NO: 7, an HCDR2 of SEQ ID NO: 8, an HCDR3 of SEQ ID NO: 9, an LCDR1 of SEQ ID NO: 10, an LCDR2 of SEQ ID NO: 11, and an LCDR3 of SEQ ID NO: 12.

[0110] The disclosure also provides a method of treating a subject having a lung cancer that lacks an EGFR activating mutation, comprising administering to the subject having a lung cancer that lacks an EGFR activating mutation a therapeutically effective amount of an isolated bispecific anti-EGFR / c-Met antibody, wherein the bispecific anti-EGFR / c-Met antibody comprises a first domain that specifically binds EGFR and a second domain that specifically binds c-Met, wherein the first domain comprises an HCDR1 of SEQ ID NO: 1, an HCDR2 of SEQ ID NO: 2, an HCDR3 of SEQ ID NO: 3, an LCDR1 of SEQ ID NO: 4, an LCDR2 of SEQ ID NO: 5, and an LCDR3 of SEQ ID NO: 6, and the second domain comprises an HCDR1 of SEQ ID NO: 7, an HCDR2 of SEQ ID NO: 8, an HCDR3 of SEQ ID NO: 9, an LCDR1 of SEQ ID NO: 10, an LCDR2 of SEQ ID NO: 11, and an LCDR3 of SEQ ID NO: 12.

[0111] The disclosure also provides a method of treating a subject having NSCLC lacking an EGFR activating mutation, comprising administering to the subject having NSCLC lacking an EGFR activating mutation a therapeutically effective amount of an isolated bispecific anti-EGFR / c-Met antibody, wherein the bispecific anti-EGFR / c-Met antibody comprises a first domain that specifically binds to EGFR and a second domain that specifically binds to c-Met, wherein the first domain comprises HCDR1 of SEQ ID NO: 1, HCDR2 of SEQ ID NO: 2, HCDR3 of SEQ ID NO: 3, LCDR1 of SEQ ID NO: 4, LCDR2 of SEQ ID NO: 5, and LCDR3 of SEQ ID NO: 6, and the second domain comprises HCDR1 of SEQ ID NO: 7, HCDR2 of SEQ ID NO: 8, HCDR3 of SEQ ID NO: 9, LCDR1 of SEQ ID NO: 10, LCDR2 of SEQ ID NO: 11, and LCDR3 of SEQ ID NO: 12.

[0112] The disclosure also provides a method of treating a subject with SCLC lacking an EGFR activating mutation, comprising administering to the subject with SCLC lacking an EGFR activating mutation a therapeutically effective amount of a bispecific anti-EGFR / c-Met antibody, wherein the bispecific anti-EGFR / c-MET antibody comprises a first domain that specifically binds EGFR and a second domain that specifically binds c-MET, wherein the first domain comprises an HCDR1 of SEQ ID NO: 1, an HCDR2 of SEQ ID NO: 2, an HCDR3 of SEQ ID NO: 3, an LCDR1 of SEQ ID NO: 4, an LCDR2 of SEQ ID NO: 5, and an LCDR3 of SEQ ID NO: 6, and the second domain comprises an HCDR1 of SEQ ID NO: 7, an HCDR2 of SEQ ID NO: 8, an HCDR3 of SEQ ID NO: 9, an LCDR1 of SEQ ID NO: 10, an LCDR2 of SEQ ID NO: 11, and an LCDR3 of SEQ ID NO: 12.

[0113] The disclosure also provides a method of treating a subject having a lung adenocarcinoma that lacks an EGFR activating mutation, comprising administering to the subject having a lung adenocarcinoma that lacks an EGFR activating mutation a therapeutically effective amount of an isolated bispecific anti-EGFR / c-Met antibody, wherein the bispecific anti-EGFR / c-Met antibody comprises a first domain that specifically binds EGFR and a second domain that specifically binds c-MET, wherein the first domain comprises an HCDR1 of SEQ ID NO: 1, an HCDR2 of SEQ ID NO: 2, an HCDR3 of SEQ ID NO: 3, an LCDR1 of SEQ ID NO: 4, an LCDR2 of SEQ ID NO: 5, and an LCDR3 of SEQ ID NO: 6, and the second domain comprises an HCDR1 of SEQ ID NO: 7, an HCDR2 of SEQ ID NO: 8, an HCDR3 of SEQ ID NO: 9, an LCDR1 of SEQ ID NO: 10, an LCDR2 of SEQ ID NO: 11, and an LCDR3 of SEQ ID NO: 12.

[0114] The disclosure provides a method of treating a subject having a cancer that lacks an EGFR activating mutation, comprising administering to the subject having a cancer that lacks an EGFR activating mutation a therapeutically effective amount of an isolated bispecific anti-EGFR / c-Met antibody, wherein the bispecific anti-EGFR / c-Met antibody comprises a first domain that specifically binds EGFR and a second domain that specifically binds c-Met, wherein the first domain comprises a VH of SEQ ID NO: 13 and a VL of SEQ ID NO: 14, and the second domain comprises a VH of SEQ ID NO: 15 and a VL of SEQ ID NO: 16.

[0115] The disclosure also provides a method of treating a subject having lung cancer that lacks an EGFR activating mutation, comprising administering to the subject having lung cancer that lacks an EGFR activating mutation a therapeutically effective amount of an isolated bispecific anti-EGFR / c-Met antibody, wherein the bispecific anti-EGFR / c-Met antibody comprises a first domain that specifically binds EGFR and a second domain that specifically binds c-Met, wherein the first domain comprises a VH of SEQ ID NO: 13 and a VL of SEQ ID NO: 14, and the second domain comprises a VH of SEQ ID NO: 15 and a VL of SEQ ID NO: 16.

[0116] The disclosure also provides a method of treating a subject having NSCLC that lacks an EGFR activating mutation, comprising administering to the subject having NSCLC that lacks an EGFR activating mutation a therapeutically effective amount of an isolated bispecific anti-EGFR / c-Met antibody, wherein the bispecific anti-EGFR / c-Met antibody comprises a first domain that specifically binds EGFR and a second domain that specifically binds c-Met, wherein the first domain comprises a VH of SEQ ID NO: 13 and a VL of SEQ ID NO: 14, and the second domain comprises a VH of SEQ ID NO: 15 and a VL of SEQ ID NO: 16.

[0117] The disclosure also provides a method of treating a subject with SCLC that lacks EGFR activating mutations, comprising administering to a subject with SCLC that is positive for EGFR that lacks activating mutations a therapeutically effective amount of an isolated bispecific anti-EGFR / c-Met antibody, wherein the bispecific anti-EGFR / c-Met antibody comprises a first domain that specifically binds EGFR and a second domain that specifically binds c-Met, wherein the first domain comprises a VH of SEQ ID NO: 13 and a VL of SEQ ID NO: 14, and the second domain comprises a VH of SEQ ID NO: 15 and a VL of SEQ ID NO: 16.

[0118] The disclosure also provides a method of treating a subject having a lung adenocarcinoma that lacks an EGFR activating mutation, comprising administering to a subject having a lung adenocarcinoma that is positive for EGFR that lacks an activating mutation a therapeutically effective amount of an isolated bispecific anti-EGFR / c-Met antibody, wherein the bispecific anti-EGFR / c-Met antibody comprises a first domain that specifically binds EGFR and a second domain that specifically binds c-Met, wherein the first domain comprises a VH of SEQ ID NO: 13 and a VL of SEQ ID NO: 14, and the second domain comprises a VH of SEQ ID NO: 15 and a VL of SEQ ID NO: 16.

[0119] The disclosure provides a method of treating a subject having a cancer that lacks an EGFR activating mutation, comprising administering to a subject having a cancer that is positive for EGFR that lacks an activating mutation a therapeutically effective amount of an isolated bispecific anti-EGFR / c-Met antibody, wherein the bispecific anti-EGFR / c-Met antibody is of IgG1 isotype and comprises a first domain that specifically binds EGFR and a second domain that specifically binds c-Met, wherein the first domain comprises a VH of SEQ ID NO: 13 and a VL of SEQ ID NO: 14, and the second domain comprises a VH of SEQ ID NO: 15 and a VL of SEQ ID NO: 16.

[0120] The disclosure also provides a method of treating a subject having lung cancer that lacks an EGFR activating mutation, comprising administering to a subject having lung cancer that is positive for EGFR that lacks an activating mutation a therapeutically effective amount of an isolated bispecific anti-EGFR / c-Met antibody, wherein the bispecific anti-EGFR / c-Met antibody is of IgG1 isotype and comprises a first domain that specifically binds EGFR and a second domain that specifically binds c-Met, wherein the first domain comprises a VH of SEQ ID NO: 13 and a VL of SEQ ID NO: 14, and the second domain comprises a VH of SEQ ID NO: 15 and a VL of SEQ ID NO: 16.

[0121] The disclosure also provides a method of treating a subject having NSCLC lacking an EGFR activating mutation, comprising administering to a subject having NSCLC positive for EGFR lacking an activating mutation a therapeutically effective amount of an isolated bispecific anti-EGFR / c-Met antibody, wherein the bispecific anti-EGFR / c-Met antibody is of IgG1 isotype and comprises a first domain that specifically binds EGFR and a second domain that specifically binds c-Met, wherein the first domain comprises a VH of SEQ ID NO: 13 and a VL of SEQ ID NO: 14, and the second domain comprises a VH of SEQ ID NO: 15 and a VL of SEQ ID NO: 16.

[0122] The disclosure also provides a method of treating a subject with SCLC that lacks EGFR activating mutations, comprising administering to a subject with SCLC that is positive for EGFR that lacks activating mutations a therapeutically effective amount of an isolated bispecific anti-EGFR / c-Met antibody, wherein the bispecific anti-EGFR / c-Met antibody is of IgG1 isotype and comprises a first domain that specifically binds EGFR and a second domain that specifically binds c-Met, wherein the first domain comprises a VH of SEQ ID NO: 13 and a VL of SEQ ID NO: 14, and the second domain comprises a VH of SEQ ID NO: 15 and a VL of SEQ ID NO: 16.

[0123] The disclosure also provides a method of treating a subject having a lung adenocarcinoma that lacks an EGFR activating mutation, comprising administering to a subject having a lung adenocarcinoma that is positive for EGFR that lacks an activating mutation a therapeutically effective amount of an isolated bispecific anti-EGFR / c-Met antibody, wherein the bispecific anti-EGFR / c-Met antibody is of IgG1 isotype and comprises a first domain that specifically binds EGFR and a second domain that specifically binds c-Met, wherein the first domain comprises a VH of SEQ ID NO: 13 and a VL of SEQ ID NO: 14, and the second domain comprises a VH of SEQ ID NO: 15 and a VL of SEQ ID NO: 16.

[0124] In some embodiments, the bispecific anti-EGFR / c-Met antibody is of the IgG1 isotype. There are some variations (e.g., known allotypes) within the IgG1 constant domain, with variations at positions 214, 356, 358, 422, 431, 435, or 436 (numbering of residues according to EU numbering) (see, e.g., IMGT Web resources; IMGT Repertoire (IG and TR); Proteins and alleles; allotypes). The bispecific anti-EGFR / c-Met antibody may be of any IgG1 allotype, such as G1m17, G1m3, G1m1, G1m2, G1m27, or G1m28.

[0125] The disclosure also provides a method of treating a subject having a cancer that lacks an EGFR activating mutation, comprising administering to a subject having a cancer that is positive for EGFR that lacks an activating mutation a therapeutically effective amount of an isolated bispecific anti-EGFR / c-Met antibody, wherein the bispecific anti-EGFR / c-Met antibody comprises an HC1 of SEQ ID NO: 17, an LC1 of SEQ ID NO: 18, an HC2 of SEQ ID NO: 19, and an LC2 of SEQ ID NO: 20.

[0126] The disclosure also provides a method of treating a subject having lung cancer that lacks an EGFR activating mutation, comprising administering to a subject having lung cancer that is positive for EGFR that lacks an activating mutation a therapeutically effective amount of an isolated bispecific anti-EGFR / c-Met antibody, wherein the bispecific anti-EGFR / c-Met antibody comprises an HC1 of SEQ ID NO: 17, an LC1 of SEQ ID NO: 18, an HC2 of SEQ ID NO: 19, and an LC2 of SEQ ID NO: 20.

[0127] The disclosure also provides a method of treating a subject having NSCLC lacking an EGFR activating mutation, comprising administering to a subject having NSCLC that is positive for EGFR lacking an activating mutation a therapeutically effective amount of an isolated bispecific anti-EGFR / c-Met antibody, wherein the bispecific anti-EGFR / c-Met antibody comprises an HC1 of SEQ ID NO: 17, an LC1 of SEQ ID NO: 18, an HC2 of SEQ ID NO: 19, and an LC2 of SEQ ID NO: 20.

[0128] The disclosure also provides a method of treating a subject having SCLC that lacks an EGFR activating mutation, comprising administering to a subject having SCLC that is positive for EGFR that lacks an activating mutation a therapeutically effective amount of an isolated bispecific anti-EGFR / c-Met antibody, wherein the bispecific anti-EGFR / c-Met antibody comprises an HC1 of SEQ ID NO: 17, an LC1 of SEQ ID NO: 18, an HC2 of SEQ ID NO: 19, and an LC2 of SEQ ID NO: 20.

[0129] The disclosure also provides a method of treating a subject having a lung adenocarcinoma that lacks an EGFR activating mutation, the method comprising administering to the subject having a lung adenocarcinoma that is positive for EGFR that lacks an activating mutation a therapeutically effective amount of an isolated bispecific anti-EGFR / c-Met antibody, wherein the bispecific anti-EGFR / c-Met antibody comprises an HC1 of SEQ ID NO: 17, an LC1 of SEQ ID NO: 18, an HC2 of SEQ ID NO: 19, and an LC2 of SEQ ID NO: 20.

[0130] In some embodiments, the subject is relapsed or refractory to treatment with one or more previous anti-cancer therapies.

[0131] In some embodiments, the one or more previous anti-cancer therapies include one or more chemotherapeutic agents, checkpoint inhibitors, targeted anti-cancer therapies, or kinase inhibitors, or any combination thereof.

[0132] In some embodiments, the kinase inhibitor is an inhibitor of EGFR, an inhibitor of c-Met, an inhibitor of HER2, an inhibitor of HER3, an inhibitor of HER4, an inhibitor of VEGFR, or an inhibitor of AXL.

[0133] In some embodiments, the kinase inhibitor is erlotinib, gefitinib, lapatinib, vandetanib, afatinib, osimertinib, lazertinib, poziotinib, cliotinib, cabozantinib, capmatinib, axitinib, lenvatinib, nintedanib, regorafenib, pazopanib, sorafenib, or sunitinib.

[0134] In some embodiments, the one or more prior anticancer therapies comprise carboplatin, paclitaxel, gemcitabine, cisplatin, vinorelbine, docetaxel, palbociclib, crizotinib, PD-(L)1 axis inhibitors, inhibitors of EGFR, inhibitors of c-Met, inhibitors of HER2, inhibitors of HER3, inhibitors of HER4, inhibitors of VEGFR, inhibitors of AXL, erlotinib, gefitinib, lapatinib, vandetanib, afatinib, osimertinib, lazertinib, poziotinib, cliotinib, cabozantinib, capmatinib, axitinib, lenvatinib, nintedanib, regorafenib, pazopanib, sorafenib, or sunitinib, or any combination thereof.

[0135] In some embodiments, the subject is resistant or has acquired resistance to an EGFR inhibitor. Exemplary EGFR inhibitors to which cancers may develop resistance include the anti-EGFR antibodies cetuximab (ERBITUX®), panchinumumab (VECTIBIX®), matuzumab, and nimotuzumab; the small molecule EGFR inhibitors erlotinib (TARCEVA®), gefitinib (IRESSA®), EKB-569 (pelitinib, an irreversible EGFR TKI); the pan-ErbB and other receptor tyrosine kinase inhibitors lapatinib (EGFR and HER2 inhibitor), pelitinib (EGFR and HER2 inhibitor), vandetanib (ZD6474, ZACTIMA™, an EGFR, VEGFR2, and RET TKI); PF00299804 (dacomitinib, an irreversible pan-ErbB TKI); CI-1033 (an irreversible pan-erbB TKI); TKI), afatinib (BIBW2992, irreversible pan-ErbB TKI), AV-412 (dual EGFR and ErbB2 inhibitor), EXEL-7647 (EGFR, ErbB2, GEVGR, and EphB4 inhibitor), CO-1686 (irreversible mutant-selective EGFR TKI), AZD9291 (irreversible mutant-selective EGFR TKI), and HKI-272 (neratinib, irreversible EGFR / ErbB2 inhibitor).

[0136] Various qualitative and / or quantitative methods can be used to determine whether a subject is resistant, develops resistance, or is prone to develop resistance to treatment with anti-cancer therapy. Symptoms that may be associated with resistance to anti-cancer therapy include a decline or plateau in the patient's health, an increase in tumor size, a halt or slowdown in tumor growth reduction, and / or the spread of cancerous cells from one location to other organs, tissues, or cells in the body. Re-establishment or worsening of various symptoms associated with cancer, such as loss of appetite, cognitive impairment, depression, dyspnea, fatigue, hormone disruption, neutropenia, pain, peripheral neuropathy, and sexual dysfunction, can also be indicators that a subject is developing or is prone to develop resistance to anti-cancer therapy. Symptoms associated with cancer can vary depending on the type of cancer. For example, symptoms associated with cervical cancer can include abnormal bleeding, abnormal heavy vaginal discharge, pelvic pain not associated with the normal menstrual cycle, bladder pain or pain during urination, and bleeding during regular menstrual periods, sexual intercourse, vaginal douching, or pelvic exam. Symptoms associated with lung cancer can include persistent cough, hemoptysis, shortness of breath, wheezing chest pain, loss of appetite, unintentional weight loss, and fatigue.Symptoms associated with liver cancer can include loss of appetite and weight, abdominal pain, especially in the upper right part of the abdomen, which may extend to the back and shoulders, nausea and vomiting, general weakness and fatigue, liver enlargement, abdominal swelling (ascites), and yellowing of the skin and whites of the eyes (jaundice).Those skilled in the art of oncology can easily identify the symptoms associated with certain types of cancer.

[0137] Exemplary PD-(L)1 axis inhibitors are antibodies that bind to PD-1, such as nivolumab (OPDIVO®), pembrolimab (KEYTRUDA®), sintilimab, cemiplimab (LIBTAYO®), tripolibamab, tislelizumab, spartalizumab, camrelizumab, dostralimab, genolimuzumab, or cetrelimab, or antibodies that bind to PD-L1, e.g., PD-L1 antibodies are embafolimab, atezolizumab (TECENTRIQ®), durvalumab (IMFINZI®), and avelumab (BAVENCIO®).

[0138] Commercially available antibodies can be purchased through authorized distributors or pharmacies. Amino acid sequence structures of small molecules can be found in the USAN and / or INN deposits by companies from the CAS registry.

[0139] In some embodiments, the subject is treatment naive.

[0140] In some embodiments, the cancer positive for EGFR lacking activating mutations is CDK4 amplified, EGFR amplified, KRAS amplified, MDM2 amplified, TERT amplified, NF1 R2450 amplified, * ;RAD50 L597Vfs * 5, MET c.3082+3A>G, positive for increased levels of circulating HGF, c-MET amplification, or any combination thereof.

[0141] In some embodiments, the cancer positive for EGFR lacking activating mutations is positive for at least one mutation in a gene selected from the group consisting of ALK, APC, BRAF, BRCA1, BRCA2, CDKN2A, CDKN2B, CTNNB1, ERBB2, ERBB3, FGFR3, KIT, LRP1B, MET, MLH1, MSH3, NOTCH1, NTRK1, RET, ROS1, STK11, TP53, and VEGFA. In some embodiments, the at least one mutation is a mutation selected from the group consisting of point mutation, deletion mutation, insertion mutation, inversion, gene amplification, and gene fusion. The mutation can be located in any part of the gene or the regulatory region associated with the gene. The mutation can be detected using methods known in the art, such as Sanger sequencing, next generation sequencing (NGS), whole exome sequencing (WES), RNA-Seq, fluorescent in situ hybridization, or immunohistochemistry.

[0142] In some embodiments, the at least one mutation in APC is S2621C, N813S, E1317Q, or R549G.

[0143] In some embodiments, the at least one mutation in BRCA1 is M128V, G275S, Y179C, F486L, or N550H.

[0144] In some embodiments, the at least one mutation in BRCA2 is S326R, R2973H, R2034C, I283V, R672X, G25X, R468X, or I1929M (wherein X is any amino acid).

[0145] In some embodiments, at least one mutation in CDKN2A is G23X, A100X, D84H, C72X, H83N, or G111X (where X is any amino acid).

[0146] In some embodiments, at least one mutation in CTNNB1 is T41A.

[0147] In some embodiments, the at least one mutation in ERBB2 is R1146W, V1180X (wherein X is any amino acid), or A386D.

[0148] In some embodiments, the at least one mutation in ERBB3 is K998R, L1177I, or G513D.

[0149] In some embodiments, the at least one mutation in FGFR3 is G639R or E85K.

[0150] In some embodiments, the at least one mutation in LRP1B is P4512A, A3816V, T3393K, Q3636H, M1V, C1554S, S1083N, T2482S, C3522Y, G1965C, P2882T, P3372A, I1266L, L4268X (wherein X is any amino acid), S449T, E4352G, C864R, F1435I, D3697Y, V2033F, A3308S, S1281N, D1807E.

[0151] In some embodiments, the at least one mutation in MET is E168D.

[0152] In some embodiments, at least one mutation in MSH3 is E1036Q.

[0153] In some embodiments, the at least one mutation in NOTCH1 is A1696V, R1279C, E1450K, Q2184R, Q2184K, T701P, or C612Y.

[0154] In some embodiments, the at least one mutation in TP53 is R280G, P278S, E198X, H193L, R379S, V172X, G245D, L194R, H179Y, L265P, R110L, R158L, R248W, I332M, G244C, R273H, Y163C, H193R, R158L, Y103X, M237I, R273L, R273H, E171X, or R249M (wherein X is any amino acid).

[0155] In some embodiments, the at least one mutation in VEGFA is R114W, R87W, or R335C.

[0156] Exemplary c-Met activating mutations include point mutations, deletion mutations, insertion mutations, inversions, or gene amplifications that increase at least one biological activity of the c-Met protein, such as increased tyrosine kinase activity, formation of receptor homodimers and heterodimers, and enhanced ligand binding. The mutations can be located in any part of the c-Met gene or in regulatory regions associated with the gene, such as mutations in the kinase domain of c-Met. Exemplary c-Met activating mutations are mutations at residue positions N375, V13, V923, R175, V136, L229, S323, R988, S1058 / T1010, and E168. Methods for detecting EGFR and c-Met mutations or gene amplifications are well known.

[0157] In some embodiments, the mutant KRAS comprises a G12V, G12C, G12A, or G12D substitution, or any combination thereof.

[0158] In some embodiments, the cancer positive for EGFR lacking activating mutations is positive for expression of at least one EGFR ligand, including, but not limited to, epidermal growth factor (EGF), amphiregulin (AREG), transforming growth factor α (TGFα), heparin-binding EGF-like growth factor (HBEGF), betacellulin (BTC), epiregulin (EREG), and epigen (EPGN).

[0159] In some embodiments, the method of treating a cancer positive for EGFR lacking activating mutations further comprises determining the level of at least one EGFR ligand and administering or providing for administration of a bispecific anti-EGFR / c-Met antibody to a subject determined to have EGFR lacking activating mutations and determined to be positive for the gene expression or protein level of at least one EGFR ligand.

[0160] In some embodiments, methods of treating cancer positive for EGFR lacking activating mutations further include determining the level of amphiregulin and administering or providing for administration of a bispecific anti-EGFR / c-Met antibody to a subject determined to have EGFR lacking activating mutations and determined to be positive for amphiregulin gene expression or protein levels. In some embodiments, the amphiregulin gene expression or protein level can be compared to a control value.

[0161] In some embodiments, the cancer lacking an EGFR activating mutation comprises lung cancer, gastric cancer, colorectal cancer, brain cancer, cancer originating from epithelial cell cancer, breast cancer, ovarian cancer, colorectal cancer, anal cancer, prostate cancer, kidney cancer, bladder cancer, head and neck cancer, pharyngeal cancer, nasal cancer, pancreatic cancer, skin cancer, oral cancer, tongue cancer, esophageal cancer, vaginal cancer, cervical cancer, spleen cancer, testicular cancer, stomach cancer, thymic cancer, colon cancer, thyroid cancer, liver cancer, hepatocellular carcinoma (HCC), or sporadic or hereditary papillary renal cell carcinoma (PRCC), or any combination thereof. In some embodiments, the cancer lacking an EGFR activating mutation comprises lung cancer. In some embodiments, the cancer lacking an EGFR activating mutation comprises gastric cancer. In some embodiments, the cancer lacking an EGFR activating mutation comprises colorectal cancer. In some embodiments, the cancer lacking an EGFR activating mutation comprises brain cancer. In some embodiments, the cancer lacking an EGFR activating mutation comprises epithelial cell cancer. In some embodiments, the cancer that lacks EGFR activating mutations comprises breast cancer. In some embodiments, the cancer that lacks EGFR activating mutations comprises ovarian cancer. In some embodiments, the cancer that lacks EGFR activating mutations comprises colorectal cancer. In some embodiments, the cancer that lacks EGFR activating mutations comprises anal cancer. In some embodiments, the cancer that lacks EGFR activating mutations comprises prostate cancer. In some embodiments, the cancer that lacks EGFR activating mutations comprises kidney cancer. In some embodiments, the cancer that lacks EGFR activating mutations comprises bladder cancer. In some embodiments, the cancer that lacks EGFR activating mutations comprises head and neck cancer. In some embodiments, the cancer that lacks EGFR activating mutations comprises pharyngeal cancer. In some embodiments, the cancer that lacks EGFR activating mutations comprises nasal cancer. In some embodiments, the cancer that lacks EGFR activating mutations comprises pancreatic cancer. In some embodiments, the cancer that lacks EGFR activating mutations comprises skin cancer. In some embodiments, the cancer that lacks EGFR activating mutations comprises oral cancer. In some embodiments, the cancer that lacks an EGFR activating mutation comprises a tongue cancer. In some embodiments, the cancer that lacks an EGFR activating mutation comprises an esophageal cancer.In some embodiments, the cancer that lacks an EGFR activating mutation comprises vaginal cancer. In some embodiments, the cancer that lacks an EGFR activating mutation comprises cervical cancer. In some embodiments, the cancer that lacks an EGFR activating mutation comprises splenic cancer. In some embodiments, the cancer that lacks an EGFR activating mutation comprises testicular cancer. In some embodiments, the cancer that lacks an EGFR activating mutation comprises gastric cancer. In some embodiments, the cancer that lacks an EGFR activating mutation comprises thymic cancer. In some embodiments, the cancer that lacks an EGFR activating mutation comprises colon cancer. In some embodiments, the cancer that lacks an EGFR activating mutation comprises thyroid cancer. In some embodiments, the cancer that lacks an EGFR activating mutation comprises liver cancer. In some embodiments, the cancer that lacks an EGFR activating mutation comprises hepatocellular carcinoma (HCC). In some embodiments, the cancer that lacks an EGFR activating mutation comprises sporadic or hereditary papillary renal cell carcinoma (PRCC).

[0162] In some embodiments, the NSCLC comprises squamous cell carcinoma, adenocarcinoma, and large cell carcinoma. In some embodiments, the cells of the NSCLC have an epithelial phenotype. In some embodiments, the NSCLC has acquired resistance to treatment with one or more EGFR inhibitors.

[0163] In some embodiments, the subject is further administered one or more anti-cancer therapies.

[0164] In some embodiments, the one or more anti-cancer therapies comprise chemotherapy, radiation therapy, surgery, a targeted anti-cancer therapy, or a kinase inhibitor, or any combination thereof.

[0165] In some embodiments, the kinase inhibitor is an inhibitor of EGFR, an inhibitor of c-Met, an inhibitor of HER2, an inhibitor of HER3, an inhibitor of HER4, an inhibitor of VEGFR, or an inhibitor of AXL. In some embodiments, the kinase inhibitor is an inhibitor of EGFR. In some embodiments, the kinase inhibitor is an inhibitor of c-Met. In some embodiments, the kinase inhibitor is an inhibitor of HER2. In some embodiments, the kinase inhibitor is an inhibitor of HER3. In some embodiments, the kinase inhibitor is an inhibitor of HER4. In some embodiments, the kinase inhibitor is an inhibitor of VEGFR. In some embodiments, the kinase inhibitor is an inhibitor of AXL.

[0166] In some embodiments, the kinase inhibitor is erlotinib, gefitinib, lapatinib, vandetanib, afatinib, osimertinib, lazertinib, poziotinib, cliotinib, cabozantinib, capmatinib, axitinib, lenvatinib, nintedanib, regorafenib, pazopanib, sorafenib, or sunitinib.

[0167] In some embodiments, the kinase inhibitor is erlotinib. In some embodiments, the kinase inhibitor is gefitinib. In some embodiments, the kinase inhibitor is lapatinib. In some embodiments, the kinase inhibitor is vandetanib. In some embodiments, the kinase inhibitor is afatinib. In some embodiments, the kinase inhibitor is osimertinib. In some embodiments, the kinase inhibitor is lazertinib. In some embodiments, the kinase inhibitor is poziotinib. In some embodiments, the kinase inhibitor is cliotinib. In some embodiments, the kinase inhibitor is cabozantinib. In some embodiments, the kinase inhibitor is capmatinib. In some embodiments, the kinase inhibitor is axitinib. In some embodiments, the kinase inhibitor is lenvatinib. In some embodiments, the kinase inhibitor is nintedanib. In some embodiments, the kinase inhibitor is regorafenib. In some embodiments, the kinase inhibitor is pazopanib. In some embodiments, the kinase inhibitor is sorafenib. In some embodiments, the kinase inhibitor is sunitinib.

[0168] Anti-cancer therapies that may be administered in combination with the bispecific anti-EGFR / c-Met antibodies in the methods of the present disclosure include any one or more of chemotherapeutic agents or other anti-cancer therapeutic agents known to those of skill in the art. Chemotherapeutic agents are chemical compounds useful in the treatment of cancer, including growth inhibitors or other cytotoxic agents, including alkylating agents, antimetabolites, anti-microtubule inhibitors, topoisomerase inhibitors, receptor tyrosine kinase inhibitors, angiogenesis inhibitors, and the like. Examples of chemotherapeutic agents include alkylating agents such as thiotepa and cyclophosphamide (CYTOXAN®); alkylsulfonates such as busulfan, improsulfan, and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, and uredopa; ethylenimines and methylamelamines, including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trimethylolmelamine; nitrogen mustards such as chlorambucil, chlornaphazine, chlorophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembitine, phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosoureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, ranimustine;Antibiotics such as aclacinomycin, actinomycin, autramycin, azaserine, bleomycin, cactinomycin, calicheamicin, carabicin, carminomycin, carzinophilin, chromomycin, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin, epirubicin, esorubicin, idarubicin, marcellomycin, mitomycin, mycophenolic acid, nogalamycin, olivomycin, peplomycin, potfiromycin, puromycin, quelamycin, rodorubicin , streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; antimetabolites such as methotrexate and 5-FU; folic acid analogues such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogues such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogues such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine, etc; androgens such as calsterone, dromostanolone propionate, epithiostanol, mepitiostane, testolactone; adrenal cortical hormone synthesis inhibitors such as aminoglutethimide, mitotane, trilostane; folic acid supplements (folic acid replenishers, such as folinic acid; aceglatone; aldophosphamide glycosides; aminolevulinic acid; amsacrine; bestravcil; bisantrene; edatraxate; defofamine; demecolcine; diaziquone; elfornithine; elliptinium acetate; etoglucide; gallium nitrate; hydroxyurea; lentinan; lonidamine; mitoguazone; mitoxantrone; mopidamol; nitracrine; pentostatin; phenamet; pirarubicin;Podophyllic acid; 2-ethylhydrazide; procarbazine; PSK®; razoxane; schizophyllan; spirogermanium; tenuazonic acid; triaziquone; 2,2',2"-trichlorotriethylamine; urethane; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside ("Ara-C"); cyclophosphamide; thiotepa; members of the taxoid or taxane family, such as paclitaxel (TAXOL®), docetaxel (TAXOTE®), RE (registered trademark) and its analogs; chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum analogs such as cisplatin and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitomycin C; mitoxantrone; vincristine; vinorelbine; navelbine; novantrone; teniposide; daunomycin; aminopterin; xeloda; ibandronate; CPT-11, topoisomerase inhibitor RFS2000; difluoromethylornithine (DMFO); retinoic acid; esperamicin; capecitabine;Inhibitors of receptor tyrosine kinase and / or angiogenesis, including sorafenib (NEXAVAR®), sunitinib (SUTENT®), pazopanib (VOTRIENT™), toceranib (PALLADIA™), vandetanib (ZACTIMA™), cediranib (RECENTIN®), regorafenib (BAY73-4506), axitinib (AG013736), lestaurtinib (CEP-701), erlotinib (TARCEVA®), gefitinib (IRESSA™), afatinib (BIBW 2992), lapatinib (TYKERB®), neratinib (HKI-272), and the like, as well as pharmaceutical acceptable salts, acids, or derivatives of any of the foregoing. Also included in this definition are anti-hormonal agents that act to regulate or inhibit hormone action on tumors, such as antiestrogens, including tamoxifen, raloxifene, aromatase-inhibiting 4(5)-imidazole, 4-hydroxytamoxifen, trioxifene, ketoxifene, LY 117018, onapristone, and toremifene (FARESTON®); and antiandrogens, such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; as well as pharma- ceutical acceptable salts, acids, or derivatives of any of the above. Other conventional cytotoxic compounds disclosed in Wiemann et al., 1985, Medical Oncology (Calabresi et al., eds.), Chapter 10, McMillan Publishing, are also applicable to the methods of the present invention.

[0169] Administration The bispecific anti-EGFR / c-Met antibody may be administered in a pharma- ceutically acceptable carrier. "Carrier" refers to a diluent, adjuvant, excipient, or vehicle with which the antibody of the present invention is administered. Such vehicles may be liquids, such as water and oils, including those of petroleum, animal, vegetable, or synthetic origin, e.g., peanut oil, soybean oil, mineral oil, sesame oil, and the like. For example, 0.4% saline and 0.3% glycine may be used to formulate the bispecific anti-EGFR / c-Met antibody. These solutions are sterile and generally free of particulate matter. They may be sterilized by conventional, well-known sterilization techniques (e.g., filtration). For parenteral administration, the carrier may comprise sterile water, with other excipients added to enhance solubility or preserve. Injectable suspensions or solutions may also be prepared utilizing aqueous carriers with appropriate additives. Suitable vehicles and formulations (including other human proteins, e.g., human serum albumin) are described, for example, in Remington: The Science and Practice of Pharmacy, 21st Edition, Troy, DB ed., Lipincott Williams and Wilkins, Philadelphia, PA 2006, Part 5, Pharmaceutical Manufacturing pp 691-1092, see in particular pp. 958-989.

[0170] The mode of administration may be any suitable route that delivers the bispecific anti-EGFR-c-Met antibody to the host, for example, parenteral administration, such as intradermal, intramuscular, intraperitoneal, intravenous, or subcutaneous, pulmonary, transmucosal (oral, intranasal, intravaginal, rectal), using tablet, capsule, liquid, powder, gel, particle formulations that may be contained in syringes, implantable devices, osmotic pumps, cartridges, micropumps, or other means well known in the art and recognized by the skilled artisan. Site-specific administration may be achieved, for example, by intratumoral, intraarticular, intrabronchial, intraabdominal, intracapsular, intrachondral, intrasinus, intracavity, intracerebellar, intraventricular, intracolonic, intracervical, intragastric, intrahepatic, intramyocardial, intraosseous, intrapelvic, intrapericardial, intraperitoneal, intrapleural, intraprostatic, intrapulmonary, intrarectal, intrarenal, intraretinal, intraspinal, intrasynovial, intrathoracic, intrauterine, intravascular, intravesical, intralesional, intravaginal, intrarectal, buccal, sublingual, intranasal, or transdermal delivery. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered intravenously (IV). In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered subcutaneously (SC). In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered using an on-body delivery device.

[0171] In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose between about 140 mg and about 2240 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose between about 140 mg and about 2240 mg.

[0172] In some embodiments, the bispecific anti-EGFR / c-Met antibody is about 200 mg, about 210 mg, about 220 mg, about 230 mg, about 240 mg, about 250 mg, about 260 mg, about 270 mg, about 280 mg, about 290 mg, about 300 mg, about 310 mg, about 320 mg, about 330 mg, about 340 mg, about 350 mg, about 360 mg, about 370 mg, about 380 mg, about 390 mg, about 400 mg, about 410 mg, about 420 mg, about 430 mg, about 440 mg, about 450 mg, about 460 mg, about 470 mg, about 480 mg, about 490 mg, about 500 mg, about 510 mg, about 520 mg, about 530 mg, about 540 mg, about 550 mg, about 560 mg, about 570 mg, about 580 mg, about 590 mg, about 600 mg, about 610 mg, about 620 mg, about 630 mg, about 640 mg, about 650 mg, about 660 mg, about 670 mg, about 680 mg, about 690 mg, about 700 mg, about 710 mg, about 720 mg, about 730 mg, about 740 mg, about 750 mg, about 760 mg, about 770 mg, about 780 mg, about 790 mg, about 800 mg, about 810 mg, about 820 mg, about 830 mg, about 840 mg, about 850 mg, about 860 mg, about 870 mg, about 880 mg, about 890 mg, about 900 mg, about 910 mg, about 920 mg, about 930 mg, about 940 mg, about 950 mg, about 960 mg, about 970 mg, about 980 mg, about 990 mg, about 1000 mg, about 1010 mg, about 1020 mg, about 1030 mg, about 1040 mg, about 1050 mg, about 1060 mg, about 1070 mg, about 1080 mg, about 1090 mg, about 1100 mg, about 1110 mg, about 1120 mg, about 1130 mg, about 1140 mg, about 1150 mg, about 1160 mg, about 1170 mg, about 1180 mg, about 1190 mg, about 1200 mg, about 1210 mg, about 1220 mg, about 1230 mg, about 1240 mg, about 1250 mg, about 1260 mg, about 1270 mg, about 1280 mg, about 1290 mg, about 1300 mg, about 1310 mg, about 1320 mg, about 1330 mg, about 1340 mg, about 1350 mg, about 1360 mg, about 1370 mg, about 1380 mg, about 1390 mg, about 1400 mg, about 1410 mg, about 1420 mg, about 1430 mg, about 1440 mg, about 1450 mg, about 1460 mg, about 1470 mg, about 1480 mg, about 1490 mg, about 1500 mg,About 1510mg, about 1520mg, about 1530mg, about 1540mg, about 1550mg, about 1560mg, about 1570mg, about 1575mg, about 1580mg, about 1590mg, about 1600mg, about 1610mg, 1620mg, about 1630mg, about 1640mg, about 1650mg, about 1660mg, about 1670mg, about 1680mg, about 1690mg, about 170 0mg, about 1710mg, about 1720mg, about 1730mg, about 1740mg, about 1750mg, about 1760mg, about 1770mg, about 1780mg, about 1790mg, about 1800m g, about 1810mg, about 1820mg, about 1830mg, about 1840mg, about 1850mg, about 1860mg, about 1870mg, about 1880mg, 1890mg, about 1900mg, about 1910mg, about 1920mg, about 1930mg, about 1940mg, about 1950mg, about 1960mg, about 1970mg, about 1980mg, about 1990mg, about 2000mg, about 20 10mg, about 2020mg, about 2030mg, about 2040mg, about 2050mg, about 2060mg, about 2070mg, about 2080mg, about 2090mg, about 2100mg, about 2110 The compound is administered in a dose of about 2120 mg, about 2130 mg, about 2140 mg, about 2150 mg, about 2160 mg, about 2170 mg, about 2180 mg, about 2190 mg, about 2200 mg, about 2210 mg, about 2220 mg, about 2230 mg, about 2240 mg, about 2250 mg, about 2260 mg, about 2270 mg, about 2280 mg, about 2290 mg, about 2300 mg.

[0173] In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 350 mg, about 700 mg, about 1050 mg, about 1400 mg, about 1575 mg, about 1600, about 2100 mg, or about 2240 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 350 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 700 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 750 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 800 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 850 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 900 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 950 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 1000 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 1050 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 1100 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 1150 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 1200 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 1250 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 1300 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 1350 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 1400 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 1575 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 1600 mg.In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 2100 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 2240 mg.

[0174] In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered once a week. In some embodiments, about 1050 mg of the bispecific anti-EGFR / c-Met antibody is administered once a week. In some embodiments, about 1400 mg of the bispecific anti-EGFR / c-Met antibody is administered once a week. In some embodiments, about 1575 mg of the bispecific anti-EGFR / c-Met antibody is administered once a week. In some embodiments, about 1600 mg of the bispecific anti-EGFR / c-Met antibody is administered once a week. In some embodiments, about 2100 mg of the bispecific anti-EGFR / c-Met antibody is administered once a week. In some embodiments, about 2240 mg of the bispecific anti-EGFR / c-Met antibody is administered once a week.

[0175] In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered once every two weeks. In some embodiments, about 1050 mg of the bispecific anti-EGFR / c-Met antibody is administered once every two weeks. In some embodiments, about 1400 mg of the bispecific anti-EGFR / c-Met antibody is administered once every two weeks. In some embodiments, about 1575 mg of the bispecific anti-EGFR / c-Met antibody is administered once every two weeks. In some embodiments, about 1600 mg of the bispecific anti-EGFR / c-Met antibody is administered once every two weeks. In some embodiments, about 2100 mg of the bispecific anti-EGFR / c-Met antibody is administered once every two weeks. In some embodiments, about 2240 mg of the bispecific anti-EGFR / c-Met antibody is administered once every two weeks.

[0176] In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered twice weekly. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered once weekly. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered once every two weeks. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered once every three weeks. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered once every four weeks.

[0177] In the case of combination therapy, one or more anti-cancer drugs may be administered using the recommended doses and administration amounts of the anti-cancer drugs.

[0178] Generation of bispecific anti-EGFR / c-Met antibodies for use in the methods of the present disclosure An exemplary bispecific anti-EGFR / c-Met antibody that can be used in the methods of the present disclosure is amivantamab. Amivantamab is characterized by the following amino acid sequence: EGFR binding arm >SEQ ID NO:1 (HCDR1, EGFR binding arm) TYGMH >SEQ ID NO:2 (HCDR2, EGFR binding arm) VIWDDGSYKYYGDSVKG >SEQ ID NO:3 (HCDR3, EGFR binding arm) DGITMVRGVMKDYFDY >SEQ ID NO:4 (LCDR1, EGFR binding arm) RASQDISSALV >SEQ ID NO:5 (LCDR2, EGFR binding arm) DASSLES >SEQ ID NO:6 (LCDR3, EGFR binding arm) QQFNSYPLT >SEQ ID NO:7 (HCDR1, c-Met binding arm) SYGIS >SEQ ID NO:8 (HCDR2, c-Met binding arm) WISAYNGYTNYAQKLQG >SEQ ID NO:9 (HCDR3, c-Met binding arm) DLRGTNYFDY >SEQ ID NO: 10 (LCDR1, c-Met binding arm) RASQGISNWLA >SEQ ID NO: 11 (LCDR2, c-Met binding arm) AASSLLS >SEQ ID NO: 12 (LCDR3, c-Met binding arm) QQANSFPIT >SEQ ID NO: 13 (VH, EGFR binding arm) QVQLVESGGGVVQPGRSLRLSCAASGFTFSTYGMHWVRQAPGKGLEWVAVIWDDGSYKYYGDSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDGITMVRGVMKDYFDYWGQGTLVTVSS >SEQ ID NO: 14 (VL, EGFR binding arm) AIQLTQSPSSLSASVGDRVTITCRASQDISSALVWYQQKPGKAPKLLIYDASSLESGVPSRFSGSESGTDFTLTISSLQPEDFATYYCQQFNSYPLTFGGGTKVEIK >SEQ ID NO: 15 (VH, c-Met binding arm) QVQLVQSGAEVKKPGASVKVSCETSGYTFTSYGISWVRQAPGHGLEWMGWISAYNGYTNYAQKLQGRVTMTTDTSTSTAYMELRSLRSDDTAVYYCARDLRGTNYFDYWGQGTLVTVSS >SEQ ID NO: 16 (VL, c-Met binding arm) DIQMTQSPSSVSASVGDRVTITCRASQGISNWLAWFQHKPGKAPKLLIYAASSLLSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANSFPITFGQGTRLEIK >Sequence number 17 HC1 QVQLVESGGGVVQPGRSLRLSCAASGFTFSTYGMHWVRQAPGKGLEWVAVIWDDGSYKYYGDSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDGITMVRGVMKDYFD YWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKS CDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIE KTISKAKGQPREPQVYTLPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFLLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK >SEQ ID NO:18 LC1 AIQLTQSPSSLSASVGDRVTITCRASQDISSALVWYQQKPGKAPKLLIYDASSLESGVPSRFSGSESGTDFTLTISSLQPEDFATYYCQQFNSYPLTFGGGTKVEIK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC >Sequence number 19 HC2 QVQLVQSGAEVKKPGASVKVSCETSGYTFTSYGISWVRQAPGHGLEWMGWISAYNGYTNYAQKLQGRVTMTTDTSTSTAYMELRSLRSDDTAVYYCARDLRGTNYFDYWGQG TLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDK THTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEK TISKAKGQPREPQVYTLPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK >Sequence number 20 LC2 DIQMTQSPSSVSASVGDRVTITCRASQGISNWLAWFQHKPGKAPKLLIYAASSLLSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANSFPITFGQGTRLEIK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0179] Other publicly available bispecific anti-EGFR / c-Met antibodies may be used in the methods of the disclosure, so long as they exhibit similar characteristics when compared to amivantamab, as described in U.S. Patent No. 9,593,164. Bispecific anti-EGFR / c-Met antibodies that may be used in the methods of the disclosure may also be generated by combining publicly available EGFR-binding VH / VL domains and c-Met-binding VH / VL domains and testing the resulting bispecific antibody for its characteristics as described in U.S. Patent No. 9,593,164.

[0180] The bispecific anti-EGFR / c-Met antibodies used in the methods of the present disclosure can be generated using Fab arm exchange (or half molecule exchange) between two monospecific bivalent antibodies, for example, by introducing substitutions in the heavy chain CH3 interface in each half molecule to favor the formation of heterodimers of two antibody half molecules with different specificities, in a cell-free environment in vitro or using co-expression. The Fab arm exchange reaction is the result of a disulfide bond isomerization reaction and dissociation-association of the CH3 domains. The heavy chain disulfide bonds in the hinge region of the parent monospecific antibodies are reduced. The resulting free cysteine ​​of one of the parent monospecific antibodies forms an intra-heavy chain disulfide bond with a cysteine ​​residue of the second parent monospecific antibody molecule, and at the same time, the CH3 domain of the parent antibody is released and reformed by dissociation-association. The CH3 domain of the Fab arm may be engineered to favor heterodimer formation over homodimer formation. The resulting product is a bispecific antibody with two Fab arms or half molecules that each bind a different epitope, i.e., an epitope in EGFR and an epitope in c-Met. For example, bispecific antibodies of the invention can be generated using the techniques described in WO 2011 / 131746. For IgG1 antibodies, the mutation F405L in one heavy chain and K409R in the other heavy chain can be used. For IgG2 antibodies, wild type IgG2 and IgG2 antibodies with F405L and R409K substitutions may be used. For IgG4 antibodies, wild type IgG4 and IgG4 antibodies with F405L and R409K substitutions may be used. To generate a bispecific antibody, a first monospecific bivalent antibody and a second monospecific bivalent antibody are engineered to have the aforementioned mutations in the Fc region, and the antibodies are incubated together under sufficient reducing conditions to allow the cysteines in the hinge region to undergo disulfide bond isomerization, thereby generating the bispecific antibody by Fab arm exchange. Incubation conditions may optimally be returned to non-reducing conditions.Exemplary reducing agents that may be used are 2-mercaptoethylamine (2-MEA), dithiothreitol (DTT), dithioerythritol (DTE), glutathione, tris(2-carboxyethyl)phosphine (TCEP), L-cysteine, and β-mercaptoethanol. For example, incubation at a temperature of at least 20° C., in the presence of at least 25 mM 2-MEA or in the presence of at least 0.5 mM dithiothreitol, at a pH of 5-8, e.g., pH 7.0 or pH 7.4, for at least 90 minutes may be used.

[0181] Bispecific anti-EGFR / c-Met antibodies for use in the methods of the disclosure can also be generated using designs such as knobs-in-holes (Genentech), CrossMAb (Roche) and electrostatically-matched (Chugai, Amgen, NovoNordisk, Oncomed), LUZ-Y (Genentech), Strand Exchange Engineered Domain body (SEEDbody) (EMD Serono), and Biclonic (Merus).

[0182] In the "knobs-in-holes" approach (see, for example, WO 2006 / 028936), selected amino acids that form the interface of the CH3 domain of human IgG can be mutated at positions that affect CH3 domain interactions to promote heterodimer formation. Amino acids with small side chains (holes) are introduced into the heavy chain of an antibody that specifically binds to a first antigen, and amino acids with large side chains (knobs) are introduced into the heavy chain of an antibody that specifically binds to a second antigen. After co-expression of the two antibodies, heterodimers are formed as a result of the preferential interaction of heavy chains with "holes" and heavy chains with "knobs". Exemplary CH3 substitution pairs that form knobs and holes are T366Y / F405A, T366W / F405W, F405W / Y407A, T394W / Y407T, T394S / Y407A, T366W / T394S, F405W / T394S, and T366W / T366S_L368A_Y407V (expressed as modification position in the first CH3 domain of the first heavy chain / modification position in the second CH3 domain of the second heavy chain).

[0183] In addition to utilizing a "knobs-in-holes" approach to promote Fab arm exchange, CrossMAb technology utilizes CH1 / CL domain swapping in one of the half-arms to ensure correct light chain pairing of the resulting bispecific antibody (see, e.g., U.S. Pat. No. 8,242,247).

[0184] Other crossover techniques may be used to generate full length bispecific antibodies of the invention by swapping variable or constant, or both, domains in one or both arms between the heavy and light chains of the bispecific antibody or within the heavy chain, including, for example, VH-CH1 and VL-CL, VH and VL, CH3 and CL, and CH3 and CH1 as described in WO 2009 / 080254, WO 2009 / 080251, WO 2009 / 018386, and WO 2009 / 080252.

[0185] Other approaches, such as promoting heavy chain heterodimerization using electrostatic interactions by substituting positively charged residues on one CH3 surface and negatively charged residues on the second CH3 surface, may be used as described in U.S. Patent Application Publication No. 2010 / 0015133, U.S. Patent Application Publication No. 2009 / 0182127, U.S. Patent Application Publication No. 2010 / 028637, or U.S. Patent Application Publication No. 2011 / 0123532. In another approach, heterodimerization can be achieved by the following substitutions: L351Y_F405A_Y407V / T394W, T366I_K392M_T394W / F405A_Y407V, T366L_K392M_T394W / F405A_Y407V, L351Y ... A / T366A_K409F, L351Y_Y407A / T366V_K409F, Y407A / T366A_K409F, or T350V_L351Y_F405A_Y407V / T350V_T366L_K392L_T394W (represented as modification position in the first CH3 domain of the first heavy chain / modification position in the second CH3 domain of the second heavy chain).

[0186] SEEDbody technology may be utilized to generate bispecific antibodies of the invention. SEEDbodies have selected IgG residues in their constant domains substituted with IgA residues to promote heterodimerization, as described in US Patent Application Publication No. 20070287170.

[0187] Mutations are typically made at the DNA level, using standard methods, on molecules such as the constant domain of an antibody.

[0188] Embodiment 1) A method of treating a subject having a cancer that is EGFR positive and lacks at least one EGFR activating mutation, comprising administering to the subject having a cancer that is EGFR positive and lacks at least one EGFR activating mutation a therapeutically effective amount of an isolated bispecific anti-epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) antibody. 2) A method of treating a subject with cancer with a bispecific anti-EGFR / c-Met antibody, comprising: a) providing a biological sample from a subject; b) determining the presence or absence of EGFR lacking activating mutations in the sample; and c) administering or providing for administration of a bispecific anti-EGFR / c-Met antibody to a subject determined to lack an EGFR activating mutation. 3) The method of embodiment 1 or 2, wherein the at least one activating mutation is a mutation that increases at least one biological activity of EGFR. 4) The method of embodiment 3, wherein at least one biological activity of EGFR is selected from the group consisting of tyrosine kinase activity, ligand-independent signaling, increased cell proliferation, signaling to the MAPK / ERK pathway, gene transcription, dimerization (EGFR:EGFR), and heterodimerization (EGFR:HER2 or EGFR:HER3). 5) At least one activating mutation that increases at least one biological activity of EGFR is selected from the group consisting of L718Q, G719A, G719X (X is any amino acid), L861X (X is any amino acid), L858R, E746K, L747S, E749Q, A750P, A755V, V765M, C797S, L858P or T790M substitutions, deletions of E746 to A750, deletions of R748 to P753, insertion of Ala (A) between M766 and A767, insertion of Ser, Val, and Ala (SVA) between S768 and V769, insertion of Asn and between P772 and H773 4. The method of embodiment 3, comprising an insertion of Ser(NS), an insertion of one or more amino acids between D761 and E762, A763 and Y764, Y764 and Y765, M766 and A767, A767 and V768, S768 and V769, V769 and D770, D770 and N771, N771 and P772, P772 and H773, H773 and V774, V774 and C775, one or more deletions in EGFR exon 20, one or more insertions in EGFR exon 20, at least one mutation selected from the group consisting of S768I, L861Q, and G719X, where X is any amino acid. 6) The method of any one of embodiments 1 to 5, wherein the method further comprises determining the presence or absence of at least one mutation in any one gene selected from the group consisting of KRAS, PIK3CA, and PTEN, and administering or providing for administration of a bispecific anti-EGFR / c-Met antibody to a subject determined to have EGFR lacking an activating mutation and determined to lack at least one mutation in any one gene selected from the group consisting of KRAS, PIK3CA, and PTEN. 7) The method of embodiment 6, wherein at least one mutation in KRAS is selected from the group consisting of G12V, G12C, G12A and G12D. 8) The method of embodiment 7, wherein at least one mutation in KRAS is G12C. 9) The method of embodiment 8, wherein at least one mutation in PI3K is selected from the group consisting of E545K, H1047L, and PI3K amplification. 10) The method of embodiment 6, wherein at least one mutation in PTEN is a PTEN deletion. 11) The method according to any one of embodiments 1 to 10, wherein the bispecific anti-EGFR / c-Met antibody comprises a first domain that specifically binds to EGFR and a second domain that specifically binds to c-Met, wherein the first domain comprises heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 1, HCDR2 of SEQ ID NO: 2, HCDR3 of SEQ ID NO: 3, light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 4, LCDR2 of SEQ ID NO: 5, and LCDR3 of SEQ ID NO: 6, and the second domain that binds to c-Met comprises HCDR1 of SEQ ID NO: 7, HCDR2 of SEQ ID NO: 8, HCDR3 of SEQ ID NO: 9, LCDR1 of SEQ ID NO: 10, LCDR2 of SEQ ID NO: 11, and LCDR3 of SEQ ID NO: 12. 12) The method according to any one of embodiments 1 to 11, wherein the first domain that specifically binds to EGFR comprises a heavy chain variable region (VH) of SEQ ID NO: 13 and a light chain variable region (VL) of SEQ ID NO: 14, and the second domain that specifically binds to c-Met comprises a VH of SEQ ID NO: 15 and a VL of SEQ ID NO: 16. 13) The method according to any one of embodiments 1 to 12, wherein the bispecific anti-EGFR / c-Met antibody is of the IgG1 isotype. 14) The method of any one of embodiments 1 to 13, wherein the bispecific anti-EGFR / c-Met antibody comprises a first heavy chain (HC1) of SEQ ID NO: 17, a first light chain (LC1) of SEQ ID NO: 18, a second heavy chain (HC2) of SEQ ID NO: 19, and a second light chain (LC2) of SEQ ID NO: 20. 15) The method according to any one of embodiments 1 to 14, wherein the bispecific anti-EGFR / c-Met antibody comprises one or more Fc silencing mutations. 16) The method of embodiment 14, wherein the one or more Fc silencing mutations reduce affinity for an Fcγ receptor. 17) The method of embodiment 15 or 16, wherein the one or more Fc silencing mutations include V234A / G237A / P238S / H268A / V309L / A330S / P331S. 18) The method of any one of embodiments 1 to 17, wherein the bispecific anti-EGFR / c-Met antibody comprises a biantennary glycan structure having a fucose content of between about 1% and about 15%. 19) The method of any one of embodiments 1 to 18, wherein the subject is recurrent or refractory to treatment with one or more prior anticancer therapies. 20) The method of embodiment 19, wherein the one or more previous anti-cancer therapies include one or more chemotherapeutic agents, checkpoint inhibitors, targeted anti-cancer therapies, or kinase inhibitors, or any combination thereof. 21) The method of embodiment 20, wherein the one or more previous anticancer therapies include carboplatin, paclitaxel, gemcitabine, cisplatin, vinorelbine, docetaxel, palbociclib, crizotinib, PD-(L)1 axis inhibitors, EGFR inhibitors, c-Met inhibitors, HER2 inhibitors, HER3 inhibitors, HER4 inhibitors, VEGFR inhibitors, AXL inhibitors, erlotinib, gefitinib, lapatinib, vandetanib, afatinib, osimertinib, lazertinib, poziotinib, cliotinib, cabozantinib, capmatinib, axitinib, lenvatinib, nintedanib, regorafenib, pazopanib, sorafenib, or sunitinib, or any combination thereof. 22) The method of any one of embodiments 1 to 18, wherein the subject is treatment-naive. 23) The method of any one of embodiments 1 to 22, wherein the cancer positive for EGFR lacking activating mutations is positive for at least one mutation in a gene selected from the group consisting of ALK, APC, BRAF, BRCA1, BRCA2, CDKN2A, CDKN2B, CTNNB1, ERBB2, ERBB3, FGFR3, KIT, LRP1B, MET, MLH1, MSH3, NOTCH1, NTRK1, RET, ROS1, STK11, TP53, and VEGFA. 24) The method according to any one of embodiments 1 to 23, wherein the cancer is lung cancer, gastric cancer, colorectal cancer, brain cancer, cancer derived from epithelial cells, breast cancer, ovarian cancer, colorectal cancer, anal cancer, prostate cancer, kidney cancer, bladder cancer, head and neck cancer, pharyngeal cancer, nasal cancer, pancreatic cancer, skin cancer, oral cancer, tongue cancer, esophageal cancer, vaginal cancer, cervical cancer, splenic cancer, testicular cancer, gastric cancer, thymic cancer, colon cancer, thyroid cancer, liver cancer, hepatocellular carcinoma (HCC), or sporadic or hereditary papillary renal cell carcinoma (PRCC), or any combination thereof. 25) The method of embodiment 24, wherein the lung cancer is non-small cell lung cancer (NSCLC), small cell lung cancer (SCLC), or lung adenocarcinoma, lung sarcomatoid carcinoma, or any combination thereof. 26) The method of any one of embodiments 1 to 25, further comprising administering to the subject one or more anticancer therapies. 27) The method of embodiment 26, wherein the one or more anti-cancer therapies include chemotherapy, radiation therapy, surgery, targeted anti-cancer therapy, kinase inhibitors, or any combination thereof. 28) The method of embodiment 20, wherein the kinase inhibitor is an inhibitor of EGFR, an inhibitor of c-Met, an inhibitor of HER2, an inhibitor of HER3, an inhibitor of HER4, an inhibitor of VEGFR, or an inhibitor of AXL. 29) The method of embodiment 28, wherein the kinase inhibitor is erlotinib, gefitinib, lapatinib, vandetanib, afatinib, osimertinib, lazertinib, poziotinib, cliotinib, cabozantinib, capmatinib, axitinib, lenvatinib, nintedanib, regorafenib, pazopanib, sorafenib, or sunitinib. 30) The method of any one of embodiments 1 to 29, wherein the bispecific anti-EGFR / c-Met antibody is administered at a dose of between about 140 mg and about 2240 mg. 31) Bispecific anti-EGFR / c-Met antibody is about 700mg, about 750mg, about 800mg, about 850mg, about 900mg, about 950mg, about 1000mg, about 1050mg mg, about 1100mg, about 1150mg, about 1200mg, about 1250mg, about 1300mg, about 1350mg, about 1400mg, about 1450mg, about 1500mg, about 15 The method of any one of embodiments 1-30, wherein the compound is administered at a dose of about 50 mg, about 1575 mg, about 1600 mg, about 1650 mg, about 1700 mg, about 1750 mg, about 1800 mg, about 1850 mg, about 1900 mg, about 1950 mg, about 2000 mg, about 2050 mg, about 2100 mg, about 2150 mg, about 2200, or about 2240 mg. 32) The method of any one of embodiments 1 to 31, wherein the bispecific anti-EGFR / c-Met antibody is administered at a dose of 1050 mg. 33) The method of any one of embodiments 1 to 31, wherein the bispecific anti-EGFR / c-Met antibody is administered at a dose of 1400 mg. 34) The method of any one of embodiments 1 to 31, wherein the bispecific anti-EGFR / c-Met antibody is administered at a dose of 1575 mg. 35) The method of any one of embodiments 1 to 31, wherein the bispecific anti-EGFR / c-Met antibody is administered at a dose of 1600 mg. 36) The method of any one of embodiments 1 to 31, wherein the bispecific anti-EGFR / c-Met antibody is administered at a dose of 2100 mg. 37) The method of any one of embodiments 1 to 31, wherein the bispecific anti-EGFR / c-Met antibody is administered at a dose of 2240 mg. 38) The method of any one of embodiments 1 to 37, wherein the bispecific anti-EGFR / c-Met antibody is administered twice a week, once a week, once every two weeks, once every three weeks, or once every four weeks. 39) The method of any one of embodiments 1 to 38, wherein the bispecific anti-EGFR / c-Met antibody is administered intravenously. 40) The method of any one of embodiments 1 to 38, wherein the bispecific anti-EGFR / c-Met antibody is administered subcutaneously. 41) The method of any one of embodiments 1-5, wherein the method further comprises determining the level of amphiregulin and administering or providing for administration of a bispecific anti-EGFR / c-Met antibody to a subject determined to have EGFR lacking an activating mutation and determined to be positive for amphiregulin.

[0189] The invention will now be described with reference to the following specific, non-limiting examples.

[0190] Example 1. Characterization of non-small cell lung cancer (NSCLC) patient-derived xenograft (PDX) tumors expressing EGFR lacking activating mutations. EGFR and MET protein levels, as determined by immunohistochemistry (IHC); signaling, as determined by proximity ligation assay (PLA); and tumor associated macrophage (TAM) content were assessed in 39 NSCLC patient-derived xenograft (PDX) models with EGFR lacking activating mutations. Absence of activating mutations in EGFR was determined by whole exome sequencing (WES). Thirty-nine formalin fixed and paraffin embedded (FFPE) NSCLC PDX tumors with EGFR lacking activating mutations were obtained from Charles River Laboratory (CRL, Freiburg, Germany). Tumors included 19 adenocarcinoma and 20 epidermoid NSCLC. Correlations between receptor protein levels and receptor signaling were assessed for both EGFR and MET in these models using IHC and PLA.

[0191] For IHC studies, tissue sections were processed as described by Smith et al. (2015) (Annotation of human cancers with EGFR signaling-associated protein complexes using proximity ligation assays. Matthew A. Smith et.al. 2015, Science Signaling, 8(359):ra4). Briefly, sections were rehydrated and antigens were retrieved. Non-specific binding was blocked by incubation with 1.5% bovine serum albumin (BSA) and incubated overnight in 0.5% BSA in PBST with rabbit antibodies targeting EGFR (Ventana, clone 5B7) or with MET clone D1C2 (Cell Signaling) and phospho-MET clone D26 (Cell Signaling). Slides were washed twice with PBST, incubated with EnVision+anti-rabbit (K400311-2, Agilent) for 1 h, and visualized by DAB (diaminobenzidine). Slides were counterstained with hematoxylin, rehydrated, and hard-mounted. To calculate the H-score, the staining intensity of cells was scored (0, 1+, 2+, 3+) and the percentage of cells at each intensity was determined. The formula [1×(% cells 1+)+2×(% cells 2+)+3×(% cells 3+)] was then used to calculate the H-score, ranging from 0 to 300, for each PDX model.

[0192] Proximity ligation assays (PLA) were performed according to published protocols (Annotation of human cancers with EGFR signaling-associated protein complexes using proximity ligation assays. Matthew A. Smith et.al. 2015, Science Signaling, 8(359):ra4). Briefly, slides containing 5 μm sections of FFPE PDX tumors were rehydrated through xylene and graded alcohols. Heat-induced epitope retrieval was performed in Tris-EDTA (pH 9) in a pressure cooker for 20 min, followed by cooling for 20 min. Nonspecific binding was blocked by incubation with 1.5% bovine serum albumin (BSA) for 30 min at room temperature. Primary antibodies were rabbit antibodies targeting EGFR (clone D38B1, Cell Signaling Technology) or MET clone D1C2 (Cell Signaling) diluted 1:300, and mouse antibodies targeting growth factor receptor-bound protein 2 (GRB2) (clone 81, BD Biosciences) were used in 1.5% BSA in 0.5% phosphate-buffered saline (PBS)-Tween 20 (PBST) overnight. PLA probes were rabbit (-) and mouse (+) and detected with the Duolink™ In Situ PLA Far Red kit (Sigma-Aldrich). Alexa Fluor488-conjugated anti-cytokeratin was used to distinguish epithelial areas (clone AE1 / AE3, eBioscience).

[0193] Confocal images were acquired on a Leica TCS SP5 AOBS (Acousto Optical Beam Splitter) laser scanning confocal microscope through a 40×1.25NA (numerical aperture) Plan-Apochromat oil immersion objective (Leica Microsystems CMS GmbH). Diode (405) and HeNe (647) laser lines were applied to excite the samples, and tunable emission was used to minimize crosstalk between fluorochromes. Z-stack (0.5 μm thick slice) images of each sample were captured with a photomultiplier detector and maximum projections were made using LAS AF software version 2.6 (Leica Microsystems). Further fluorescent images were acquired on a fully automated upright Zeiss AxioImagerZ.1 microscope equipped with a 40×1.25NA oil immersion objective and DAPI and Cy5 filter cubes. Images were generated using an AxioCam MRm CCD (charge-coupled device) camera and the Axiovision version 4.6 software suite (Carl Zeiss Inc.) All tissue-based PLA and AQUA analysis images were acquired using a 20× objective (dry) on an AQUA workstation (PM-2000, HistoRx) equipped with a fully motorized stage and DAPI, Cy3, FITC (fluorescein isothiocyanate), and Cy5 filter cubes.Images were saved as individual channels and exported as a merged RGB TIFF image. (Annotation of human cancers with EGFR signaling-associated protein complexes using proximity ligation assays. Matthew A. Smith et.al. 2015, Science Signaling, 8(359):8(359):ra4; MET-GRB2 Signaling-Associated Complexes Correlate with Oncogenic MET Signaling and Sensitivity to MET Kinase Inhibitors. Matthew A. Smith et.al. 2017, Clin Cancer Res. 2017,23(22):7084-7096).

[0194] PLA scores were determined as previously described (Smith et al. 2015). Briefly, PLA was manually quantified using a scoring criterion based on foci per cell (0, not detectable; 1+, 1–5 foci per cell; 2+, >5–20 foci per cell; 3+, >20 foci per cell) and annotated as “high” (2+–3+ in both cores) or “low” (0–1+).

[0195] The results suggested that a statistically significant correlation between IHC score and PLA score was observed for both EGFR (r=0.63, p<0.0001, see FIG. 1A) and MET (r=0.83, p<0.0001, see FIG. 2A). Spearman's correlation coefficient was used to calculate r values, and two-tailed tests were used to calculate p values ​​(Prism Graphpad v.7.0.0). Specifically, if there were less than 17 XY pairs, a permutation test was used to calculate the p value of the Spearman correlation coefficient. Otherwise, the statistical value

[0196]

number

[0197] Example 2. Fc receptor engagement is not required for amivantamab efficacy in NSCLC models with EGFR lacking activating mutations. Next, we tested the efficacy of amivantamab in vivo in an NSCLC model that harbors EGFR lacking activating mutations.

[0198] In vivo studies were performed at Charles River Laboratory (CRL, Freiburg, Germany) in accordance with the policies and procedures of the Janssen Animal Care and Use Committee. Fourteen NSCLC PDX tumors were implanted subcutaneously into the flanks of NMRI nu / nu mice (CRL) until tumors reached 50–200 mm 3 Treatment was administered by intraperitoneal injection of either isotype control, or amivantamab, or EGFR / MET bispecific antibody with silent Fc at 10 mg / kg twice weekly for 3 weeks. The EGFR / MET-silent Fc antibody retains the EGFR and MET arms of amivantamab and has substitutions V234A / G237A / P238S / H268A / V309L / A330S / P331S made to the heavy chain to statistically significantly reduce affinity for Fcγ receptors and Cq1 complement. Tumors were measured with calipers and were calculated using the formula: (length x (width) 2 Tumor volumes were calculated using the formula: {1-[(treatment t -treatment i ) / (Control t - Control i )]} × 100 was used to calculate 7 days after the last dose.

[0199] Results suggested that amivantamab inhibited tumor growth in 13 of the 14 PDX models tested (representative efficacy plots are shown in Figures 2A and 2B). Of these 13 models in which amivantamab showed activity, EGFR / MET-silent Fc antibodies lacking binding to Fc receptors had comparable efficacy (Figures 2A and 2B). LXFA2158, a model with MET gene amplification, was the only PDX model in which amivantamab Fc binding was important for efficacy (Figure 2C). This is consistent with previously reported data (Smruthi Vijayaraghavan et.al. 2020, Molecular Cancer Therapeutics, 19(10):2044-2056) that demonstrated the requirement for amivantamab interaction with Fc in MET amplified and EGFR mutant model systems.

[0200] Example 3. Relationship between receptor expression and signaling and in vivo efficacy of amivantamab. Next, the relationship between receptor expression or signaling and the in vivo efficacy of amivantamab was evaluated.

[0201] IHC and PLA assays were performed as described in Example 1. Amivantamab efficacy (%TGI) in PDX tumors with EGFR lacking activating mutations was plotted with respect to EGFR and MET IHC H-scores and PLA scores (see Figures 3A-D). For EGFR, a positive correlation was determined between efficacy and both IHC and PLA (IHC: r=0.397, PLA: r=0.22), but none of the correlations were statistically significant (p>0.05). No positive correlation was observed between tumor efficacy and MET expression or signaling.

[0202] Example 4. Genomic analysis of PDX tumors. Next, the association between expression and mutation status of PDX tumors with EGFR lacking activating mutations and the tumor growth inhibition % obtained in Example 2 was evaluated. The expression data and mutation status of common cancer genes in PDX tumors with EGFR lacking activating mutations used in Example 2 were provided by Charles River Laboratory (Freiburg, Germany) and are wholly owned. Expression of the common cancer genes listed in Figure 4A was determined by RNA-Seq and mutation status was determined by whole exome analysis (Figure 4B). Amivantamab was highly effective in many of these NSCLC PDX models with EGFR lacking activating mutations (Figure 4C), but mutations in the KRAS and PI3K pathways were identified as biomarkers of reduced anti-tumor activity (see Figures 4A-C).

[0203] When assessing EGFR correlation to efficacy, we observed a subset of models in which the observed %TGI tended to fall below the linear fit of the data set, but with relatively high EGFR levels (Figure 3A).Interestingly, this subset of models was found to have mutations in the KRAS or PI3K pathways, suggesting that aberrant signaling in these pathways may reduce the activity of amivantamab.

[0204] Example 5. PDX tumors lacking KRAS and PI3K pathway mutations were associated with increased efficacy of amivantamab. Next, the association of amivantamab efficacy (%TGI) with the combined H-score and PLA-score (see Table 1) was evaluated in a subgroup of PDX tumors from Example 2, where tumors positive for known tumorigenic driver mutations downstream of EGFR and MET, such as KRAS and PI3K, were removed (see Table 2). Correlation coefficients and p-values ​​were calculated using Spearman's correlation coefficient and two-tailed tests (Prism Graphpad 7.00) as described in Example 1.

[0205] Results showed that elimination of tumors with KRAS or PI3K pathway mutations resulted in a statistically significant correlation between amivantamab efficacy and EGFR expression and signaling (IHC+PLA-r=0.88, p=0.0032, see FIG. 5A). Indeed, models with EGFR H-score≧170 and no alternative driver mutations achieved long-term tumor stasis or complete tumor regression upon treatment with amivantamab (representative tumor growth curves are shown in FIG. 2A and FIG. 2B). No correlation between amivantamab efficacy and MET expression and signaling was observed (FIG. 5B).

[0206] [Table 1]

[0207] [Table 2]

[0208] Example 6. Analysis of the association between EGFR ligand expression and amivantamab efficacy in PDX tumors with EGFR lacking activating mutations. Next, the expression of EGFR ligands in PDX tumors with EGFR lacking activating mutations was evaluated in relation to the tumor growth inhibition % obtained in Example 2. The expression of epidermal growth factor (EGF), amphiregulin (AREG), transforming growth factor alpha (TGFα), heparin-binding EGF-like growth factor (HBEGF), betacellulin (BTC), epiregulin (EREG), and epigen (EPGN) in PDX tumors with EGFR lacking activating mutations used in Example 2 was determined using RNA-Seq. These data were provided and fully owned by Charles River Laboratory (Freiburg, Germany). Amphiregulin expression was only available for 11 of the 14 NSCLC models in which in vivo efficacy was tested (no data was available for LXFA2158, LXFA2165, and LXFA2201). A statistically significant correlation (r=0.66, p=0.03) was found between amphiregulin expression and the in vivo efficacy of amivantamab (Figure 6). This association between amphiregulin expression and the efficacy of amivantamab was independent of KRAS and PI3K pathway mutations. Thus, amphiregulin expression can identify patients who are likely to derive clinical benefit from treatment with amivantamab.

Claims

1. 1. A formulation for use in a method of treating a subject having a cancer that is positive for EGFR and lacks at least one EGFR activating mutation, comprising: an isolated bispecific anti-epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) antibody; The method comprises administering to the subject having a cancer that is positive for EGFR and lacks at least one EGFR activating mutation a therapeutically effective amount of the bispecific anti-EGFR / c-Met antibody.

2. 1. A formulation for use in a method of treating a subject with cancer with a bispecific anti-EGFR / c-Met antibody, comprising: comprising a bispecific anti-EGFR / c-Met antibody, The method comprises: a) providing a biological sample from said subject; b) determining the presence or absence of an EGFR activating mutation in the sample; and c) administering or providing for administration of said bispecific anti-EGFR / c-Met antibody to said subject determined to lack an EGFR activating mutation.

3. The formulation of claim 1 or 2, wherein the at least one activating mutation is a mutation that increases at least one biological activity of EGFR.

4. 4. The formulation of claim 3, wherein the at least one biological activity of EGFR is selected from the group consisting of tyrosine kinase activity, ligand-independent signaling, increased cell proliferation, signaling to the MAPK / ERK pathway, gene transcription, dimerization (EGFR:EGFR), and heterodimerization (EGFR:HER2 or EGFR:HER3).

5. The at least one activating mutation that increases at least one biological activity of EGFR is selected from the group consisting of L718Q, G719A, G719X (X is any amino acid), L861X (X is any amino acid), L858R, E746K, L747S, E749Q, A750P, A755V, V765M, C797S, L858P, and T790M substitutions, deletions of E746 to A750, deletions of R748 to P753, insertion of Ala (A) between M766 and A767, insertion of Ser, Val, and Ala (SVA) between S768 and V769, insertion of Asn and / or Ala (A) between P772 and H773, insertion of Ser, Val, and / or Ala (SVA ...

4. The formulation of claim 3, comprising an insertion of Ser(NS), an insertion of one or more amino acids between D761 and E762, A763 and Y764, Y764 and Y765, M766 and A767, A767 and V768, S768 and V769, V769 and D770, D770 and N771, N771 and P772, P772 and H773, H773 and V774, V774 and C775, one or more deletions in EGFR exon 20, one or more insertions in EGFR exon 20, at least one mutation selected from the group consisting of S768I, L861Q, and G719X (wherein X is any amino acid).

6. 6. The formulation of any one of claims 1 to 5, wherein the method further comprises determining the presence or absence of at least one mutation in any one gene selected from the group consisting of KRAS, PIK3CA, and PTEN, and administering or providing for administration of the bispecific anti-EGFR / c-Met antibody to the subject determined to have an EGFR lacking an activating mutation and determined to lack at least one mutation in any one gene selected from the group consisting of KRAS, PIK3CA, and PTEN.

7. 7. The formulation of claim 6, wherein the at least one mutation in KRAS is selected from the group consisting of G12V, G12C, G12A and G12D.

8. 8. The formulation of claim 7, wherein the at least one mutation in KRAS is G12C.

9. 9. The formulation of claim 8, wherein the at least one mutation in PI3K is selected from the group consisting of E545K, H1047L, and PI3K amplification.

10. 7. The formulation of claim 6, wherein the at least one mutation in PTEN is a PTEN deletion.

11. 11. The formulation of any one of claims 1 to 10, wherein the bispecific anti-EGFR / c-Met antibody comprises a first domain that specifically binds EGFR and a second domain that specifically binds c-Met, wherein the first domain comprises heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 1, HCDR2 of SEQ ID NO: 2, HCDR3 of SEQ ID NO: 3, light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 4, LCDR2 of SEQ ID NO: 5, and LCDR3 of SEQ ID NO: 6, and the second domain that binds c-Met comprises HCDR1 of SEQ ID NO: 7, HCDR2 of SEQ ID NO: 8, HCDR3 of SEQ ID NO: 9, LCDR1 of SEQ ID NO: 10, LCDR2 of SEQ ID NO: 11, and LCDR3 of SEQ ID NO:

12.

12. 12. The formulation of any one of claims 1 to 11, wherein the first domain that specifically binds EGFR comprises a heavy chain variable region (VH) of SEQ ID NO: 13 and a light chain variable region (VL) of SEQ ID NO: 14, and the second domain that specifically binds c-Met comprises a VH of SEQ ID NO: 15 and a VL of SEQ ID NO:

16.

13. The formulation of any one of claims 1 to 12, wherein the bispecific anti-EGFR / c-Met antibody is of the IgG1 isotype.

14. The formulation of any one of claims 1 to 13, wherein the bispecific anti-EGFR / c-Met antibody comprises a first heavy chain (HC1) of SEQ ID NO: 17, a first light chain (LC1) of SEQ ID NO: 18, a second heavy chain (HC2) of SEQ ID NO: 19, and a second light chain (LC2) of SEQ ID NO:

20.

15. The formulation of any one of claims 1 to 14, wherein the bispecific anti-EGFR / c-Met antibody comprises one or more Fc-silencing mutations.

16. The formulation of claim 14, wherein the one or more Fc silencing mutations reduce affinity for an Fcγ receptor.

17. The formulation of claim 15 or 16, wherein the one or more Fc silencing mutations include V234A / G237A / P238S / H268A / V309L / A330S / P331S.

18. The formulation of any one of claims 1 to 17, wherein the bispecific anti-EGFR / c-Met antibody comprises a biantennary glycan structure having a fucose content of between about 1% and about 15%.

19. The formulation of any one of claims 1 to 18, wherein the subject is relapsed or refractory to treatment with one or more prior anti-cancer therapies.

20. 20. The formulation of claim 19, wherein the one or more prior anti-cancer therapies comprise one or more chemotherapeutic agents, checkpoint inhibitors, targeted anti-cancer therapies, or kinase inhibitors, or any combination thereof.

21. 21. The formulation of claim 20, wherein the one or more prior anticancer therapies comprise carboplatin, paclitaxel, gemcitabine, cisplatin, vinorelbine, docetaxel, palbociclib, crizotinib, PD-(L)1 axis inhibitors, EGFR inhibitors, c-Met inhibitors, HER2 inhibitors, HER3 inhibitors, HER4 inhibitors, VEGFR inhibitors, AXL inhibitors, erlotinib, gefitinib, lapatinib, vandetanib, afatinib, osimertinib, lazertinib, poziotinib, cliotinib, cabozantinib, capmatinib, axitinib, lenvatinib, nintedanib, regorafenib, pazopanib, sorafenib, or sunitinib, or any combination thereof.

22. The formulation of any one of claims 1 to 18, wherein the subject is treatment naive.

23. 23. The formulation of any one of claims 1 to 22, wherein the cancer positive for EGFR lacking activating mutations is positive for at least one mutation in a gene selected from the group consisting of ALK, APC, BRAF, BRCA1, BRCA2, CDKN2A, CDKN2B, CTNNB1, ERBB2, ERBB3, FGFR3, KIT, LRP1B, MET, MLH1, MSH3, NOTCH1, NTRK1, RET, ROS1, STK11, TP53, and VEGFA.

24. 24. The formulation of any one of claims 1 to 23, wherein the cancer is lung cancer, gastric cancer, colorectal cancer, brain cancer, cancer originating from epithelial cells, breast cancer, ovarian cancer, colorectal cancer, anal cancer, prostate cancer, kidney cancer, bladder cancer, head and neck cancer, pharyngeal cancer, nasal cancer, pancreatic cancer, skin cancer, oral cancer, tongue cancer, esophageal cancer, vaginal cancer, cervical cancer, spleen cancer, testicular cancer, stomach cancer, thymic cancer, colon cancer, thyroid cancer, liver cancer, hepatocellular carcinoma (HCC) or sporadic or hereditary papillary renal cell carcinoma (PRCC), or any combination thereof.

25. 25. The formulation of claim 24, wherein the lung cancer is non-small cell lung cancer (NSCLC), small cell lung cancer (SCLC), or lung adenocarcinoma, lung sarcomatoid carcinoma, or any combination thereof.

26. The formulation of any one of claims 1 to 25, wherein the method further comprises administering to the subject one or more anti-cancer therapies.

27. 27. The formulation of claim 26, wherein the one or more anti-cancer therapies comprise chemotherapy, radiation therapy, surgery, targeted anti-cancer therapy, a kinase inhibitor, or any combination thereof.

28. 21. The formulation of claim 20, wherein the kinase inhibitor is an inhibitor of EGFR, an inhibitor of c-Met, an inhibitor of HER2, an inhibitor of HER3, an inhibitor of HER4, an inhibitor of VEGFR, or an inhibitor of AXL.

29. 29. The formulation of claim 28, wherein the kinase inhibitor is erlotinib, gefitinib, lapatinib, vandetanib, afatinib, osimertinib, lazertinib, poziotinib, cliotinib, cabozantinib, capmatinib, axitinib, lenvatinib, nintedanib, regorafenib, pazopanib, sorafenib, or sunitinib.

30. 30. The formulation of any one of claims 1 to 29, wherein the bispecific anti-EGFR / c-Met antibody is administered at a dose of between about 140 mg to about 2240 mg.

31. The bispecific anti-EGFR / c-Met antibody is about 700 mg, about 750 mg, about 800 mg, about 850 mg, about 900 mg, about 950 mg, about 1000 mg, about 1050 mg g, about 1100mg, about 1150mg, about 1200mg, about 1250mg, about 1300mg, about 1350mg, about 1400mg, about 1450mg, about 1500mg, about 155 31. The formulation of any one of claims 1 to 30, wherein the formulation is administered in a dose of about 0 mg, about 1575 mg, about 1600 mg, about 1650 mg, about 1700 mg, about 1750 mg, about 1800 mg, about 1850 mg, about 1900 mg, about 1950 mg, about 2000 mg, about 2050 mg, about 2100 mg, about 2150 mg, about 2200 mg, or about 2240 mg.

32. The formulation of any one of claims 1 to 31, wherein the bispecific anti-EGFR / c-Met antibody is administered at a dose of 1050 mg.

33. The formulation of any one of claims 1 to 31, wherein the bispecific anti-EGFR / c-Met antibody is administered at a dose of 1400 mg.

34. The formulation of any one of claims 1 to 31, wherein the bispecific anti-EGFR / c-Met antibody is administered at a dose of 1575 mg.

35. The formulation of any one of claims 1 to 31, wherein the bispecific anti-EGFR / c-Met antibody is administered at a dose of 1600 mg.

36. The formulation of any one of claims 1 to 31, wherein the bispecific anti-EGFR / c-Met antibody is administered at a dose of 2100 mg.

37. 32. The formulation of any one of claims 1 to 31, wherein the bispecific anti-EGFR / c-Met antibody is administered at a dose of 2240 mg.

38. 38. The formulation of any one of claims 1 to 37, wherein the bispecific anti-EGFR / c-Met antibody is administered twice weekly, once weekly, once every two weeks, once every three weeks, or once every four weeks.

39. The formulation of any one of claims 1 to 38, wherein the bispecific anti-EGFR / c-Met antibody is administered intravenously.

40. The formulation of any one of claims 1 to 38, wherein the bispecific anti-EGFR / c-Met antibody is administered subcutaneously.

Images