HK40081424A - Treatment of patients having c-met exon 14 skipping mutations - Google Patents

Description

Treatment of patients with c-MET exon 14 skipping mutations

Cross Reference to Related Applications

This patent application claims priority to U.S. provisional application serial No. 62/975,406, filed on 12/2/2020. The disclosures in the above applications are incorporated herein by reference in their entirety.

Electronically submitted reference sequence Listing

This application contains a sequence listing electronically submitted via EFS-Web as an ASCII formatted sequence listing with a file name of "jbit6242usnpp 1 sequilist. txt" and a creation date of 2021 year, 1 month, 19 days, and a size of 19 kb. This sequence listing, filed via EFS-Web, is part of this specification and is incorporated herein by reference in its entirety.

Technical Field

The present invention relates to the treatment of subjects having a c-Met exon 14 skipping mutation.

Background

The separate roles of both EGFR and 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); thus, jointly suppressing this pair may limit the possibility of compensating path activation, thereby improving overall efficacy.

Mutations in c-Met have been associated with many cancers, including renal, gastric, nervous system, sarcoma, and lung cancers. Mutations that result in higher activity or loss of expression or negative regulatory sites are often implicated in these cancers. For example, deletion of exon 14 and the negative regulatory site at Tyr 1003 is associated with a significant percentage of non-small cell lung cancer (NSCLC) and adenocarcinoma.

Relapse or resistance to existing therapeutics is common. Thus, there is a need for improved therapeutics or combinations of therapeutics and patient stratification biomarkers to develop more effective treatments for diseases such as EGFR or c-Met positive cancers.

Disclosure of Invention

The present disclosure provides a method of treating a subject having a cancer positive for a c-Met exon 14 skipping mutation comprising administering to a subject having a cancer positive for a c-Met exon 14 skipping mutation a therapeutically effective amount of an isolated bispecific anti-Epidermal Growth Factor Receptor (EGFR)/hepatocyte growth factor receptor (c-Met) antibody.

The present disclosure also provides a method of treating a subject having cancer with a bispecific anti-EGFR/c-Met antibody, the method comprising:

providing a biological sample from the subject;

determining the presence or absence of a c-Met exon 14 skipping mutation in said sample;

administering or providing administration of the bispecific anti-EGFR/c-Met antibody to the subject determined to have a c-Met exon 14 skipping mutation.

The present disclosure also provides a method of treating a subject having a cancer positive for a c-Met exon 14 skipping mutation comprising administering to a subject having a cancer positive for a c-Met exon 14 skipping mutation a therapeutically effective amount of an isolated bispecific anti-Epidermal Growth Factor Receptor (EGFR)/hepatocyte growth factor receptor (c-Met) antibody.

In one embodiment, the present disclosure provides a method for treating a subject having cancer with a bispecific anti-EGFR/c-Met antibody, the method comprising:

a) providing a biological sample from the subject;

b) determining the presence or absence of a c-Met exon 14 skipping mutation in said sample;

c) administering or providing administration of the bispecific anti-EGFR/c-Met antibody to the subject determined to have a c-Met exon 14 skipping mutation.

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 of SEQ ID NO:1(HCDR 1), HCDR2 of SEQ ID NO:2, HCDR3 of SEQ ID NO:3, light chain complementarity determining region 1 of SEQ ID NO:4(LCDR 1), LCDR2 of SEQ ID NO:5, and LCDR3 of SEQ ID NO: 6; a second domain that binds c-Met, wherein 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.

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

In one embodiment, the bispecific anti-EGFR/c-Met antibody is an IgG1 isotype.

In one embodiment, the bispecific anti-EGFR/c-Met antibody comprises the first heavy chain of SEQ ID NO:17 (HC1), the first light chain of SEQ ID NO:18 (LC1), the second heavy chain of SEQ ID NO:19 (HC2), and the second light chain of SEQ ID NO:20 (LC 2).

In one embodiment, the bispecific anti-EGFR/c-Met antibody comprises a double branched glycan structure with a fucose content of between about 1% to about 15%.

In one embodiment, the subject has relapsed or refractory to treatment with one or more previous anti-cancer therapies.

In one embodiment, 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.

In one embodiment, the one or more prior anti-cancer therapies comprise carboplatin, paclitaxel, gemcitabine, cisplatin, vinorelbine, docetaxel, palbociclib, crizotinib, PD- (L)1 axis inhibitor, EGFR inhibitor, c-Met inhibitor, HER2 inhibitor, HER3 inhibitor, HER4 inhibitor, VEGFR inhibitor, erlotinib, gefitinib, lapatinib, vandetanib, afatinib, oxitinib, lazatinib, crirtitinib, cabozantinib, camatinib, axitinib, lenvatinib, nilatinib, regorafenib, pazopanib, sorafenib, or sunitinib, or any combination thereof.

In one embodiment, the subject is treatment naive.

In one embodiment, a cancer positive for a c-Met exon 14 skipping mutation is selected from CDK4 amplification, EGFR amplification, KRAS amplification, MDM2 amplification, TERT amplification, NF 1R 2450; RAD50L597Vfs 5, MET c.3082+3A > G, EGFR, wild type EGFR, EGFR activating mutation, circulating HGF, c-MET amplification or increased levels of mutant KRAS or any combination thereof.

In one embodiment, the EGFR activating mutation comprises: 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; deletion of E746-A750; absence of R748-P753; insertion of ala (a) between M766 and a 767; insertion of Ser, Val and Ala (SVA) between S768 and V769; insertion of Asn and ser (ns) between P772 and H773; insertion of one or more amino acids between D761 and E762, between a763 and Y764, between Y764 and Y765, between M766 and a767, between a767 and V768, between S768 and V769, between V769 and D770, between D770 and N771, between N771 and P772, between P772 and H773, between H773 and V774, between V774 and C775; one or more deletions in EGFR exon 20; or one or more insertions in EGFR exon 20; or any combination thereof.

In one embodiment, the mutant KRAS comprises a G12V, G12C, G12A, or G12D substitution, or any combination thereof.

In one embodiment, the cancer is lung cancer, stomach cancer, colorectal cancer, brain cancer, epithelial cell-derived cancer, breast cancer, ovarian cancer, colorectal cancer, anal cancer, prostate cancer, kidney cancer, bladder cancer, head and neck cancer, pharynx cancer, nose cancer, pancreatic cancer, skin cancer, oral cancer, tongue cancer, esophageal cancer, vaginal cancer, cervical cancer, spleen cancer, testicular cancer, stomach cancer, thymus cancer, colon cancer, thyroid cancer, liver cancer, hepatocellular carcinoma (HCC), or sporadic or hereditary Papillary Renal Cell Carcinoma (PRCC), or any combination thereof.

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.

In one embodiment, the method comprises further administering to the subject one or more anti-cancer therapies.

In one embodiment, the one or more anti-cancer therapies comprise chemotherapy, radiation therapy, surgery, targeted anti-cancer therapy, or a kinase inhibitor, or any combination thereof.

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

In one embodiment, the kinase inhibitor is erlotinib, gefitinib, lapatinib, vandetanib, afatinib, oxitinib, lacitinib, pozitinib, critinib, cabozantinib, camatinib, axitinib, lenvatinib, nidanib, regorafenib, pazopanib, sorafenib, or sunitinib.

In one embodiment, the c-Met exon 14 skipping mutation is a de novo (de novo) mutation.

In one embodiment, the c-Met exon 14 skipping mutation is an acquired mutation.

In one embodiment, the bispecific anti-EGFR/c-Met antibody is administered at a dose of between about 140mg and about 1750 mg.

In one embodiment, the bispecific anti-EGFR/c-Met antibody is administered at a dose of about 700mg, about 750mg, about 800mg, about 850mg, 900mg, 950mg, 1000mg, 1050mg, 1100mg, 1150mg, 1200mg, 1250mg, 1300mg, 1350mg, or 1400 mg.

In one embodiment, the bispecific anti-EGFR/c-Met antibody is administered twice weekly, once biweekly, once weekly, or once four weeks.

Embodiments of the present disclosure provide methods of treating a subject having a cancer positive for a c-Met exon 14 skipping mutation, the method comprising administering to the subject a combination therapy, wherein the combination therapy comprises a therapeutically effective amount of an isolated bispecific anti-Epidermal Growth Factor Receptor (EGFR)/hepatocyte growth factor receptor (c-Met) antibody and a therapeutically effective amount of a compound of formula (II)

Or a solvate, hydrate, tautomer or pharmaceutically acceptable salt thereof.

In one embodiment, the bispecific anti-EGFR/c-Met antibody comprises: a first domain that binds EGFR comprising heavy chain complementarity determining region 1 of SEQ ID NO:1(HCDR 1), HCDR2 of SEQ ID NO:2, HCDR3 of SEQ ID NO:3, light chain complementarity determining region 1 of SEQ ID NO:4(LCDR 1), LCDR2 of SEQ ID NO:5, and LCDR3 of SEQ ID NO: 6; and a second domain that binds c-Met, the second domain comprising 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.

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

In one embodiment, the bispecific anti-EGFR/c-Met antibody is an IgG1 isotype.

In one embodiment, the bispecific anti-EGFR/c-Met antibody comprises the first heavy chain of SEQ ID NO:17 (HC1), the first light chain of SEQ ID NO:18 (LC1), the second heavy chain of SEQ ID NO:19 (HC2), and the second light chain of SEQ ID NO:20 (LC 2).

In one embodiment, the bispecific anti-EGFR/c-Met antibody has a double branched glycan structure with a fucose content of between about 1% to about 15%.

In one embodiment, the compound of formula (II) or a solvate, hydrate, tautomer or pharmaceutically acceptable salt thereof is 2-fluoro-N-methyl-4- [7- (quinolin-6-ylmethyl) imidazo [1,2-b ] [1,2,4] triazin-2-yl ] benzamide-hydrogen chloride-water (1/2/1).

In one embodiment, the subject has relapsed or refractory to treatment with one or more previous anti-cancer therapies.

In one embodiment, 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.

In one embodiment, the one or more prior anti-cancer therapies comprise carboplatin, paclitaxel, gemcitabine, cisplatin, vinorelbine, docetaxel, palbociclib, crizotinib, PD- (L)1 axis inhibitor, EGFR inhibitor, c-Met inhibitor, HER2 inhibitor, HER3 inhibitor, HER4 inhibitor, VEGFR inhibitor, erlotinib, gefitinib, lapatinib, vandetanib, afatinib, oxitinib, lazatinib, poetinib, crirtitinib, cabozantinib, axitinib, lenvatinib, nidabib, regorafenib, pazopanib, sorafenib, or sunitinib, or any combination thereof.

In one embodiment, the subject is untreated.

In one embodiment, a cancer positive for a c-Met exon 14 skipping mutation is selected from CDK4 amplification, EGFR amplification, KRAS amplification, MDM2 amplification, TERT amplification, NF 1R 2450; RAD50L597Vfs 5, MET c.3082+3A > G, EGFR, wild type EGFR, EGFR activating mutation, circulating HGF, c-MET amplification or increased levels of mutant KRAS or any combination thereof are positive.

In one embodiment, the EGFR activating mutation comprises: 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; deletion of E746-A750; absence of R748-P753; insertion of Ala (A) between M766 and A767; insertion of Ser, Val and Ala (SVA) between S768 and V769; insertion of Asn and ser (ns) between P772 and H773; insertion of one or more amino acids between D761 and E762, between a763 and Y764, between Y764 and Y765, between M766 and a767, between a767 and V768, between S768 and V769, between V769 and D770, between D770 and N771, between N771 and P772, between P772 and H773, between H773 and V774, between V774 and C775; one or more deletions in EGFR exon 20; or one or more insertions in EGFR exon 20; or any combination thereof.

In one embodiment, the mutant KRAS comprises a G12V, G12C, G12A, or G12D substitution, or any combination thereof.

In one embodiment, the cancer is lung cancer, stomach cancer, colorectal cancer, brain cancer, epithelial cell-derived cancer, breast cancer, ovarian cancer, colorectal cancer, anal cancer, prostate cancer, kidney cancer, bladder cancer, head and neck cancer, pharynx cancer, nose cancer, pancreatic cancer, skin cancer, oral cancer, tongue cancer, esophageal cancer, vaginal cancer, cervical cancer, spleen cancer, testicular cancer, stomach cancer, thymus cancer, colon cancer, thyroid cancer, liver cancer, hepatocellular carcinoma (HCC), or sporadic or hereditary Papillary Renal Cell Carcinoma (PRCC), or any combination thereof.

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.

In one embodiment, the method comprises further administering to the subject one or more anti-cancer therapies.

In one embodiment, the one or more anti-cancer therapies comprise chemotherapy, radiation therapy, surgery, targeted anti-cancer therapy, or a kinase inhibitor, or any combination thereof.

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

In one embodiment, the kinase inhibitor is erlotinib, gefitinib, lapatinib, vandetanib, afatinib, oxitinib, lacitinib, pozitinib, critinib, cabozantinib, camatinib, axitinib, lenvatinib, nidanib, regorafenib, pazopanib, sorafenib, or sunitinib.

In one embodiment, the c-Met exon 14 skipping mutation is a neogenetic mutation.

In one embodiment, the c-Met exon 14 skipping mutation is an acquired mutation.

In one embodiment, the bispecific anti-EGFR/c-Met antibody is administered at a dose of between about 140mg and about 1750 mg.

In one embodiment, the bispecific anti-EGFR/c-Met antibody is administered at a dose of about 700mg, about 750mg, about 800mg, about 850mg, 900mg, 950mg, 1000mg, 1050mg, 1100mg, 1150mg, 1200mg, 1250mg, 1300mg, 1350mg, or 1400 mg.

In one embodiment, the bispecific anti-EGFR/c-Met antibody is administered twice weekly, once biweekly, once weekly, or once four weeks.

Drawings

Figure 1 shows the efficacy of the EGFR/c-Met antibody (JNJ-372) in tumor models with c-Met exon 14 skipping mutations. Tumor shrinkage was evident in JNJ-372 treated mice, whereas tumors were amplified in erlotinib or cetuximab treated animals.

Fig. 2 shows CT scans at baseline (top panel), 6 weeks of treatment with 1050mg JNJ-372 (middle panel), and 12 weeks of treatment (bottom panel). Tumor size is indicated in each figure.

Fig. 3A-3B show the average tumor volume (fig. 3A) and average body weight (fig. 3B) in mice treated with JNJ-61186372(JNJ-372), carbamatinib, or isotype control with tumors carrying the c-Met exon 14 skipping mutation (LU 2503). Arrows indicate termination of individual animals due to tumor size; the vertical dashed line indicates the end of dosing.

Fig. 4A-4B show the average tumor volume (fig. 4A) and average body weight (fig. 4B) in mice with tumors carrying the c-Met exon 14 skipping mutation (LU2503), treated with JNJ-61186372(JNJ-372), carbamatinib, JNJ-61186372 and carbamatinib, or isotype controls. Arrows indicate termination of individual animals due to tumor size; the vertical dashed line indicates the end of dosing.

Figure 5 shows protein levels determined by western blot in LU2503 tumors grown in mice treated with JNJ-61186372(JNJ-372), carbamatinib, JNJ-61186372 and carbamatinib, or isotype controls.

Fig. 6A-6B show LU2503 tumors grown in mice treated with JNJ-61186372 ("372"), carbamatinib ("cap"), JNJ-61186372 and carbamatinib ("372 cap"), or isotype control ("Iso Ctrl"), as determined by western blot and quantified by using image J software and normalized to the protein level of β -tubulin.

Fig. 7A-7C show LU2503 tumors grown in mice treated with JNJ-61186372 ("372"), carbamatinib ("cap"), JNJ-61186372 and carbamatinib ("372 cap"), or isotype control ("Iso Ctrl"), as determined by western blot and quantified by using image J software and normalized to the protein level of β -tubulin.

Figure 8 shows the mean tumor volume in mice with tumors carrying the c-Met exon 14 skipping mutation (DFCI-440), treated with JNJ-61186372(JNJ-372), carbamatinib, JNJ-61186372 and carbamatinib, or isotype controls.

Detailed Description

Definition of

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 herein.

It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting. Unless defined otherwise, 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.

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

When a list is provided, it is to be understood that each individual element of the list, and each combination of elements in the list, is a separate embodiment, unless otherwise indicated. For example, a list of embodiments presented as "A, B or C" will be understood to include embodiments "a", "B", "C", "a or B", "a or C", "B or C" or "A, B or C".

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.

The connecting term "and/or" between a plurality of enumerated elements is understood to encompass both individual and combined options. For example, where two elements are connected by "and/or," a first option refers to the first element being applied without the second element. The second option means that the second element is applied without the first element. A third option refers to the suitability of using the first and second elements together. Any of these options is understood to fall within the meaning and thus meet the requirements of the term "and/or" as used herein. Parallel applicability of more than one option is also understood to fall within the meaning and thus meet the requirements of the term "and/or".

The transitional terms "comprising," "consisting essentially of … …," and "consisting of … …" are intended to imply their accepted meanings in patent parlance; that is, (i) "comprising" is synonymous with "including", "containing", or "characterized by", and is inclusive or open-ended and does not exclude additional, unrecited elements or method steps; (ii) "consisting of … …" excludes any element, step, or ingredient not specified in the claims; and (iii) "consisting essentially of … …" limits the scope of the claims to the specified materials or steps "and materials or steps that do not materially affect the basic and novel characteristics of the claimed invention. Also provided are embodiments described by the phrase "comprising" (or equivalents thereof), such as those embodiments described independently by "consisting of … …" and "consisting essentially of … …".

"co-administration," with.

"isolated" refers to a homogeneous population of molecules (such as synthetic polynucleotides, polypeptide vectors, or viruses) that have been substantially isolated and/or purified from other components of a system in which the molecules are produced (such as recombinant cells), as well as proteins that have been subjected to at least one purification or isolation step. "isolated" refers to a molecule that is substantially free of other cellular material and/or chemicals, and encompasses molecules that are isolated to a higher degree of purity (such as 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% purity).

"treating (Treat/treating)" a disease or disorder, such as cancer, refers to achieving one or more of the following: reducing the severity and/or duration of a disorder, inhibiting worsening of symptoms characteristic of the disorder being treated, limiting or preventing recurrence of the disorder in a subject previously suffering from the disorder, or limiting or preventing recurrence of symptoms in a subject previously suffering from symptoms of the disorder.

"preventing (previous/predicting/promoting)" disease or disorder means preventing the occurrence of the disorder in a subject.

"Diagnosing" refers to a method of determining whether a subject has a given disease or disorder or is likely to develop a given disease or disorder in the future or is likely to respond to treatment for a previously diagnosed disease or disorder (i.e., stratifying a patient population according to the likelihood of responding to treatment). Diagnosis is typically performed by a physician based on general guidelines for the disease to be diagnosed or other criteria indicating that the subject is likely to respond to a particular treatment.

"response," "responsiveness," or "likely response" refers to any type of amelioration or positive response, whether detectable or undetectable, such as alleviation or amelioration of one or more symptoms, diminishment of extent of disease, stabilization (i.e., not worsening) of the disease state, prevention of spread of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, and remission (whether partial or total).

"newly diagnosed" refers to a subject who has been diagnosed with an EGFR-or c-Met-expressing cancer but has not received multiple myeloma treatment.

"therapeutically effective amount" means an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic result. The therapeutically effective amount may vary depending on the following factors: such as the disease state, age, sex, and weight of the individual, and 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, improving the health of a patient.

"refractory" refers to a disease that is not responsive to treatment. Refractory diseases can be resistant to treatment before or at the beginning of treatment, or refractory diseases can become resistant during treatment.

By "recurrence" is meant the recurrence of the disease or signs and symptoms of the disease after a period of improvement following prior treatment with a therapeutic agent.

"subject" includes any human or non-human animal. "non-human animal" includes all vertebrates, e.g., mammals and non-mammals, such as non-human primates, sheep, dogs, cats, horses, cattle, chickens, amphibians, reptiles, and the like. The terms "subject" and "patient" are used interchangeably herein.

"about" means within an acceptable error range for the particular value as determined by one of ordinary skill in the art, which will depend in part on how the value is measured or determined, i.e., the limitations of the measurement system. In the context of a particular assay, result, or embodiment, "about" means within one standard deviation or up to 5% (whichever is larger) of the standard deviation as is customary in the art, unless explicitly stated otherwise in the example or elsewhere in the specification.

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

"EGFR-or c-Met-expressing cancer" refers to a cancer that has detectable expression of EGFR or c-Met or has mutation or amplification of EGFR or c-Met. EGFR or c-Met expression, amplification and mutation status can be detected using known methods such as sequencing, fluorescence in situ hybridization, immunohistochemistry, flow cytometry or western blotting using tumor biopsy or blood samples. Expression can also be detected by sequencing circulating tumor dna (ctdna).

"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 GenBank accession NP-005219, as well as naturally occurring variants thereof.

As used herein, "hepatocyte growth factor receptor" or "c-Met" refers to human c-Met and its native variants having the amino acid sequence set forth in GenBank accession No. NP _ 001120972.

By "bispecific anti-EGFR/c-Met antibody" or "bispecific EGFR/c-Met antibody" is meant 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 to EGFR and c-Met are typically VH/VL pairs, and bispecific anti-EGFR/c-Met antibodies are monovalent in binding to EGFR and c-Met.

By "specifically binds" or "binding" is meant that the antibody binds to an antigen or epitope within an antigen with a higher affinity than to other antigens. Typically, an antibody binds to an antigen or epitope within an antigen, balancing the dissociation constant (K) _D ) Is about 5X 10 ^-8 M or less, e.g. about 1X 10 ^-9 M or less, about 1X 10 ^-10 M or less, about 1X 10 ^-11 M or less, or about 1X 10 ^-12 M or less, usually K _D At least one hundred-fold lower than its KD for binding to non-specific antigens (e.g., BSA, casein). The dissociation constant can be measured using known protocols. However, antibodies that bind to an antigen or an epitope within an antigen may be cross-reactive to other related antigens, e.g., from other sourcesThe same antigen of a species (homologous), such as a human or monkey, e.g. cynomolgus monkey (Macaca fascicularis, cyno) or chimpanzee (chimpanzee, chimp), is cross-reactive.

"antibody" refers broadly to and includes immunoglobulin molecules, including in particular monoclonal antibodies (including murine monoclonal antibodies, human monoclonal antibodies, humanized monoclonal antibodies, and chimeric monoclonal antibodies), antigen binding fragments, multispecific antibodies (such as bispecific antibodies, trispecific antibodies, tetraspecific antibodies, etc.), dimeric, tetrameric or multimeric antibodies, single chain antibodies, domain antibodies, and any other modified configuration of an immunoglobulin molecule comprising an antigen binding site with the desired specificity. A "full-length antibody" comprises two Heavy Chains (HC) and two Light Chains (LC) interconnected by disulfide bonds, and multimers thereof (e.g., IgM). 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 CH 3). Each light chain is composed of a light chain variable region (VL) and a light chain constant region (CL). The VH and VL regions can be further subdivided into hypervariable regions, termed Complementarity Determining Regions (CDRs) with intervening Framework Regions (FRs). Each VH and VL is composed of three CDRs and four FR segments, arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3 and FR 4.

"complementarity determining regions" (CDRs) are antibody regions that bind antigen. CDRs may be defined using various delineations, such as Kabat (Wu et al, (1970) J Exp Med 132:211-50) (Kabat et al, Sequences of Proteins of Immunological Interest, published Health Service 5 th edition, 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). The correspondence between the various delineations and the variable region numbering is described (see, e.g., Lefranc et al, (2003) Dev Comp Immunol 27: 55-77; Honegger and Pluckthun, (2001) J Mol Biol, 309: 657-70; International Immunogenetics (IMGT) database; Web resources, http:// www _ IMGT _ org). Available programs (such as abYsis of UCL Business PLC) can be used to delineate CDRs. As used herein, the terms "CDR," "HCDR 1," "HCDR 2," "HCDR 3," "LCDR 1," "LCDR 2," and "LCDR 3" include CDRs defined by any of the above methods (Kabat, Chothia, IMGT, or AbM), unless the specification expressly indicates otherwise.

Immunoglobulins can be assigned to five major classes, namely IgA, IgD, IgE, IgG and IgM, depending on the heavy chain constant domain amino acid sequence. IgA and IgG are further sub-classified into isotypes IgA1, IgA2, IgG1, IgG2, IgG3 and IgG 4. The light chain of an antibody of any vertebrate species can be assigned to one of two completely different types, κ and λ, based on the amino acid sequence of its constant domains.

An "antigen-binding fragment" refers to the antigen-binding portion of an immunoglobulin molecule. The antigen-binding fragment may be a synthetic, enzymatically obtainable or genetically engineered polypeptide and comprises 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, minimal recognition units consisting of amino acid residues that mimic the CDRs of an antibody, such as FR3-CDR3-FR4 portions, HCDR1, HCDR2 and/or HCDR3, and LCDR1, LCDR2 and/or LCDR 3. The VH and VL domains may be linked together via a synthetic linker to form various types of single chain antibody designs, where the VH/VL domains may pair intramolecularly or intermolecularly in the case where the VH and VL domains are expressed from separate single chain antibody constructs, to form a monovalent antigen binding site, such as single chain fv (scfv) or diabodies; for example, as described in International patent publication Nos. WO1998/44001, WO1988/01649, WO1994/13804, and WO 1992/01047.

"monoclonal antibody" refers to an antibody obtained from a substantially homogeneous population of antibody molecules, i.e., the individual antibodies comprising the population are identical except for possible well-known changes such as removal of the C-terminal lysine from the heavy chain of the antibody or post-translational modifications such as amino acid isomerization or deamidation, methionine oxidation or asparagine or glutamine deamidation. Monoclonal antibodies typically bind to an epitope. Bispecific monoclonal antibodies bind to two different epitopes. Monoclonal antibodies can have heterogeneous glycosylation within the antibody population. Monoclonal antibodies can be monospecific or multispecific, such as bispecific, monovalent, bivalent, or multivalent.

"recombinant" refers to DNA, antibodies and other proteins that are prepared, expressed, formed or isolated by recombinant means when segments from different sources are joined to produce recombinant DNA, antibodies or proteins.

"bispecific" refers to an antibody that specifically binds to two different antigens or two different epitopes within the same antigen. Bispecific antibodies may be cross-reactive to other relevant antigens, e.g. to the same antigen from other species (homologues), such as humans or monkeys, e.g. cynomolgus Macaca (cynomolgus), cyno or chimpanzee (Pan troglodytes), or may bind to an epitope shared between two or more different antigens.

An "antagonist" or "inhibitor" refers to a molecule that, when bound to a cellular protein, inhibits at least one response or activity induced by the natural ligand of the protein. The molecule is an antagonist when at least one response or activity is inhibited to 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 that is inhibited in the absence of the antagonist (e.g., a negative control), or when the inhibition is statistically significant compared to the inhibition in the absence of the antagonist.

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

"biological sample" refers to collections of similar fluids, cells, or tissues isolated from a subject, as well as fluids, cells, or tissues present in a subject. Exemplary samples are biological fluids such as blood, serum and serosal fluids, plasma, lymph, urine, saliva, cyst fluid, tears, excreta, sputum, mucosal secretions of secreted tissues and organs, vaginal secretions, ascites, pleural fluid, pericardial fluid, peritoneal fluid and other bodily cavity fluids, fluids collected from bronchial lavage, synovial fluid, liquid solutions in contact with a subject or biological source (e.g., cell and organ culture media (including cell or organ conditioned media), lavage fluids, etc.), tissue biopsies, tumor tissue samples, fine needle aspirates, surgically excised tissues, organ cultures, or cell cultures.

As used herein, "low fucose" or "low fucose content" refers to antibodies having a fucose content of about 1% to 15%.

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

By "c-Met exon 14 deletion" or "c-Met exon 14 skipping mutation" is meant a c-Met gene of a c-Met transcript that is mutated to remove at least a portion of exon 14 of c-Met or spliced to remove at least a portion of exon 14 of c-Met. The deleted part may include a part encoding the negative regulatory site Tyr 1003 in the membrane-proximal region of the c-Met protein. The exon 14 region of the c-Met gene encompasses nucleotides 3284 to 3424 in the full length nucleotide sequence of GenBank accession No. nm _000245 or residues 964 to 1009 in the full length c-Met amino acid sequence of GenBank accession No. np _ 000236. Various mutations at the DNA level may lead to exon 14 skipping (see, e.g., Kong-Beltran et al (2006) Cancer Res.66; Dhanasekharan et al (2014) Nature Communication 10: 1038; Awad et al, J Clin Oncology 34:721,2016). Exon 14 of c-Met encodes 47 amino acids.

By "pharmaceutical composition" is meant a composition comprising an active ingredient such as a bispecific EGFR/c-Met antibody and one or more pharmaceutically acceptable carriers or a pharmaceutically acceptable carrier or excipients such as the EGFR TK inhibitor carbamatinib, or a solvate, hydrate, tautomer, or pharmaceutically acceptable salt thereof.

By "pharmaceutically acceptable carrier" or "excipient" is meant an ingredient of a pharmaceutical composition other than the active ingredient, which is not toxic to the subject. Pharmaceutically acceptable carriers include, but are not limited to, buffers, stabilizers, or preservatives. Pharmaceutically acceptable carriers include, but are not limited to, diluents, disintegrants, or glidants; or diluents, disintegrants, wetting agents, glidants or lubricants.

"solvates" and "hydrates" are solvent addition forms that the compounds of the present invention are able to form, whereby the multicomponent compound comprises a primary molecule (e.g. a compound of formula (I) or a salt thereof) and a guest molecule (water ("hydrate") or another solvent ("solvate") incorporated into the structure).

"tautomer" or "tautomeric form" refers to structural isomers of different energies that can be interconverted via a low energy barrier. For example, proton tautomers (also referred to as prototropic tautomers) include interconversion via proton migration, such as keto-enol and imine-enamine isomerizations. Valence tautomers include interconversion by recombination of some of the bonding electrons.

Methods of the present disclosure

JNJ-61186372(JNJ-372) is an IgG1 anti-EGFR/c-Met bispecific antibody described in U.S. Pat. No. 9,593,164, also known as Epavantamab (Amivantamab).

The present disclosure is based, at least in part, on the following findings: JNJ-372 is effective in treating subjects having c-Met exon skipping mutations.

The c-Met exon 14 skipping mutation includes point mutations, insertions, deletions and complex mutations, such as a combination of insertions and deletions, that affect the splice acceptor or donor site and induce in-frame exon 14 skipping, resulting in deletion of the membrane-proximal domain of c-Met (a known c-Met negative regulatory domain) (see, e.g., Descatentries et al, J Thoracic oncology 13: 1873-. More than 160 mutations have been described that affect c-Met exon 14 (see, e.g., Cortot et al, J Natl Cancer Insti 109: djw262,2017). Next Generation Sequencing (NGS) of patient samples can be used to identify C-Met exon 14 skipping mutations. The exon 14 skipping mutation may be generated either newly or as a resistant mutation to a previous treatment (such as a TKI generation 3). c-Met exon 14 encodes the amino acid sequence DLGSELVRYDARVHTPHLDRLVSARSVSPTTEMVSNESVDYRATFPE (SEQ ID NO: 21).

The present disclosure provides a method of treating a subject having a cancer positive for a c-Met exon 14 skipping mutation comprising administering to a subject having a cancer positive for a c-Met exon 14 skipping mutation a therapeutically effective amount of an isolated bispecific anti-Epidermal Growth Factor Receptor (EGFR)/hepatocyte growth factor receptor (c-Met) antibody.

Cancers in which the c-Met exon 14 skipping mutation has been identified include lung, gastric, colorectal and brain cancers such as non-small cell lung cancer (NSCLC), Small Cell Lung Cancer (SCLC), lung adenocarcinoma and lung sarcomatoid cancer (PSC). Any other cancer carrying a c-Met exon skipping mutation may also be treated with the bispecific EGFR/c-Met antibodies of the present disclosure.

The present disclosure also provides a method of treating a subject having a lung cancer positive for a c-Met exon 14 skipping mutation, comprising administering to a subject having a lung cancer positive for a c-Met exon 14 skipping mutation a therapeutically effective amount of an isolated bispecific anti-EGFR/c-Met antibody.

The present disclosure also provides a method of treating a subject having NSCLC positive for a c-Met exon 14 skipping mutation comprising administering to a subject having NSCLC positive for a c-Met exon 14 skipping mutation a therapeutically effective amount of an isolated bispecific anti-EGFR/c-Met antibody.

The present disclosure also provides a method of treating a subject having an SCLC positive for a c-Met exon 14 skipping mutation comprising administering to a subject having an SCLC positive for a c-Met exon 14 skipping mutation a therapeutically effective amount of an isolated bispecific anti-EGFR/c-Met antibody.

The present disclosure also provides a method of treating a subject having a lung adenocarcinoma positive for a c-Met exon 14 skipping mutation comprising administering to a subject having a lung adenocarcinoma positive for a c-Met exon 14 skipping mutation a therapeutically effective amount of an isolated bispecific EGFR/c-Met antibody.

The present disclosure also provides a method of treating a subject having a PSC positive for a c-Met exon 14 skipping mutation comprising administering to a subject having a PSC positive for a c-Met exon 14 skipping mutation a therapeutically effective amount of an isolated bispecific anti-EGFR/c-Met antibody.

The present disclosure also provides a method of treating a subject having gastric cancer positive for a c-Met exon 14 skipping mutation comprising administering to a subject having gastric cancer positive for a c-Met exon 14 skipping mutation a therapeutically effective amount of an isolated bispecific anti-EGFR/c-Met antibody.

The present disclosure also provides a method of treating a subject having colorectal cancer that is positive for a c-Met exon 14 skipping mutation, comprising administering to a subject having colorectal cancer that is positive for a c-Met exon 14 skipping mutation a therapeutically effective amount of an isolated bispecific anti-EGFR/c-Met antibody.

The present disclosure also provides a method of treating a subject having a brain cancer positive for a c-Met exon 14 skipping mutation, comprising administering to a subject having a brain cancer positive for a c-Met exon 14 skipping mutation a therapeutically effective amount of an isolated bispecific anti-EGFR/c-Met antibody.

The present disclosure also provides a method of treating a subject having cancer with a bispecific anti-EGFR/c-Met antibody, the method comprising:

providing a biological sample from the subject;

determining the presence or absence of a c-Met exon 14 skipping mutation in said sample;

administering or providing administration of the bispecific anti-EGFR/c-Met antibody to the subject determined to have a c-Met exon 14 skipping mutation.

In some embodiments, the biological sample is a blood sample.

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

In some embodiments, 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 of SEQ ID NO:1(HCDR 1), HCDR2 of SEQ ID NO:2, HCDR3 of SEQ ID NO:3, light chain complementarity determining region 1 of SEQ ID NO:4(LCDR 1), 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.

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

In some embodiments, the bispecific anti-EGFR/c-Met antibody is an IgG1 isotype.

In some embodiments, the bispecific anti-EGFR/c-Met antibody comprises the first heavy chain of SEQ ID NO:17 (HC1), the first light chain of SEQ ID NO:18 (LC1), the second heavy chain of SEQ ID NO:19 (HC2), and the second light chain of SEQ ID NO:20 (LC 2).

In some embodiments, the bispecific anti-EGFR/c-Met antibody comprises a double-branched glycan structure with a fucose content of between about 1% to about 15%.

Antibodies with reduced fucose content can be made using different reported methods for causing successful expression of relatively higher defucosylated antibodies with dual-branched complex-type Fc oligosaccharides, such as controlling the culture osmolality (Konno et al, Cytotechnology 64(:249-65,2012), using the variant CHO cell line Lec13 as host cell line (Shields et al, J Biol Chem 277: 733 and 26740,2002), using the variant CHO cell line EB 2 as host cell line (Olivier et al, MAbs; 2(4), 2010; Epub ahead of print; PMID:20562582), using the large mouse hybridoma cell line YB2/0 as host cell line (Shinkawa et al, J Biol Chem 278:3466-, or co-expressing beta-1, 4-N-acetylglucosaminyltransferase III and golgi α -mannosidase II or a potent α -mannosidase I inhibitor, kifunorine (Ferrara et al, J Biol Chem281: 5032. sup. 5036,2006, Ferrara et al, Biotechnol Bioeng93: 851. sup. 861, 2006; Xhou et al, Biotechnol Bioeng 99: 652. sup. 65, 2008). Generally, reducing the fucose content in the glycans of an antibody enhances antibody-mediated cytotoxicity (ADCC).

The present disclosure also provides a method of treating a subject having a cancer positive for a c-Met exon 14 skipping mutation, comprising administering to a subject having a cancer positive for a c-Met exon 14 skipping 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 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.

The present disclosure also provides a method of treating a subject having a lung cancer positive for a c-Met exon 14 skipping mutation, comprising administering to a subject having a lung cancer positive for a c-Met exon 14 skipping 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 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.

The present disclosure also provides a method of treating a subject having NSCLC positive for a c-Met exon 14 skipping mutation, comprising administering to a subject having NSCLC positive for a c-Met exon 14 skipping 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 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.

The present disclosure also provides a method of treating a subject having an SCLC positive for a c-Met exon 14 skipping mutation comprising administering to a subject having an SCLC positive for a c-Met exon 14 skipping 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 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.

The present disclosure also provides a method of treating a subject having lung adenocarcinoma positive for a c-Met exon 14 skipping mutation comprising administering to a subject having lung adenocarcinoma positive for a c-Met exon 14 skipping 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 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.

The present disclosure also provides a method of treating a subject having a PSC positive for a c-Met exon 14 skipping mutation, comprising administering to a subject having a PSC positive for a c-Met exon 14 skipping 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 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.

The present disclosure also provides a method of treating a subject having a gastric cancer positive for a c-Met exon 14 skipping mutation, comprising administering to a subject having a gastric cancer positive for a c-Met exon 14 skipping 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 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.

The present disclosure also provides a method of treating a subject having colorectal cancer positive for a c-Met exon 14 skipping mutation, comprising administering to a subject having colorectal cancer positive for a c-Met exon 14 skipping 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 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.

The present disclosure also provides a method of treating a subject having a brain cancer positive for a c-Met exon 14 skipping mutation, comprising administering to a subject having a brain cancer positive for a c-Met exon 14 skipping 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 HCDR1 of SEQ ID No. 1, dr2 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.

The present disclosure provides a method of treating a subject having a cancer positive for a c-Met exon 14 skipping mutation, comprising administering to a subject having a cancer positive for a c-Met exon 14 skipping 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 the VH of SEQ ID NO:13 and the VL of SEQ ID NO: 14; and the second domain comprises the VH of SEQ ID NO. 15 and the VL of SEQ ID NO. 16.

The present disclosure also provides a method of treating a subject having a lung cancer positive for a c-Met exon 14 skipping mutation, comprising administering to a subject having a lung cancer positive for a c-Met exon 14 skipping 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 the VH of SEQ ID NO:13 and the VL of SEQ ID NO: 14; and the second domain comprises the VH of SEQ ID NO. 15 and the VL of SEQ ID NO. 16.

The present disclosure also provides a method of treating a subject having NSCLC positive for a c-Met exon 14 skipping mutation, comprising administering to a subject having NSCLC positive for a c-Met exon 14 skipping 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 the VH of SEQ ID No. 13 and the VL of SEQ ID No. 14; and the second domain comprises the VH of SEQ ID NO. 15 and the VL of SEQ ID NO. 16.

The present disclosure also provides a method of treating a subject having an SCLC positive for a c-Met exon 14 skipping mutation, comprising administering to a subject having an SCLC positive for a c-Met exon 14 skipping 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 the VH of SEQ ID No. 13 and the VL of SEQ ID No. 14; and the second domain comprises the VH of SEQ ID NO. 15 and the VL of SEQ ID NO. 16.

The present disclosure also provides a method of treating a subject having a lung adenocarcinoma positive for a c-Met exon 14 skipping mutation, comprising administering to a subject having a lung adenocarcinoma positive for a c-Met exon 14 skipping 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 the VH of SEQ ID No. 13 and the VL of SEQ ID No. 14; and the second domain comprises the VH of SEQ ID NO. 15 and the VL of SEQ ID NO. 16.

The present disclosure also provides a method of treating a subject having a PSC positive for a c-Met exon 14 skipping mutation, comprising administering to a subject having a PSC positive for a c-Met exon 14 skipping 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 the VH of SEQ ID NO:13 and the VL of SEQ ID NO: 14; and the second domain comprises the VH of SEQ ID NO. 15 and the VL of SEQ ID NO. 16.

The present disclosure also provides a method of treating a subject having gastric cancer positive for a c-Met exon 14 skipping mutation, comprising administering to a subject having gastric cancer positive for a c-Met exon 14 skipping 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 the VH of SEQ ID NO:13 and the VL of SEQ ID NO: 14; and the second domain comprises the VH of SEQ ID NO. 15 and the VL of SEQ ID NO. 16.

The present disclosure also provides a method of treating a subject having colorectal cancer that is positive for a c-Met exon 14 skipping mutation, comprising administering to a subject having colorectal cancer that is positive for a c-Met exon 14 skipping 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 the VH of SEQ ID NO:13 and the VL of SEQ ID NO: 14; and the second domain comprises the VH of SEQ ID NO. 15 and the VL of SEQ ID NO. 16.

The present disclosure also provides a method of treating a subject having a brain cancer positive for a c-Met exon 14 skipping mutation, comprising administering to a subject having a brain cancer positive for a c-Met exon 14 skipping 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 the VH of SEQ ID No. 13 and the VL of SEQ ID No. 14; and the second domain comprises the VH of SEQ ID NO. 15 and the VL of SEQ ID NO. 16.

The present disclosure provides a method of treating a subject having a cancer positive for a c-Met exon 14 skipping mutation, comprising administering to a subject having a cancer positive for a c-Met exon 14 skipping 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 specifically binding to EGFR and a second domain specifically binding to c-Met, wherein the first domain comprises the VH of SEQ ID NO:13 and the VL of SEQ ID NO: 14; and the second domain comprises the VH of SEQ ID NO. 15 and the VL of SEQ ID NO. 16.

The present disclosure also provides a method of treating a subject having a lung cancer positive for a c-Met exon 14 skipping mutation, comprising administering to a subject having a lung cancer positive for a c-Met exon 14 skipping mutation a therapeutically effective amount of an isolated bispecific anti-EGFR/c-Met antibody, wherein the bispecific anti-EGFR/c-Met antibody is of the 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 the VH of SEQ ID NO:13 and the VL of SEQ ID NO: 14; and the second domain comprises the VH of SEQ ID NO. 15 and the VL of SEQ ID NO. 16.

The present disclosure also provides a method of treating a subject having NSCLC positive for a c-Met exon 14 skipping mutation, comprising administering to a subject having NSCLC positive for a c-Met exon 14 skipping mutation a therapeutically effective amount of an isolated bispecific anti-EGFR/c-Met antibody, wherein the bispecific anti-EGFR/c-Met antibody is of the 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 the VH of SEQ ID NO:13 and the VL of SEQ ID NO: 14; and the second domain comprises the VH of SEQ ID NO. 15 and the VL of SEQ ID NO. 16.

The present disclosure also provides a method of treating a subject having an SCLC positive for a c-Met exon 14 skipping mutation, comprising administering to a subject having an SCLC positive for a c-Met exon 14 skipping mutation a therapeutically effective amount of an isolated bispecific anti-EGFR/c-Met antibody, wherein the bispecific anti-EGFR/c-Met antibody is of the 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 the VH of SEQ ID NO 13 and the VL of SEQ ID NO 14; and the second domain comprises the VH of SEQ ID NO. 15 and the VL of SEQ ID NO. 16.

The present disclosure also provides a method of treating a subject having a lung adenocarcinoma positive for a c-Met exon 14 skipping mutation, comprising administering to a subject having a lung adenocarcinoma positive for a c-Met exon 14 skipping mutation a therapeutically effective amount of an isolated bispecific anti-EGFR/c-Met antibody, wherein the bispecific anti-EGFR/c-Met antibody is of the 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 the VH of SEQ ID NO:13 and the VL of SEQ ID NO: 14; and the second domain comprises the VH of SEQ ID NO. 15 and the VL of SEQ ID NO. 16.

The present disclosure also provides a method of treating a subject having gastric cancer positive for a c-Met exon 14 skipping mutation, comprising administering to a subject having gastric cancer positive for a c-Met exon 14 skipping mutation a therapeutically effective amount of an isolated bispecific anti-EGFR/c-Met antibody, wherein the bispecific anti-EGFR/c-Met antibody is of the 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 the VH of SEQ ID NO:13 and the VL of SEQ ID NO: 14; and the second domain comprises the VH of SEQ ID NO. 15 and the VL of SEQ ID NO. 16.

The present disclosure also provides a method of treating a subject having colorectal cancer that is positive for a c-Met exon 14 skipping mutation, comprising administering to a subject having colorectal cancer that is positive for a c-Met exon 14 skipping mutation a therapeutically effective amount of an isolated bispecific anti-EGFR/c-Met antibody, wherein the bispecific anti-EGFR/c-Met antibody is of the 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 the VH of SEQ ID NO:13 and the VL of SEQ ID NO: 14; and the second domain comprises the VH of SEQ ID NO. 15 and the VL of SEQ ID NO. 16.

The present disclosure also provides a method of treating a subject having a brain cancer positive for a c-Met exon 14 skipping mutation, comprising administering to a subject having a brain cancer positive for a c-Met exon 14 skipping mutation a therapeutically effective amount of an isolated bispecific anti-EGFR/c-Met antibody, wherein the bispecific anti-EGFR/c-Met antibody is of the 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 the VH of SEQ ID NO:13 and the VL of SEQ ID NO: 14; and the second domain comprises the VH of SEQ ID NO. 15 and the VL of SEQ ID NO. 16.

In some embodiments, the bispecific anti-EGFR/c-Met antibody is an IgG1 isotype. There are variations (e.g., well known allotypes) within the constant domain of IgG1, where the variations are located at positions 214, 356, 358, 422, 431, 435 or 436 (residue numbering according to EU numbering) (see, e.g., IMGT Web resources; IMGT repertoires (IG and TR); proteins and alleles; allotypes). The bispecific anti-EGFR/c-Met antibody may be any IgG1 allotype, such as G1m17, G1m3, G1m1, G1m2, G1m27, or G1m 28.

The present disclosure also provides a method of treating a subject having a cancer positive for a c-Met exon 14 skipping mutation, comprising administering to a subject having a cancer positive for a c-Met exon 14 skipping mutation a therapeutically effective amount of an isolated bispecific anti-EGFR/c-Met antibody, wherein the bispecific anti-EGFR/c-Met antibody comprises HC1 of SEQ ID NO:17, LC1 of SEQ ID NO:18, HC2 of SEQ ID NO:19, and LC2 of SEQ ID NO: 20.

The present disclosure also provides a method of treating a subject having a lung cancer positive for a c-Met exon 14 skipping mutation, comprising administering to a subject having a lung cancer positive for a c-Met exon 14 skipping mutation a therapeutically effective amount of an isolated bispecific anti-EGFR/c-Met antibody, wherein the bispecific anti-EGFR/c-Met antibody comprises HC1 of SEQ ID NO:17, LC1 of SEQ ID NO:18, HC2 of SEQ ID NO:19, and LC2 of SEQ ID NO: 20.

The present disclosure also provides a method of treating a subject having NSCLC positive for a c-Met exon 14 skipping mutation, comprising administering to a subject having NSCLC positive for a c-Met exon 14 skipping mutation a therapeutically effective amount of an isolated bispecific anti-EGFR/c-Met antibody, wherein the bispecific anti-EGFR/c-Met antibody comprises HC1 of SEQ ID No. 17, LC1 of SEQ ID No. 18, HC2 of SEQ ID No. 19, and LC2 of SEQ ID No. 20.

The present disclosure also provides a method of treating a subject having an SCLC positive for a c-Met exon 14 skipping mutation, comprising administering to a subject having an SCLC positive for a c-Met exon 14 skipping mutation a therapeutically effective amount of an isolated bispecific anti-EGFR/c-Met antibody, wherein the bispecific anti-EGFR/c-Met antibody comprises HC1 of SEQ ID NO:17, LC1 of SEQ ID NO:18, HC2 of SEQ ID NO:19, and LC2 of SEQ ID NO: 20.

The present disclosure also provides a method of treating a subject having a lung adenocarcinoma positive for a c-Met exon 14 skipping mutation comprising administering to a subject having a lung adenocarcinoma positive for a c-Met exon 14 skipping mutation a therapeutically effective amount of an isolated bispecific anti-EGFR/c-Met antibody, wherein the bispecific anti-EGFR/c-Met antibody comprises HC1 of SEQ ID NO:17, LC1 of SEQ ID NO:18, HC2 of SEQ ID NO:19, and LC2 of SEQ ID NO: 20.

The present disclosure also provides a method of treating a subject having a PSC positive for a c-Met exon 14 skipping mutation, comprising administering to a subject having a PSC positive for a c-Met exon 14 skipping mutation a therapeutically effective amount of an isolated bispecific anti-EGFR/c-Met antibody, wherein the bispecific anti-EGFR/c-Met antibody comprises HC1 of SEQ ID NO:17, LC1 of SEQ ID NO:18, HC2 of SEQ ID NO:19, and LC2 of SEQ ID NO: 20.

The present disclosure also provides a method of treating a subject having gastric cancer positive for a c-Met exon 14 skipping mutation comprising administering to a subject having gastric cancer positive for a c-Met exon 14 skipping mutation a therapeutically effective amount of an isolated bispecific anti-EGFR/c-Met antibody, wherein the bispecific anti-EGFR/c-Met antibody comprises HC1 of SEQ ID NO:17, LC1 of SEQ ID NO:18, HC2 of SEQ ID NO:19, and LC2 of SEQ ID NO: 20.

The present disclosure also provides a method of treating a subject having colorectal cancer that is positive for a c-Met exon 14 skipping mutation, comprising administering to a subject having colorectal cancer that is positive for a c-Met exon 14 skipping mutation a therapeutically effective amount of an isolated bispecific anti-EGFR/c-Met antibody, wherein the bispecific anti-EGFR/c-Met antibody comprises HC1 of SEQ ID NO:17, LC1 of SEQ ID NO:18, HC2 of SEQ ID NO:19, and LC2 of SEQ ID NO: 20.

The present disclosure also provides a method of treating a subject having a brain cancer positive for a c-Met exon 14 skipping mutation, comprising administering to a subject having a brain cancer positive for a c-Met exon 14 skipping mutation a therapeutically effective amount of an isolated bispecific anti-EGFR/c-Met antibody, wherein the bispecific anti-EGFR/c-Met antibody comprises HC1 of SEQ ID NO:17, LC1 of SEQ ID NO:18, HC2 of SEQ ID NO:19, and LC2 of SEQ ID NO: 20.

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

In some embodiments, the subject obtains a c-Met exon 14 skipping mutation as a result of treatment with one or more prior anti-cancer therapies.

In some embodiments, the subject has acquired a c-Met exon 14 skipping mutation as a result of treatment with a kinase inhibitor.

In some embodiments, the subject has acquired a c-Met exon 14 skipping mutation as a result of treatment with an EGFR kinase inhibitor.

In some embodiments, the subject obtains a c-Met exon 14 skipping mutation as a result of treatment with a c-Met kinase inhibitor.

In some embodiments, 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.

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

In some embodiments, the kinase inhibitor is erlotinib, gefitinib, lapatinib, vandetanib, afatinib, oxitinib, lazertinib, britinib, criertinib, cabozantinib, camatinib, axitinib, lenvatinib, nintedanib, regorafenib, pazopanib, sorafenib, or sunitinib.

In some embodiments, the one or more prior anti-cancer therapies comprise carboplatin, paclitaxel, gemcitabine, cisplatin, vinorelbine, docetaxel, palbociclib, crizotinib, PD- (L)1 axis inhibitor, EGFR inhibitor, c-Met inhibitor, HER2 inhibitor, HER3 inhibitor, HER4 inhibitor, VEGFR inhibitor, erlotinib, gefitinib, lapatinib, vandetanib, afatinib, oxitinib, lazatinib, crirtitinib, cabozantinib, camatinib, axitinib, lenvatinib, nilatinib, regorafenib, pazopanib, sorafenib, or sunitinib, or any combination thereof.

In some embodiments, the subject is resistant to or has acquired resistance to an EGFR inhibitor. An exemplary EGFR inhibitor to which cancer may acquire resistance is the anti-EGFR antibody cetuximabPanitumumabMatuzumab, nimotuzumab, small molecule EGFR inhibitor erlotinibGefitinibEKB-569 (pelitinib, irreversible EGFR TKI), pan ErbB and other receptor tyrosine kinase inhibitors, lapatinib (EGFR and HER2 inhibitors), pelitinib (EGFR and HER2 inhibitors), vandetanib (ZD6474, ZACTIMA) ^TM EGFR, VEGFR2 and RET TKI), PF00299804 (Dacrotinib, irreversible pan-ErbB TKI), CI-1033 (irreversible pan-ErbB TKI), Afatinib (BIBW2992, irreversible pan-ErbB TKI), AV-412 (dual EGFR and ErbB2 inhibitors), EXEL-7647(EGFR, ErbB2, GEVGR and EphB4 inhibitors), CO-1686 (irreversible mutant selective EGFR TKI), AZD9291 (irreversible mutant selective EGFR TKI) and HKI-272 (lenatinib, irreversible EGFR/ErbB2 inhibitors).

Various qualitative and/or quantitative methods can be used to determine whether a subject is resistant to treatment with an anti-cancer therapy or has developed or is predisposed to developing resistance to such treatment. Symptoms that may be associated with resistance to anticancer therapy include a decrease or stasis in the patient's health status, an increase in the size of the tumor, a suppression or slowing of the decrease in tumor growth, and/or the spread of cancer cells from one location to other organs, tissues, or cells in the body. Reconstitution or worsening of various symptoms associated with cancer may also be indicative of: the subject has developed or is predisposed to developing resistance to anticancer therapy, such as anorexia, cognitive dysfunction, depression, dyspnea, fatigue, hormonal imbalance, neutropenia, pain, peripheral neuropathy, and sexual dysfunction. The symptoms associated with cancer may vary depending on the type of cancer. For example, symptoms associated with cervical cancer can include abnormal bleeding, abnormally large amounts of vaginal secretions, pelvic pain not associated with normal menstrual cycles, bladder pain or pain during urination, and bleeding between normal menstrual periods, after sexual intercourse, irrigation, or pelvic examination. Symptoms associated with lung cancer may include persistent cough, hemoptysis, shortness of breath, chest pain from wheezing, loss of appetite, involuntary weight loss, and fatigue. Symptoms of liver cancer may include loss of appetite and weight, abdominal pain (especially the upper right portion of the abdomen that may extend to the back and shoulders), nausea and vomiting, general weakness and fatigue, increased liver, abdominal swelling (ascites), and white yellowing of the skin and eyes (jaundice). The oncological technician can readily identify the symptoms associated with a particular cancer type.

An exemplary PD- (L)1 axis inhibitor is an antibody that binds PD-1 (such as nivolumab)PembrolizumabCentilizumab, Ceimipril monoclonal antibodyRibosizumab (tripolibamab), tirezumab, sibatuzumab, carpriclizumab, dolastazumab, gemuzumab or cerlizumab) or bound to PD-L1Antibodies (such as PD-L1 antibodies are Envorlizumab, Antilizumab)DuorufuzumabAnd Abameluumab)。

Commercially available antibodies can be purchased via an authorized reseller or pharmacy. The amino acid sequence structure of the small molecules can be found in USAN and/or INN filed by CAS registry.

In some embodiments, the subject is untreated.

In some embodiments, the c-Met exon 14 skipping mutation is a neogenetic mutation.

In some embodiments, a cancer positive for a c-Met exon 14 skipping mutation is more resistant to CDK4 amplification, EGFR amplification, KRAS amplification, MDM2 amplification, TERT amplification, NF 1R 2450; RAD50L597Vfs 5, MET c.3082+3A > G, EGFR, wild type EGFR, EGFR activating mutation, circulating HGF, c-MET amplification or increased levels of mutant KRAS or any combination thereof are positive.

EGFR activating mutations that may be associated with cancer include point mutations, deletion mutations, insertion mutations, inversion or gene amplification that result in an increase in at least one biological activity of EGFR (such as increased tyrosine kinase activity, receptor homodimer and heterodimer formation, enhanced ligand binding, etc.). The mutation may be located in the EGFR gene or any part of the regulatory region associated with the EGFR gene and includes a mutation in exon 18, 19, 20 or 21 or a mutation in the kinase domain. Other examples of EGFR activating mutations are those known in the art (see, e.g., U.S. patent publication No. US 2005/0272083). Information on EGFR and other ErbB receptors, including receptor homodimers and heterodimers, receptor ligands, autophosphorylation sites, and signaling molecules involved in ErbB-mediated signaling, is known in the art (see, e.g., Hynes and Lane, Nature Reviews Cancer 5:341-354, 2005).

In some embodiments, the EGFR activating mutation comprises: 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; deletion of E746-A750; absence of R748-P753; insertion of Ala (A) between M766 and A767; insertion of Ser, Val and Ala (SVA) between S768 and V769; insertion of Asn and ser (ns) between P772 and H773; insertion of one or more amino acids between D761 and E762, between a763 and Y764, between Y764 and Y765, between M766 and a767, between a767 and V768, between S768 and V769, between V769 and D770, between D770 and N771, between N771 and P772, between P772 and H773, between H773 and V774, between V774 and C775; 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 typically resistant to EGFR Tyrosine Kinase Inhibitors (TKIs) (see, e.g., international patent publication No. WO 2018/094225).

Exemplary c-Met activating mutations include point mutations, deletion mutations, insertion mutations, inversions, or gene amplifications that result in an increase in at least one biological activity of the c-Met protein (such as increased tyrosine kinase activity, receptor homodimer and heterodimer formation, enhanced ligand binding, etc.). The mutation may be located in any part of the c-Met gene or regulatory region associated with the gene, such as in the kinase domain of c-Met. Exemplary c-Met activating mutations are 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 amplification are well known.

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

In some embodiments, cancers that are positive for a c-Met exon 14 skipping mutation include 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, spleen cancer, testicular cancer, gastric cancer, thymus 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 that is positive for a c-Met exon 14 skipping mutation comprises lung cancer. In some embodiments, cancers that are positive for a c-Met exon 14 skipping mutation include gastric cancer. In some embodiments, the cancer that is positive for a c-Met exon 14 skipping mutation comprises colorectal cancer. In some embodiments, the cancer that is positive for a c-Met exon 14 skipping mutation comprises a brain cancer. In some embodiments, the cancer that is positive for a c-Met exon 14 skipping mutation comprises an epithelial cell cancer. In some embodiments, the cancer that is positive for a c-Met exon 14 skipping mutation comprises breast cancer. In some embodiments, the cancer that is positive for a c-Met exon 14 skipping mutation comprises ovarian cancer. In some embodiments, cancers that are positive for a c-Met exon 14 skipping mutation include colorectal cancer. In some embodiments, the cancer that is positive for a c-Met exon 14 skipping mutation comprises an anal cancer. In some embodiments, the cancer that is positive for a c-Met exon 14 skipping mutation comprises prostate cancer. In some embodiments, the cancer that is positive for a c-Met exon 14 skipping mutation comprises a renal cancer. In some embodiments, the cancer that is positive for a c-Met exon 14 skipping mutation comprises bladder cancer. In some embodiments, the cancer that is positive for a c-Met exon 14 skipping mutation comprises a head and neck cancer. In some embodiments, the cancer that is positive for a c-Met exon 14 skipping mutation comprises a pharyngeal cancer. In some embodiments, the cancer that is positive for a c-Met exon 14 skipping mutation comprises a nasal cancer. In some embodiments, the cancer that is positive for a c-Met exon 14 skipping mutation comprises pancreatic cancer. In some embodiments, the cancer that is positive for a c-Met exon 14 skipping mutation comprises a skin cancer. In some embodiments, the cancer that is positive for a c-Met exon 14 skipping mutation comprises an oral cancer. In some embodiments, the cancer that is positive for a c-Met exon 14 skipping mutation comprises a tongue cancer. In some embodiments, the cancer that is positive for a c-Met exon 14 skipping mutation comprises esophageal cancer. In some embodiments, the cancer that is positive for a c-Met exon 14 skipping mutation comprises a vaginal cancer. In some embodiments, the cancer that is positive for a c-Met exon 14 skipping mutation comprises cervical cancer. In some embodiments, the cancer that is positive for a c-Met exon 14 skipping mutation comprises a spleen cancer. In some embodiments, the cancer that is positive for a c-Met exon 14 skipping mutation comprises testicular cancer. In some embodiments, cancers that are positive for a c-Met exon 14 skipping mutation include gastric cancer. In some embodiments, the cancer that is positive for a c-Met exon 14 skipping mutation comprises a cancer of the thymus. In some embodiments, the cancer that is positive for a c-Met exon 14 skipping mutation comprises colon cancer. In some embodiments, the cancer that is positive for a c-Met exon 14 skipping mutation comprises thyroid cancer. In some embodiments, the cancer that is positive for a c-Met exon 14 skipping mutation comprises liver cancer. In some embodiments, the cancer that is positive for the c-Met exon 14 skipping mutation comprises hepatocellular carcinoma (HCC). In some embodiments, the cancer that is positive for a c-Met exon 14 skipping mutation comprises sporadic or hereditary Papillary Renal Cell Carcinoma (PRCC).

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

In NSCLC, specific mutations in the EGFR gene are associated with a high response rate (70-80%) to EGFR tyrosine kinase inhibitors (EGFR-TKI). Deletion of 5 amino acids in exon 19 or the point mutation L858R in EGFR correlated with EGFR-TKI sensitivity (Nakata and Gotoh, Expert Opin Ther Targets 16:771-781,2012). These mutations allow ligand-independent activation of EGFR kinase activity. Activating EGFR mutations occur in 10-30% of NSCLC patients and are significantly more common in eastern Asians, women, never-smokers and patients with adenocarcinoma histology (Janne and Johnson Clin Cancer Res 12(14Suppl):4416s-4420s, 2006). EGFR gene amplification is also strongly correlated with response following EGFR-TKI treatment (CappUzzo et al, J Natl Cancer Inst97: 643-. EGFR exon 20 insertion has been associated with EGFR TKI resistance.

Although most NSCLC patients with EGFR mutations initially responded to EGFR TKI treatment, virtually all patients acquired resistance that prevented a durable response. 50-60% of patients acquire resistance due to the second site mutation in the kinase domain of EGFR (T790M). Almost 60% of all tumors that become resistant to EGFR tyrosine kinase inhibitors increase c-Met expression, amplify the c-Met gene, or increase its unique known ligand, HGF (Turke et al, Cancer Cell,17:77-88,2010).

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

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

In some embodiments, the kinase inhibitor is an EGFR inhibitor, a c-Met inhibitor, a HER2 inhibitor, a HER3 inhibitor, a HER4 inhibitor, a VEGFR inhibitor, or an AXL inhibitor. 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 HER 2. In some embodiments, the kinase inhibitor is an inhibitor of HER 3. In some embodiments, the kinase inhibitor is an inhibitor of HER 4. In some embodiments, the kinase inhibitor is an inhibitor of VEGFR. In some embodiments, the kinase inhibitor is an inhibitor of AXL.

In some embodiments, the kinase inhibitor is erlotinib, gefitinib, lapatinib, vandetanib, afatinib, oxitinib, lazertinib, britinib, criertinib, cabozantinib, camatinib, axitinib, lenvatinib, nintedanib, regorafenib, pazopanib, sorafenib, or sunitinib.

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 oxitinib. In some embodiments, the kinase inhibitor is lacitinib. In some embodiments, the kinase inhibitor is bosutinib. In some embodiments, the kinase inhibitor is critinib. In some embodiments, the kinase inhibitor is cabozantinib. In some embodiments, the kinase inhibitor is carbamatinib. 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.

The anti-cancer therapies that can be administered in combination with the bispecific anti-EGFR/c-Met antibody in the methods of the present disclosure include any one or more of chemotherapeutic drugs or other anti-cancer therapeutics known to those of skill in the art. Chemotherapeutic agents are chemical compounds useful in the treatment of cancer and include growth inhibitors or other cytotoxic agents, and include alkylating agents, antimetabolites, antimicrotubule inhibitors, topoisomerase inhibitors, receptor tyrosine kinase inhibitors, angiogenesis inhibitors, and the like. Examples of chemotherapeutic agents include: alkylating agents, such as thiotepa and cyclophosphamideAlkyl sulfonates such as busulfan, endosulfan, and azinam; aziridines such as benzodidopa (benzodipa), carboquone (carboquone), metridopa (meteedopa), and ulidopa (uredopa); ethyleneimine and methylmelamine including hexamethylmelamine, triethylenemelamineTetramethylene melamine, triethylene phosphoramide (triethylenethiophosphoramide), triethylene melamine (trimethylolmelamine); nitrogen mustards (nitrosgen mustards), such as chlorambucil, chlorambucil (chlorenaphazine), cholorophosphamide (cholorophosphamide), estramustine (estramustine), ifosfamide, mechlorethamine hydrochloride (mechlorethamine oxide hydrochloride), melphalan, neomustard (novembichin), benzene mustard cholesterol (phenesterine), prednimustine (prednimustine), trofosfamide (trofosfamide), uracil mustard (uracilmustard); nitrosoureas (nitroureas) such as carmustine, chlorourethrine, fotemustine (fotemustine), lomustine (lomustine), nimustine (nimustine), ramustine (ranimustine); antibiotics such as aclacinomycins (aclacinomycins), actinomycins, anthranomycin (antrramycin), azaserin (azaserine), bleomycin, actinomycin (cactinomycin), calicheamicin (calicheamicin), carabicin, carminomycin (carminomycin), carzinophilin (carzinophilin), tryptomycin, dactinomycin, daunorubicin, ditorexin (deubixin), 6-diaza-5-oxo-L-norleucine, doxorubicin, epirubicin, eprinocin, idarubicin (marcellomycin), mitomycins (mitomycins), mycophenolic acid (mycophenolic acid), norramycin (nogalamycin), olivomycin (olivomycin), aureomycin (polypeomycin), potomycins (uromycin), puromycin, mycins (gentamycin), streptomycins (streptomycins), tuberculin (tuberculin), tuberculin (tuberculin), and streptomycins (streptomycins), and a, Zorubicin (zorubicin); antimetabolites such as methotrexate and 5-FU; folic acid analogs such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogs such as fludarabine (fludarabine), 6-mercaptopurine, thiamiprine (thiamiprine), thioguanine; pyrimidine analogs such as, for example, ancitabine (ancitabine), azacitidine (azacitidine), 6-azauridine (azauridine), carmofur (carmofur), cytarabine, dideoxyuridine (dideoxyuridine), doxifluridine(doxifluridine), enocitabine (enocitabine), floxuridine; androgens such as carotinone (calusterone), dromostanolone propionate, epitioandrostanol (epitiostanol), mepiquitane (mepiquitane), testolactone (testolactone); anti-adrenal species such as aminoglutethimide, mitotane, trostane (trilostane); folic acid supplements such as folinic acid (frilic acid); acetoglucurolactone (acegultone); (ii) an aldophosphamide glycoside; aminolevulinic acid (aminolevulinic acid); amsacrine; bestrabuucil; bisantrene; edatrexae; desphosphamide (defofamide); dimecorsine (demecolcine); diazaquinone; elfornithine; ammonium etitanium acetate; etoglut (etoglucid); gallium nitrate; a hydroxyurea; lentinan (lentinan); lonidamine (lonidamine); mitoguazone (mitoguzone); mitoxantrone; mopidanol (mopidanmol); diaminenitracridine (nitrarine); pentostatin (pentostatin); methionine mustard (phenamett); pirarubicin (pirarubicin); podophyllinic acid (podophyllic acid); 2-ethyl hydrazide; procarbazine;razoxane (rizoxane); sizofuran (sizofiran); helical germanium (spirogermanium); tenuazonic acid (tenuazonic acid); triimine quinone (triaziquone); 2,2' -trichlorotriethylamine; urethane (urethan); vindesine; dacarbazine; mannomustine (mannomustine); dibromomannitol (mitobronitol); dibromodulcitol (mitolactol); pipobromane (pipobroman); a polycytidysine; cytarabine (arabine) ("Ara-C"); cyclophosphamide; thiotepa; novel taxanes or members of the taxane family, e.g. paclitaxel (TDocetaxel) And the like; chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum analogs such as cisplatin and carboplatin; vinblastine(ii) a Platinum; etoposide (VP-16); ifosfamide; mitomycin C; mitoxantrone; vincristine; vinorelbine; catharanthine removal; nuantro (novantrone); (ii) teniposide; daunomycin; aminopterin (aminopterin); (ii) Hirodad; ibandronate (ibandronate); CPT-11; topoisomerase inhibitor RFS 2000; difluoromethyl ornithine (DMFO); retinoic acid; epothilones (esperamicins); capecitabine; inhibitors of receptor tyrosine kinases and/or angiogenesis, including sorafenibSunitinibPazopanib (VOTRIENT) ^TM ) Toxicillin (PALLADIA) ^TM ) Vandetanib (ZACTIMA) ^TM ) XidinibRegorafenib (BAY73-4506), axitinib (AG013736), lestaurtinib (CEP-701), erlotinibGefitinibAfatinib (BIBW 2992), lapatinibNeratinib (HKI-272), and the like, and pharmaceutically acceptable salts, acids, or derivatives of any of the above. Also included in this definition are anti-hormonal agents, such as anti-estrogenic agents, for modulating or inhibiting the effects of hormones on tumors, including, for example, tamoxifen, raloxifene, aromatase inhibiting 4(5) -imidazole, 4-hydroxyttamoxifen, trovaxifene, keoxifene, LY117018, onapristone, and toremifeneAnd antiandrogenDrugs such as flutamide, nilutamide, bicalutamide, leuprolide and goserelin; and a pharmaceutically acceptable salt, acid or derivative of any of the above. Other conventional cytotoxic chemical compounds such as those disclosed in Wiemann et al, 1985, in Medical Oncology (edited by Calabresi et al), Chapter 10, McMillan Publishing, are also suitable for use in the methods of the invention.

Administration of

The bispecific anti-EGFR/c-Met antibody can be administered in a pharmaceutically acceptable carrier. By "carrier" is meant a diluent, adjuvant, excipient, or vehicle with which an antibody of the invention is administered. Such vehicles may be liquids, such as water and oils, including those of petroleum, animal, vegetable origin, or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil, and the like. For example, 0.4% saline and 0.3% glycine can be used to formulate bispecific anti-EGFR/c-Met antibodies. These solutions are sterile and generally free of particulate matter. They may be sterilized by conventional well-known sterilization techniques, such as filtration. For parenteral administration, the carrier may include sterile water, and other excipients may be added to increase solubility or preservation. Injectable suspensions or solutions may also be prepared using water-based carriers along with suitable additives. Suitable vehicles and formulations comprising other human proteins (e.g., human serum albumin) are described, for example, in Remington: The Science and Practice of Pharmacy, 21 st edition, Troy, D.B. eds, Lipincott Williams and Wilkins, Philadelphia, PA, 2006, part 5, Pharmaceutical Manufacturing, page 691-.

The mode of administration may be any suitable route of delivery of the bispecific anti-EGFR-c-Met antibody to the host, such as parenteral administration, e.g. intradermal, intramuscular, intraperitoneal, intravenous or subcutaneous, pulmonary, transmucosal (buccal, intranasal, intravaginal, rectal), using formulations in the form of tablets, capsules, solutions, powders, gels, granules; and contained in syringes, implant devices, osmotic pumps, cassettes, micropumps; or other means known in the art as understood by the skilled artisan. Site-specific administration can be achieved, for example, by: intratumoral, parenteral, intrabronchial, intraabdominal, intracapsular, intracartilaginous, intracavitary, intracerebroventricular, intracolic, intracervical, intragastric, intrahepatic, intracardiac, intraosteal, intrapelvic, intrapericardiac, intraperitoneal, intrapleural, intraprostatic, intrapulmonary, intrarectal, intrarenal, intraretinal, intraspinal, intrasynovial, intrathoracic, intrauterine, intravascular, intravesical, intralesional, vaginal, rectal, buccal, sublingual, intranasal, or transdermal delivery.

In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered at a dose of between about 140mg to about 1750 mg. In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered at a dose of between about 140mg to about 1750 mg.

In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered in an amount of about 200mg, about 210mg, about 220mg, about 230mg, about 240mg, about 250mg, about 260mg, about 270mg, about 280mg, about 290mg, about 300mg, about 310mg, about 320mg, about 330mg, about 340mg, about 350mg, about 360mg, about 370mg, about 380mg, about 390mg, about 400mg, about 410mg, about 420mg, about 430mg, about 440mg, about 450mg, about 460mg, about 470mg, about 480mg, about 490mg, about 500mg, about 510mg, about 520mg, about 530mg, about 540mg, about 550mg, about 560mg, about 570mg, about 580mg, about 590mg, about 600mg, about 610mg, about 620mg, about 630mg, about 640mg, about 650mg, about 660mg, about 680mg, about 690mg, about 700mg, about 740mg, about 720mg, about 730mg, about 750mg, About 780mg, about 790mg, about 800mg, about 810mg, about 820mg, about 830mg, about 840mg, about 850mg, about 860mg, about 870mg, about 880mg, about 890mg, about 900mg, about 910mg, about 920mg, about 930mg, about 940mg, about 950mg, about 960mg, about 970mg, about 980mg, about 990mg, about 1000mg, about 1010mg, about 1020mg, about 1030mg, about 1040mg, about 1050mg, about 1060mg, about 1070mg, about 1080mg, about 1090mg, about 1100mg, about 1110mg, about 1120mg, about 1330mg, about 1140mg, about 1150mg, about 1160mg, about 1170mg, about 1180mg, about 1190mg, about 1200mg, about 1210mg, about 1220mg, about 1230mg, about 1240mg, about 1250mg, about 1260mg, about 1270mg, about 1280mg, about 1290mg, about 1310mg, about 1380mg, about 1410mg, about 1380mg, about 1370mg, about 1410mg, about 1380mg, about 1370mg, about 1380mg, about 1410mg, about 1380mg, about 1370mg, about 1410mg, about 1370mg, about 10mg, about 1370mg, about 10mg, About 1430mg, about 1440mg, about 1450mg, about 1460mg, about 1470mg, about 1480mg, about 1490mg, about 1500mg, about 1510mg, about 1520mg, about 1530mg, about 1540mg, about 1550mg, about 1560mg, about 1570mg, about 1580mg, about 1590mg, about 1600mg, about 1610mg, 1620mg, about 1630mg, about 1640mg, about 1650mg, about 1660mg, about 1670mg, about 1680mg, about 1690mg, about 1700mg, about 1710mg, about 1720mg, about 1730mg, about 1740mg, about 1750mg, about 1760mg, about 1770mg, about 1780mg, about 1790mg, about 1810mg, about 1820mg, about 1830mg, about 1840mg, about 1860mg, about 1870mg, about 1910mg, 1890mg, about 1950mg, about 1940mg, about 1920mg, about 2000mg, about 1920mg, about 1930mg, about 19310 mg, about 1930mg, about 1990mg, about 1920mg, about 2000mg, about 1920mg or about 1920 mg.

In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered at a dose of about 350mg, about 700mg, about 1050mg, or about 1400 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 once weekly. In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered about 1050mg once weekly. In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered about 1400mg once per week.

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 about 1050mg once every two weeks. In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered about 1400mg once every two weeks.

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.

For combination therapy, one or more anti-cancer agents may be administered using recommended doses and dosage forms of the anti-cancer agents.

Production 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 JNJ-372. JNJ-372 is characterized by the following amino acid sequence:

EGFR binding arms

1(HCDR1, EGFR binding arm)

TYGMH

2(HCDR2, EGFR binding arm)

VIWDDGSYKYYGDSVKG

3(HCDR3, EGFR binding arm)

DGITMVRGVMKDYFDY

[ SEQ ID NO:4(LCDR1, EGFR binding arm)

RASQDISSALV

5(LCDR2, EGFR binding arm)

DASSLES

6(LCDR3, EGFR binding arm)

QQFNSYPLT

7(HCDR1, c-Met binding arm)

SYGIS

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

12(LCDR3, c-Met binding arm)

QQANSFPIT

SEQ ID NO 13(VH, EGFR binding arm)

QVQLVESGGGVVQPGRSLRLSCAASGFTFSTYGMHWVRQAPGKGLEWVAVIWDDGSYKYYGDSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDGITMVRGVMKDYFDYWGQGTLVTVSS

14(VL, EGFR binding arm)

AIQLTQSPSSLSASVGDRVTITCRASQDISSALVWYQQKPGKAPKLLIYDASSLESGVPSRFSGSESGTDFTLTISSLQPEDFATYYCQQFNSYPLTFGGGTKVEIK

15(VH, c-Met binding arm)

QVQLVQSGAEVKKPGASVKVSCETSGYTFTSYGISWVRQAPGHGLEWMGWISAYNGYTNYAQKLQGRVTMTTDTSTSTAYMELRSLRSDDTAVYYCARDLRGTNYFDYWGQGTLVTVSS

[ SEQ ID NO:16(VL, c-Met binding arm)

DIQMTQSPSSVSASVGDRVTITCRASQGISNWLAWFQHKPGKAPKLLIYAASSLLSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANSFPITFGQGTRLEIK

>SEQ ID NO:17HC1

QVQLVESGGGVVQPGRSLRLSCAASGFTFSTYGMHWVRQAPGKGLEWVAVIWDDGSYKYYGDSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDGITMVRGVMKDYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFLLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

>SEQ ID NO:18LC1

AIQLTQSPSSLSASVGDRVTITCRASQDISSALVWYQQKPGKAPKLLIYDASSLESGVPSRFSGSESGTDFTLTISSLQPEDFATYYCQQFNSYPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

>SEQ ID NO:19HC2

QVQLVQSGAEVKKPGASVKVSCETSGYTFTSYGISWVRQAPGHGLEWMGWISAYNGYTNYAQKLQGRVTMTTDTSTSTAYMELRSLRSDDTAVYYCARDLRGTNYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

>SEQ ID NO:20LC2

DIQMTQSPSSVSASVGDRVTITCRASQGISNWLAWFQHKPGKAPKLLIYAASSLLSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANSFPITFGQGTRLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

Other bispecific anti-EGFR/c-Met antibodies that are publicly available may also be used in the methods of the present disclosure, provided that they exhibit similar properties when compared to JNJ-372, as described in U.S. patent No. 9,593,164. Bispecific anti-EGFR/c-Met antibodies useful in the methods of the invention can also be produced by combining publicly available EGFR-binding VH/VL domains and c-Met-binding VH/VL domains and testing the resulting bispecific antibody for characteristics as described in U.S. patent No. 9,593,164.

Bispecific anti-EGFR/c-Met antibodies for use in the methods of the present disclosure can be generated, for example, using Fab arm exchange (or half molecule exchange) between two monospecific bivalent antibodies by: substitutions are introduced at the heavy chain CH3 interface in each half molecule to facilitate the formation of heterodimers of two antibody halves with different specificities in an in vitro cell-free environment or using co-expression. The Fab arm exchange reaction is the result of disulfide bond isomerization and dissociation-association of the CH3 domain. The heavy chain disulfide bonds in the hinge region of the parent monospecific antibody are reduced. The resulting free cysteine of one of the parent monospecific antibodies forms an inter-heavy chain disulfide bond with the cysteine residue of a second parent monospecific antibody molecule, while the CH3 domain of the parent antibody is released and reformed by dissociation-association. The CH3 domain of the Fab arm can be engineered to favor heterodimerization rather than homodimerization. The resulting product is a bispecific antibody with two Fab arms or half-molecules each binding to a different epitope (i.e. an epitope on EGFR and an epitope on c-Met). For example, bispecific antibodies of the invention can be generated using the techniques described in international patent publication No. WO 2011/131746. In the case of IgG1 antibodies, the mutation F405L in one heavy chain and K409R in the other heavy chain can be used. For the IgG2 antibody, wild-type IgG2 and IgG2 antibodies with F405L and R409K substitutions may be used. For the IgG4 antibody, wild-type IgG4 and IgG4 antibodies with F405L and R409K substitutions may be used. To produce 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, the antibodies being incubated together under reducing conditions sufficient to allow disulfide isomerization of the cysteines in the hinge region; thereby generating bispecific antibodies by Fab arm exchange. The incubation conditions are optimally restored to non-reducing conditions. Exemplary reducing agents that can be used are 2-mercaptoethylamine (2-MEA), Dithiothreitol (DTT), Dithioerythritol (DTE), glutathione, tris (2-carboxyethyl) phosphine (TCEP), L-cysteine, and β -mercaptoethanol. For example, the following conditions may be used: incubating at a pH of 5-8, e.g., pH7.0 or pH7.4, in the presence of at least 25mM 2-MEA or in the presence of at least 0.5mM dithiothreitol at a temperature of at least 20 ℃ for at least 90 minutes.

Bispecific anti-EGFR/c-Met antibodies for use in the methods of the present disclosure can also be generated using designs such as Knob-in-Hole (Genentech), crossmabs (roche), and electrostatic matching (Chugai, Amgen, NovoNordisk, unitized), LUZ-y (Genentech), Strand Exchange engineered Domain body (SEEDbody) (EMD Serono), and biniclc (merus).

In the "knob and hole" strategy (see, e.g., international publication WO 2006/028936), selected amino acids that form the boundary of the CH3 domain in human IgG may be mutated at positions that affect the CH3 domain interaction, thereby promoting 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. Upon co-expression of both antibodies, heterodimers are formed due to the preferential interaction of heavy chains with a "hole" with heavy chains with a "knob". An exemplary CH3 substitution pair (denoted as modified position in the first CH3 domain of the first heavy chain/modified position in the second CH3 domain of the second heavy chain) that forms a knob and a hole is: T366Y/F405A, T366W/F405W, F405W/Y407A, T394W/Y407T, T394S/Y407A, T366W/T394S, F405W/T394S and T366W/T366S _ L368A _ Y407V.

In addition to using the "knob and hole" strategy to facilitate Fab wall exchange, CrossMAb technology also utilizes a CH1/CL domain replacement in one half-arm to ensure proper light chain pairing of the resulting bispecific antibody (see, e.g., U.S. patent 8,242,247).

Other crossover strategies can be used to generate full-length bispecific antibodies of the invention as follows: in one or both arms of the bispecific antibody, the variable domain or constant domain, or both domains, are exchanged between the heavy and light chains or within the heavy chain. These exchanges include, for example, VH-CH1 and VL-CL, VH and VL, CH3 and CL, and CH3 and CH1, as described in international patent publications WO2009/080254, WO2009/080251, WO2009/018386, and WO 2009/080252.

Other strategies may also be used, such as promoting heavy chain heterodimerization using electrostatic interactions by replacing positively charged residues on one CH3 surface and negatively charged residues on the second CH3 surface, as described in U.S. patent publication nos. US 2010/0015133; U.S. patent publication nos. US 2009/0182127; U.S. patent publication US2010/028637 or U.S. patent publication No. US 2011/0123532. In other strategies, heterodimerization may be promoted by the following substitutions (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): L351Y _ F405A _ Y407V/T394W, T366I _ K392M _ T394W/F405A _ Y407V, T366L _ K392M _ T394W/F405A _ Y407V, L351Y _ Y407A/T366A _ K409F, L351Y _ Y407A/T366V _ K409F, Y407A/T366A _ K409F or T350V _ L351Y _ F405A _ Y407V/T350V _ T366 _ V _ K392V _ T394 363672, as described in US patent publication US 2012/V or US patent publication No. US 2013/V.

SEEDbody technology can be used to generate bispecific antibodies of the invention. SEEDbodies have IgG residues substituted with IgA residues in their constant domains selected to promote heterodimerization as described in US patent US 20070287170.

Mutations are typically performed at the DNA level to the molecular level (such as the constant domains of antibodies) using standard methods.

Embodiments of the present disclosure provide methods of treating a subject having cancer comprising administering to the subject a combination therapy, wherein the combination therapy comprises a therapeutically effective amount of an isolated bispecific anti-Epidermal Growth Factor Receptor (EGFR)/hepatocyte growth factor receptor (c-Met) antibody and a therapeutically effective amount of a compound of formula (I)

Or a solvate, hydrate, tautomer or pharmaceutically acceptable salt thereof.

Embodiments of the present disclosure also provide methods of treating a subject having an EGFR-or c-Met expressing cancer comprising administering to the subject a combination therapy, wherein the combination therapy comprises a therapeutically effective amount of an isolated bispecific anti-EGFR/c-Met antibody and a therapeutically effective amount of a compound of formula (I)

Or a solvate, hydrate, tautomer or pharmaceutically acceptable salt thereof.

Embodiments of the present disclosure provide pharmaceutical combinations comprising a therapeutically effective amount of an isolated bispecific anti-Epidermal Growth Factor Receptor (EGFR)/hepatocyte growth factor receptor (c-Met) antibody and a therapeutically effective amount of a compound of formula (I)

Or a solvate, hydrate, tautomer or pharmaceutically acceptable salt thereof, for use as a medicament, in particular for use as a medicament in a subject.

Embodiments of the present disclosure provide pharmaceutical combinations comprising a therapeutically effective amount of an isolated bispecific anti-Epidermal Growth Factor Receptor (EGFR)/hepatocyte growth factor receptor (c-Met) antibody and a therapeutically effective amount of a compound of formula (I)

Or a solvate, hydrate, tautomer or pharmaceutically acceptable salt thereof, for use in the treatment of cancer, in particular for use in the treatment of cancer in a subject.

Embodiments of the present disclosure provide pharmaceutical combinations comprising a therapeutically effective amount of an isolated bispecific anti-Epidermal Growth Factor Receptor (EGFR)/hepatocyte growth factor receptor (c-Met) antibody and a therapeutically effective amount of a compound of formula (I)

Or a solvate, hydrate, tautomer or pharmaceutically acceptable salt thereof, for use in treating a subject suffering from a cancer comprising a c-Met exon 14 skipping mutation.

Embodiments of the present disclosure provide for the use of a combination comprising a therapeutically effective amount of an isolated bispecific anti-Epidermal Growth Factor Receptor (EGFR)/hepatocyte growth factor receptor (c-Met) antibody and a therapeutically effective amount of a compound of formula (I)

Or a solvate, hydrate, tautomer or pharmaceutically acceptable salt thereof, for use in the manufacture of a medicament for the treatment of cancer, in particular for the treatment of cancer in a subject.

Embodiments of the present disclosure provide for the use of a combination comprising a therapeutically effective amount of an isolated bispecific anti-Epidermal Growth Factor Receptor (EGFR)/hepatocyte growth factor receptor (c-Met) antibody and a therapeutically effective amount of a compound of formula (I)

Or a solvate, hydrate, tautomer or pharmaceutically acceptable salt thereof, for use in the manufacture of a medicament for the treatment of cancer, in particular for the treatment of a cancer comprising a c-Met exon 14 skipping mutation in a subject.

Embodiments of the present disclosure provide pharmaceutical combinations comprising a therapeutically effective amount of an isolated bispecific anti-Epidermal Growth Factor Receptor (EGFR)/hepatocyte growth factor receptor (c-Met) antibody and a therapeutically effective amount of a compound of formula (I)

Or a solvate, hydrate, tautomer or pharmaceutically acceptable salt thereof.

Embodiments of the present disclosure provide products comprising a therapeutically effective amount of an isolated bispecific anti-Epidermal Growth Factor Receptor (EGFR)/hepatocyte growth factor receptor (c-Met) antibody and a therapeutically effective amount of a compound of formula (I)

Or a solvate, hydrate, tautomer or pharmaceutically acceptable salt thereof, as a combined preparation for simultaneous, separate or sequential use in the treatment of cancer, in particular in the treatment of cancer in a subject.

Embodiments of the present disclosure provide products comprising a therapeutically effective amount of an isolated bispecific anti-Epidermal Growth Factor Receptor (EGFR)/hepatocyte growth factor receptor (c-Met) antibody and a therapeutically effective amount of a compound of formula (I)

Or a solvate, hydrate, tautomer or pharmaceutically acceptable salt thereof, as a combined preparation for simultaneous, separate or sequential use in the treatment of cancer, in particular in the treatment of a cancer comprising a c-Met exon 14 skipping mutation in a subject.

Embodiments of the present disclosure provide isolated bispecific anti-Epidermal Growth Factor Receptor (EGFR)/hepatocyte growth factor receptor (c-Met) antibodies, particularly therapeutically effective amounts of isolated bispecific anti-Epidermal Growth Factor Receptor (EGFR)/hepatocyte growth factor receptor (c-Met) antibodies, for use in combination with a compound of formula (I), particularly in combination with a therapeutically effective amount of a compound of formula (I),

or a solvate, hydrate, tautomer or pharmaceutically acceptable salt thereof, for use in the treatment of cancer, in particular for use in the treatment of cancer in a subject.

Embodiments of the present disclosure provide isolated bispecific anti-Epidermal Growth Factor Receptor (EGFR)/hepatocyte growth factor receptor (c-Met) antibodies, particularly therapeutically effective amounts of isolated bispecific anti-Epidermal Growth Factor Receptor (EGFR)/hepatocyte growth factor receptor (c-Met) antibodies, for use in combination with a compound of formula (I), particularly in combination with a therapeutically effective amount of a compound of formula (I),

or a solvate, hydrate, tautomer or pharmaceutically acceptable salt thereof, for use in the treatment of an EGFR-or c-Met-expressing cancer, in particular for use in the treatment of a cancer comprising a c-Met exon 14 skipping mutation in a subject.

In each embodiment, the bispecific anti-EGFR/c-Met antibody and the carbamatinib compound, or a solvate, hydrate, tautomer, or pharmaceutically acceptable salt thereof, may be administered simultaneously (e.g., as part of the same pharmaceutical composition, or in separate pharmaceutical compositions) or at different times, as described herein.

Pharmaceutically acceptable salt forms include pharmaceutically acceptable acidic salts/anionic salts or basic salts/cationic salts. Pharmaceutically acceptable acid/anion salts include acetate, benzenesulfonate, benzoate, bicarbonate, bitartrate, bromide, calcium edetate, camphorsulfonate, carbonate, chloride, citrate, dihydrochloride, edetate, edisylate, etonate, ethanesulfonate, fumarate, glucoheptonate, gluconate, glutamate, p-hydroxyacetaminophenylarsonate, hexylresorcinate, hydrobromide, hydrochloride, hydroxynaphthoate, iodide, isethionate, lactate, lactobionate, malate, maleate, malonate, mandelate, methanesulfonate, methylsulfate, mucate, naphthalenesulfonate, nitrate, pamoate, pantothenate, phosphate/diphosphate, polygalacturonate, salicylate, stearate, calcium edetate, picrate, acetate, or salt, and/diphosphate, acetate, and/or a salt, Hypoacetates, succinates, sulfates, bisulfates, tannates, tartrates, theachlorates, tosylates and triethiodide salts. Pharmaceutically acceptable basic/cationic salts include sodium, potassium, calcium, magnesium, diethanolamine, N-methyl-D-glucamine, L-lysine, L-arginine, ammonium, ethanolamine, piperazine, and triethanolamine salts.

Pharmaceutically acceptable acid salts are formed by reacting the free base form of the compound of formula (I) with a suitable inorganic or organic acid including, but not limited to, hydrobromic acid, hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, succinic acid, maleic acid, formic acid, acetic acid, propionic acid, fumaric acid, citric acid, tartaric acid, lactic acid, benzoic acid, salicylic acid, glutamic acid, aspartic acid, p-toluenesulfonic acid, benzenesulfonic acid, methanesulfonic acid, ethanesulfonic acid, naphthalenesulfonic acid (such as 2-naphthalenesulfonic acid) or hexanoic acid. Pharmaceutically acceptable acid addition salts of compounds of formula (I) may include or be, for example, hydrobromide, hydrochloride, sulphate, nitrate, phosphate, succinate, maleate, formate, acetate, propionate, fumarate, citrate, tartrate, lactate, benzoate, salicylate, glutamate, aspartate, p-toluenesulphonate, benzenesulphonate, methanesulphonate, ethanesulphonate, naphthalenesulphonate (e.g. 2-naphthalenesulphonate) or hexanoate.

The free acid or free base form of the compounds of formula (I) may be prepared from the corresponding base addition salt or acid addition salt form, respectively. For example, a compound of the invention in acid addition salt form can be converted to the corresponding free base form by treatment with a suitable base (e.g., ammonium hydroxide solution, sodium hydroxide, etc.). The compounds of the present invention in base addition salt form can be converted to the corresponding free acids by treatment with a suitable acid (e.g., hydrochloric acid, etc.).

In some embodiments, the compound of formula (I) or a solvate, hydrate, tautomer, or pharmaceutically acceptable salt thereof is represented by a compound of formula (II)

Solvates, hydrates or tautomers thereof.

In some embodiments, the cancer is a cancer that expresses c-Met.

In some embodiments, the cancer comprises a c-Met exon 14 skipping mutation.

Exemplary c-Met exon 14 skipping mutations include mutations in the c-Met gene in which at least a portion of exon 14 of c-Met is removed, or the c-Met transcript is spliced to remove at least a portion of c-Met exon 14. The deleted part may include a part encoding the negative regulatory site Tyr 1003 in the membrane-proximal region of the c-Met protein. The exon 14 region of the c-Met gene encompasses nucleotides 3284 to 3424 in the full-length nucleotide sequence of GenBank accession No. nm _000245 or residues 964 to 1009 in the full-length c-Met amino acid sequence of GenBank accession No. np _ 000236. Various mutations at the DNA level may lead to exon 14 skipping (see, e.g., Kong-Beltran et al (2006) Cancer Res.66; Dhanasekharan et al (2014) Nature Communication 10: 1038; Awad et al, J Clin Oncology 34:721,2016). Exon 14 of c-Met encodes 47 amino acids.

Methods for detecting c-Met mutations are well known.

In some embodiments, the bispecific anti-EGFR/c-Met antibody comprises: a first domain that binds EGFR comprising heavy chain complementarity determining region 1 of SEQ ID NO:1(HCDR 1), HCDR2 of SEQ ID NO:2, HCDR3 of SEQ ID NO:3, light chain complementarity determining region 1 of SEQ ID NO:4(LCDR 1), LCDR2 of SEQ ID NO:5, and LCDR3 of SEQ ID NO: 6; and a second domain that binds c-Met, the second domain comprising 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.

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

In some embodiments, the bispecific anti-EGFR/c-Met antibody is an IgG1 isotype. There are variations (e.g., well known allotypes) within the constant domain of IgG1, where the variations are located at positions 214, 356, 358, 422, 431, 435 or 436 (residue numbering according to EU numbering) (see, e.g., IMGT Web resources; IMGT repertoires (IG and TR); proteins and alleles; allotypes). The bispecific anti-EGFR/c-Met antibody may be any IgG1 allotype, such as G1m17, G1m3, G1m1, G1m2, G1m27, or G1m 28.

In some embodiments, the bispecific anti-EGFR/c-Met antibody comprises HC1 of SEQ ID No. 17, LC1 of SEQ ID No. 18, HC2 of SEQ ID No. 19, and LC2 of SEQ ID No. 20.

In some embodiments, the subject has a newly diagnosed cancer.

In some embodiments, the subject has a newly diagnosed c-Met expressing cancer.

In some embodiments, the subject has a newly diagnosed cancer comprising a c-Met exon 14 skipping mutation.

In some embodiments, the subject has not been treated with a Tyrosine Kinase Inhibitor (TKI).

In some embodiments, the subject has not received treatment with an EGFR Tyrosine Kinase Inhibitor (TKI).

In some embodiments, the subject is resistant to treatment with an EGFR TKI or will relapse.

In some embodiments, the subject is resistant to or has acquired resistance to treatment with a prior anti-cancer therapy.

In some embodiments, the prior anti-cancer therapy is chemotherapy, a targeted anti-cancer therapy, or a kinase inhibitor.

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

In some embodiments, the TKI is erlotinib, gefitinib, lapatinib, vandetanib, afatinib, oxitinib, povatinib, canertinib, regorafenib, pazopanib, sorafenib, or sunitinib.

In some embodiments, the cancer is non-small cell lung cancer (NSCLC), epithelial cell cancer, breast cancer, ovarian cancer, lung adenocarcinoma, squamous cell lung cancer, small cell lung 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, gastric cancer, thymus cancer, colon cancer, thyroid cancer, liver cancer, hepatocellular carcinoma (HCC), or sporadic or hereditary Papillary Renal Cell Carcinoma (PRCC). In some embodiments, the cancer is metastatic cancer.

In some embodiments, the cancer is NSCLC. In some embodiments, the cancer is an epithelial cell cancer. In some embodiments, the cancer is breast cancer. In some embodiments, the cancer is ovarian cancer. In some embodiments, the cancer is lung cancer. In some embodiments, the cancer is lung adenocarcinoma. In some embodiments, the cancer is squamous cell lung cancer. In some embodiments, the cancer is small cell lung cancer. In some embodiments, the cancer is colorectal cancer. In some embodiments, the cancer is anal cancer. In some embodiments, the cancer is prostate cancer. In some embodiments, the cancer is renal cancer. In some embodiments, the cancer is bladder cancer. In some embodiments, the cancer is a head and neck cancer. In some embodiments, the cancer is pharyngeal cancer. In some embodiments, the cancer is a nasal cancer. In some embodiments, the cancer is pancreatic cancer. In some embodiments, the cancer is a skin cancer. In some embodiments, the cancer is an oral cancer. In some embodiments, the cancer is tongue cancer. In some embodiments, the cancer is esophageal cancer. In some embodiments, the cancer is a vaginal cancer. In some embodiments, the cancer is cervical cancer. In some embodiments, the cancer is a cancer of the spleen. In some embodiments, the cancer is testicular cancer. In some embodiments, the cancer is gastric cancer. In some embodiments, the cancer is a thymus carcinoma. In some embodiments, the cancer is colon cancer. In some embodiments, the cancer is thyroid cancer. In some embodiments, the cancer is liver cancer. In some embodiments, the cancer is HCC. In some embodiments, the cancer is PRCC.

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

In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered at a dose of between about 200mg/kg and about 2000 mg/kg.

In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered at a dose of between about 350mg/kg and about 1400 mg/kg.

In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered in an amount of about 200mg, about 210mg, about 220mg, about 230mg, about 240mg, about 250mg, about 260mg, about 270mg, about 280mg, about 290mg, about 300mg, about 310mg, about 320mg, about 330mg, about 340mg, about 350mg, about 360mg, about 370mg, about 380mg, about 390mg, about 400mg, about 410mg, about 420mg, about 430mg, about 440mg, about 450mg, about 460mg, about 470mg, about 480mg, about 490mg, about 500mg, about 510mg, about 520mg, about 530mg, about 540mg, about 550mg, about 560mg, about 570mg, about 580mg, about 590mg, about 600mg, about 610mg, about 620mg, about 630mg, about 640mg, about 650mg, about 660mg, about 680mg, about 690mg, about 700mg, about 740mg, about 720mg, about 730mg, about 750mg, About 780mg, about 790mg, about 800mg, about 810mg, about 820mg, about 830mg, about 840mg, about 850mg, about 860mg, about 870mg, about 880mg, about 890mg, about 900mg, about 910mg, about 920mg, about 930mg, about 940mg, about 950mg, about 960mg, about 970mg, about 980mg, about 990mg, about 1000mg, about 1010mg, about 1020mg, about 1030mg, about 1040mg, about 1050mg, about 1060mg, about 1070mg, about 1080mg, about 1090mg, about 1100mg, about 1110mg, about 1120mg, about 1330mg, about 1140mg, about 1150mg, about 1160mg, about 1170mg, about 1180mg, about 1190mg, about 1200mg, about 1210mg, about 1220mg, about 1230mg, about 1240mg, about 1250mg, about 1260mg, about 1270mg, about 1280mg, about 1290mg, about 1310mg, about 1380mg, about 1410mg, about 1380mg, about 1370mg, about 1410mg, about 1380mg, about 1370mg, about 1380mg, about 1410mg, about 1380mg, about 1370mg, about 1410mg, about 1370mg, about 10mg, about 1370mg, about 10mg, About 1430mg, about 1440mg, about 1450mg, about 1460mg, about 1470mg, about 1480mg, about 1490mg, about 1500mg, about 1510mg, about 1520mg, about 1530mg, about 1540mg, about 1550mg, about 1560mg, about 1570mg, about 1580mg, about 1590mg, about 1600mg, about 1610mg, 1620mg, about 1630mg, about 1640mg, about 1650mg, about 1660mg, about 1670mg, about 1680mg, about 1690mg, about 1700mg, about 1710mg, about 1720mg, about 1730mg, about 1740mg, about 1750mg, about 1760mg, about 1770mg, about 1800mg, about 1790mg, about 1810mg, about 1820mg, about 1830mg, about 1840mg, about 1940mg, about 1860mg, about 1870mg, about 1910mg, about 890mg, about 1961880 mg, about 1951880 mg, about 19510 mg, about 1920mg, about 1950mg, about 19310 mg, about 1930mg, about 19310 mg, about 1990mg, about 1920mg, about 980mg, about 1920mg or about 1850 mg.

In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered at a dose of about 350mg, about 700mg, about 1050mg, or about 1400 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 1050 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 once weekly.

In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered once every two weeks.

In some embodiments, a compound of formula (I) or a solvate, hydrate, tautomer or pharmaceutically acceptable salt thereof (e.g., a compound of formula (II), carbamatinib hydrochloride) is administered at a dose of between about 50mg and about 500 mg. The dosage of a compound of formula (I) or a solvate, hydrate, tautomer or pharmaceutically acceptable salt thereof as described herein refers to the amount of free base of the compound of formula (I) in a dosage. For example, according to an embodiment wherein the dose comprises carbamatinib hydrochloride (compound of formula (II)), the dose refers to the amount of carbamatinib free base (compound of formula (I)).

In some embodiments, a compound of formula (I) or a solvate, hydrate, tautomer, or pharmaceutically acceptable salt thereof is administered at a dose of between about 50mg and about 400 mg. In some embodiments, a compound of formula (I) or a solvate, hydrate, tautomer, or pharmaceutically acceptable salt thereof is administered at a dose of between about 100mg and about 500 mg. In some embodiments, a compound of formula (I) or a solvate, hydrate, tautomer, or pharmaceutically acceptable salt thereof is administered at a dose of between about 200mg and about 450 mg. In some embodiments, a compound of formula (I) or a solvate, hydrate, tautomer, or pharmaceutically acceptable salt thereof is administered at a dose of between about 250mg and about 300 mg. In some embodiments, a compound of formula (I) or a solvate, hydrate, tautomer, or pharmaceutically acceptable salt thereof is administered at a dose of between about 100mg and about 400 mg.

In some embodiments, the compound of formula (II) or a solvate, hydrate, tautomer, or pharmaceutically acceptable salt thereof is administered at a dose of between about 50mg and about 400 mg. In some embodiments, the compound of formula (II) or a solvate, hydrate, tautomer, or pharmaceutically acceptable salt thereof is administered at a dose of between about 100mg and about 500 mg. In some embodiments, the compound of formula (II) or a solvate, hydrate, tautomer, or pharmaceutically acceptable salt thereof is administered at a dose of between about 200mg and about 450 mg. In some embodiments, the compound of formula (II) or a solvate, hydrate, tautomer, or pharmaceutically acceptable salt thereof is administered at a dose of between about 250mg and about 300 mg. In some embodiments, the compound of formula (II) or a solvate, hydrate, tautomer, or pharmaceutically acceptable salt thereof is administered at a dose of between about 100mg and about 400 mg.

In some embodiments, the compound of formula (I) or a solvate, hydrate, tautomer, or pharmaceutically acceptable salt thereof (e.g., the compound of formula (II), carbamotinib hydrochloride) is administered at a dose of at least about 50mg, at least about 100mg, at least about 150mg, at least about 200mg, at least about 250mg, at least about 300mg, at least about 350mg, at least about 400mg, at least about 450mg, or at least about 500 mg.

In some embodiments, the compound of formula (I) or a solvate, hydrate, tautomer, or pharmaceutically acceptable salt thereof (e.g., a compound of formula (II), carbamatinib hydrochloride) is administered once daily.

In some embodiments, the compound of formula (I) or a solvate, hydrate, tautomer, or pharmaceutically acceptable salt thereof (e.g., a compound of formula (II), carbamatinib hydrochloride) is administered once daily.

In some embodiments, the subject is further administered a third anti-cancer therapy.

In some embodiments, the third anti-cancer therapy is chemotherapy, a targeted anti-cancer therapy, or a kinase inhibitor.

The anti-cancer therapies that can be administered in combination with the bispecific anti-EGFR/c-Met antibody and carbamatinib in the methods of the present disclosure include any one or more of chemotherapeutic drugs or other anti-cancer therapeutics known to those of skill in the art. Chemotherapeutic agents are chemical compounds useful in the treatment of cancer and include growth inhibitors or other cytotoxic agents, and include alkylating agents, antimetabolites, antimicrotubule inhibitors,Topoisomerase inhibitors, receptor tyrosine kinase inhibitors, angiogenesis inhibitors, and the like. Examples of chemotherapeutic agents include: alkylating agents, such as thiotepa and cyclophosphamideAlkyl sulfonates such as busulfan, endosulfan, and azinam; aziridines such as benzodidopa (benzodipa), carboquone (carboquone), metridopa (meteedopa), and ulidopa (uredopa); ethyleneimine and methylmelamine, including hexamethylmelamine, triethylenemelamine, triethylenephosphoramide (triethylenephosphoramide), triethylenethiophosphorylamide (triethylenephosphoramide), and trimethylolmelamine (trimethylemelamine); nitrogen mustards (nitrosgen mustards), such as chlorambucil, chlorambucil (chlorenaphazine), cholorophosphamide (cholorophosphamide), estramustine (estramustine), ifosfamide, mechlorethamine hydrochloride (mechlorethamine oxide hydrochloride), melphalan, neomustard (novembichin), benzene mustard cholesterol (phenesterine), prednimustine (prednimustine), trofosfamide (trofosfamide), uracil mustard (uracilmustard); nitrosoureas (nitroureas) such as carmustine, chlorourethrine, fotemustine (fotemustine), lomustine (lomustine), nimustine (nimustine), ramustine (ranimustine); antibiotics such as aclacinomycin (aclacinomysins), actinomycin, anthranomycin (antrramycin), azaserine (azaserine), bleomycin, actinomycin (cactinomycin), calicheamicin (calicheamicin), carabicin, carminomycin (carminomycin), carzinophilin (carzinophilin), tryptomycin, dactinomycin, daunorubicin, ditorexin (deubixin), 6-diaza-5-oxo-L-norleucine, doxorubicin, epirubicin, orbifloxacin, idarubicin, marijumycin (marcellomycin), mitomycins (mitomycins), mycophenolic acid (mycophenolic acid), norramycin (nogalamycin), olivomycin (olivomycin), lomycin (polypeomycin), potricin, puromycin (gentamycin), streptomycin (streptomycin), streptomycin, and streptomycin, and streptomycin, or streptomycin, or streptomycin(ubenimex), neat (zinostatin), zorubicin (zorubicin); antimetabolites such as methotrexate and 5-FU; folic acid analogs such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogs such as fludarabine (fludarabine), 6-mercaptopurine, thioguanine (thiamirine), thioguanine; pyrimidine analogs such as ancitabine (ancitabine), azacitidine (azacitidine), 6-azauridine (azauridine), carmofur (carmofur), cytarabine, dideoxyuridine (dideoxyuridine), doxifluridine (doxifluridine), enocitabine (enocitabine), floxuridine; androgens such as carotinone (calusterone), dromostanolone propionate, epitioandrostanol (epitiostanol), mepiquitane (mepiquitane), testolactone (testolactone); anti-adrenal species such as aminoglutethimide, mitotane, trostane (trilostane); folic acid supplements such as folinic acid (frilic acid); acetoglucurolactone (acegultone); (ii) an aldophosphamide glycoside; aminolevulinic acid (aminolevulinic acid); amsacrine; bestrabuucil; bisantrene; edatrexae; desphosphamide (defofamide); dimecorsine (demecolcine); diazaquinone; elfornithine; ammonium etitanium acetate; etoglut (etoglucid); gallium nitrate; a hydroxyurea; lentinan (lentinan); lonidamine (lonidamine); mitoguazone (mitoguzone); mitoxantrone; mopidanol (mopidanmol); diaminenitracridine (nitrarine); pentostatin (pentostatin); methionine mustard (phenamett); pirarubicin (pirarubicin); podophyllinic acid (podophyllic acid); 2-ethyl hydrazide; procarbazine;razoxane (rizoxane); sizofuran (sizofiran); helical germanium (spirogermanium); tenuazonic acid (tenuazonic acid); triimine quinone (triaziquone); 2,2' -trichlorotriethylamine; urethane (urethan); vindesine; dacarbazine; mannomustine (mannomustine); dibromomannitol (mitobronitol); dibromodulcitol (mitolactotol); pipobromane (pipobroman); a polycytidysine; cytarabine (arabino)side) ("Ara-C"); cyclophosphamide; thiotepa; novel taxanes or members of the taxane family, e.g. paclitaxel (TDocetaxel) And the like; chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum analogs such as cisplatin and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitomycin C; mitoxantrone; vincristine; vinorelbine; catharanthine removal; nuantro (novantrone); (ii) teniposide; daunomycin; aminopterin (aminopterin); (ii) Hirodar; ibandronate (ibandronate); CPT-11; topoisomerase inhibitor RFS 2000; difluoromethyl ornithine (DMFO); retinoic acid; epothilones (esperamicins); capecitabine; inhibitors of receptor tyrosine kinases and/or angiogenesis, including sorafenibSunitinibPazopanib (VOTRIENT) ^TM ) Toxicillin (PALLADIA) ^TM ) Vandetanib (ZACTIMA) ^TM ) XidinibRegorafenib (BAY73-4506), axitinib (AG013736), lestaurtinib (CEP-701), erlotinibGefitinibAfatinib (BIBW 2992), lapatinibNeratinib (HKI-272), and the like, and pharmaceutically acceptable salts, acids, or derivatives of any of the above. Also included in this definition are anti-hormonal agents, such as anti-estrogenic agents, for modulating or inhibiting the effects of hormones on tumors, including, for example, tamoxifen, raloxifene, aromatase inhibiting 4(5) -imidazole, 4-hydroxyttamoxifen, trovaxifene, keoxifene, LY117018, onapristone, and toremifeneAnd antiandrogens such as flutamide, nilutamide, bicalutamide, leuprolide and goserelin; and a pharmaceutically acceptable salt, acid or derivative of any of the above. Other conventional cytotoxic chemical compounds such as those disclosed in Wiemann et al, 1985, in Medical Oncology (edited by Calabresi et al), Chapter 10, McMillan Publishing, are also suitable for use in the methods of the invention.

In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered prior to administration of the compound of formula (I) or a solvate, hydrate, tautomer, or pharmaceutically acceptable salt thereof.

In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered prior to administration of the compound of formula (II) or a solvate, hydrate, or tautomer thereof.

In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered after administration of the compound of formula (I) or a solvate, hydrate, tautomer, or pharmaceutically acceptable salt thereof.

In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered after administration of the compound of formula (II) or a solvate, hydrate, or tautomer thereof.

In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered one or more times after administration of the compound of formula (I) or a solvate, hydrate, tautomer, or pharmaceutically acceptable salt thereof.

In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered one or more times after the compound of formula (II) or a solvate, hydrate, or tautomer thereof.

In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered two, three, four, five, six, seven, eight, nine, ten or more times after the administration of the compound of formula (I) or a solvate, hydrate, tautomer or pharmaceutically acceptable salt thereof.

In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered two, three, four, five, six, seven, eight, nine, ten or more times after administration of the compound of formula (II) or a solvate, hydrate or tautomer thereof.

In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered intermittently after the compound of formula (I) or a solvate, hydrate, tautomer, or pharmaceutically acceptable salt thereof is administered.

In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered intermittently after the compound of formula (II) or solvate, hydrate, or tautomer thereof is administered.

The length of time between administration of the bispecific anti-EGFR/c-Met antibody and the compound of formula (I) or a solvate, hydrate, tautomer, or pharmaceutically acceptable salt thereof, or the compound of formula (II) or a solvate, hydrate, or tautomer thereof, or the third anti-cancer therapy may be several minutes (such as about 1 minute, 2 minutes, 5 minutes, 10 minutes, 30 minutes, or 60 minutes) or several hours (such as about 2 hours, 4 hours, 6 hours, 10 hours, 12 hours, 24 hours, or 36 hours) or such as about 2 days, 4 days, 7 days, 14 days, 21 days, 28 days, 35 days, 42 days, 49 days, 56 days or more.

The bispecific anti-EGFR/c-Met antibody and the compound of formula (I) or a solvate, hydrate, tautomer or pharmaceutically acceptable salt thereof or a third anti-cancer agent may be administered as a pharmaceutical composition.

The bispecific anti-EGFR/c-Met antibody and the compound of formula (II) or a solvate, hydrate, or tautomer thereof, or the third anti-cancer agent, can be administered as a pharmaceutical composition.

The bispecific anti-EGFR/c-Met antibody can be formulated as a pharmaceutical composition comprising the bispecific anti-EGFR/c-Met antibody and a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier may be one or more diluents, adjuvants, excipients, vehicles, and the like. Such vehicles may be liquids, such as water and oils, including those of petroleum, animal, vegetable origin, or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil, and the like. For example, 0.4% saline and 0.3% glycine can be used to formulate bispecific anti-EGFR/c-Met antibodies. These solutions are sterile and generally free of particulate matter. They may be sterilized by conventional well-known sterilization techniques, such as filtration.

In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered by intravenous injection. In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered by subcutaneous injection.

In some embodiments, the compound of formula (I) or a solvate, hydrate, tautomer or pharmaceutically acceptable salt thereof, or the compound of formula (II) or a solvate, hydrate or tautomer thereof, is administered as an oral formulation, such as, for example, a solid oral formulation such as a powder, capsule and tablet.

For solid oral formulations such as powders, capsules and tablets such as, for example, for the compound of formula (I) or the compound of formula (II), suitable carriers and additives include starches, sugars, diluents, granulating agents, lubricants, binders, disintegrating agents and the like. Solid oral formulations may also be coated with substances such as sugars or with enteric coatings to regulate the primary site of absorption. For parenteral administration, the carrier may include sterile water, and other excipients may be added to increase solubility or preservative properties. Injectable suspensions or solutions may also be prepared using water-based carriers along with suitable additives. Suitable vehicles and formulations comprising other human proteins (e.g., human serum albumin) are described, for example, in Remington: The Science and Practice of Pharmacy, 21 st edition, Troy, D.B. eds, Lipincott Williams and Wilkins, Philadelphia, PA, 2006, part 5, Pharmaceutical Manufacturing, page 691-.

The composition may contain pharmaceutically acceptable auxiliary substances as necessary to approximate physiological conditions, such as pH adjusting and buffering agents, stabilizing agents, thickening agents, lubricants, and coloring agents, and the like. The concentration of the bispecific anti-EGFR/c-Met antibody in the pharmaceutical formulation can vary from less than about 0.5% by weight, typically to at least about 1% to as much as 15%, 20%, 30%, 40% or 50% by weight, and can be selected based primarily on the desired dose, fluid volume, viscosity, etc., depending on the particular mode of administration selected. Pharmaceutical compositions comprising solid forms may comprise from about 0.1mg to about 2000mg (such as about 1mg, about 5mg, about 10mg, about 25mg, about 50mg, about 100mg, about 150mg, about 200mg, about 300mg, about 500mg, about 600mg or about 1000mg) of the active ingredient.

The mode of administration may be any suitable route of delivery of the antibody to the host, such as parenteral administration, e.g., intradermal, intramuscular, intraperitoneal, intravenous or subcutaneous, pulmonary, transmucosal (buccal, intranasal, intravaginal, rectal), using formulations in the form of tablets, capsules, solutions, powders, gels, granules; and contained in syringes, implant devices, osmotic pumps, cassettes, micropumps; or other means known in the art as understood by the skilled artisan. Site-specific administration can be achieved, for example, by: intratumoral, parenteral, intrabronchial, intraabdominal, intracapsular, intracartilaginous, intracavitary, intracerebroventricular, intracolic, intracervical, intragastric, intrahepatic, intracardiac, intraosteal, intrapelvic, intrapericardiac, intraperitoneal, intrapleural, intraprostatic, intrapulmonary, intrarectal, intrarenal, intraretinal, intraspinal, intrasynovial, intrathoracic, intrauterine, intravascular, intravesical, intralesional, vaginal, rectal, buccal, sublingual, intranasal, or transdermal delivery.

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

Detailed description of the preferred embodiments

The following clauses describe specific embodiments of the present invention.

1) A method of treating a subject having a cancer positive for a c-Met exon 14 skipping mutation, comprising administering to the subject having a cancer positive for a c-Met exon 14 skipping 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 having cancer with a bispecific anti-EGFR/c-Met antibody, the method comprising:

a) providing a biological sample from the subject;

b) determining the presence or absence of a c-Met exon 14 skipping mutation in said sample;

c) administering or providing administration of the bispecific anti-EGFR/c-Met antibody to the subject determined to have a c-Met exon 14 skipping mutation.

3) The method according to embodiment 1 or 2, 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 of SEQ ID NO:1(HCDR 1), HCDR2 of SEQ ID NO:2, HCDR3 of SEQ ID NO:3, light chain complementarity determining region 1 of SEQ ID NO:4(LCDR 1), LCDR2 of SEQ ID NO:5, and LCDR3 of SEQ ID NO: 6; a second domain that binds c-Met, wherein said 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.

4) The method of embodiment 3, wherein said first domain that specifically binds EGFR comprises the heavy chain variable region (VH) of SEQ ID NO:13 and the light chain variable region (VL) of SEQ ID NO:14 and said second domain that specifically binds c-Met comprises the VH of SEQ ID NO:15 and the VL of SEQ ID NO: 16.

5) The method according to any one of embodiments 1-4, wherein the bispecific anti-EGFR/c-Met antibody is an IgG1 isotype.

6) The method according to any one of embodiments 1-5, wherein the bispecific anti-EGFR/c-Met antibody comprises a first heavy chain of SEQ ID NO:17 (HC1), a first light chain of SEQ ID NO:18 (LC1), a second heavy chain of SEQ ID NO:19 (HC2), and a second light chain of SEQ ID NO:20 (LC 2).

7) The method of any one of embodiments 1-6, wherein the bispecific anti-EGFR/c-Met antibody comprises a double-branched glycan structure with a fucose content of between about 1% to about 15%.

8) The method according to any one of embodiments 1-7, wherein the subject is relapsed or resistant to treatment with one or more previous anti-cancer therapies.

9) The method of embodiment 8, 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.

10) The method of embodiment 8, wherein the one or more prior anti-cancer therapies comprise carboplatin, paclitaxel, gemcitabine, cisplatin, vinorelbine, docetaxel, palbociclib, crizotinib, PD- (L)1 axis inhibitor, EGFR inhibitor, c-Met inhibitor, HER2 inhibitor, HER3 inhibitor, HER4 inhibitor, VEGFR inhibitor, erlotinib, gefitinib, lapatinib, vandetanib, afatinib, oxitinib, laquiniib, critinib, caboztinib, camatinib, axitinib, lenvatinib, regorafenib, pazopanib, sorafenib, or sunitinib, or any combination thereof.

11) The method according to any one of embodiments 1-7, wherein the subject is untreated.

12) The method according to any one of embodiments 1-11, wherein the cancer positive for a c-Met exon 14 skipping mutation is amplified for CDK4, EGFR amplification, KRAS amplification, MDM2 amplification, TERT amplification, NF 1R 2450; RAD50L597Vfs 5, MET c.3082+3A > G, EGFR, wild type EGFR, EGFR activating mutation, circulating HGF, c-MET amplification or increased levels of mutant KRAS or any combination thereof are positive.

13) The method of embodiment 12, wherein the EGFR activating mutation comprises: 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; deletion of E746-A750; absence of R748-P753; insertion of ala (a) between M766 and a 767; insertion of Ser, Val and Ala (SVA) between S768 and V769; insertion of Asn and ser (ns) between P772 and H773; insertion of one or more amino acids between D761 and E762, between a763 and Y764, between Y764 and Y765, between M766 and a767, between a767 and V768, between S768 and V769, between V769 and D770, between D770 and N771, between N771 and P772, between P772 and H773, between H773 and V774, between V774 and C775; one or more deletions in EGFR exon 20; or one or more insertions in EGFR exon 20; or any combination thereof.

14) The method of embodiment 12, wherein the mutant KRAS comprises a G12V, G12C, G12A, or G12D substitution, or any combination thereof.

15) The method of any one of embodiments 1-14, wherein the cancer is lung cancer, gastric cancer, colorectal cancer, brain cancer, epithelial cell-derived 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, gastric cancer, thymus cancer, colon cancer, thyroid cancer, liver cancer, hepatocellular carcinoma (HCC), or sporadic or hereditary Papillary Renal Cell Carcinoma (PRCC), or any combination thereof.

16) The method of embodiment 15, 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.

17) The method of any one of embodiments 1-16, further comprising administering to the subject one or more anti-cancer therapies.

18) The method of embodiment 17, wherein the one or more anti-cancer therapies comprise chemotherapy, radiation therapy, surgery, targeted anti-cancer therapy, or a kinase inhibitor, or any combination thereof.

19) The method of embodiment 18, wherein the kinase inhibitor is an EGFR inhibitor, a c-Met inhibitor, a HER2 inhibitor, a HER3 inhibitor, a HER4 inhibitor, a VEGFR inhibitor, or an AXL inhibitor.

20) The method of embodiment 19, wherein the kinase inhibitor is erlotinib, gefitinib, lapatinib, vandetanib, afatinib, oxitinib, lacitinib, pozitinib, critinib, cabozinib, camatinib, axitinib, lenvatinib, nintedanib, regrafenib, pazopanib, sorafenib, or sunitinib.

21) The method according to any one of embodiments 1-20, wherein said c-Met exon 14 skipping mutation is a de novo mutation.

22) The method according to any one of embodiments 1-21, wherein said c-Met exon 14 skipping mutation is an acquired mutation.

23) The method of any one of embodiments 1-22, wherein the bispecific anti-EGFR/c-Met antibody is administered at a dose of between about 140mg to about 1750 mg.

24) The method according to any one of embodiments 1-23, wherein the bispecific anti-EGFR/c-Met antibody is administered at a dose of about 700mg, about 750mg, about 800mg, about 850mg, 900mg, 950mg, 1000mg, 1050mg, 1100mg, 1150mg, 1200mg, 1250mg, 1300mg, 1350mg or 1400 mg.

25) The method of any one of embodiments 1-24, wherein the bispecific anti-EGFR/c-Met antibody is administered twice weekly, once biweekly, once every three weeks, or once four weeks.

26) A method of treating a subject having a cancer positive for a c-Met exon 14 skipping mutation, comprising administering to the subject a combination therapy, wherein the combination therapy comprises a therapeutically effective amount of an isolated bispecific anti-Epidermal Growth Factor Receptor (EGFR)/hepatocyte growth factor receptor (c-Met) antibody and a therapeutically effective amount of a compound of formula (II)

Or a solvate, hydrate, tautomer or pharmaceutically acceptable salt thereof.

27) The method of embodiment 26, wherein the bispecific anti-EGFR/c-Met antibody comprises: a first domain that binds EGFR comprising heavy chain complementarity determining region 1 of SEQ ID NO. 1(HCDR1), HCDR2 of SEQ ID NO. 2, HCDR3 of SEQ ID NO. 3, light chain complementarity determining region 1 of SEQ ID NO. 4(LCDR 1), LCDR2 of SEQ ID NO. 5, and LCDR3 of SEQ ID NO. 6; and a second domain that binds c-Met, said second domain comprising 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.

28) The method of embodiment 27, wherein said first domain that binds EGFR comprises the heavy chain variable region (VH) of SEQ ID NO:13 and the light chain variable region (VL) of SEQ ID NO:14 and said second domain that binds c-Met comprises the VH of SEQ ID NO:15 and the VL of SEQ ID NO: 16.

29) The method according to any one of embodiments 26-28, wherein the bispecific anti-EGFR/c-Met antibody is an IgG1 isotype.

30) The method of any one of embodiments 26-29, wherein the bispecific anti-EGFR/c-Met antibody comprises a first heavy chain of SEQ ID NO:17 (HC1), a first light chain of SEQ ID NO:18 (LC1), a second heavy chain of SEQ ID NO:19 (HC2), and a second light chain of SEQ ID NO:20 (LC 2).

31) The method of any one of embodiments 26-30, wherein the bispecific anti-EGFR/c-Met antibody has a double-branched glycan structure with a fucose content of between about 1% to about 15%.

32) The method according to any one of embodiments 26-31, wherein the compound of formula (II) or a solvate, hydrate, tautomer or pharmaceutically acceptable salt thereof is 2-fluoro-N-methyl-4- [7- (quinolin-6-ylmethyl) imidazo [1,2-b ] [1,2,4] triazin-2-yl ] benzamide-hydrogen chloride-water (1/2/1).

33) The method according to any one of embodiments 26-32, wherein the subject is relapsed or resistant to treatment with one or more previous anti-cancer therapies.

34) The method of embodiment 33, 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.

35) The method of embodiment 33, wherein the one or more prior anti-cancer therapies comprises carboplatin, paclitaxel, gemcitabine, cisplatin, vinorelbine, docetaxel, palbociclib, crizotinib, PD- (L)1 axis inhibitor, EGFR inhibitor, c-Met inhibitor, HER2 inhibitor, HER3 inhibitor, HER4 inhibitor, VEGFR inhibitor, erlotinib, gefitinib, lapatinib, vandetanib, afatinib, oxitinib, lazatinib, boritinib, cabozitinib, axitinib, lenvatinib, nilatinib, regorafenib, pazopanib, sorafenib, or sunitinib, or any combination thereof.

36) The method according to any one of embodiments 26-32, wherein the subject is untreated.

37) The method of any one of embodiments 26-36, wherein a cancer positive for a c-Met exon 14 skipping mutation is amplified for CDK4, EGFR amplification, KRAS amplification, MDM2 amplification, TERT amplification, NF 1R 2450; RAD50L597Vfs 5, MET c.3082+3A > G, EGFR, wild type EGFR, EGFR activating mutation, circulating HGF, c-MET amplification or increased levels of mutant KRAS or any combination thereof are positive.

38) The method of embodiment 37, wherein the EGFR activating mutation comprises: 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; deletion of E746-A750; absence of R748-P753; insertion of ala (a) between M766 and a 767; insertion of Ser, Val and Ala (SVA) between S768 and V769; insertion of Asn and ser (ns) between P772 and H773; insertion of one or more amino acids between D761 and E762, between a763 and Y764, between Y764 and Y765, between M766 and a767, between a767 and V768, between S768 and V769, between V769 and D770, between D770 and N771, between N771 and P772, between P772 and H773, between H773 and V774, between V774 and C775; one or more deletions in EGFR exon 20; or one or more insertions in EGFR exon 20; or any combination thereof.

39) The method of embodiment 37, wherein the mutant KRAS comprises a G12V, G12C, G12A, or G12D substitution, or any combination thereof.

40) The method of any one of embodiments 26-39, wherein the cancer is lung cancer, gastric cancer, colorectal cancer, brain cancer, epithelial cell-derived 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, gastric cancer, thymus cancer, colon cancer, thyroid cancer, liver cancer, hepatocellular carcinoma (HCC), or sporadic or hereditary Papillary Renal Cell Carcinoma (PRCC), or any combination thereof.

41) The method of embodiment 40, 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.

42) The method of any one of embodiments 26-41, further comprising administering to the subject one or more anti-cancer therapies.

43) The method of embodiment 42, wherein said one or more anti-cancer therapies comprise chemotherapy, radiation therapy, surgery, targeted anti-cancer therapy, or a kinase inhibitor, or any combination thereof.

44) The method of embodiment 43, wherein the kinase inhibitor is an EGFR inhibitor, c-Met inhibitor, HER2 inhibitor, HER3 inhibitor, HER4 inhibitor, VEGFR inhibitor, or AXL inhibitor.

45) The method of embodiment 44, wherein the kinase inhibitor is erlotinib, gefitinib, lapatinib, vandetanib, afatinib, oxitinib, lacitinib, pozitinib, critinib, cabozinib, camatinib, axitinib, lenvatinib, nintedanib, regrafenib, pazopanib, sorafenib, or sunitinib.

46) The method according to any one of embodiments 26-45, wherein said c-Met exon 14 skipping mutation is a de novo mutation.

47) The method according to any one of embodiments 26-46, wherein said c-Met exon 14 skipping mutation is an acquired mutation.

48) The method of any one of embodiments 26-47, wherein the bispecific anti-EGFR/c-Met antibody is administered at a dose of between about 140mg to about 1750 mg.

49) The method according to any one of embodiments 26-48, wherein the bispecific anti-EGFR/c-Met antibody is administered at a dose of about 700mg, about 750mg, about 800mg, about 850mg, 900mg, 950mg, 1000mg, 1050mg, 1100mg, 1150mg, 1200mg, 1250mg, 1300mg, 1350mg or 1400 mg.

50) The method of any one of embodiments 26-49, wherein the bispecific anti-EGFR/c-Met antibody is administered twice weekly, once biweekly, once every three weeks, or once four weeks.

Example 1 JNJ-372 is effective in a c-Met driven PDX model

The efficacy of JNJ-372 was tested in a PDX model carrying the c-Met exon 14 skipping mutation. Erlotinib and cetuximab were used as a comparator. For these experiments, JNJ-372 was expressed in wild-type CHO cells and thus exhibited fucose content characteristics of wild-type CHO cells.

Tumor debris was collected from stock mice inoculated with selected primary human NSCLC tissues and used to inoculate BALB/c nude mice. Each mouse was inoculated subcutaneously on the right side with a fragment of primary human NSCLC model (LU2503) (passage 5, 2-4mm in diameter) for tumor development. When the mean tumor size reached about 151.5mm ³ At that time, treatment is initiated. Mice were randomly assigned to four entities according to their tumor sizeIn the experimental group. Each group consisted of 10 mice, 5 mice per cage. This day is denoted as day 0. The test substance was administered to tumor-bearing mice from day 0 to day 25 according to the predetermined protocol shown in table 1.

TABLE 1.

The primary endpoint was to see if tumor growth could be delayed or if tumor-bearing mice could be cured. Tumor size was measured twice per week in two dimensions using calipers, and can be measured in mm using the following formula ³ Volume is expressed in units: v is 0.5a × b ² Wherein a and b are the long and short diameters of the tumor, respectively. The tumor size was then used to calculate both the T-C value and the T/C value. T-C is calculated as where T is the mean tumor size of the treated group to a predetermined size (e.g., 1000 mm) ³ ) The time required (in days) and C is the time (in days) for the mean tumor size of the control group to reach the same size. The T/C value (in percent) is an indicator of the effectiveness of the anti-tumor; t and C are the average volumes of the treatment and control groups, respectively, on a given day. Mean tumor burden in control group reached 2000mm when scheduled dosing was complete and vehicle treated ³ The study was terminated at value (v). The significance of the difference between the mean values of tumor sizes of the comparison groups was analyzed using SPSS software. P<0.05 was considered statistically significant.

Tumor size (expressed as mean) within treatment group at different time points during treatment+SEM) are shown in table 2. Tumor growth inhibition is summarized in table 3. T-C was calculated as the tumor reached a predetermined size (1000 mm) ³ ). Fig. 1 shows tumor volume over time.

TABLE 2.

TABLE 3.

In group 2 (erlotinib 50mg/kg, 2 days without), group 3 (cetuximab 10mg/kg, BIW × 4) and group 4 (JNJ-37210 mg/kg, BIW × 4) the weight change on day 27 of treatment was-8%, 3.6% and 6.6%, respectively (data not shown).

On day 27, the mean tumor size of vehicle-treated mice reached 2160.5mm ³ And the study was terminated on day 28. Treatment with 10mg/kg BIW × 4 JNJ-372 produced a significant anti-tumor response (P) compared to vehicle treatment<0.0001). JNJ-372 treatment with 10mg/kg BIW 4 resulted in an average tumor size of 48.35mm ³ And its T-C cannot be calculated due to shrinkage of the tumor. In summary, in this study, JNJ-372 produced significant anti-tumor activity against primary human NSCLC tumor xenograft model LU2503 at 10mg/kg BIW × 4.

Example 2 JNJ-372 inhibition of tumor growth in patients with c-Met exon 14 skipping mutations

The potential clinical benefit of JNJ-372 in NSCLC with MET driver changes was evaluated. JNJ-372 was produced in cell lines for clinical studies, resulting in antibody fucose content below 15%.

Eligible patients with metastatic NSCLC received increasing doses (140mg to 1750mg) of JNJ-372 in RP2D in the expanded cohort of part 1 or part 2.JNJ-372 was administered Intravenously (IV) on days 1,2, 8, 15, and 22 of cycle 1 and on days 1 and 15 during the subsequent cycles in a 28-day cycle. Disease response was assessed by investigator assessment every 6 weeks according to RECIST v.1.1 criteria.

Patient 1: a76 year old patient with severe pre-treated metastatic gonadal squamous carcinoma NSCLC and having a MET exon 14 skipping mutation was first diagnosed with metastatic disease. Next Generation Sequencing (NGS) revealed a MET ex14 deletion following multiple chemotherapy regimens (carboplatin and paclitaxel, carboplatin and gemcitabine, cisplatin and vinorelbine, docetaxel), topical irradiation, and nivolumab; NF 1R 2450; CDK4 amplification; MDM2 amplification; EPHB1 amplification; FRS2 amplification; RAD 50N 598fs x 4. Patients received palbociclib without response in the trial followed by crizotinib for 11 months with stable disease as the best response. Post-crizotinib biopsy of the left axillary lymph node revealed metastatic adenosquamous carcinoma of the lung, and next generation sequencing (HopeSeq) demonstrated CDK4 amplification; amplification of EGFR; KRAS amplification; MET ex14 deletion, MET c.3082+3A > G; MDM2 amplification; NF 1R 2450; RAD50L597Vfs 5; TERT amplification; PD-L15%. As has been previously reported, this suggests EGFR and KRAS amplification as a potential mechanism for resistance to crizotinib. Patients started JNJ-372 at 1050mg in the study phase 1 part. The re-staging CT scan at 6 weeks showed a Partial Response (PR) of 32% reduction in tumor (9.1 to 6.2cm) compared to the baseline CT scan. CT scan at 12 weeks confirmed a sustained PR at 41% (5.4cm) reduction of the tumor. At the last visit, patients tolerated the treatment well and the toxicity was mild for 4 cycles and remained at the 1050mg dose. Fig. 2 shows CT scans indicating partial response at 6 and 12 weeks post-treatment.

A first primary MET ex14 deletion of NSCLC, resistant to crizotinib, demonstrated partial response to JNJ-372 in phase 1 studies after multi-line treatment is reported herein. This suggests a potential new treatment option for patients with MET ex14 loss.

Example 3 JNJ-372 is effective in inhibiting c-Met driven growth of NSCLC PDX tumors in LU2503。

The efficacy of JNJ-372 and the small molecule c-Met inhibitor camatinib was evaluated in NSCLC PDX model LU2503 carrying the c-Met exon 14 skipping mutation (seleck, S2788). LU2503PDX model established by CrownBio and described in Yang M, Shann B, Li Q, Song X, Cai J, Deng J, et al, overlying dependence resistance with a targetated linear region in a path-derived from non-reactive NSCLC path, International journal of cancer.2013; 132: E74-84. LU2503 tumor fragments were harvested from mice bearing stock tumors (passage R17P12) and used for inoculation into BALB/c nude mice. Each mouse was inoculated subcutaneously in the right flank with PDX LU2503 tumor fragments (approximately 2-3mm in diameter) for tumor development.

Tumor growth was measured twice a week after establishment of palpable lesions. Once the tumor volume reached about 200mm3, animals were randomly assigned to relevant study groups, each group having 8 mice. Randomization was performed according to the tumor size of each group, and the day of randomization was denoted as day 0. In table 4, treatment was initiated according to study design on the same day of randomization.

TABLE 4.

i.p.: intraperitoneal administration; p.o.: oral administration; BIW: once every two weeks; BID: twice daily

The study endpoint was a comparison of tumor growth in each group at the end of treatment with subsequent tumor growth after dosing had ceased. Tumor size was measured twice weekly in two dimensions using calipers, and in mm using the following equation ³ Volume is expressed in units: v ═ 2(L × W)/where V is tumor volume, L is tumor length (longest tumor size) and W is tumor width (longest tumor size perpendicular to L). Tumor sizes (expressed as mean + SEM) within the treatment groups at different time points during treatment are shown in table 5, and tumor growth curves over time are shown in fig. 3A.

TABLE 5.

a: data represent mean tumor volume ± Standard Error of Mean (SEM).

Tumor growth inhibition (TGI%) is an indicator of anti-tumor activity and is calculated as% Δ inhibition of 100 × ((C-C0) - (T-T0))/(C-C0), T and C are the average tumor volumes of the treatment and control groups, respectively, on the day when the average Tumor Volume (TV) of the vehicle group reached the renci endpoint (>2000mm3, day 13). Tumor growth inhibition is summarized in table 6. To compare the mean tumor volume of the treatment groups to the vehicle control group, all groups were first examined for the hypothesis of homogeneity of variance using the Bartlett test. The Bartlett test has a p-value <0.05, and for overall equality of the medians of all groups, the Kruskal-Wallis test (<0.05) was run. Post hoc tests were then further performed by running the Conover nonparametric test using a single step P-value adjustment (P <0.05 was considered statistically significant). Body weight changes were monitored and shown in fig. 3B.

TABLE 6.

a: data represent mean tumor volume ± SEM.

b: Δ% inhibition was calculated as follows:

c (13 or 0): mean tumor volume of control group at indicated study day.

T (13 or 0): mean tumor volume in treatment group at indicated study day.

c: the P value is calculated by performing a Conover nonparametric many-to-one comparison test.

As shown in figure 3A, 10mg/kg BIW x 3 JNJ-372 and 10mg/kg BID x 21 carbamtinib produced significant antitumor activity against primary LU2503 human NSCLC xenografts in this study (108.44% and 109.78% TGI, respectively). During the no dose monitoring period after day 21, all 8 carbamatinib-treated animals showed faster tumor regrowth compared to JNJ-372-treated animals (6 out of 8 animals had no measurable tumor at the end of the study).

Example 4 combination of JNJ-372 with MET-TKI (Caratinib) shows deeper and more persistent evidence in LU2503 Tumor inhibition of。

The follow-up study in LU2503 included combination treatment of JNJ-372 and carbamatinib to see if there were any additional benefits when JNJ-372 was combined with MET-TKI. Tumor fragments from stock mice (passage R18P2) were implanted subcutaneously into the right flank of BALB/c nude mice for tumor propagation.

When the average tumor size reaches about 150-200mm ³ At that time, treatment was initiated in the efficacy study according to the following design (table 7). Animals were randomly assigned to four experimental groups of 8 mice each, depending on tumor size. Tumor size was measured twice weekly and body weight was monitored as described in example 3. Tumor sizes (expressed as mean + SEM) within the treatment groups at different time points during treatment are shown in table 8, tumor growth curves over time are shown in fig. 4A, and body weight changes are plotted in fig. 4B.

TABLE 7.

i.p.: intraperitoneally; p.o.: oral administration; BIW: once every two weeks; BID: twice daily

TABLE 8.

a: data represent mean tumor volume ± SEM; BIW: once every two weeks; BID: twice daily

When the mean tumor size reached 1400mm on day 14 ³ When, isotype and vehicle control groups were terminated. Tumor growth inhibition (TGI%) in each treatment group was calculated using the equation Δ inhibition of 100 × ((C-C0) - (T-T0))/(C-C0) and shown in table 9, T and C are the average tumor volumes of the treatment group and the control group, respectively. Statistical analysis was performed using the same method as example 3.

TABLE 9.

a: data represent mean tumor volume ± SEM.

b: Δ% inhibition was calculated as follows:

c (14 or 0): mean tumor volume of control group at indicated study day.

T (14 or 0): mean tumor volume in treatment group at indicated study day.

c: the P value is calculated by performing a Conover nonparametric many-to-one comparison test.

Although all three treatment groups induced rapid tumor regression, the combination of JNJ-372 and carbamatinib produced a longer and more sustained response compared to monotherapy in this study. 2 of the 8 animals in the JNJ-372 group grew again on day 70 and all animals in the combination group remained tumor free until the end of the study (more than 120 days).

A concomitant study to assess pharmacodynamic parameters in LU2503 was performed and treatment was initiated when tumors reached approximately 450-. Samples were collected at time points according to the design in table 10 in all treatment groups. The snap-frozen tumors were homogenized in RIPA buffer containing a mixture of protease and phosphatase inhibitors. Each lane was loaded with 50mg total protein (100 mg for phosphorylated EGFR) for western blotting (see figure 5). Total MET (Cell Signaling,8198S), EGFR (Cell Signaling,4267S), MET-Tyr1234/1235(Cell Signaling,3077S), MET-Tyr1349(Cell Signaling,3133S), EGFR-Tyr1068(Cell Signaling,3777S), and EGFR-117Tyr 3(Cell Signaling,4407S) protein levels were detected using the indicated primary antibodies, and the following secondary antibodies were detected using the manufacturer' S recommended methods: IRDye800CW goat-anti-rabbit (Li-Cor,925-32211) and IRDye680RD goat-anti-mouse (Li-Cor, 925-68070). Protein levels were quantified using Image J software and normalized to load control β -tubulin in fig. 6A-6B and 7A-7C.

Watch 10.

i.p.: intraperitoneal administration; p.o.: oral administration; BIW: once every two weeks; BID: twice daily

Consistent with published mechanisms of action, JNJ-372 causes downregulation of the total EGFR and MET receptors. Unexpectedly, the JNJ-372/Caratinib combination group further reduced EGFR and MET receptor levels. Similar synergy in inhibition of phosphoegfr and phosphomet signaling was also observed in the combination group. In conclusion, more robust PD marker inhibition in combination therapy is consistent with the observed efficacy and warrants further investigation.

Example 5 JNJ-372 effective inhibition of tumorigenesis in a DFCI-440MET exon 14 skipping NSCLC PDX model Long and long。

The efficacy of JNJ-372, the small molecule MET inhibitor camatinib (Advanced ChemBlocks, Burlingame, CA), and combinations thereof was compared in NSCLC patient explant (PDX) model NSCLC PDX model DFCI-440, developed at Dana Farber Institute (Boston, MA), which carries a MET exon 14 skipping mutation. Harvesting from tumor-bearing miceTumor fragments were pooled and used for inoculation into female NSG ^TM In mice. Each mouse was inoculated subcutaneously in the right flank with PDX DFCI-440 tumor fragments (approximately 2-3mm in diameter) for tumor development.

Tumor growth was measured twice a week after establishment of palpable lesions. Once the tumor volume reached 150-250mm3, animals were randomly assigned to the relevant study groups, each group having 8 mice. Day of randomization was denoted as day 0; treatment was started on day 0 and the dosing schedule in table 11 was followed.

TABLE 11.

LFI: a low fucose isoform; HPMC: hydroxypropyl methylcellulose; i.p.: intraperitoneal administration; p.o.: oral administration;

the study endpoint was a comparison of tumor growth in each group at the end of treatment with subsequent tumor growth after dosing had ceased. Tumor size was measured twice weekly in two dimensions using calipers, and volume was expressed in mm3 using the following equation: v ═ 2(L × W)/where V is tumor volume, L is tumor length (longest tumor size) and W is tumor width (longest tumor size perpendicular to L). Tumor sizes (expressed as mean + SEM) over time for each group are shown in figure 8.

As shown in fig. 8, both JNJ-372 and carbamatinib, as single agents, significantly inhibited DFCI-440 tumor growth and caused recovery of these tumors. After treatment cessation, tumors in mice treated with JNJ-372 or carbamatinib as single agents restored growth. However, combination treatment with JNJ-372+ carbamatinib was also effective in inhibiting tumor growth and leading to tumor regression, however, combination treatment eliminated tumors in 8 out of 8 mice, even after treatment was discontinued.

Sequence listing

<110> Janssen Biotech, Inc.

Knoblauch, Roland

Laquerre, Sylvie

Moores, Sheri

<120> treatment of patients with c-MET exon 14 skipping mutations

<130> JBI6242WOPCT1

<140> pending transfer

<141> 2021-02-12

<150> US 62/975,406

<151> 2020-02-12

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<212> PRT

<213> Artificial sequence

<220>

<223> heavy chain 1

<400> 17

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Thr Tyr

20 25 30

Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Val Ile Trp Asp Asp Gly Ser Tyr Lys Tyr Tyr Gly Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Gly Ile Thr Met Val Arg Gly Val Met Lys Asp Tyr Phe

100 105 110

Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr

115 120 125

Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser

130 135 140

Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu

145 150 155 160

Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His

165 170 175

Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser

180 185 190

Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys

195 200 205

Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu

210 215 220

Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro

225 230 235 240

Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys

245 250 255

Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val

260 265 270

Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp

275 280 285

Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr

290 295 300

Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp

305 310 315 320

Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu

325 330 335

Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg

340 345 350

Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys

355 360 365

Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp

370 375 380

Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys

385 390 395 400

Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Leu Leu Tyr Ser

405 410 415

Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser

420 425 430

Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser

435 440 445

Leu Ser Leu Ser Pro Gly Lys

450 455

<210> 18

<211> 214

<212> PRT

<213> Artificial sequence

<220>

<223> light chain 1

<400> 18

Ala Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Ser Ala

20 25 30

Leu Val Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Glu Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Asn Ser Tyr Pro Leu

85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala

100 105 110

Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly

115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala

130 135 140

Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln

145 150 155 160

Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser

165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr

180 185 190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser

195 200 205

Phe Asn Arg Gly Glu Cys

210

<210> 19

<211> 449

<212> PRT

<213> Artificial sequence

<220>

<223> heavy chain 2

<400> 19

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Glu Thr Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Gly Ile Ser Trp Val Arg Gln Ala Pro Gly His Gly Leu Glu Trp Met

35 40 45

Gly Trp Ile Ser Ala Tyr Asn Gly Tyr Thr Asn Tyr Ala Gln Lys Leu

50 55 60

Gln Gly Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Leu Arg Gly Thr Asn Tyr Phe Asp Tyr Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe

115 120 125

Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu

130 135 140

Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp

145 150 155 160

Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu

165 170 175

Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser

180 185 190

Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro

195 200 205

Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys

210 215 220

Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro

225 230 235 240

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser

245 250 255

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp

260 265 270

Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn

275 280 285

Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val

290 295 300

Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu

305 310 315 320

Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys

325 330 335

Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr

340 345 350

Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr

355 360 365

Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu

370 375 380

Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu

385 390 395 400

Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys

405 410 415

Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu

420 425 430

Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

435 440 445

Lys

<210> 20

<211> 214

<212> PRT

<213> Artificial sequence

<220>

<223> light chain 2

<400> 20

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Asn Trp

20 25 30

Leu Ala Trp Phe Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Ala Ala Ser Ser Leu Leu Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ala Asn Ser Phe Pro Ile

85 90 95

Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Arg Thr Val Ala Ala

100 105 110

Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly

115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala

130 135 140

Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln

145 150 155 160

Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser

165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr

180 185 190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser

195 200 205

Phe Asn Arg Gly Glu Cys

210

<210> 21

<211> 47

<212> PRT

<213> Intelligent (Homo sapiens)

<400> 21

Asp Leu Gly Ser Glu Leu Val Arg Tyr Asp Ala Arg Val His Thr Pro

1 5 10 15

His Leu Asp Arg Leu Val Ser Ala Arg Ser Val Ser Pro Thr Thr Glu

20 25 30

Met Val Ser Asn Glu Ser Val Asp Tyr Arg Ala Thr Phe Pro Glu

35 40 45

Claims (50)

1. A method of treating a subject having a cancer positive for a c-Met exon 14 skipping mutation, comprising administering to the subject having a cancer positive for a c-Met exon 14 skipping 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 having cancer with a bispecific anti-EGFR/c-Met antibody, the method comprising:

a. providing a biological sample from the subject;

b. determining the presence or absence of a c-Met exon 14 skipping mutation in said sample;

c. administering or providing administration of the bispecific anti-EGFR/c-Met antibody to the subject determined to have a c-Met exon 14 skipping mutation.

3. The method of claim 1 or 2, 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 of SEQ ID NO:1(HCDR 1), HCDR2 of SEQ ID NO:2, HCDR3 of SEQ ID NO:3, light chain complementarity determining region 1 of SEQ ID NO:4(LCDR 1), LCDR2 of SEQ ID NO:5, and LCDR3 of SEQ ID NO: 6; a second domain that binds c-Met, wherein said 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.

4. The method of claim 3, wherein the first domain that specifically binds EGFR comprises the heavy chain variable region (VH) of SEQ ID NO 13 and the light chain variable region (VL) of SEQ ID NO 14, and the second domain that specifically binds c-Met comprises the VH of SEQ ID NO 15 and the VL of SEQ ID NO 16.

5. The method of any one of claims 1-4, wherein the bispecific anti-EGFR/c-Met antibody is an IgG1 isotype.

6. The method of any one of claims 1-5, wherein the bispecific anti-EGFR/c-Met antibody comprises a first heavy chain of SEQ ID NO 17 (HC1), a first light chain of SEQ ID NO 18 (LC1), a second heavy chain of SEQ ID NO 19 (HC2), and a second light chain of SEQ ID NO 20 (LC 2).

7. The method of any one of claims 1-6, wherein the bispecific anti-EGFR/c-Met antibody comprises a double-branched glycan structure with a fucose content of between about 1% to about 15%.

8. The method of any one of claims 1-7, wherein the subject is relapsed or resistant to treatment with one or more previous anti-cancer therapies.

9. The method of claim 8, 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.

10. The method of claim 8, wherein the one or more prior anti-cancer 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, erlotinib, gefitinib, lapatinib, vandetanib, afatinib, oxitinib, lazatinib, critinib, cabozantinib, camatinib, axitinib, lenvatinib, nilatinib, regrafenib, pazopanib, felinib, or sunitinib, or any combination thereof.

11. The method of any one of claims 1-7, wherein the subject is untreated.

12. The method of any one of claims 1-11, wherein a cancer positive for a c-Met exon 14 skipping mutation is amplified for CDK4, EGFR amplification, KRAS amplification, MDM2 amplification, TERT amplification, NF 1R 2450; RAD50L597Vfs 5, MET c.3082+3A > G, EGFR, wild type EGFR, EGFR activating mutation, circulating HGF, c-MET amplification or increased levels of mutant KRAS or any combination thereof are positive.

13. The method of claim 12, wherein the EGFR activating mutation comprises: 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; deletion of E746-A750; absence of R748-P753; insertion of ala (a) between M766 and a 767; insertion of Ser, Val and Ala (SVA) between S768 and V769; insertion of Asn and ser (ns) between P772 and H773; insertion of one or more amino acids between D761 and E762, between a763 and Y764, between Y764 and Y765, between M766 and a767, between a767 and V768, between S768 and V769, between V769 and D770, between D770 and N771, between N771 and P772, between P772 and H773, between H773 and V774, between V774 and C775; one or more deletions in EGFR exon 20; or one or more insertions in EGFR exon 20; or any combination thereof.

14. The method of claim 12, wherein the mutant KRAS comprises a G12V, G12C, G12A, or G12D substitution, or any combination thereof.

15. The method of any one of claims 1-14, wherein the cancer is lung cancer, gastric cancer, colorectal cancer, brain cancer, epithelial cell-derived 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, gastric cancer, thymus cancer, colon cancer, thyroid cancer, liver cancer, hepatocellular carcinoma (HCC), or sporadic or hereditary Papillary Renal Cell Carcinoma (PRCC), or any combination thereof.

16. The method of claim 15, wherein lung cancer is non-small cell lung cancer (NSCLC), Small Cell Lung Cancer (SCLC), or lung adenocarcinoma, lung sarcomatoid carcinoma, or any combination thereof.

17. The method of any one of claims 1-16, further comprising administering one or more anti-cancer therapies to the subject.

18. The method of claim 17, wherein the one or more anti-cancer therapies comprise chemotherapy, radiation therapy, surgery, targeted anti-cancer therapy, or a kinase inhibitor, or any combination thereof.

19. The method of claim 18, wherein the kinase inhibitor is an EGFR inhibitor, a c-Met inhibitor, a HER2 inhibitor, a HER3 inhibitor, a HER4 inhibitor, a VEGFR inhibitor, or an AXL inhibitor.

20. The method of claim 19, wherein the kinase inhibitor is erlotinib, gefitinib, lapatinib, vandetanib, afatinib, oxitinib, lacitinib, pozitinib, critinib, cabozinib, camatinib, axitinib, lenvatinib, nintedanib, regrafenib, pazopanib, sorafenib, or sunitinib.

21. The method of any one of claims 1-20, wherein the c-Met exon 14 skipping mutation is a de novo mutation.

22. The method of any one of claims 1-21, wherein the c-Met exon 14 skipping mutation is an acquired mutation.

23. The method of any one of claims 1-22, wherein the bispecific anti-EGFR/c-Met antibody is administered at a dose of between about 140mg to about 1750 mg.

24. The method of any one of claims 1-23, wherein the bispecific anti-EGFR/c-Met antibody is administered at a dose of about 700mg, about 750mg, about 800mg, about 850mg, 900mg, 950mg, 1000mg, 1050mg, 1100mg, 1150mg, 1200mg, 1250mg, 1300mg, 1350mg, or 1400 mg.

25. The method of any one of claims 1-24, wherein the bispecific anti-EGFR/c-Met antibody is administered twice weekly, once biweekly, once every three weeks, or once four weeks.

26. A method of treating a subject having a cancer positive for a c-Met exon 14 skipping mutation, comprising administering to the subject a combination therapy, wherein the combination therapy comprises a therapeutically effective amount of an isolated bispecific anti-Epidermal Growth Factor Receptor (EGFR)/hepatocyte growth factor receptor (c-Met) antibody and a therapeutically effective amount of a compound of formula (II)

（II），

Or a solvate, hydrate, tautomer or pharmaceutically acceptable salt thereof.

27. The method of claim 26, wherein the bispecific anti-EGFR/c-Met antibody comprises: a first domain that binds EGFR comprising heavy chain complementarity determining region 1 of SEQ ID NO:1(HCDR 1), HCDR2 of SEQ ID NO:2, HCDR3 of SEQ ID NO:3, light chain complementarity determining region 1 of SEQ ID NO:4(LCDR 1), LCDR2 of SEQ ID NO:5, and LCDR3 of SEQ ID NO: 6; and a second domain that binds c-Met, said second domain comprising 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.

28. The method of claim 27, wherein the first domain that binds EGFR comprises the heavy chain variable region (VH) of SEQ ID NO:13 and the light chain variable region (VL) of SEQ ID NO:14, and the second domain that binds c-Met comprises the VH of SEQ ID NO:15 and the VL of SEQ ID NO: 16.

29. The method of any one of claims 26-28, wherein the bispecific anti-EGFR/c-Met antibody is an IgG1 isotype.

30. The method of any one of claims 26-29, wherein the bispecific anti-EGFR/c-Met antibody comprises a first heavy chain of SEQ ID No. 17 (HC1), a first light chain of SEQ ID No. 18 (LC1), a second heavy chain of SEQ ID No. 19 (HC2), and a second light chain of SEQ ID No. 20 (LC 2).

31. The method of any one of claims 26-30, wherein the bispecific anti-EGFR/c-Met antibody has a double-branched glycan structure with a fucose content of between about 1% to about 15%.

32. The method of any one of claims 26-31, wherein the compound of formula (II) or a solvate, hydrate, tautomer, or pharmaceutically acceptable salt thereof is 2-fluoro-N-methyl-4- [7- (quinolin-6-ylmethyl) imidazo [1,2-b ] [1,2,4] triazin-2-yl ] benzamide-hydrogen chloride-water (1/2/1).

33. The method of any one of claims 26-32, wherein the subject is relapsed or resistant to treatment with one or more previous anti-cancer therapies.

34. The method of claim 33, 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.

35. The method of claim 33, wherein the one or more prior anti-cancer therapies comprise carboplatin, paclitaxel, gemcitabine, cisplatin, vinorelbine, docetaxel, palbociclib, crizotinib, PD- (L)1 axis inhibitor, EGFR inhibitor, c-Met inhibitor, HER2 inhibitor, HER3 inhibitor, HER4 inhibitor, VEGFR inhibitor, erlotinib, gefitinib, lapatinib, vandetanib, afatinib, oxitinib, lazatinib, boritinib, cabozitinib, axitinib, lenvatinib, nilatinib, regorafenib, pazopanib, sorafenib, or sunitinib, or any combination thereof.

36. The method of any one of claims 26-32, wherein the subject is untreated.

37. The method of any one of claims 26-36, wherein a cancer positive for a c-Met exon 14 skipping mutation is amplified for CDK4, EGFR amplification, KRAS amplification, MDM2 amplification, TERT amplification, NF 1R 2450; RAD50L597Vfs 5, MET c.3082+3A > G, EGFR, wild type EGFR, EGFR activating mutation, circulating HGF, c-MET amplification or increased levels of mutant KRAS or any combination thereof are positive.

38. The method of claim 37, wherein the EGFR activating mutation comprises: 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; deletion of E746-A750; absence of R748-P753; insertion of ala (a) between M766 and a 767; insertion of Ser, Val and Ala (SVA) between S768 and V769; insertion of Asn and ser (ns) between P772 and H773; insertion of one or more amino acids between D761 and E762, between a763 and Y764, between Y764 and Y765, between M766 and a767, between a767 and V768, between S768 and V769, between V769 and D770, between D770 and N771, between N771 and P772, between P772 and H773, between H773 and V774, between V774 and C775; one or more deletions in EGFR exon 20; or one or more insertions in EGFR exon 20; or any combination thereof.

39. The method of claim 37, wherein the mutant KRAS comprises a G12V, G12C, G12A, or G12D substitution, or any combination thereof.

40. The method of any one of claims 26-39, wherein the cancer is lung cancer, gastric cancer, colorectal cancer, brain cancer, epithelial cell-derived 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, gastric cancer, thymus cancer, colon cancer, thyroid cancer, liver cancer, hepatocellular carcinoma (HCC), or sporadic or hereditary Papillary Renal Cell Carcinoma (PRCC), or any combination thereof.

41. The method of claim 40, wherein lung cancer is non-small cell lung cancer (NSCLC), Small Cell Lung Cancer (SCLC), or lung adenocarcinoma, lung sarcomatoid carcinoma, or any combination thereof.

42. The method of any one of claims 26-41, further comprising administering to the subject one or more anti-cancer therapies.

43. The method of claim 42, wherein the one or more anti-cancer therapies comprise chemotherapy, radiation therapy, surgery, targeted anti-cancer therapy, or a kinase inhibitor, or any combination thereof.

44. The method of claim 43, wherein the kinase inhibitor is an EGFR inhibitor, a c-Met inhibitor, a HER2 inhibitor, a HER3 inhibitor, a HER4 inhibitor, a VEGFR inhibitor, or an AXL inhibitor.

45. The method of claim 44, wherein the kinase inhibitor is erlotinib, gefitinib, lapatinib, vandetanib, afatinib, oxitinib, Lazetinib, pozitinib, critinib, cabozantinib, camatinib, axitinib, lenvatinib, nintedanib, regrafenib, pazopanib, sorafenib, or sunitinib.

46. The method of any one of claims 26-45, wherein the c-Met exon 14 skipping mutation is a de novo mutation.

47. The method of any one of claims 26-46, wherein the c-Met exon 14 skipping mutation is an acquired mutation.

48. The method of any one of claims 26-47, wherein the bispecific anti-EGFR/c-Met antibody is administered at a dose of between about 140mg to about 1750 mg.

49. The method of any one of claims 26-48, wherein the bispecific anti-EGFR/c-Met antibody is administered at a dose of about 700mg, about 750mg, about 800mg, about 850mg, 900mg, 950mg, 1000mg, 1050mg, 1100mg, 1150mg, 1200mg, 1250mg, 1300mg, 1350mg, or 1400 mg.

50. The method of any one of claims 26-49, wherein the bispecific anti-EGFR/c-Met antibody is administered twice weekly, once biweekly, once every three weeks, or once four weeks.