Methods for identifying cancer patients for combination treatment

Abstract

The present disclosure provides methods and kits for determining whether a cancer in a subject is susceptible to treatment with a combination therapy comprising a bispecific anti-epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) antibody and an EGFR tyrosine kinase inhibitor (TKI). The present disclosure also provides methods for treating cancer in a subject based on the susceptibility of the cancer to treatment with a combination therapy comprising a bispecific EGFR / c-Met antibody and an EGFR TKI.

Description

[Technical field]

[0001] (CROSS REFERENCE TO RELATED APPLICATIONS) This application claims the benefit of U.S. Provisional Patent Application No. 63 / 190,004, filed May 18, 2021, the entire disclosure of which is incorporated herein by reference.

[0002] (Sequence Listing) The sequence listing of the present application has been submitted electronically as an ASCII sequence listing with the file name "JBI6555WOPCT1SEQLIST.TXT" (created on May 16, 2022 and size: 19 kilobytes (KB)). This submitted sequence listing is a part of the present specification and is incorporated herein by reference in its entirety.

[0003] FIELD OF THEINVENTION The present disclosure relates to methods and kits for identifying and treating cancer patients who would benefit from treatment with a combination therapy comprising a bispecific anti-epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) antibody and an EGFR tyrosine kinase inhibitor (TKI). [Background technology]

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

[0005] Molecular segmentation of advanced non-small cell lung cancer (NSCLC) based on cancer driver mutations has improved overall survival and quality of life for patients with treatable driver mutations, as well as consolidation solid tumor targeted therapy. In NSCLC, specific mutations in the EGFR gene are associated with high response rates to EGFR tyrosine kinase inhibitors (EGFR TKIs). Although the majority of NSCLC patients with EGFR mutations respond initially to EGFR TKI therapy, virtually all acquire resistance that prevents a sustained response. Nearly 60% of all tumors that become resistant to EGFR tyrosine kinase inhibitors have increased c-Met expression, amplification of the c-Met gene, or increased expression of its only known ligand, hepatocyte growth factor (Turke et al., Cancer Cell, 17:77-88, 2010).

[0006] The development of acquired resistance to EGFR-TKIs such as osimertinib in epidermal growth factor receptor mutant (EGFRm) NSCLC likely results from complex and heterogeneous patterns of resistance, with simultaneous occurrence of multiple resistance mechanisms, and thus, the details of such mechanisms remain unclear.

[0007] Amivantamab is a fully human bispecific antibody that targets both EGFR and the receptor tyrosine kinase mesenchymal-epithelial transition factor (c-Met) and contains a fragment crystallizable (Fc) region that has been shown to exhibit immune cell-targeting activity (Vijayaraghavan et al., Mol Cancer Ther 19:2044, 2020). Amivantamab has demonstrated clinical activity across a range of EGFRm NSCLC and has been granted breakthrough therapy designation for post-chemotherapy EGFRm exon 20 insertion NSCLC in the United States and China (Haura et al., JCO 37:9009, 2019; Park et al., JCO 38:9512, 2020; Sabari et al., JTO 16:S108, 2021).

[0008] Lazertinib is a potential third-generation tyrosine kinase inhibitor (TKI) with efficacy in activating EGFR mutation T790M and central nervous system (CNS) diseases (Ahn et al., Lancet Oncol 20: P1681, 2019; Kim et al., JCO 38: 9571, 2020). Lazertinib is associated with a low rate of EGFR-related toxicities, such as rash and diarrhea, and low cardiovascular risk, thus having a safety profile that supports its combination with other anti-EGFR molecules (Ahn et al., Lancet Oncol 20: P1681, 2019; Haddish-Berhane et al., JTO 16: S677, 2022). Summary of the Invention

[0009] As identified in the Background section above, there is a need in the art to identify and treat cancer patients who would benefit from treatment with a combination therapy comprising a bispecific anti-epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) antibody and an EGFR tyrosine kinase inhibitor (TKI).

[0010] In one aspect, provided herein is a method for determining whether a cancer in a subject is susceptible to treatment with a combination therapy comprising a bispecific anti-epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) bispecific antibody and an EGFR tyrosine kinase inhibitor (TKI), comprising: a) determining the presence of one or more mutations in tumor DNA obtained from the subject, wherein the one or more mutations are selected from a mutation in one or more genes from the RAS / RAF / MEK pathway and a mutation in PIK3CA; b) (i) identifying the cancer in the subject as susceptible to treatment with the combination therapy if tumor DNA from the subject does not have the mutation, or (ii) identifying the cancer in the subject as not susceptible to treatment with the combination therapy if tumor DNA from the subject has one or more of the mutations.

[0011] In one aspect, provided herein is a method for treating cancer in a subject in need thereof, comprising: a) determining the presence of one or more mutations in tumor DNA obtained from the subject, wherein the one or more mutations are selected from a mutation in one or more genes from the RAS / RAF / MEK pathway and a mutation in PIK3CA; b) (i) administering to the subject a therapeutically effective amount of a combination therapy comprising a bispecific anti-epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) bispecific antibody and an EGFR tyrosine kinase inhibitor (TKI) if the tumor DNA from the subject does not have the mutations, or (ii) administering to the subject a cancer therapy that does not include the combination therapy used in (i) if the tumor DNA from the subject has one or more of the mutations.

[0012] In some embodiments of the above diagnostic or therapeutic methods, the one or more genes from the RAS / RAF / MEK pathway are FGFR3, KRAS, BRAF, ERBB2, ALK, NRAS, PDGFRA and / or RET. In some embodiments, the mutations in the one or more genes from the RAS / RAF / MEK pathway include FGFR3 fusions, BRAF G469A, BRAF V600E, ERBB2 copy number changes, ALK fusions, ERBB2 I767M, ERBB2 V777L, KRAS A18V, KRAS copy number changes, KRAS G12X (X is any amino acid), NRAS Q61R, PDGFRA copy number changes, and RET fusions. In some embodiments, the KRAS G12X mutations are KRAS G12D, KRAS G12A, KRAS G12C, and KRAS G12V.

[0013] In some embodiments of the above diagnostic or therapeutic methods, the mutation in PIK3CA comprises PIK3CA E545K.

[0014] In some embodiments of the above diagnostic or therapeutic methods, the one or more mutations are further selected from mutations in one or more genes from the WNT / b-catenin pathway. In some embodiments, the one or more genes from the WNT / b-catenin pathway are APC and CTNNB1. In some embodiments, the mutations in one or more genes from the WNT / b-catenin pathway include APC Q1469, APC R405, APC S713, CTNNB1 S33P, CTNNB1 S37C, CTNNB1 S37F, and CTNNB1 S45P.

[0015] In another aspect, provided herein is a method for determining whether a cancer in a subject is susceptible to treatment with a combination therapy comprising a bispecific anti-epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) bispecific antibody and an EGFR tyrosine kinase inhibitor (TKI), comprising: a) determining the presence of one or more mutations in tumor DNA obtained from the subject, the one or more mutations falling into one of the following two groups: (1) PIK3CA E545K, PIK3CA E542K / V, PIK3CA H1047R, PIK3CA amplification, KRAS G12V / C / D / X (X is any amino acid other than G, V, C, and D), KRAS amplification, BRAF V600E, BRAF amplification, CCND1 amplification, CCND2 amplification, CCNE1 amplification, CDK4 amplification, CDK6 amplification, HER2 amplification, HER2 oncogenic alteration, PTEN deletion, PTEN N48K, CDKN2A G101W, CDKN2B mutation, ALK fusion, FGFR3-TACC3 fusion, TPM3-NTRK1 fusion, RET fusion, BRAF fusion, and other oncogenic fusion events; (2) selected from EGFR C797S, EGFR L792H, EGFR amplification, EGFR G796S, EGFR L718X (wherein X is any amino acid), EGFR E709K, EGFR G724S, MET amplification, and MET exon 14 skipping (METex14) mutation; b) (i) identifying the cancer in the subject as susceptible to treatment with the combination therapy if the tumor DNA from the subject either has no mutations from group (1), or has one or more mutations from group (1) and one or more mutations from group (2); or (ii) identifying the cancer in the subject as not susceptible to treatment with the combination therapy if the tumor DNA from the subject has one or more mutations from group (1) and no mutations from group (2).

[0016] In another aspect, provided herein is a method for treating cancer in a subject in need thereof, comprising: a) determining the presence of one or more mutations in tumor DNA obtained from the subject, the one or more mutations falling into one of the following two groups: (1) PIK3CA E545K, PIK3CA E542K / V, PIK3CA H1047R, PIK3CA amplification, KRAS G12V / C / D / X, KRAS amplification, BRAF V600E, BRAF amplification, CCND1 amplification, CCND2 amplification, CCNE1 amplification, CDK4 amplification, CDK6 amplification, HER2 amplification, HER2 oncogenic alterations, PTEN deletion, PTEN N48K, CDKN2A G101W, CDKN2B, ALK fusions, FGFR3-TACC3 and other fusions, RET fusions, BRAF fusions, and other oncogenic fusion events; (2) selected from EGFR C797S, EGFR L792H, EGFR amplification, EGFR G796S, EGFR L718X (wherein X is any amino acid), EGFR E709K, EGFR G724S, MET amplification, and MET exon 14 skipping (METex14) mutation; b) (i) administering to the subject a therapeutically effective amount of a combination therapy comprising a bispecific anti-epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) bispecific antibody and an EGFR tyrosine kinase inhibitor (TKI), if the tumor DNA from the subject either does not have a mutation from group (1) or has one or more mutations from group (1) and one or more mutations from group (2); or (ii) administering to the subject a cancer therapy that does not include the combination therapy used in (i), if the tumor DNA from the subject has one or more mutations from group (1) and does not have a mutation from group (2).

[0017] In some embodiments of the above diagnostic or therapeutic methods, the HER2 oncogenic alterations include HER2 Y772_A775 duplication, HER2 L755M / S / W, and HER2 S310F / Y. In some embodiments, the PTEN deletions include PTEN I33del and PTEN I14del. In some embodiments, the ALK fusions include SQSTM1-ALK fusions and EML4-ALK fusions. In some embodiments, the RET fusions include CCDC6-RET fusions, KIF5B-RET fusions, and NCOA4-RET fusions.

[0018] In some embodiments of the above diagnostic or therapeutic methods, the cancer is lung cancer, such as non-small cell lung cancer (NSCLC).

[0019] In some embodiments of the above diagnostic or therapeutic methods, the cancer in the subject is resistant to treatment with an EGFR TKI that is not the same as the EGFR TKI used in the combination therapy.In some embodiments, the EGFR TKI to which the cancer is resistant is selected from osimertinib, erlotinib, afatinib, rociletinib, olmutinib, and any combination thereof.In one embodiment, the EGFR TKI to which the cancer is resistant is osimertinib.

[0020] In some embodiments of the above diagnostic or therapeutic methods, the subject is chemotherapy naive.

[0021] In some embodiments of the above diagnostic or therapeutic methods, the tumor DNA from the subject has at least one EGFR activating mutation. In some embodiments, the EGFR activating mutation is selected from exon 19 deletion, L858R, and T790M.

[0022] In some embodiments of the above diagnostic or therapeutic methods, the tumor DNA is circulating tumor DNA (ctDNA). In some embodiments, the ctDNA is present in a biological sample isolated from a subject. In some embodiments, the biological sample is a blood sample or a plasma sample. In some embodiments, the ctDNA is isolated from the biological sample prior to mutation identification.

[0023] In some embodiments of the above diagnostic or therapeutic methods, the tumor DNA is present in a tumor sample isolated from the subject. In some embodiments, the tumor DNA is isolated from the tumor sample prior to mutation identification.

[0024] In some embodiments of the above diagnostic or therapeutic methods, the one or more mutations are determined by sequencing. In some embodiments, the one or more mutations are determined using next generation sequencing (NGS).

[0025] In some embodiments of the above diagnostic or therapeutic methods, the bispecific anti-EGFR / c-Met antibody comprises a first domain that specifically binds to EGFR and a second domain that specifically binds to c-Met, the first domain comprises heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 1, HCDR2 of SEQ ID NO: 2, HCDR3 of SEQ ID NO: 3, light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 4, LCDR2 of SEQ ID NO: 5, and LCDR3 of SEQ ID NO: 6, and the second domain that binds to c-Met comprises HCDR1 of SEQ ID NO: 7, HCDR2 of SEQ ID NO: 8, HCDR3 of SEQ ID NO: 9, LCDR1 of SEQ ID NO: 10, LCDR2 of SEQ ID NO: 11, and LCDR3 of SEQ ID NO: 12. In some embodiments, the first domain that specifically binds to EGFR comprises a heavy chain variable region (VH) of SEQ ID NO: 13 and a light chain variable region (VL) of SEQ ID NO: 14, and the second domain that specifically binds to c-Met comprises a VH of SEQ ID NO: 15 and a VL of SEQ ID NO: 16. In some embodiments, the bispecific anti-EGFR / c-Met antibody is of the IgG1 isotype.

[0026] In some embodiments of the above diagnostic or therapeutic methods, the bispecific anti-EGFR / c-Met antibody comprises a first heavy chain (HC1) of SEQ ID NO: 17, a first light chain (LC1) of SEQ ID NO: 18, a second heavy chain (HC2) of SEQ ID NO: 19, and a second light chain (LC2) of SEQ ID NO: 20.

[0027] In some embodiments of the above diagnostic or therapeutic methods, the bispecific anti-EGFR / c-Met antibody comprises a biantennary glycan structure having a fucose content of about 1% to about 15%.

[0028] In some embodiments of the above diagnostic or therapeutic methods, the bispecific anti-EGFR / c-Met antibody is administered intravenously to the subject. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 140 mg to about 2240 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 700 mg, about 750 mg, about 800 mg, about 850 mg, 900 mg, 950 mg, 1000 mg, 1050 mg, 1100 mg, 1150 mg, 1200 mg, 1250 mg, 1300 mg, 1350 mg, 1400 mg, 1575 mg, 1600 mg, 2100 mg, or 2240 mg. In one embodiment, if the subject has a body weight of less than 80 kg, the bispecific anti-EGFR / c-Met antibody is administered at a dose of 1050 mg. In one embodiment, the bispecific anti-EGFR / c-Met antibody is administered at a dose of 1400 mg if the subject weighs 80 kg or more.

[0029] In some embodiments of the above diagnostic or therapeutic methods, the bispecific anti-EGFR / c-Met antibody is administered subcutaneously or intradermally to the subject. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered subcutaneously or intradermally at a dose sufficient to achieve a therapeutic effect in the subject.

[0030] In some embodiments of the above diagnostic or therapeutic methods, the bispecific anti-EGFR / c-Met antibody is administered twice a week, once a week, once every two weeks, once every three weeks, or once every four weeks. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered once a week. In other embodiments, the bispecific anti-EGFR / c-Met antibody is administered once every two weeks.

[0031] In some embodiments of the above diagnostic or therapeutic methods, the EGFR TKI administered in combination with the bispecific anti-EGFR / c-Met antibody is lazertinib. In some embodiments, the EGFR TKI administered in combination with the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 20 to about 320 mg. In some embodiments, the EGFR TKI administered in combination with the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 240 mg. In some embodiments, the EGFR TKI administered in combination with the bispecific anti-EGFR / c-Met antibody is administered daily, every other day, twice a week, or once a week. In some embodiments, the EGFR TKI administered in combination with the bispecific anti-EGFR / c-Met antibody is administered daily. In some embodiments, the EGFR TKI administered in combination with the bispecific anti-EGFR / c-Met antibody is administered orally.

[0032] In some embodiments of the above methods of treatment, the cancer therapy not including a combination therapy used in (i) is a platinum-based chemotherapy. In some embodiments, the platinum-based chemotherapy includes carboplatin and / or cisplatin.

[0033] In some embodiments of the above diagnostic or therapeutic methods, the method further includes obtaining a biological sample from the subject prior to step (a), wherein the biological sample contains tumor DNA (e.g., ctDNA), and optionally purifying the tumor DNA (e.g., ctDNA) from the biological sample.

[0034] In another aspect, provided herein is a method for determining whether a cancer in a subject is susceptible to treatment with a combination therapy comprising a bispecific anti-epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) bispecific antibody and an EGFR tyrosine kinase inhibitor (TKI), comprising: a) determining the expression level of EGFR or MET in a tumor sample obtained from the subject using immunohistochemistry (IHC); b) determining a staining intensity score on a scale of 0 to 3+ based on the expression level of EGFR or MET determined in step (a); c) (i) identifying the cancer in the subject as susceptible to treatment with the combination therapy if the staining intensity score is 3+, or (ii) identifying the cancer in the subject as not susceptible to treatment with the combination therapy if the staining intensity score is less than 3+.

[0035] In some embodiments of the above method, step (c) comprises identifying the cancer in the subject as susceptible to treatment with the combination therapy if 25% or more of the cells in the tumor sample have a staining intensity score of 3+; or (ii) identifying the cancer in the subject as not susceptible to treatment with the combination therapy if less than 25% of the cells in the tumor sample have a staining intensity score of 3+.

[0036] In another aspect, provided herein is a method for treating cancer in a subject in need thereof, comprising: a) determining the expression level of EGFR or MET in a tumor sample obtained from the subject using immunohistochemistry (IHC); b) determining a staining intensity score on a scale of 0 to 3+ based on the expression level of EGFR or MET determined in step (a); c) (i) administering to the subject a therapeutically effective amount of a combination therapy comprising a bispecific anti-epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) bispecific antibody and an EGFR tyrosine kinase inhibitor (TKI) if the staining intensity score is 3+; or (ii) not administering to the subject the combination therapy used in (i), or administering to the subject a cancer therapy that does not include the combination therapy used in (i), if the staining intensity score is less than 3+.

[0037] In some embodiments of the above method, step (c) comprises (i) administering a therapeutically effective amount of the combination therapy to the subject if 25% or more of the cells in the tumor sample have a staining intensity score of 3+; or (ii) not administering to the subject the combination therapy used in (i), or administering to the subject a cancer therapy that does not include the combination therapy used in (i), if less than 25% of the cells in the tumor sample have a staining intensity score of 3+.

[0038] In another aspect, provided herein is a method for determining whether a cancer in a subject is susceptible to treatment with a combination therapy comprising a bispecific anti-epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) bispecific antibody and an EGFR tyrosine kinase inhibitor (TKI), comprising: a) determining the expression levels of EGFR and MET in a tumor sample obtained from the subject using immunohistochemistry (IHC); b) calculating a combined H-score based on the expression levels of EGFR and MET determined in step (a); and c) (i) identifying the cancer in the subject as susceptible to treatment with the combination therapy if the combined H-score is 400 or greater, or (ii) identifying the cancer in the subject as not susceptible to treatment with the combination therapy if the combined H-score is less than 400.

[0039] In another aspect, provided herein is a method for treating cancer in a subject in need thereof, comprising: a) determining the expression levels of EGFR and MET in a tumor sample obtained from the subject using immunohistochemistry (IHC); b) calculating a combined H-score based on the expression levels of EGFR and MET determined in step (a); and c) (i) administering to the subject a therapeutically effective amount of a combination therapy comprising a bispecific anti-epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) bispecific antibody and an EGFR tyrosine kinase inhibitor (TKI) if the combined H-score is 400 or greater; and (ii) not administering to the subject the combination therapy used in (i), or administering to the subject a cancer therapy that does not include the combination therapy used in (i), if the combined H-score is less than 400.

[0040] In some embodiments of the above diagnostic or therapeutic methods, the cancer is lung cancer, such as non-small cell lung cancer (NSCLC).

[0041] In some embodiments of the above diagnostic or therapeutic methods, the cancer in the subject is resistant to treatment with an EGFR TKI that is not the same as the EGFR TKI used in the combination therapy.In some embodiments, the EGFR TKI to which the cancer is resistant is selected from osimertinib, erlotinib, afatinib, rociletinib, olmutinib, and any combination thereof.In one embodiment, the EGFR TKI to which the cancer is resistant is osimertinib.

[0042] In some embodiments of the above diagnostic or therapeutic methods, the subject is chemotherapy naive.

[0043] In some embodiments of the above diagnostic or therapeutic methods, the subject's tumor has at least one EGFR activating mutation. In some embodiments, the EGFR activating mutation is selected from exon 19 deletion, L858R, and T790M.

[0044] In some embodiments of the above diagnostic or therapeutic methods, the bispecific anti-EGFR / c-Met antibody comprises a first domain that specifically binds to EGFR and a second domain that specifically binds to c-Met, the first domain comprises heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 1, HCDR2 of SEQ ID NO: 2, HCDR3 of SEQ ID NO: 3, light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 4, LCDR2 of SEQ ID NO: 5, and LCDR3 of SEQ ID NO: 6, and the second domain that binds to c-Met comprises HCDR1 of SEQ ID NO: 7, HCDR2 of SEQ ID NO: 8, HCDR3 of SEQ ID NO: 9, LCDR1 of SEQ ID NO: 10, LCDR2 of SEQ ID NO: 11, and LCDR3 of SEQ ID NO: 12. In some embodiments, the first domain that specifically binds to EGFR comprises a heavy chain variable region (VH) of SEQ ID NO: 13 and a light chain variable region (VL) of SEQ ID NO: 14, and the second domain that specifically binds to c-Met comprises a VH of SEQ ID NO: 15 and a VL of SEQ ID NO: 16. In some embodiments, the bispecific anti-EGFR / c-Met antibody is of the IgG1 isotype.

[0045] In some embodiments of the above diagnostic or therapeutic methods, the bispecific anti-EGFR / c-Met antibody comprises a first heavy chain (HC1) of SEQ ID NO: 17, a first light chain (LC1) of SEQ ID NO: 18, a second heavy chain (HC2) of SEQ ID NO: 19, and a second light chain (LC2) of SEQ ID NO: 20.

[0046] In some embodiments of the above diagnostic or therapeutic methods, the bispecific anti-EGFR / c-Met antibody comprises a biantennary glycan structure having a fucose content of about 1% to about 15%.

[0047] In some embodiments of the above diagnostic or therapeutic methods, the bispecific anti-EGFR / c-Met antibody is administered intravenously to the subject. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 140 mg to about 2240 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 700 mg, about 750 mg, about 800 mg, about 850 mg, 900 mg, 950 mg, 1000 mg, 1050 mg, 1100 mg, 1150 mg, 1200 mg, 1250 mg, 1300 mg, 1350 mg, 1400 mg, 1575 mg, 1600 mg, 2100 mg, or 2240 mg. In one embodiment, if the subject has a body weight of less than 80 kg, the bispecific anti-EGFR / c-Met antibody is administered at a dose of 1050 mg. In one embodiment, the bispecific anti-EGFR / c-Met antibody is administered at a dose of 1400 mg if the subject weighs 80 kg or more.

[0048] In some embodiments of the above diagnostic or therapeutic methods, the bispecific anti-EGFR / c-Met antibody is administered subcutaneously or intradermally to the subject. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered subcutaneously or intradermally at a dose sufficient to achieve a therapeutic effect in the subject.

[0049] In some embodiments of the above diagnostic or therapeutic methods, the bispecific anti-EGFR / c-Met antibody is administered twice a week, once a week, once every two weeks, once every three weeks, or once every four weeks. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered once a week. In other embodiments, the bispecific anti-EGFR / c-Met antibody is administered once every two weeks.

[0050] In some embodiments of the above diagnostic or therapeutic methods, the EGFR TKI administered in combination with the bispecific anti-EGFR / c-Met antibody is lazertinib. In some embodiments, the EGFR TKI administered in combination with the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 20 to about 320 mg. In some embodiments, the EGFR TKI administered in combination with the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 240 mg. In some embodiments, the EGFR TKI administered in combination with the bispecific anti-EGFR / c-Met antibody is administered daily, every other day, twice a week, or once a week. In some embodiments, the EGFR TKI administered in combination with the bispecific anti-EGFR / c-Met antibody is administered daily. In some embodiments, the EGFR TKI administered in combination with the bispecific anti-EGFR / c-Met antibody is administered orally.

[0051] In some embodiments of the above methods of treatment, the cancer therapy not including a combination therapy used in (i) is a platinum-based chemotherapy. In some embodiments, the platinum-based chemotherapy includes carboplatin and / or cisplatin.

[0052] In some embodiments of the above diagnostic or therapeutic methods, the method further comprises a tumor sample from the subject prior to step (a).

[0053] In another aspect, provided herein is a diagnostic kit comprising: (i) one or more reagents for determining the presence of one or more mutations in tumor DNA from a subject having cancer; and (ii) optionally, packaging and / or instructions for use, wherein the one or more mutations are selected from mutations in one or more genes from the RAS / RAF / MEK pathway and a mutation in PIK3CA.

[0054] In some embodiments of the above kit, the one or more genes from the RAS / RAF / MEK pathway are FGFR3, KRAS, BRAF, ERBB2, ALK, NRAS, PDGFRA and / or RET. In some embodiments, the mutations in the one or more genes from the RAS / RAF / MEK pathway include FGFR3 fusions, BRAF G469A, BRAF V600E, ERBB2 copy number alterations, ALK fusions, ERBB2 I767M, ERBB2 V777L, KRAS A18V, KRAS copy number alterations, KRAS G12X (X is any amino acid), NRAS Q61R, PDGFRA copy number alterations, and RET fusions. In some embodiments, the KRAS G12X mutations are KRAS G12D, KRAS G12A, KRAS G12C, and KRAS G12V.

[0055] In some embodiments of the above kit, the mutation in PIK3CA comprises PIK3CA E545K.

[0056] In some embodiments of the above kit, the one or more mutations are further selected from mutations in one or more genes from the WNT / b-catenin pathway. In some embodiments, the one or more genes from the WNT / b-catenin pathway are APC and CTNNB1. In some embodiments, the mutations in one or more genes from the WNT / b-catenin pathway include APC Q1469, APC R405, APC S713, CTNNB1 S33P, CTNNB1 S37C, CTNNB1 S37F, and CTNNB1 S45P.

[0057] In another aspect, provided herein is a diagnostic kit comprising: (i) one or more reagents for determining the presence of one or more mutations in tumor DNA from a subject having cancer; and (ii) optionally, packaging and / or instructions for use, wherein the one or more mutations fall into one of the following two groups: (1) PIK3CA E545K, PIK3CA E542K / V, PIK3CA H1047R, PIK3CA amplification, KRAS G12V / C / D / X (X is any amino acid other than G, V, C, and D), KRAS amplification, BRAF V600E, BRAF amplification, CCND1 amplification, CCND2 amplification, CCNE1 amplification, CDK4 amplification, CDK6 amplification, HER2 amplification, HER2 oncogenic alteration, PTEN deletion, PTEN N48K, CDKN2A G101W, CDKN2B mutation, ALK fusion, FGFR3-TACC3 fusion, TPM3-NTRK1 fusion, RET fusion, BRAF fusion, and other oncogenic fusion events; (2) A diagnostic kit for a gene that is selected from EGFR C797S, EGFR L792H, EGFR amplification, EGFR G796S, EGFR L718X (X is any amino acid), EGFR E709K, EGFR G724S, MET amplification, and MET exon 14 skipping (METex14) mutation.

[0058] In some embodiments of the above kits, the HER2 oncogenic alterations include HER2 Y772_A775 duplication, HER2 L755M / S / W, and HER2 S310F / Y. In some embodiments, the PTEN deletions include PTEN I33del and PTEN I14del. In some embodiments, the ALK fusions include SQSTM1-ALK fusions and EML4-ALK fusions. In some embodiments, the RET fusions include CCDC6-RET fusions, KIF5B-RET fusions, and NCOA4-RET fusions.

[0059] In some embodiments of the above kit, the tumor DNA is circulating tumor DNA (ctDNA). In some embodiments, the ctDNA is present in a biological sample isolated from a subject. In some embodiments, the biological sample is a blood sample or a plasma sample. In some embodiments, the tumor DNA is present in a tumor sample isolated from a subject.

[0060] In some embodiments of the above kits, the kit further comprises one or more reagents for purifying said tumor DNA from said biological sample from a subject.

[0061] In some embodiments of the above kits, one or more reagents may be used in conjunction with sequencing techniques, such as next generation sequencing (NGS), to determine one or more mutations.

[0062] In another aspect, provided herein is a diagnostic kit comprising (i) one or more reagents for determining expression levels of EGFR and / or MET in a tumor sample from a subject having cancer, and (ii) optionally, packaging and / or instructions for use. In some embodiments, the one or more reagents can be used in conjunction with immunohistochemistry (IHC) to determine expression levels of EGFR and / or MET.

[0063] These and other aspects described herein will become apparent to those of ordinary skill in the art in the following description, claims, and drawings. [Brief description of the drawings]

[0064] [Figure 1] A schematic diagram of the structures of amivantamab and lazertinib (left) and a detailed description of the mechanism of action (MOA) of amivantamab (right). [Diagram 2] A schematic diagram of the progression of acquired resistance to osimertinib in epidermal growth factor receptor mutant (EGFRm) non-small cell lung cancer (NSCLC). Sequencing of single tumor sections may not reveal heterogeneous patterns in resistance or co-occurring mutations. In this sense, next-generation sequencing (NGS) of circulating tumor DNA (ctDNA) from plasma samples may be more useful (Papadimitrakopoulou et al., Annals of Oncol 29:VIII741, 2018; Ramalingam et al., Annals of Oncol 29:VIII740, 2018). ctDNA, circulating tumor DNA; exon 19del, exon 19 deletion. [Diagram 3] The study design for the CHRYSALIS Phase 1 trial corresponds to the combination cohort (Cho et al., Ann Oncol 31:S813, 2020). a One or more alterations were detected in 42 / 44 ctDNA and 29 / 45 tumor NGS analyses. C, cycle; IHC, immunohistochemistry; QW, weekly; Q2W, every 2 weeks; RP2CD, recommended phase 2 combination dose. [Figure 4] Summary chart of patient demographics and baseline disease characteristics is shown. a Based on local testing, central testing identified an exon 19 deletion. [Figure 5A] The results show durable responses with manageable safety observed with the amivantamab + lazertinib combination (amivantamab / lazertinib combination treatment). Figure 5A shows a plot of the percentage change from baseline in the sum of total diameters (SoD) of target lesions over the months of the study. Four patients did not have post-baseline disease assessments and are not included in the plot. Figure 5B shows a summary chart of investigator-assessed responses (N=45 patients). The safety profile demonstrated by these data is consistent with our previous experience with amivantamab + lazertinib (Cho et al., Ann Oncol 31:S813, 2020). The most common adverse events (AEs) were infusion-related reactions (IRR; 78%), rash (acneiform dermatitis 51% + rash 27%), and paronychia (49%), the majority of which were grade 1-2. Of treatment-related events, 16% were grade ≥3 AEs, 4% were discontinuations, and 18% were dose reductions.CBR, clinical benefit rate (CR, PR, or SD ≥11 weeks);CR, complete response;IRR, infusion-related response;mDOR, median duration of response;mDOT, median duration of treatment;mF / U, median duration of follow-up;mPFS, median progression-free survival;NE, not evaluable, not reached;ORR, overall response rate;PD, progressive disease;PR, partial response;SD, stable disease;UNK, unknown. [Figure 5B]The results show durable responses with manageable safety observed with the amivantamab + lazertinib combination (amivantamab / lazertinib combination treatment). Figure 5A shows a plot of the percentage change from baseline in the sum of total diameters (SoD) of target lesions over the months of the study. Four patients did not have post-baseline disease assessments and are not included in the plot. Figure 5B shows a summary chart of investigator-assessed responses (N=45 patients). The safety profile demonstrated by these data is consistent with our previous experience with amivantamab + lazertinib (Cho et al., Ann Oncol 31:S813, 2020). The most common adverse events (AEs) were infusion-related reactions (IRR; 78%), rash (acneiform dermatitis 51% + rash 27%), and paronychia (49%), the majority of which were grade 1-2. Of treatment-related events, 16% were grade ≥3 AEs, 4% were discontinuations, and 18% were dose reductions.CBR, clinical benefit rate (CR, PR, or SD ≥11 weeks);CR, complete response;IRR, infusion-related response;mDOR, median duration of response;mDOT, median duration of treatment;mF / U, median duration of follow-up;mPFS, median progression-free survival;NE, not evaluable, not reached;ORR, overall response rate;PD, progressive disease;PR, partial response;SD, stable disease;UNK, unknown. [Figure 6A] Responses among patients with identified EGFR / MET resistance are shown. Plots of best percentage change in tumor volume for EGFR-, MET-, and EGFR+MET (EGFR+MET)-resistant groups are shown. RAS / RAF pathway (†), mTOR pathway (Δ), cell cycle (¥), and fusion events (

[0065]

number

[0066]

number

[0067] definition All publications, including but not limited to patents and patent applications, cited in this specification are herein incorporated by reference as if fully set forth.

[0068] It should be understood that the terms used herein are used only to describe particular embodiments and are not intended to be limiting. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0069] Although any methods and materials similar or equivalent to those described herein can be used in the practice for testing of the present invention, exemplary materials and methods are described herein. In describing and claiming the present invention, the following terminology is used.

[0070] When lists are presented, unless otherwise stated, it is to be understood that each individual element of that list and every combination of that list is a separate embodiment. For example, a list of embodiments presented as "A, B, or C" should be interpreted to include the embodiments "A," "B," "C," "A or B," "A or C," "B or C," or "A, B, or C."

[0071] 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.

[0072] The conjunction phrase "and / or" between multiple listed elements is understood to encompass both individual and combined options. For example, when two elements are connected by "and / or," the first option refers to the first element being applicable without the second element. The second option refers to the second element being applicable without the first element. The third option refers to the first and second elements being applicable together. Any one of these options is understood to be within the meaning and thus meets the requirements of the term "and / or" as used herein. The simultaneous applicability of two or more of the options is also understood to be within the meaning and thus meets the requirements of the term "and / or."

[0073] The transitional phrases "comprising," "consisting essentially of," and "consisting" are intended to connote their generally accepted meanings in patent language, i.e., (i) "comprising" is synonymous with "comprising," "containing," or "characterized by" and is inclusive or open-ended and does not exclude other unrecited elements or method steps, (ii) "consisting of" excludes any element, step, or ingredient not specified in the claim, and (iii) "consisting essentially of" limits the claim to the materials or steps specified and those that do not materially affect the "basic and novel characteristics" of the claimed invention. Embodiments described with the phrase "comprising" (or its equivalents) are also provided as embodiments described independently with "consisting of" and "consisting essentially of."

[0074] "Co-administration," "administered with," "administered in combination," "in combination with," and the like are intended to encompass administration of selected therapeutic agents or drugs to a single patient, and include therapeutic regimens in which the therapeutic agents or drugs are administered by the same or different routes of administration or at the same or different times.

[0075] "Isolated" refers to a homogenous population of molecules (e.g., synthetic polynucleotides, polypeptides, vectors, or viruses) that have been substantially separated and / or purified away from other components of the system in which they are produced, such as recombinant cells, as well as to proteins that have been subjected to at least one purification or isolation step. "Isolated" refers to molecules that are substantially free of other cellular material and / or chemicals, and includes molecules that have been isolated to greater degrees of purity, e.g., 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% purity.

[0076] "Treating," "treating," or "treatment" of a disease or disorder, such as cancer, refers to achieving one or more of the following: reducing the severity and / or duration of the disorder, inhibiting the worsening of symptoms characteristic of the disorder being treated, limiting or preventing the recurrence of the disorder in a subject who previously had the disorder, or limiting or preventing the recurrence of symptoms in a subject who was previously symptomatic for the disorder.

[0077] "Prevent," "preventing," "prevention," or "prophylaxis" of a disease or disorder means barring the occurrence of the disorder in a subject.

[0078] "Diagnosing" or "diagnosis" refers to a method of determining whether a subject is afflicted with a given disease or condition, or may develop a given disease or condition in the future, or is likely to respond to treatment for a previously diagnosed disease or condition, i.e., stratifying a patient population for the likelihood of responding to treatment. Diagnosis is typically made by a physician based on a general guide to the disease being diagnosed, or other criteria that indicate that the subject is likely to respond to a particular treatment.

[0079] "Responsive," "responsive," or "likely to respond" refers to any type of improvement or positive response, whether detectable or undetectable, such as reduction or amelioration of one or more symptoms, reduction in the extent of disease, stabilized (i.e., not worsening) disease state, prevention of the spread of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, and remission (whether partial or total).

[0080] "Newly diagnosed" refers to a subject who has been diagnosed with cancer (e.g., an EGFR or c-Met expressing cancer) but has not yet received treatment (e.g., treatment for lung cancer).

[0081] "Therapeutically effective amount" refers to an amount effective to achieve the desired therapeutic result at the dosage and duration required. Therapeutically effective amounts may vary depending on factors such as the individual's condition, age, sex, and weight, as well as the ability of the therapeutic agent or combination of therapeutic agents to elicit a desired response in the individual. Exemplary indicators of an effective therapeutic agent or combination of therapeutic agents include, for example, improved health of the patient.

[0082] "Refractory" refers to a disease that does not respond to treatment. A refractory disease may be resistant to treatment before or at the start of treatment, or a refractory disease may become resistant during treatment.

[0083] "Relapse" refers to the return of a disease or signs and symptoms of a disease after a period of improvement following previous treatment with a therapeutic agent.

[0084] A "subject" includes any human or non-human animal. A "human animal" includes all vertebrates, e.g., mammals and non-mammals, such as non-human primates, sheep, dogs, cats, horses, cows, chickens, amphibians, reptiles, etc. The terms "subject" and "patient" are used interchangeably herein.

[0085] "About" means within an acceptable range of error for a particular value as determined by one of ordinary skill in the art, which will depend in part on the limitations of the method by which the value is measured or determined, i.e., the measurement system. In the context of a particular assay, result, or embodiment, unless expressly stated otherwise in the Examples or elsewhere herein, "about" means within one standard deviation, or up to 5%, whichever is greater, as per practice in the art.

[0086] "Cancer" refers to an abnormal growth of cells that tend to proliferate uncontrolled and, in some cases, metastasize (spread) to other areas of the patient's body.

[0087] "EGFR or c-Met expressing cancer" refers to a cancer that has detectable expression of EGFR or c-Met, or has mutations or amplifications of EGFR or c-Met. EGFR or c-Met expression, amplification, and mutation status can be detected using known methods such as sequencing, fluorescent in situ hybridization, immunohistochemistry, flow cytometry, or western blotting.

[0088] "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 No. NP_005219), as well as naturally occurring variants thereof.

[0089] As used herein, "hepatocyte growth factor receptor" or "c-Met" refers to human c-Met having the amino acid sequence set forth in GenBank Accession No. NP_001120972 and naturally occurring variants thereof.

[0090] A "bispecific anti-EGFR / c-Met antibody" or "bispecific EGFR / c-Met antibody" refers to a bispecific antibody having a first domain that specifically binds EGFR and a second domain that specifically binds c-Met. The domains that specifically bind EGFR and c-Met are typically VH / VL paired, and the bispecific anti-EGFR / c-Met antibody is monovalent with respect to binding to EGFR and c-Met.

[0091] "Specific binding" or "specifically binds" or "specific binding" or "binds" refers to an antibody binding to an antigen or an epitope within an antigen with higher affinity than other antigens. Typically, antibodies bind to an antigen with an equilibrium dissociation constant (K D ) is the K for binding to non-specific antigens (e.g., BSA, casein) D At least 100 times smaller than about 5 × 10 -8 M or less, for example, about 1×10 -9 M or less, approximately 1×10 -10 M or less, approximately 1×10 -11 M or less, approximately 1×10 -12 K below M D The dissociation constant can be measured using known protocols. However, an antibody that binds to an antigen or an epitope within an antigen may have cross-reactivity to other related antigens, e.g., the same antigen (homolog) from other species, such as humans or monkeys, e.g., Macaca fascicularis (cynomolgus monkey, cyno) or Pan troglodytes (chimp). A monospecific antibody binds to one antigen or one epitope, whereas a bispecific antibody binds to two different antigens or two different epitopes.

[0092] "Antibody" has a broad meaning and includes monoclonal antibodies, including murine, human, humanized, and chimeric monoclonal antibodies, antigen-binding fragments, multispecific antibodies, such as bispecific, trispecific, tetraspecific, dimeric, tetrameric, or multimeric antibodies, single-chain antibodies, domain antibodies, and immunoglobulin molecules, including any other modified form of an immunoglobulin molecule that contains an antigen-binding site of the required specificity. A "full-length antibody" is composed of two heavy chains (HC) and two light chains (LC), 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 CH3). Each light chain is composed of a light chain variable region (VL) and a light chain constant region (CL). The VH and VL regions can be further subdivided into regions of hypervariability called complementarity determining regions (CDRs) interspersed with framework regions (FRs). Each VH and VL is composed of three CDR and four FR segments, arranged from the amino terminus to the carboxy terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4.

[0093] "Complementarity determining region (CDR)" is the region of an antibody that binds to an antigen. CDRs can be defined using various descriptions, such as Kabat (Wu et al. (1970) J Exp Med 132:211-50) (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md., 1991), Chothia (Chothia et al. (1987) J Mol Biol 196:901-17), IMGT (Lefranc et al. (2003) Dev Comp Immunol 27:55-77), and AbM (Martin and Thornton (1996) J Bmol Biol 263:800-15). Correspondence between the various descriptions and numbering of the variable regions has been described (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 resource, http: / / imgt_org). Available programs such as abYsis by UCL Business PLC can be used to describe the CDRs. As used herein, the terms "CDR", "HCDR1", "HCDR2", "HCDR3", "LCDR1", "LCDR2" and "LCDR3" include CDRs defined by any of the methods of Kabat, Chothia, IMGT or AbM as described above, unless otherwise expressly stated herein.

[0094] Immunoglobulins can be assigned to five major classes, IgA, IgD, IgE, IgG, and IgM, depending on the amino acid sequence of the heavy chain constant domain. IgA and IgG are further subdivided into isotypes IgA1, IgA2, IgG1, IgG2, IgG3, and IgG4. The antibody light chain of any vertebrate species can be assigned to one of two clearly distinct types, kappa (κ) and lambda (λ), based on the amino acid sequence of its constant domain.

[0095] "Antigen-binding fragment" refers to a portion of an immunoglobulin molecule that binds to an antigen. Antigen-binding fragments may be synthetic, enzymatically obtainable, or recombinant polypeptides, and include VH, VL, VH and VL, Fab, F(ab')2, Fd, and Fv fragments, domain antibodies (dAbs) consisting of one VH domain or one VL domain, shark variable IgNAR domains, camelized VH domains, and the minimum recognition unit consisting of amino acid residues reproducing the CDRs of an antibody, such as the FR3-CDR3-FR4 portion, HCDR1, HCDR2, and / or HCDR3, and LCDR1, LCDR2, and / or LCDR3. The VH and VL domains can be linked to each other via synthetic linkers to form various types of single chain antibody designs; when the VH and VL domains are expressed as separate single chain antibody constructs, the VH / VL domains can pair intramolecularly or intermolecularly to form monovalent antigen binding sites, such as single chain Fvs (scFvs) or diabodies, as described, for example, in WO 1998 / 44001, WO 1988 / 01649, WO 1994 / 13804, and WO 1992 / 01047.

[0096] By "monoclonal antibody" is meant an antibody obtained from a substantially homogeneous population of antibody molecules, i.e., individual antibodies that make up the population, that are identical except for possible well-known alterations, such as removal of the C-terminal lysine from the antibody heavy chain, or post-translational modifications such as isomerization or deamidation of amino acids, oxidation of methionine, or deamidation of asparagine or glutamine. A monoclonal antibody typically binds to one antigenic epitope. A bispecific monoclonal antibody binds to two different antigenic epitopes. A monoclonal antibody may have heterogeneous glycosylation within the antibody population. A monoclonal antibody may be monospecific or multispecific, such as bispecific, and may be monovalent, bivalent, or multivalent.

[0097] "Recombinant" refers to DNA, antibodies, and other proteins that are prepared, expressed, produced, or isolated by recombinant means when segments from different sources are joined to produce the recombinant DNA, antibody, or protein.

[0098] "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 related antigens, e.g., the same antigen (homologs) from other species such as humans or monkeys, e.g., Macaca cynomolgus (cyno) or Pan troglodytes, or may bind to epitopes shared between two or more different antigens.

[0099] "Antagonist" or "inhibitor" refers to a molecule that, when bound to a cellular protein, inhibits at least one response or activity induced by the protein's natural ligand. A molecule is an antagonist when at least one response or activity is inhibited by at least about 20%, 30%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% more than the at least one response or activity inhibited in the absence of the antagonist (e.g., a negative control), or when the inhibition is statistically significant compared to inhibition in the absence of the antagonist.

[0100] A "PD-(L)1 axis inhibitor" refers to a molecule that inhibits PD-1 downstream signaling. A PD-(L)1 axis inhibitor may be a molecule that binds to PD-1, PD-L1, or PD-L2.

[0101] "Biological sample" refers to a collection of fluids, cells, or tissues present within a subject, in addition to similar fluids, cells, or tissues isolated from a subject. Exemplary samples are biological fluids, such as blood, serum and serous fluid, plasma, lymphatic fluid, urine, saliva, cyst fluid, tears, feces, sputum, mucosal secretions of secretory tissues and organs, vaginal secretions, ascites, fluids of the pleural, pericardial, peritoneal, abdominal cavities, and other body cavities, fluids collected by bronchial lavage, synovial fluid, liquid solutions in contact with a subject or biological source, such as cell and organ culture media, including conditioned media of cells or organs, lavage fluids, etc., tissue biopsies, tumor tissue biopsies, tumor tissue samples, fine needle aspirations, surgically excised tissues, organ cultures, or cell cultures. As a non-limiting example, the biological sample is a blood sample. As another non-limiting example, the biological sample is a plasma sample. As yet another non-limiting example, the biological sample is a tumor sample. In some embodiments, the biological sample is circulating tumor DNA (ctDNA), which may be isolated from a variety of other biological samples disclosed herein, such as, but not limited to, a blood or plasma sample. In some embodiments, the biological sample is tumor DNA, which may be isolated, for example, from a tumor sample.

[0102] As used herein, "low fucose" or "low fucose content" refers to an antibody having a fucose content of between about 1% and 15%.

[0103] As used herein, "normal fucose" or "normal fucose content" refers to an antibody having a fucose content of greater than about 50%, typically greater than about 80% or greater than 85%.

[0104] Methods of the Disclosure Diagnostic methods In one aspect, the disclosure provides a method for determining whether a cancer in a subject is susceptible to treatment with a combination therapy comprising a bispecific anti-epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) bispecific antibody and an EGFR tyrosine kinase inhibitor (TKI).

[0105] In some embodiments, the method includes: a) determining the presence of one or more mutations in tumor DNA obtained from the subject, where the one or more mutations are selected from a mutation in one or more genes from the RAS / RAF / MEK pathway and a mutation in phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA); and b) (i) identifying the cancer in the subject as sensitive to treatment with the combination therapy if the tumor DNA from the subject does not have the mutations, or (ii) identifying the cancer in the subject as not sensitive to treatment with the combination therapy if the tumor DNA from the subject has the one or more mutations.

[0106] In some embodiments, the one or more mutations are further selected from a mutation in one or more genes from the WNT / b-catenin pathway.

[0107] Mutations associated with the RAS / RAF / MEK pathway or the WNT / b-catenin pathway, or mutations in PIK3CA can include pathogenic mutations known in the art.

[0108] In some embodiments, mutations may be found in one or more genes from the RAS / RAF / MEK pathway, such as, but not limited to, fibroblast growth factor receptor 3 (FGFR3), Kirsten rat sarcoma viral oncogene (KRAS), v-raf murine sarcoma viral oncogene homolog B1 (BRAF), Erb-B2 receptor tyrosine kinase 2 (ERBB2), anaplastic lymphoma receptor tyrosine kinase (ALK), neuroblastoma-RAS (NRAS), platelet-derived growth factor receptor A (PDGFRA), and / or Ret proto-oncogene (RET).

[0109] In some embodiments, mutations in one or more genes from the RAS / RAF / MEK pathway can include, but are not limited to, FGFR3 fusions, BRAF G469A, BRAF V600E, ERBB2 copy number alterations, ALK fusions, ERBB2 I767M, ERBB2 V777L, KRAS A18V, KRAS copy number alterations, KRAS G12X (wherein X is any amino acid), NRAS Q61R, PDGFRA copy number alterations, and RET fusions.

[0110] In some embodiments, the KRAS G12X mutation is KRAS G12D, KRAS G12A, KRAS G12C, and KRAS G12V.

[0111] In some embodiments, the mutation in BRAF is as described in R. Yaeger et al., Targeting Alterations in the RAF-MEK Pathway. Cancer Discov (2019) 9(3):329-341, which is incorporated by reference in its entirety, including, but not limited to, V600E / K / D / R / M, P367L / S, G464V / E, L485W, N486_A489delinsK, N486_P490del, E586K, L597Q / R / S / V, T599TT / TS, T599I / K, K601E / N / T, K601_S602delinsNT, BRAF kinase duplication, fusion of BRAF kinase domain, D287H, V459L, G466A / E / V, S467L, G469E, N581I / S / T, D594A / G / H / N, F595L, G596D / R.

[0112] In some embodiments, mutations in BRAF may include, but are not limited to, those described in H. Yang et al., New Horizons in KRAS-Mutant Lung Cancer: Dawn After Darkness. Front. Oncol., 25 September 2019 (incorporated herein by reference in its entirety), such as E3K, G12C / V / D / A / S / R / F, G13C / D / E / V / R, V14I, Q61L / E / H / R, F61L, L19F, D33E, T58I, A59T, A146P / V / T, C118S, A59_G60delinsGV.

[0113] In some embodiments of the above diagnostic or therapeutic methods, the mutation in PIK3CA comprises PIK3CA E545K.

[0114] In some embodiments, mutations may be found in one or more genes from the WNT / b-catenin pathway, such as, but not limited to, APC and CTNNB1.

[0115] In some embodiments, the mutations in one or more genes from the WNT / b-catenin pathway include APC Q1469, APC R405, APC S713, CTNNB1 S33P, CTNNB1 S37C, CTNNB1 S37F, and CTNNB1 S45P.

[0116] In some embodiments, the present disclosure also provides a method for determining whether a cancer in a subject is susceptible to treatment with a combination therapy comprising a bispecific anti-epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) bispecific antibody and an EGFR tyrosine kinase inhibitor (TKI), comprising: a) determining the presence of one or more mutations in tumor DNA (e.g., circulating tumor DNA (ctDNA)) obtained from the subject, wherein the one or more mutations fall into one of the following two groups: (1) phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA) E545K, PIK3CA E542K / V, PIK3CA H1047R, PIK3CA amplification, Kirsten rat sarcoma viral oncogene (KRAS) G12V / C / D / X (X is any amino acid other than G, V, C, or D), KRAS amplification, v-raf murine sarcoma viral oncogene homolog B1 (BRAF) V600E, BRAF amplification, cyclin D1 (CCND1) amplification, cyclin D2 (CCND2) amplification, cyclin E1 (CCNE1) amplification, cyclin-dependent kinase 4 (CDK4) amplification, cyclin-dependent kinase 6 (CDK6) amplification, Erb-B2 receptor tyrosine kinase 2 (HER2) amplification, HER2 oncogenic alterations, phosphatase and tensin homolog (PTEN) deletion, PTEN N48K, cyclin-dependent kinase inhibitor 2A (CDKN2A) G101W, CDKN2B mutations, anaplastic lymphoma receptor tyrosine kinase (ALK) fusions, fibroblast growth factor receptor 3-transforming acidic coiled-coil-containing protein 3 (FGFR3-TACC3) fusions and other fusions (e.g., TPM3-NTRK1 fusions), Ret proto-oncogene (RET) fusions, v-raf murine sarcoma viral oncogene homolog B1 (BRAF) fusions, and other oncogenic fusion events;(2) selected from EGFR C797S, EGFR L792H, EGFR amplification, EGFR G796S, EGFR L718X (wherein X is any amino acid), EGFR E709K, EGFR G724S, MET amplification, and MET exon 14 skipping (METex14) mutations; and b) (i) identifying the cancer in the subject as susceptible to treatment with the combination therapy if the tumor DNA (e.g., ctDNA) from the subject does not have a mutation from group (1) or has one or more mutations from group (1) and one or more mutations from group (2), or (ii) identifying the cancer in the subject as not susceptible to treatment with the combination therapy if the tumor DNA (e.g., ctDNA) from the subject has one or more mutations from group (1) and does not have a mutation from group (2);

[0117] Non-limiting examples of HER2 oncogenic alterations include HER2 Y772_A775 duplication, HER2 L755M / S / W, and HER2 S310F / Y. Non-limiting examples of PTEN deletions include PTEN I33del and PTEN I14del. Non-limiting examples of ALK fusions include SQSTM1-ALK fusion and EML4-ALK fusion. Non-limiting examples of RET fusions include CCDC6-RET fusion, KIF5B-RET fusion, and NCOA4-RET fusion. Non-limiting examples of BRAF fusions include those described by Ross et al. al., Int. J. Cancer: 138, 881-890 (2016), which is incorporated by reference in its entirety, e.g., KIAA1549-BRAF, MKRN1-BRAF, TRIM24-BRAF, AGAP3-BRAF, ZC3HAV1-BRAF, AKAP9-BRAF, CCDC6-BRAF, AGK-BRAF, EPS15-BRAF, NUP214-BRAF, ARMC10-BRAF, BTF3L4-BRAF, GHR-BRAF, ZNF767 -BRAF, CCDC91-BRAF, DYNC1I2-BRAF, ZKSCAN1-BRAF, GTF2I-BRAF, MZT1-BRAF, RAD18-BRAF, CUX1-BRAF, SLC12A7-BRAF, MYRIP-BRAF, SND1-BRAF, NUB1-BRAF, KLHL7-BRAF, TANK-BRAF, RBMS3-BRAF, STRN3-BRAF, STK35-BRAF, ETFA-BRAF, SVOPL-BRAF, and JHDM1D-BRAF. Other oncogenic fusion events include, but are not limited to, those described in Figure 1 of Gao et al., Cell Rep. 2018 April 03; 23(1): 227-238. e3. (incorporated herein by reference in its entirety).

[0118] In addition to the mutations specifically mentioned, mutations can also be selected as follows. - Annotate genes as oncogenes or tumor suppressor genes based on the COSMIC cancer gene census (Sondk et al., Nature Reviews Cancer volume 18, 696-705 (2018) (hereby incorporated by reference in its entirety). -For oncogenes, activating short variants are identified in the following cases: o Listed as carcinogenic or possibly carcinogenic in OncoKb (Chakravarty et al., JCO Precision Oncology. 2017:1, 1-16, which is incorporated herein by reference in its entirety); o found in a cancer hotspot, i.e., mutated statistically significantly more frequently than expected by chance, as listed in a cancer hotspot (Chang et al., Cancer Discov. 2018 Feb;8(2):174-183, which is incorporated herein by reference in its entirety); o If there is an explicitly known activating mutation (i.e., KRAS G12C). -A limited set of cancer genes are assessed for copy number and fusions on the Guardant 360 panel. These are also classified as activating if the copy number is greater than 3 or if any fusions are detected. For tumor suppressors, inactivating short variants are identified in the following cases: o Listed as carcinogenic or possibly carcinogenic in OncoKb; ○ When found in a cancer hotspot within a cancer hotspot; o Truncating mutations, i.e. resulting in nonsense, frameshift or splice site mutations; o If it is an explicitly known inactivating mutation.

[0119] In another aspect, the disclosure provides a method for determining whether a cancer in a subject is sensitive to treatment with a combination therapy comprising a bispecific anti-epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) bispecific antibody and an EGFR tyrosine kinase inhibitor (TKI), the method comprising: a) determining expression levels of EGFR and MET in a tumor sample obtained from the subject using immunohistochemistry (IHC); b) calculating a composite H-score based on the expression levels of EGFR and MET determined in step (a); and c) (i) identifying the cancer in the subject as sensitive to treatment with the combination therapy if the composite H-score is 400 or greater, or (ii) identifying the cancer in the subject as not sensitive to treatment with the combination therapy if the composite H-score is less than 400.

[0120] Staining intensity values ​​can be assigned to tumor samples as a semi-quantitative approach useful for analyzing immunohistochemical results. Such approaches are well known in the art. Staining intensity can be assigned on a scale of 0 to 3+ (0, 1+, 2+, or 3+), with 0 being assigned when no staining is visible or detectable and 3+ being assigned to the most intense staining, and can be determined for each cell in a fixed field. Tumor samples can be fixed in formalin paraffin-embedded tissue (FFPE).

[0121] As a semi-quantitative approach useful for analyzing immunohistochemical results, an H-score (or tissue score) can be assigned to the tumor sample (Hirsch FR et al., J Clin Oncol 21:3798-3807, 2003; John T et al., Oncogene 28:S14-S23, 2009). As a non-limiting example, membrane staining intensity (0, 1+, 2+, or 3+) can be determined for each cell in the fixed field. The tumor sample can be fixed in formalin paraffin embedded tissue (FFPE). In some embodiments, the H-score can be based on the predominant staining intensity. In some embodiments, the H-score can include the sum of the individual H-scores for each intensity level found. As a non-limiting example, the percentage of cells at each staining intensity level can be calculated, and finally, an H-score can be assigned using the following exemplary formula: [1×(% cells 1+)+2×(% cells 2+)+3×(% cells 3+)]. The final calculated H-score, which ranges from 0 to 300, may give more relative weight to more intense membrane staining in a given tumor sample. In some embodiments, tumor samples may be considered to be either positive or negative based on a particular discrimination threshold.

[0122] A "composite H-score" as referred to herein can be generated by adding an H-score calculated from the analysis of one biomarker (e.g., EGFR expression) to an H-score calculated from the analysis of a second biomarker (e.g., MET expression). Thus, a composite H-score can have a range of 0-600.

[0123] In another aspect, the disclosure provides a method for determining whether a cancer in a subject is sensitive to treatment with a combination therapy comprising a bispecific anti-epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) bispecific antibody and an EGFR tyrosine kinase inhibitor (TKI), the method comprising: a) determining an expression level of EGFR or MET in a tumor sample obtained from the subject using immunohistochemistry (IHC); b) determining a staining intensity score on a scale of 0 to 3+ based on the expression level of EGFR or MET determined in step (a); and c) (i) identifying the cancer in the subject as sensitive to treatment with the combination therapy if the staining intensity score is 3+, or (ii) identifying the cancer in the subject as not sensitive to treatment with the combination therapy if the staining intensity score is less than 3+.

[0124] In some embodiments, the method comprises in step c) identifying the cancer in the subject as susceptible to treatment with the combination therapy if 5%, 7.5%, 10%, 12.5%, 15%, 17.5%, 20%, 22.5%, 25%, 27.5%, 30%, 32.5%, 35%, 37.5%, 40%, 42.5%, 45%, 47.5% or 50% or more of the cells of the tumor sample have an intensity score of 3+. Thus, step c) may include (ii) identifying the cancer in the subject as not sensitive to treatment with the combination therapy if the intensity score is 3+ in less than 5%, 7.5%, 10%, 12.5%, 15%, 17.5%, 20%, 22.5%, 25%, 27.5%, 30%, 32.5%, 35%, 37.5%, 40%, 42.5%, 45%, 47.5% or 50% of the cells in the tumor sample.

[0125] In one embodiment, the method comprises, in step c), identifying the cancer in the subject as sensitive to treatment with the combination therapy if 25% or more of the cells in the tumor sample have an intensity score of 3+; or (ii) identifying the cancer in the subject as not sensitive to treatment with the combination therapy if less than 25% of the cells in the tumor sample have an intensity score of 3+.

[0126] In various embodiments, the cancer evaluated by the method of the present disclosure is lung cancer. In some embodiments, the lung cancer is non-small cell lung cancer (NSCLC). In some embodiments, the cancer in the subject is resistant to treatment with an EGFR TKI that is not the same as the EGFR TKI used in the combination therapy. Non-limiting examples of the EGFR TKI that the cancer can be resistant to are osimertinib, erlotinib, afatinib, rociletinib, olmutinib, and any combination thereof. In some embodiments, the EGFR TKI that the cancer can be resistant to is osimertinib.

[0127] In some embodiments, the subject is resistant or has acquired resistance to an EGFR inhibitor. Exemplary EGFR inhibitors to which cancers may develop resistance include the anti-EGFR antibodies cetuximab (ERBITUX®), paninumumab (VECTIBIX®), matuzumab, and nimotuzumab; the small molecule EGFR inhibitors erlotinib (TARCEVA®), gefitinib (IRESSA®), EKB-569 (pelitinib, an irreversible EGFR TKI); the pan-ErbB and other receptor tyrosine kinase inhibitors lapatinib (EGFR and HER2 inhibitor), pelitinib (EGFR and HER2 inhibitor), vandetanib (ZD6474, ZACTIMA™, an EGFR, VEGFR2, and RET TKI); PF00299804 (dacomitinib, an irreversible pan-ErbB TKI); CI-1033 (an irreversible pan-erbB TKI); TKI), afatinib (BIBW2992, irreversible pan-ErbB TKI), AV-412 (dual EGFR and ErbB2 inhibitor), EXEL-7647 (EGFR, ErbB2, GEVGR, and EphB4 inhibitor), CO-1686 (irreversible mutant-selective EGFR TKI), AZD9291 (irreversible mutant-selective EGFR TKI), mobocertinib (TAK788, irreversible EGFR TKI), poziotinib (irreversible pan-EGFR or HER TKI), and HKI-272 (neratinib, irreversible EGFR / ErbB2 inhibitor).

[0128] Various qualitative and / or quantitative methods can be used to determine whether a subject is resistant, develops resistance, or is prone to develop resistance to treatment with anti-cancer therapy. Symptoms that may be associated with resistance to anti-cancer therapy include a decline or plateau in the patient's health, an increase in tumor size, a halt or slowdown in tumor growth reduction, and / or the spread of cancerous cells from one location to other organs, tissues, or cells in the body. Re-establishment or worsening of various symptoms associated with cancer, such as loss of appetite, cognitive impairment, depression, dyspnea, fatigue, hormone disruption, neutropenia, pain, peripheral neuropathy, and sexual dysfunction, can also be indicators that a subject is developing or is prone to develop resistance to anti-cancer therapy. Symptoms associated with cancer can vary depending on the type of cancer. For example, symptoms associated with cervical cancer can include abnormal bleeding, abnormal heavy vaginal discharge, pelvic pain not associated with the normal menstrual cycle, bladder pain or pain during urination, and bleeding during regular menstrual periods, sexual intercourse, vaginal douching, or pelvic exam. Symptoms associated with lung cancer can include persistent cough, hemoptysis, shortness of breath, wheezing chest pain, loss of appetite, unintentional weight loss, and fatigue.Symptoms associated with liver cancer can include loss of appetite and weight, abdominal pain, especially in the upper right part of the abdomen, which may extend to the back and shoulders, nausea and vomiting, general weakness and fatigue, liver enlargement, abdominal swelling (ascites), and yellowing of the skin and whites of the eyes (jaundice).Those skilled in the art of oncology can easily identify the symptoms associated with certain types of cancer.

[0129] In some embodiments, the subject is chemotherapy naive.

[0130] In some embodiments, the subject has at least one activating EGFR mutation.

[0131] EGFR activating mutations that may be associated with cancer include point mutations, deletion mutations, insertion mutations, inversions, or gene amplifications that result in an increase in at least one biological activity of EGFR (such as increased tyrosine kinase activity), enhanced ligand binding, ligand-independent signaling, increased cell proliferation, signaling to the MAPK / ERK pathway, gene transcription, receptor homodimer and heterodimer formation, dimerization (EGFR:EGFR), heterodimerization (EGFR:HER2 or EGFR:HER3). The mutations may be located in any part of the EGFR gene or the regulatory region associated with the EGFR gene, including mutations in exons 18, 19, 20, or 21, or mutations in the kinase domain. Other examples of EGFR activating mutations are known in the art (see, for example, U.S. Patent Application Publication No. 2005 / 0272083, which is incorporated herein by reference in its entirety). Information regarding EGFR and other ErbB receptors, including receptor homo- and heterodimers, receptor ligands, autophosphorylation sites, and signaling molecules involved in ErbB-mediated signal transduction, is known in the art (see, e.g., Hynes and Lane, Nature Reviews Cancer 5:341-354, 2005, which is incorporated herein by reference in its entirety).

[0132] In some embodiments, the EGFR activating mutation is a substitution of G719A, G719X (wherein X is any amino acid), L861X (wherein X is any amino acid), L858R, E746K, L747S, E749Q, A750P, A755V, V765M, L858P, or T790M, a deletion of E746 to A750, a deletion of R748 to P753, an insertion of Ala (A) between M766 and A767, an insertion of Ser, Val, and Ala (SVA) between S768 and V769, an insertion of Asn and H773 between P772 and H773, or a deletion of R748 to P753. These include an insertion of Ser(NS), an insertion of one or more amino acids between D761 and E762, A763 and Y764, Y764 and Y765, M766 and A767, A767 and V768, S768 and V769, V769 and D770, D770 and N771, N771 and P772, P772 and H773, H773 and V774, V774 and C775, one or more deletions in EGFR exon 19, one or more deletions in EGFR exon 20, or one or more insertions in EGFR exon 20, or any combination thereof.

[0133] In some embodiments, the EGFR activating mutations include one or more rare EGFR activating mutations, such as S768I, L861Q, and G719X (wherein X is any amino acid).

[0134] In some embodiments, the EGFR activating mutation may be selected from one or more deletions in exon 19, L858R, and T790M. In some embodiments, the EGFR activating mutation is one or more deletions in exon 19. In some embodiments, the EGFR activating mutation is L858R. In some embodiments, the EGFR activating mutation is selected from one or more deletions in exon 19 and L858R. A five amino acid deletion in exon 19 or the point mutation L858R in EGFR may be associated with EGFR TKI sensitivity (Nakata and Gotoh, Expert Opin Ther Targets 16:771-781, 2012, which is incorporated herein by reference in its entirety). In tumor models driven by TKI-sensitive EGFR mutations such as L858R or exon 19 deletions, amivantamab has several proposed mechanisms of action (MOAs), including blocking ligand binding, reducing receptor expression, inhibiting downstream signaling, and eliciting immune-directed anti-cancer activity (Commins et al., J Allergy Clin Immun 2010;125(2):S53-S72, which is incorporated herein by reference in its entirety).

[0135] In some embodiments, exon 19 deletions include E746_A750del, L747_P753delinsS, E746_S752delinsV, L747_A750delinsP, L747_T751 deletion, E746_P753delinsVS, E746_T751delinsA, E746_T751delinsL, L747_E749 deletion, L747_K754delinsATSPE, L747_K754delinsSN, L747_S752del, L747-T751delinsP, and T751-I759delinsN.

[0136] The presence or absence of any of the mutations disclosed herein, including but not limited to those listed in groups (1) and (2), may be detected using methods known in the art, such as, for example, Sanger sequencing, next generation sequencing (NGS), whole exome sequencing (WES), RNA-Seq, fluorescent in situ hybridization, or immunohistochemistry.

[0137] In some embodiments, the presence or absence of one or more mutations in the biological sample disclosed herein can be detected using next generation sequencing (NGS).Non-limiting examples of biological samples are blood samples, plasma samples, and tumor samples.Another non-limiting example of biological sample is circulating tumor DNA (ctDNA) isolated from blood or plasma samples. In such embodiments, the one or more mutations are selected from the group consisting of PIK3CA E545K, PIK3CA E542K / V, PIK3CA H1047R, PIK3CA amplification, mutations in one or more genes from the RAS / RAF / MEK pathway as described herein, mutations in one or more genes from the WNT / b-catenin pathway as described herein, KRAS G12V / C / D / X (wherein X is any amino acid other than G, V, C, and D), KRAS amplification, BRAF V600E, BRAF amplification, CCND1 amplification, CCND2 amplification, CCNE1 amplification, CDK4 amplification, CDK6 amplification, HER2 amplification, HER2 oncogenic alterations, PTEN deletion, PTEN N48K, CDKN2A G101W, CDKN2B mutations, ALK fusions, FGFR3-TACC3 fusions, TPM3-NTRK1 fusions, RET fusions, BRAF fusions and other oncogenic fusion events, EGFR C797S, EGFR The mutation is selected from L792H, EGFR amplification, EGFR G796S, EGFR L718X (wherein X is any amino acid), EGFR E709K, EGFR G724S, MET amplification, MET exon 14 skipping (METex14) mutations, and EGFR activating mutations described herein.

[0138] In some embodiments, the methods of the present disclosure may include determining the presence or absence of one or more mutations in tumor DNA (e.g., ctDNA) obtained from a subject. In some embodiments, the methods of the present disclosure may include determining the presence or absence of one or more mutations in ctDNA. In some embodiments, the tumor DNA (e.g., ctDNA) is present in a biological sample isolated from a subject. As non-limiting examples, the biological sample is a blood sample, a plasma sample, or a tumor sample. In some embodiments, the tumor DNA (e.g., ctDNA) may be isolated from the biological sample prior to mutation identification. In some embodiments, any of the tumor DNA (e.g., ctDNA) obtained from any of the various biological samples disclosed herein may be optionally purified from the biological sample.

[0139] 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, the first domain comprising heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 1, HCDR2 of SEQ ID NO: 2, HCDR3 of SEQ ID NO: 3, light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 4, LCDR2 of SEQ ID NO: 5, and LCDR3 of SEQ ID NO: 6, and the second domain that binds c-Met 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 specifically binds EGFR comprises a heavy chain variable region (VH) of SEQ ID NO: 13 and a light chain variable region (VL) of SEQ ID NO: 14, and the second domain that specifically binds c-Met comprises a VH of SEQ ID NO: 15 and a VL of SEQ ID NO: 16.

[0140] In some embodiments, the bispecific anti-EGFR / c-Met antibody is of the IgG1 isotype.

[0141] In some embodiments, the bispecific anti-EGFR / c-Met antibody comprises a first heavy chain (HC1) of SEQ ID NO: 17, a first light chain (LC1) of SEQ ID NO: 18, a second heavy chain (HC2) of SEQ ID NO: 19, and a second light chain (LC2) of SEQ ID NO: 20.

[0142] In some embodiments, the bispecific anti-EGFR / c-Met antibody comprises a biantennary glycan structure having a fucose content of about 1% to about 15%. In some embodiments, the bispecific anti-EGFR / c-Met antibody comprises a biantennary glycan structure having a fucose content of about 2% to about 14%. In some embodiments, the bispecific anti-EGFR / c-Met antibody comprises a biantennary glycan structure having a fucose content of about 3% to about 13%. In some embodiments, the bispecific anti-EGFR / c-Met antibody comprises a biantennary glycan structure having a fucose content of about 4% to about 12%. In some embodiments, the bispecific anti-EGFR / c-Met antibody comprises a biantennary glycan structure having a fucose content of about 5% to about 11%. In some embodiments, the bispecific anti-EGFR / c-Met antibody comprises a biantennary glycan structure having a fucose content of about 1%. In some embodiments, the bispecific anti-EGFR / c-Met antibody comprises a biantennary glycan structure with a fucose content of about 2%. In some embodiments, the bispecific anti-EGFR / c-Met antibody comprises a biantennary glycan structure with a fucose content of about 3%. In some embodiments, the bispecific anti-EGFR / c-Met antibody comprises a biantennary glycan structure with a fucose content of about 4%. In some embodiments, the bispecific anti-EGFR / c-Met antibody comprises a biantennary glycan structure with a fucose content of about 5%. In some embodiments, the bispecific anti-EGFR / c-Met antibody comprises a biantennary glycan structure with a fucose content of about 6%. In some embodiments, the bispecific anti-EGFR / c-Met antibody comprises a biantennary glycan structure with a fucose content of about 7%. In some embodiments, the bispecific anti-EGFR / c-Met antibody comprises a biantennary glycan structure with a fucose content of about 8%. In some embodiments, the bispecific anti-EGFR / c-Met antibody comprises a biantennary glycan structure with a fucose content of about 9%. In some embodiments, the bispecific anti-EGFR / c-Met antibody comprises a biantennary glycan structure with a fucose content of about 10%. In some embodiments, the bispecific anti-EGFR / c-Met antibody comprises a biantennary glycan structure with a fucose content of about 11%.In some embodiments, the bispecific anti-EGFR / c-Met antibody comprises a biantennary glycan structure with a fucose content of about 12%. In some embodiments, the bispecific anti-EGFR / c-Met antibody comprises a biantennary glycan structure with a fucose content of about 13%. In some embodiments, the bispecific anti-EGFR / c-Met antibody comprises a biantennary glycan structure with a fucose content of about 14%. In some embodiments, the bispecific anti-EGFR / c-Met antibody comprises a biantennary glycan structure with a fucose content of about 15%.

[0143] In some embodiments, the bispecific anti-EGFR / c-Met antibodies disclosed herein may be administered in combination with a tyrosine kinase inhibitor (TKI), such as, but not limited to, an epidermal growth factor receptor (EGFR TKI). Non-limiting examples of TKIs are kinase inhibitors such as erlotinib, gefitinib, lapatinib, vandetanib, afatinib, osimertinib, lazertinib, poziotinib, cliotinib, cabozantinib, capmatinib, axitinib, lenvatinib, nintedanib, regorafenib, pazopanib, sorafenib, or sunitinib. In some embodiments, the bispecific anti-EGFR / c-Met antibodies disclosed herein may be administered in combination with lazertinib.

[0144] therapeutic method In one aspect, the disclosure provides methods for treating cancer in a subject in need thereof based on the biomarker strategies described herein.

[0145] In some embodiments, methods of treatment include methods comprising: a) determining the presence of one or more mutations in tumor DNA obtained from the subject, where the one or more mutations are selected from one or more genes from the RAS / RAF / MEK pathway and a mutation in PIK3CA; and b) (i) administering to the subject a therapeutically effective amount of a combination therapy comprising a bispecific anti-epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) bispecific antibody and an EGFR tyrosine kinase inhibitor (TKI) if the tumor DNA from the subject does not have the mutations, or (ii) administering to the subject a cancer therapy that does not include the combination therapy used in (i) if the tumor DNA from the subject does have the one or more mutations.

[0146] In some embodiments, the one or more mutations are further selected from a mutation in one or more genes from the WNT / b-catenin pathway.

[0147] Mutations associated with the RAS / RAF / MEK pathway or the WNT / b-catenin pathway, or mutations in PIK3CA include pathogenic mutations known in the art.

[0148] In some embodiments, mutations may be found in one or more genes from the RAS / RAF / MEK pathway, such as, but not limited to, FGFR3, KRAS, BRAF, ERBB2, ALK, NRAS, PDGFRA, and / or RET.

[0149] In some embodiments, mutations in one or more genes from the RAS / RAF / MEK pathway can include, but are not limited to, FGFR3 fusions, BRAF G469A, BRAF V600E, ERBB2 copy number alterations, ALK fusions, ERBB2 I767M, ERBB2 V777L, KRAS A18V, KRAS copy number alterations, KRAS G12X (wherein X is any amino acid), NRAS Q61R, PDGFRA copy number alterations, and RET fusions.

[0150] In some embodiments, the KRAS G12X mutation is KRAS G12D, KRAS G12A, KRAS G12C, and KRAS G12V.

[0151] In some embodiments, the mutation in BRAF is as described in R. Yaeger et al., Targeting Alterations in the RAF-MEK Pathway. Cancer Discov (2019) 9(3):329-341 (incorporated herein by reference in its entirety), e.g., V600E / K / D / R / M, P367L / S, G464V / E, L485W, N486_A489delinsK, N486_P490del, E586K, L597Q / R / S / V, T599TT / TS, T599I / K, K601E / N / T, K601_S602delinsNT, BRAF kinase duplication, fusion of BRAF kinase domain, D287H, V459L, G466A / E / V, S467L, G469E, N581I / S / T, D594A / G / H / N, F595L, G596D / R.

[0152] In some embodiments, mutations in BRAF may include those described in H. Yang et al., New Horizons in KRAS-Mutant Lung Cancer: Dawn After Darkness. Front. Oncol., 25 September 2019 (incorporated herein by reference in its entirety), e.g., E3K, G12C / V / D / A / S / R / F, G13C / D / E / V / R, V14I, Q61L / E / H / R, F61L, L19F, D33E, T58I, A59T, A146P / V / T, C118S, A59_G60delinsGV.

[0153] In some embodiments of the above diagnostic or therapeutic methods, the mutation in PIK3CA comprises PIK3CA E545K.

[0154] In some embodiments, mutations may be found in one or more genes from the WNT / b-catenin pathway, such as, but not limited to, APC and CTNNB1.

[0155] In some embodiments, the mutations in one or more genes from the WNT / b-catenin pathway include APC Q1469, APC R405, APC S713, CTNNB1 S33P, CTNNB1 S37C, CTNNB1 S37F, and CTNNB1 S45P.

[0156] In some embodiments, the method of treatment includes a) determining the presence of one or more mutations in tumor DNA (e.g., circulating tumor DNA (ctDNA)) obtained from the subject, wherein the one or more mutations fall into one of two groups: (1) PIK3CA E545K, PIK3CA E542K / V, PIK3CA H1047R, PIK3CA amplification, KRAS G12V / C / D / X, KRAS amplification, BRAF V600E, BRAF amplification, CCND1 amplification, CCND2 amplification, CCNE1 amplification, CDK4 amplification, CDK6 amplification, HER2 amplification, HER2 oncogenic alterations, PTEN deletion, PTEN N48K, CDKN2A G101W, CDKN2B, ALK fusions, FGFR3-TACC3 and other fusions, RET fusions, BRAF fusions, and other oncogenic fusion events; (2) EGFR C797S, EGFR L792H, EGFR amplification, EGFR G796S, EGFR and b) (i) administering to the subject an effective amount of a combination therapy comprising a bispecific anti-epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) bispecific antibody and an EGFR tyrosine kinase inhibitor (TKI), if the tumor DNA from the subject has either no mutations from group (1) or has one or more mutations from group (1) and one or more mutations from group (2); or (ii) administering to the subject a cancer therapy that does not include the combination therapy used in (i), if the tumor DNA (e.g., ctDNA) from the subject has one or more mutations from group (1) and no mutations from group (2).

[0157] Non-limiting examples of HER2 oncogenic alterations include HER2 Y772_A775 duplication, HER2 L755M / S / W, and HER2 S310F / Y. Non-limiting examples of PTEN deletions include PTEN I33del and PTEN I14del. Non-limiting examples of ALK fusions include SQSTM1-ALK fusion and EML4-ALK fusion. Non-limiting examples of RET fusions include CCDC6-RET fusion, KIF5B-RET fusion, and NCOA4-RET fusion. Non-limiting examples of BRAF fusions include those described by Ross et al. al., Int. J. Cancer: 138, 881-890 (2016), which is incorporated by reference in its entirety, e.g., KIAA1549-BRAF, MKRN1-BRAF, TRIM24-BRAF, AGAP3-BRAF, ZC3HAV1-BRAF, AKAP9-BRAF, CCDC6-BRAF, AGK-BRAF, EPS15-BRAF, NUP214-BRAF, ARMC10-BRAF, BTF3L4-BRAF, GHR-BRAF, ZNF767 -BRAF, CCDC91-BRAF, DYNC1I2-BRAF, ZKSCAN1-BRAF, GTF2I-BRAF, MZT1-BRAF, RAD18-BRAF, CUX1-BRAF, SLC12A7-BRAF, MYRIP-BRAF, SND1-BRAF, NUB1-BRAF, KLHL7-BRAF, TANK-BRAF, RBMS3-BRAF, STRN3-BRAF, STK35-BRAF, ETFA-BRAF, SVOPL-BRAF, and JHDM1D-BRAF. Other oncogenic fusion events include, but are not limited to, those described in Figure 1 of Gao et al., Cell Rep. 2018 April 03; 23(1): 227-238. e3. (incorporated herein by reference in its entirety).

[0158] In another aspect, the disclosure is a method for treating cancer in a subject in need of such treatment, comprising: a) determining expression levels of EGFR and MET in a tumor sample obtained from the subject using immunohistochemistry (IHC); b) calculating a composite H-score based on the expression levels of EGFR and MET determined in step (a); and c) (i) administering to the subject an effective amount of a combination therapy comprising a bispecific anti-epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) bispecific antibody and an EGFR tyrosine kinase inhibitor (TKI), if the combined H-score is 400 or greater; and (ii) not administering to the subject the combination therapy used in (i), or administering to the subject a cancer therapy that does not include the combination therapy used in (i), if the combined H-score is less than 400.

[0159] In another aspect, the disclosure provides a method for treating cancer in a subject in need of such treatment, comprising: a) determining expression levels of EGFR and MET in a tumor sample obtained from the subject using immunohistochemistry (IHC); b) determining a staining intensity score on a scale of 0 to 3+ based on the expression levels of EGFR or MET determined in step (a); and c) (i) administering to the subject a therapeutically effective amount of a combination therapy comprising a bispecific anti-epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) bispecific antibody and an EGFR tyrosine kinase inhibitor (TKI) if the staining intensity score is 3+, or (ii) not administering to the subject the combination therapy used in (i), or administering to the subject a cancer therapy that does not include the combination therapy used in (i), if the staining intensity score is less than 3+.

[0160] In some embodiments, the method further comprises, in step (c), (i) administering a therapeutically effective amount of the combination therapy to the subject if 5%, 7.5%, 10%, 12.5%, 15%, 17.5%, 20%, 22.5%, 25%, 27.5%, 30%, 32.5%, 35%, 37.5%, 40%, 42.5%, 45%, 47.5% or 50% of the cells of the tumor sample have a staining intensity score of 3+; or (ii) administering a therapeutically effective amount of the combination therapy to the subject if 5%, 7.5%, 10%, 12.5%, 15%, 17.5%, 20%, 22.5%, 25%, 27.5%, 30%, 32.5%, 35%, 37.5%, 40%, 42.5%, 45%, 47.5% or 50% of the cells of the tumor sample have a staining intensity score of 3+. %, 7.5%, 10%, 12.5%, 15%, 17.5%, 20%, 22.5%, 25%, 27.5%, 30%, 32.5%, 35%, 37.5%, 40%, 42.5%, 45%, 47.5% or 50% of the cells have a staining intensity score of 3+, then not administering the combination therapy used in (i) to the subject, or administering a cancer therapy that does not include the combination therapy used in (i) to the subject.

[0161] In one embodiment, the method comprises, in step (c), (i) administering a therapeutically effective amount of the combination therapy to the subject if 25% or more of the cells in the tumor sample have a staining intensity score of 3+; or (ii) not administering to the subject the combination therapy used in (i), or administering to the subject a cancer therapy that does not include the combination therapy used in (i), if less than 25% of the cells in the tumor sample have a staining intensity score of 3+.

[0162] In various embodiments, the cancer is a solid tumor, a brain tumor, or a hematological malignancy. In certain embodiments, the hematological malignancy is AML, ALL, B-ALL, T-ALL, or lymphoma. Examples of tumors include, but are not limited to, soft tissue tumors (e.g., lymphomas), and tumors of the blood and blood-forming organs (e.g., leukemias), as well as solid tumors (e.g., carcinomas) that grow at anatomical sites outside the bloodstream. Examples of cancer include, but are not limited to, carcinomas, lymphomas, blastomas, sarcomas (e.g., Ewing's sarcoma and other tumors of the Ewing's sarcoma family, osteosarcoma, or rhabdomyosarcoma), and leukemia or lymphoid malignancies. More specific examples of such cancers include squamous cell carcinoma (e.g., epithelial squamous cell carcinoma), adenosquamous cell carcinoma, lung cancer (including, e.g., small cell lung cancer, non-small cell lung cancer, lung adenocarcinoma, and squamous cell carcinoma of the lung), peritoneal cancer, hepatocellular carcinoma, gastric or stomach cancer (including, e.g., gastrointestinal cancer and pancreatic cancer), cervical cancer, ovarian cancer, liver cancer, bladder cancer, urinary tract cancer, liver cancer, breast cancer, colon cancer, rectal cancer, colorectal cancer, endometrial cancer or uterine cancer, salivary gland cancer, and kidney cancer. cancer), prostate cancer, vulvar cancer, thyroid cancer, liver cancer, anal cancer, penile cancer, primary or metastatic melanoma, multiple myeloma and B-cell lymphoma, non-Hodgkin's lymphoma, Hodgkin's lymphoma, brain (e.g., high-grade glioma, diffuse pontine glioma, ependymoma, neuroblastoma, or glioblastoma), and head and neck cancer and associated metastases.Further examples of tumors can be found in The Merck Manual of Diagnosis and Therapy, 19th Edition, § on Hematology and Oncology, published by Merck Sharp & Dohme Corp., 2011 (ISBN 978-0-911910-19-3), The Merck Manual of Diagnosis and Therapy, 20th Edition, § on Hematology and Oncology, published by Merck Sharp & Dohme Corp., 2018 (ISBN 978-0-911-91042-1) (2018 digital online version at the Merck Manuals internet website), and the SEER Program Coding and Staging Manual 2016 (each of which is incorporated by reference in its entirety for all purposes).

[0163] In various embodiments, the tumor is selected from the group consisting of osteosarcoma, rhabdomyosarcoma, Ewing sarcoma and other tumors of the Ewing sarcoma family, neuroblastoma, ganglioneuroblastoma, desmoplastic small round cell tumor, malignant peripheral nerve sheath tumor, synovial sarcoma, undifferentiated sarcoma, adrenocortical carcinoma, hepatoblastoma, Wilms tumor, rhabdoid tumor, high-grade glioma (glioblastoma multiforme), medulloblastoma, astrocytoma, glioma, ependymoma, atypical malformation tumor, and / or sarcoma. The tumor is selected from rhabdoid tumor, meningioma, craniopharyngioma, primitive neuroectodermal tumor, diffuse intrinsic pontine glioma and other brain tumors, acute myeloid leukemia, multiple myeloma, lung cancer, mesothelioma, breast cancer, bladder cancer, gastric cancer, prostate cancer, colorectal cancer, endometrial cancer, uterine cancer, kidney cancer, esophageal cancer, ovarian cancer, pancreatic cancer, hepatocellular carcinoma and other liver cancers, head and neck cancer, leiomyosarcoma, and melanoma. In some embodiments, the tumor is a solid tumor. In various embodiments, the solid tumor is Ewing's sarcoma, lung adenocarcinoma, osteosarcoma, breast cancer, or prostate cancer. In certain embodiments, the tumor is a brain tumor. In some embodiments, the brain tumor is a glioblastoma or neuroblastoma.

[0164] In some embodiments, the methods of the present disclosure are directed to the treatment of squamous cell carcinoma, adenosquamous carcinoma, lung cancer, peritoneal cancer, hepatocellular carcinoma, gastric cancer, cervical cancer, ovarian cancer, liver cancer, bladder cancer, urinary tract cancer, hepatoma, breast cancer, colon cancer, colorectal cancer, endometrial cancer, salivary gland cancer, kidney cancer, prostate cancer, vulvar cancer, thyroid cancer, liver cancer, anal cancer, penile cancer, skin cancer, multiple myeloma, and acute lymphoblastic leukemia. (ALL), acute myeloid leukemia (AML), chronic myeloid leukemia (CML) and chronic lymphocytic leukemia (CLL), lymphomas such as Hodgkin's lymphoma (HL) and non-Hodgkin's lymphoma (NHL), follicular lymphoma, chronic lymphocytic leukemia / small lymphocytic lymphoma (CLL / SLL), mantle cell lymphoma (MCL), marginal zone B-cell lymphoma, primary mediastinal large B-cell lymphoma, lymphoma), Burkitt's lymphoma, lymphoplasmacytic lymphoma, immunoblastic large cell lymphoma, hairy cell leukemia (HCL), precursor B-lymphoblastic lymphoma and primary central nervous system (CNS) lymphoma, T-cell NHL (e.g., precursor T-lymphoblastic lymphoma / leukemia, peripheral T-cell lymphoma (PTCL), angioimmunoblastic T-cell lymphoma, extranodal natural killer T-cell lymphoma, enteropathy-type T-cell lymphoma, In some embodiments, the present invention may be useful for treating a cancer selected from the group consisting of pulmonary leukemia, panniculitis-like T-cell lymphoma, anaplastic large cell lymphoma, etc., a mixture of one or more of the above leukemias / lymphomas, brain and head and neck cancer, bile duct cancer, bronchial cancer, chordoma, choriocarcinoma, epithelial carcinoma, endothelial carcinoma, esophageal cancer, Ewing's sarcoma, heavy chain disease, hematopoietic cancer, immunocytic amyloidosis, monoclonal gammopathy of undetermined significance, myelodysplastic syndrome, myeloproliferative disorder, agnogenic myeloid metaplasia (AMM) or myelofibrosis (MF), chronic idiopathic myelofibrosis, myeloproliferative neoplasm, polycythemia vera, rectal adenocarcinoma, essential thrombocytosis, chronic neutrophilic leukemia, hypereosinophilic syndrome, or soft tissue sarcoma, as well as metastases thereof.

[0165] In some embodiments, the methods of the present disclosure may be useful for treating lung cancer. In some embodiments, the lung cancer is non-small cell lung cancer (NSCLC).

[0166] In some embodiments, the methods of the present disclosure may be useful for treating cancer in a subject in need of cancer treatment, the subject having relapsed or is resistant to treatment with one or more previous anti-cancer therapies. In some embodiments, the one or more previous anti-cancer therapies include one or more EGFR TKIs, and the EGFR TKIs are not the same as the EGFR TKIs used in the combination therapy of the present disclosure. In some embodiments, the one or more EGFR TKIs include osimertinib, erlotinib, afatinib, rociletinib, olmutinib, or any combination thereof.

[0167] In some embodiments, a method of the present disclosure useful for treating cancer in a subject in need thereof may comprise administering to the subject an effective amount of a combination therapy comprising a bispecific anti-epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) bispecific antibody and an EGFR tyrosine kinase inhibitor (TKI). In some embodiments, the bispecific anti-EGFR / c-Met antibody comprises a first domain that specifically binds to EGFR and a second domain that specifically binds to c-Met, the first domain comprising a heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 1, a HCDR2 of SEQ ID NO: 2, a HCDR3 of SEQ ID NO: 3, a light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 4, a LCDR2 of SEQ ID NO: 5, and a LCDR3 of SEQ ID NO: 6, and the second domain that binds to c-Met comprises a HCDR1 of SEQ ID NO: 7, a HCDR2 of SEQ ID NO: 8, a HCDR3 of SEQ ID NO: 9, a LCDR1 of SEQ ID NO: 10, a LCDR2 of SEQ ID NO: 11, and a LCDR3 of SEQ ID NO: 12. In some embodiments, the first domain that specifically binds to EGFR comprises a heavy chain variable region (VH) of SEQ ID NO: 13 and a light chain variable region (VL) of SEQ ID NO: 14, and the second domain that specifically binds to c-Met comprises a VH of SEQ ID NO: 15 and a VL of SEQ ID NO: 16. 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 a first heavy chain (HC1) of SEQ ID NO: 17, a first light chain (LC1) of SEQ ID NO: 18, a second heavy chain (HC2) of SEQ ID NO: 19, and a second light chain (LC2) of SEQ ID NO: 20. In some embodiments, the bispecific anti-EGFR / c-Met antibody comprises a biantennary glycan structure having a fucose content of about 1% to about 15%.

[0168] In some embodiments, the bispecific anti-EGFR / c-Met antibodies disclosed herein may be administered in combination with a tyrosine kinase inhibitor (TKI), such as, but not limited to, an epidermal growth factor receptor (EGFR TKI). Non-limiting examples of TKIs are kinase inhibitors such as erlotinib, gefitinib, lapatinib, vandetanib, afatinib, osimertinib, lazertinib, poziotinib, cliotinib, cabozantinib, capmatinib, axitinib, lenvatinib, nintedanib, regorafenib, pazopanib, sorafenib, or sunitinib. In some embodiments, the bispecific anti-EGFR / c-Met antibodies disclosed herein may be administered in combination with lazertinib.

[0169] In some embodiments, the methods of the disclosure may be useful for treating cancer in a subject in need thereof, the methods comprising administering to the subject a cancer therapy that does not comprise a combination therapy comprising a bispecific anti-EGFR / c-Met bispecific antibody disclosed herein and an EGFR TKI.

[0170] In some embodiments, the one or more anti-cancer therapies comprise one or more chemotherapeutic agents, checkpoint inhibitors, targeted cancer therapies, or kinase inhibitors, or any combination thereof.

[0171] In some embodiments, the kinase inhibitor is an inhibitor of EGFR, an inhibitor of MET, an inhibitor of HER2, an inhibitor of HER3, an inhibitor of HER4, an inhibitor of VEGFR, or an inhibitor of AXL. In some embodiments, the kinase inhibitor is an inhibitor of EGFR. In some embodiments, the kinase inhibitor is an inhibitor of MET. In some embodiments, the kinase inhibitor is an inhibitor of HER2. In some embodiments, the kinase inhibitor is an inhibitor of HER3. In some embodiments, the kinase inhibitor is an inhibitor of HER4. In some embodiments, the kinase inhibitor is an inhibitor of VEGFR. In some embodiments, the kinase inhibitor is an inhibitor of AXL.

[0172] In some embodiments, the one or more anticancer therapies comprise carboplatin, paclitaxel, gemcitabine, cisplatin, vinorelbine, docetaxel, palbociclib, crizotinib, PD-(L)1 axis inhibitors, EGFR inhibitors, MET inhibitors, HER2 inhibitors, HER3 inhibitors, HER4 inhibitors, VEGFR inhibitors, AXL inhibitors, erlotinib, gefitinib, lapatinib, vandetanib, afatinib, osimertinib, lazertinib, poziotinib, cliotinib, cabozantinib, capmatinib, axitinib, lenvatinib, nintedanib, regorafenib, pazopanib, sorafenib, or sunitinib, or any combination thereof. Exemplary PD-(L)1 axis inhibitors are antibodies that bind to PD-1, such as nivolumab (OPDIVO®), pembrolimab (KEYTRUDA®), sintilimab, cemiplimab (LIBTAYO®), tripolibamab, tislelizumab, spartalizumab, camrelizumab, dostralimab, genolimuzumab, or cetrelimab, or antibodies that bind to PD-L1, e.g., PD-L1 antibodies are embafolimab, atezolizumab (TECENTRIQ®), durvalumab (IMFINZI®), and avelumab (BAVENCIO®). Commercially available antibodies can be purchased through authorized distributors or pharmacies. Amino acid sequence structures of small molecules can be found in the USAN and / or INN deposits by companies from the CAS registry.

[0173] In some embodiments, the cancer therapy not including the combination therapy of the present disclosure can be a platinum-based chemotherapy, such as, but not limited to, carboplatin, cisplatin, or a combination thereof.

[0174] In some embodiments, the methods of the present disclosure may be useful for treating cancer in a subject in need of such treatment, wherein the subject is chemotherapy naive.

[0175] In some embodiments, the methods of the present disclosure may be useful for treating cancer in a subject in need thereof, the subject having at least one activating EGFR mutation. Non-limiting examples of activating EGFR mutations are exon 19 deletion, L858R, and T790M.

[0176] Administration The bispecific anti-EGFR / c-Met antibody may be administered in a pharma- ceutically acceptable carrier. "Carrier" refers to a diluent, adjuvant, excipient, or vehicle with which the antibody of the present invention is administered. Such vehicles may be liquids, such as water and oils, including those of petroleum, animal, vegetable, or synthetic origin, e.g., peanut oil, soybean oil, mineral oil, sesame oil, and the like. For example, 0.4% saline and 0.3% glycine may be used to formulate the bispecific anti-EGFR / c-Met antibody. These solutions are sterile and generally free of particulate matter. They may be sterilized by conventional, well-known sterilization techniques (e.g., filtration). For parenteral administration, the carrier may comprise sterile water, with other excipients added to enhance solubility or preserve. Injectable suspensions or solutions may also be prepared utilizing aqueous carriers with appropriate additives. Suitable vehicles and formulations (including other human proteins such as human serum albumin) are described, for example, in Remington: The Science and Practice of Pharmacy, 21st Edition, Troy, DBed., Lipincott Williams and Wilkins, Philadelphia, PA 2006, Part 5, Pharmaceutical Manufacturing pp 691-1092, see in particular pp. 958-989.

[0177] The mode of administration may be any suitable route that delivers the bispecific anti-EGFR-c-Met antibody to the host, for example, parenteral administration, such as intradermal, intramuscular, intraperitoneal, intravenous, or subcutaneous, pulmonary, transmucosal (oral, intranasal, intravaginal, rectal), using tablet, capsule, liquid, powder, gel, particle formulations that may be contained in syringes, implantable devices, osmotic pumps, cartridges, micropumps, or other means well known in the art and recognized by the skilled artisan. Site-specific administration can be achieved, for example, by intratumoral, intraarticular, intrabronchial, intraabdominal, intracapsular, intrachondral, intrasinus, intracavity, intracerebellar, intraventricular, intracolonic, intracervical, intragastric, intrahepatic, intramyocardial, intraosseous, intrapelvic, intrapericardial, intraperitoneal, intrapleural, intraprostatic, intrapulmonary, intrarectal, intrarenal, intraretinal, intraspinal, intrasynovial, intrathoracic, intrauterine, intravascular, intravesical, intralesional, intravaginal, intrarectal, buccal, sublingual, intranasal, or transdermal delivery.

[0178] In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered intravenously.

[0179] In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered subcutaneously or intradermally to the subject. The bispecific anti-EGFR / c-Met antibody may be administered subcutaneously or intradermally at a dose sufficient to achieve a therapeutic effect in the subject.

[0180] In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose between about 140 mg and about 2240 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose between about 140 mg and about 1750 mg.

[0181] In some embodiments, the bispecific anti-EGFR / c-Met antibody is about 200 mg, about 210 mg, about 220 mg, about 230 mg, about 240 mg, about 250 mg, about 260 mg, about 270 mg, about 280 mg, about 290 mg, about 300 mg, about 310 mg, about 320 mg, about 330 mg, about 340 mg, about 350 mg, about 360 mg, about 370 mg, about 380 mg, about 390 mg, about 400 mg, about 410 mg, about 420 mg, about 430 mg, about 440 mg, about 450 mg, about 460 mg, about 470 mg, about 480 mg, about 490 mg, about 500 mg, about 510 mg, about 520 mg, about 530 mg, about 540 mg, about 550 mg, about 560 mg, about 570 mg, about 580 mg, about 590 mg, about 600 mg, about 610 mg, about 620 mg, about 630 mg, about 640 mg, about 650 mg, about 660 mg, about 670 mg, about 680 mg, about 690 mg, about 700 mg, about 710 mg, about 720 mg, about 730 mg, about 740 mg, about 750 mg, about 760 mg, about 770 mg, about 780 mg, about 790 mg, about 800 mg, about 810 mg, about 820 mg, about 830 mg, about 840 mg, about 850 mg, about 860 mg, about 870 mg, about 880 mg, about 890 mg, about 900 mg, about 910 mg, about 920 mg, about 930 mg, about 940 mg, about 950 mg, about 960 mg, about 970 mg, about 980 mg, about 990 mg, about 1000 mg, about 1010 mg, about 1020 mg, about 1030 mg, about 1040 mg, about 1050 mg, about 1060 mg, about 1070 mg, about 1080 mg, about 1090 mg, about 1100 mg, about 1110 mg, about 1120 mg, about 1130 mg, about 1140 mg, about 1150 mg, about 1160 mg, about 1170 mg, about 1180 mg, about 1190 mg, about 1200 mg, about 1210 mg, about 1220 mg, about 1230 mg, about 1240 mg, about 1250 mg, about 1260 mg, about 1270 mg, about 1280 mg, about 1290 mg, about 1300 mg, about 1310 mg, about 1320 mg, about 1330 mg, about 1340 mg, about 1350 mg, about 1360 mg, about 1370 mg, about 1380 mg, about 1390 mg, about 1400 mg, about 1410 mg, about 1420 mg, about 1430 mg, about 1440 mg, about 1450 mg, about 1460 mg, about 1470 mg, about 1480 mg, about 1490 mg, about 1500 mg,Approximately 1510mg, approximately 1520mg, approximately 1530mg, approximately 1540mg, approximately 1550mg, approximately 1560mg, approximately 1570mg, 1575mg, approximately 15 80mg, 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 1 750mg, about 1760mg, about 1770mg, about 1780mg, about 1790mg, about 1800mg, about 1810mg, about 1820mg, about 1830 The compound is administered in a dose of about 1800 mg, about 1840 mg, about 1850 mg, about 1860 mg, about 1870 mg, about 1880 mg, 1890 mg, about 1900 mg, about 1910 mg, about 1920 mg, about 1930 mg, about 1940 mg, about 1950 mg, about 1960 mg, about 1970 mg, about 1980 mg, about 1990 mg, about 2000 mg, 2100 mg, 2110 mg, 2120 mg, 2130 mg, 2140 mg, 2150 mg, 2160 mg, 2170 mg, 2180 mg, 2190 mg, 2200 mg, 2210 mg, 2220 mg, 2230 mg, 2240 mg, 2250 mg, or 2260 mg.

[0182] In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 350 mg, about 700 mg, about 1050 mg, 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.

[0183] In some embodiments, if the subject weighs less than 80 kg, the bispecific anti-EGFR / c-Met antibody is administered at a dose of 1050 mg. In some embodiments, if the subject weighs 80 kg or more, the bispecific anti-EGFR / c-Met antibody is administered at a dose of 1400 mg.

[0184] In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered once a week. In some embodiments, about 1050 mg of the bispecific anti-EGFR / c-Met antibody is administered once a week. In some embodiments, about 1400 mg of the bispecific anti-EGFR / c-Met antibody is administered once a week.

[0185] In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered once every two weeks. In some embodiments, about 1050 mg of the bispecific anti-EGFR / c-Met antibody is administered once every two weeks. In some embodiments, about 1400 mg of the bispecific anti-EGFR / c-Met antibody is administered once every two weeks.

[0186] 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.

[0187] In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered twice a week, once a week, once every two weeks, once every three weeks, or once every four weeks.

[0188] In some embodiments, the bispecific anti-EGFR / c-Met antibodies disclosed herein may be administered in combination with a tyrosine kinase inhibitor TKI, such as, but not limited to, an epidermal growth factor receptor (EGFR TKI). Non-limiting examples of TKIs are kinase inhibitors such as erlotinib, gefitinib, lapatinib, vandetanib, afatinib, osimertinib, lazertinib, poziotinib, cliotinib, cabozantinib, capmatinib, axitinib, lenvatinib, nintedanib, regorafenib, pazopanib, sorafenib, or sunitinib. In certain embodiments, the EGFR TKI is lazertinib.

[0189] For combination therapy, the EGFR TKI may be administered using the recommended dose and dosage of the EGFR TKI.

[0190] The mode of administration may be any suitable route that delivers the EGFR TKI to the host, e.g., parenteral, e.g., intradermal, intramuscular, intraperitoneal, intravenous, or subcutaneous, pulmonary, transmucosal (oral, intranasal, intravaginal, rectal), using tablet, capsule, liquid, powder, gel, particle formulations, which may be contained in syringes, implantable devices, osmotic pumps, cartridges, micropumps, or other means well known in the art and recognized by the skilled artisan. Site-specific administration can be achieved, for example, by intratumoral, intraarticular, intrabronchial, intraabdominal, intracapsular, intrachondral, intrasinus, intracavity, intracerebellar, intraventricular, intracolonic, intracervical, intragastric, intrahepatic, intramyocardial, intraosseous, intrapelvic, intrapericardial, intraperitoneal, intrapleural, intraprostatic, intrapulmonary, intrarectal, intrarenal, intraretinal, intraspinal, intrasynovial, intrathoracic, intrauterine, intravascular, intravesical, intralesional, intravaginal, intrarectal, buccal, sublingual, intranasal, or transdermal delivery.

[0191] In some embodiments, a mode of administration, which may be a suitable route of delivery of lazertinib to a subject, may be oral administration.

[0192] In some embodiments, the EGFR TKI is administered at a dose of about 10 mg to about 400 mg. In some embodiments, the EGFR TKI is administered at a dose of about 20 mg to about 320 mg. In some embodiments, the EGFR TKI is administered at a dose of about 50 mg to about 300 mg. In some embodiments, the EGFR TKI is administered at a dose of about 100 mg to about 300 mg. In some embodiments, the EGFR TKI is administered at a dose of about 150 mg to about 280 mg. In some embodiments, the EGFR TKI is administered at a dose of about 200 mg to about 250 mg. In some embodiments, the EGFR TKI is administered at a dose of about 220 mg to about 250 mg.

[0193] In some embodiments, the EGFR TKI is administered at a dose of about 20 mg, about 50 mg, about 100 mg, about 110 mg, about 120 mg, about 130 mg, about 140 mg, about 150 mg, about 160 mg, about 170 mg, about 180 mg, about 190 mg, about 200 mg, about 210 mg, about 220 mg, about 230 mg, about 240 mg, about 250 mg, about 260 mg, about 270 mg, about 280 mg, about 290 mg, about 300 mg, about 310 mg, about 320 mg, about 330 mg, about 340 mg, about 350 mg, about 360 mg, about 370 mg, about 380 mg, about 390 mg, or about 400 mg. In some embodiments, the EGFR TKI is administered at a dose of about 240 mg.

[0194] In some embodiments, the EGFR TKI is administered daily. In some embodiments, the EGFR TKI is administered twice a week. In some embodiments, the EGFR TKI is administered once a week. In some embodiments, the lazertinib is administered once every two weeks. In some embodiments, the lazertinib is administered once every three weeks. In some embodiments, the EGFR TKI is administered once every four weeks.

[0195] In some embodiments, the bispecific anti-EGFR / c-Met antibodies disclosed herein may be administered in combination with lazertinib, which may be administered using any of the doses and dosages disclosed herein. In some embodiments, lazertinib is administered at a dose of about 10 mg to about 400 mg. In some embodiments, lazertinib is administered at a dose of about 20 mg to about 320 mg. In some embodiments, lazertinib is administered at a dose of about 20 mg, about 50 mg, about 100 mg, about 110 mg, about 120 mg, about 130 mg, about 140 mg, about 150 mg, about 160 mg, about 170 mg, about 180 mg, about 190 mg, about 200 mg, about 210 mg, about 220 mg, about 230 mg, about 240 mg, about 250 mg, about 260 mg, about 270 mg, about 280 mg, about 290 mg, about 300 mg, about 310 mg, about 320 mg, about 330 mg, about 340 mg, about 350 mg, about 360 mg, about 370 mg, about 380 mg, about 390 mg, or about 400 mg. In some embodiments, lazertinib is administered at a dose of about 240 mg.

[0196] In some embodiments, the bispecific anti-EGFR / c-Met antibodies disclosed herein may be administered in any of these doses and dosages disclosed herein in combination with lazertinib, which may be administered in any of these doses and dosages disclosed herein. As a non-limiting example, 700 mg of amivantamab may be administered in combination with 240 mg of lazertinib. As a non-limiting example, 1050 mg of amivantamab may be administered in combination with 240 mg of lazertinib. As a non-limiting example, 1050 mg of amivantamab may be administered in combination with 240 mg of lazertinib. As a non-limiting example, 1400 mg of amivantamab may be administered in combination with 240 mg of lazertinib.

[0197] In some embodiments, the bispecific anti-EGFR / c-Met antibodies disclosed herein may be administered in combination with lazertinib, where lazertinib is administered daily, every other day, twice weekly, or once weekly. In some embodiments, the bispecific anti-EGFR / c-Met antibodies disclosed herein may be administered in combination with lazertinib, where lazertinib is administered daily. In some embodiments, the bispecific anti-EGFR / c-Met antibodies disclosed herein may be administered in combination with lazertinib, where lazertinib is administered orally.

[0198] In some embodiments, the combination therapy comprising a bispecific anti-EGFR / c-Met bispecific antibody and an EGFR TKI may further comprise one or more additional anti-cancer therapies.

[0199] In some embodiments, the methods of the present disclosure include administering to a subject a cancer therapy that does not include a combination therapy comprising a bispecific anti-EGFR / c-Met bispecific antibody disclosed herein and an EGFR TKI. In some embodiments, the cancer therapy may include any one of those described herein. As non-limiting examples, the cancer therapy that may be administered in the methods of the present disclosure may include any number of different platinum-based chemotherapy or combinations thereof. As non-limiting examples, platinum-based chemotherapy includes carboplatin, cisplatin, or combinations thereof.

[0200] Anti-cancer therapies that may be administered in the methods of the present disclosure include any one or more of chemotherapeutic agents or other anti-cancer therapeutic agents known to those of skill in the art. Chemotherapeutic agents are chemical compounds useful in the treatment of cancer, including growth inhibitors or other cytotoxic agents, including alkylating agents, antimetabolites, anti-microtubule inhibitors, topoisomerase inhibitors, receptor tyrosine kinase inhibitors, angiogenesis inhibitors, and the like. Examples of chemotherapeutic agents include alkylating agents such as thiotepa and cyclophosphamide (CYTOXAN®); alkylsulfonates such as busulfan, improsulfan, and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, and uredopa; ethylenimines and methylamelamines, including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trimethylolmelamine; nitrogen mustards such as chlorambucil, chlornaphazine, chlorophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembitine, phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosoureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, ranimustine; antibiotics such as aclacinomycin, actinomycin, autramycin, azaserine, bleomycin, cactinomycin, calicheamicin, carabicin, carminomycin, carzinophilin, chromomycin, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin, epirubicin, esorubicin, idarubicin, marcellomycin, mitomycin, mycophenolic acid, nogalamycin, olivomycin, peplomycin, potfiromycin, puromycin, quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin;antimetabolites such as methotrexate and 5-FU; folic acid analogues such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogues such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogues such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine, etc.; androgens such as calsterone, dromostanolone propionate, epithiostanol, mepitiostane, testolactone; adrenal cortical hormone synthesis inhibitors such as aminoglutethimide, mitotane, trilostane; folic acid replenishers such as folinic acid acid;aceglatone;aldophosphamide glycoside;aminolevulinic acid;amsacrine;bestrabucil;bisantrene;edatraxate;defofamine;demecolcine;diaziquone;elfornithine;elliptinium acetate acetate);Etoglucide;Gallium nitrate;Hydroxyurea;Lentinan;Lonidamine;Mitoguazone;Mitoxantrone;Mopidamol;Nitracrine;Pentostatin;Phenamet;Pirarubicin;Podophyllic acid;2-Ethylhydrazide;Procarbazine;PSK(R);Razoxane;Sizofiran;Spirogermanium;Tenuazonic acid;Triaziquone;2,2',2"-Trichlorotriethylamine;Urethane;Vindesine;Dacarbazine;Mannomustine;Mitobronitrol;Mitolactol ;pipobroman;gacytosine;arabinoside ("Ara-C");cyclophosphamide;thiotepa;members of the taxoid or taxane family, such as paclitaxel (TAXOL®), docetaxel (TAXOTERE®) and their analogues;chlorambucil;gemcitabine;6-thioguanine;mercaptopurine;methotrexate;platinum analogues, such as cisplatin and carboplatin;vinblastine;platinum;etoposide (VP-16);ifosfamide;mitomycin C;mitoxantrone;Vincristine; Vinorelbine; Navelbine; Novantrone; Teniposide; Daunomycin; Aminopterin; Xeloda; Ibandronate; CPT-11; Topoisomerase inhibitor RFS2000; Difluoromethylornithine (DMFO); Retinoic acid; Esperamicin; Capecitabine; Sorafenib (NEXAVAR®), Sunitinib (SUTENT®), Pazopanib (V and inhibitors of receptor tyrosine kinase and / or angiogenesis, including inhibitors of gliomas such as gliomas with leukemia (ALK), ... Also included in this definition are anti-hormonal agents that act to regulate or inhibit hormone action on tumors, such as antiestrogens, including tamoxifen, raloxifene, aromatase-inhibiting 4(5)-imidazole, 4-hydroxytamoxifen, trioxifene, ketoxifene, LY 117018, onapristone, and toremifene (FARESTON®); and antiandrogens, such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; as well as pharma- ceutical acceptable salts, acids, or derivatives of any of the above. Other conventional cytotoxic compounds disclosed in Wiemann et al., 1985, Medical Oncology (Calabresi et al., eds.), Chapter 10, McMillan Publishing, are also applicable to the methods of the present invention.

[0201] Generation of bispecific anti-EGFR / c-Met antibodies for use in the methods of the present disclosure An exemplary bispecific anti-EGFR / c-Met antibody that can be used in the methods of the present disclosure is amivantamab. Amivantamab or JNJ-61186372 (JNJ-372) is an IgG1 anti-EGFR / c-Met bispecific antibody described in U.S. Patent No. 9,593,164, which is incorporated herein by reference in its entirety. Amivantamab is characterized by the following amino acid sequence: EGFR binding arm

[0202] >SEQ ID NO:1 (HCDR1, EGFR binding arm) TYGMH

[0203] >SEQ ID NO:2 (HCDR2, EGFR binding arm) VIWDDGSYKYYGDSVKG

[0204] >SEQ ID NO:3 (HCDR3, EGFR binding arm) DGITMVRGVMKDYFDY

[0205] >SEQ ID NO:4 (LCDR1, EGFR binding arm) RASQDISSALV

[0206] >SEQ ID NO:5 (LCDR2, EGFR binding arm) DASSLES

[0207] >SEQ ID NO:6 (LCDR3, EGFR binding arm) QQFNSYPLT

[0208] >SEQ ID NO:7 (HCDR1, c-Met binding arm) SYGIS

[0209] >SEQ ID NO:8 (HCDR2, c-Met binding arm) WISAYNGYTNYAQKLQG

[0210] >SEQ ID NO:9 (HCDR3, c-Met binding arm) DLRGTNYFDY

[0211] >SEQ ID NO: 10 (LCDR1, c-Met binding arm) RASQGISNWLA

[0212] >SEQ ID NO: 11 (LCDR2, c-Met binding arm) AASSLLS

[0213] >SEQ ID NO: 12 (LCDR3, c-Met binding arm) QQANSFPIT

[0214] >SEQ ID NO: 13 (VH, EGFR binding arm) QVQLVESGGGVVQPGRSLRLSCAASGFTFSTYGMHWVRQAPGKGLEWVAVIWDDGSYKYYGDSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDGITMVRGVMKDYFDYWGQGTLVTVSS

[0215] >SEQ ID NO: 14 (VL, EGFR binding arm) AIQLTQSPSSLSASVGDRVTITCRASQDISSALVWYQQKPGKAPKLLIYDASSLESGVPSRFSGSESGTDFTLTISSLQPEDFATYYCQQFNSYPLTFGGGTKVEIK

[0216] >SEQ ID NO: 15 (VH, c-Met binding arm) QVQLVQSGAEVKKPGASVKVSCETSGYTFTSYGISWVRQAPGHGLEWMGWISAYNGYTNYAQKLQGRVTMTTDTSTSTAYMELRSLRSDDTAVYYCARDLRGTNYFDYWGQGTLVTVSS

[0217] >SEQ ID NO: 16 (VL, c-Met binding arm) DIQMTQSPSSVSASVGDRVTITCRASQGISNWLAWFQHKPGKAPKLLIYAASSLLSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANSFPITFGQGTRLEIK

[0218] >Sequence number 17 HC1 QVQLVESGGGVVQPGRSLRLSCAASGFTFSTYGMHWVRQAPGKGLEWVAVIWDDGSYKYYGDSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDGITMVRGVMKDYFD YWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKS CDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIE KTISKAKGQPREPQVYTLPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFLLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0219] >SEQ ID NO:18 LC1 AIQLTQSPSSLSASVGDRVTITCRASQDISSALVWYQQKPGKAPKLLIYDASSLESGVPSRFSGSESGTDFTLTISSLQPEDFATYYCQQFNSYPLTFGGGTKVEIK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0220] >Sequence number 19 HC2 QVQLVQSGAEVKKPGASVKVSCETSGYTFTSYGISWVRQAPGHGLEWMGWISAYNGYTNYAQKLQGRVTMTTDTSTSTAYMELRSLRSDDTAVYYCARDLRGTNYFDYWGQG TLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDK THTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEK TISKAKGQPREPQVYTLPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0221] >Sequence number 20 LC2 DIQMTQSPSSVSASVGDRVTITCRASQGISNWLAWFQHKPGKAPKLLIYAASSLLSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANSFPITFGQGTRLEIK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0222] 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 (HCDR1) of SEQ ID NO: 1, HCDR2 of SEQ ID NO: 2, HCDR3 of SEQ ID NO: 3, light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 4, LCDR2 of SEQ ID NO: 5, and LCDR3 of SEQ ID NO: 6, and the second domain comprises HCDR1 of SEQ ID NO: 7, HCDR2 of SEQ ID NO: 8, HCDR3 of SEQ ID NO: 9, LCDR1 of SEQ ID NO: 10, LCDR2 of SEQ ID NO: 11, and LCDR3 of SEQ ID NO: 12.

[0223] In some embodiments, a first domain that specifically binds EGFR comprises a heavy chain variable region (VH) of SEQ ID NO: 13 and a light chain variable region (VL) of SEQ ID NO: 14, and a second domain that specifically binds c-Met comprises a VH of SEQ ID NO: 15 and a VL of SEQ ID NO: 16.

[0224] In some embodiments, the bispecific anti-EGFR / c-Met antibody is of the IgG1 isotype.

[0225] In some embodiments, the bispecific anti-EGFR / c-Met antibody comprises a first heavy chain (HC1) of SEQ ID NO: 17, a first light chain (LC1) of SEQ ID NO: 18, a second heavy chain (HC2) of SEQ ID NO: 19, and a second light chain (LC2) of SEQ ID NO: 20.

[0226] In one embodiment, the bispecific anti-EGFR / c-Met antibody comprises one or more Fc silencing mutations.

[0227] In one embodiment, the one or more Fc silencing mutations reduce affinity for an Fcγ receptor.

[0228] In one embodiment, the one or more Fc silencing mutations include V234A / G237A / P238S / H268A / V309L / A330S / P331S.

[0229] In one embodiment, the bispecific anti-EGFR / c-Met antibody comprises a biantennary glycan structure with a fucose content between about 1% and about 15%. Antibodies with low fucose content can be produced using a variety of methods that have been reported to successfully express relatively highly defucosylated antibodies with biantennary complex type Fc oligosaccharides, such as: controlling the osmolarity of the culture (Konno et al., Cytotechnology 64(:249-65, 2012), applying the variant CHO line Lec13 as a host cell line (Shields et al., J Biol Chem 277:26733-26740, 2002), applying the variant CHO line EB66 as a host cell line (Olivier et al., MAbs;2(4), 2010; Epub ahead of print; PMID:20562582), applying the rat hybridoma cell line YB2 / 0 as a host cell line (Shinkawa et al., J Biol Chem 278:3466-3473, 2003), introduction of small interfering RNA specific for the 1,6-fucosyltransferase (FUT8) gene (Mori et al., Biotechnol Bioeng 88:901-908, 2004), or co-expression of β-1,4-N-acetylglucosaminyltransferase III and Golgi α-mannosidase II or kifunensine, a potent α-mannosidase I inhibitor (Ferrara et al., J Biol Chem 281:5032-5036, 2006; Ferrara et al., Biotechnol Bioeng 93:851-861, 2006; Xhou et al., Biotechnol Bioeng 99:652-65, 2008). In general, reducing the fucose content in the glycans of antibodies enhances antibody-mediated cellular cytotoxicity (ADCC).

[0230] Other publicly available bispecific anti-EGFR / c-Met antibodies may be used in the methods of the disclosure, so long as they exhibit similar characteristics when compared to amivantamab, as described in U.S. Patent No. 9,593,164. Bispecific anti-EGFR / c-Met antibodies that may be used in the methods of the disclosure may also be generated by combining publicly available EGFR-binding VH / VL domains and c-Met-binding VH / VL domains and testing the resulting bispecific antibody for its characteristics as described in U.S. Patent No. 9,593,164.

[0231] The bispecific anti-EGFR / c-Met antibodies used in the disclosed methods can be generated using Fab arm exchange (or half molecule exchange) between two monospecific bivalent antibodies, for example, by introducing substitutions in the heavy chain CH3 interface in each half molecule to favor the formation of heterodimers of two antibody half molecules with different specificities, in a cell-free environment in vitro or using co-expression. The Fab arm exchange reaction is the result of a disulfide bond isomerization reaction and dissociation-association of the CH3 domains. The heavy chain disulfide bonds in the hinge region of the parent monospecific antibodies are reduced. The resulting free cysteine ​​of one of the parent monospecific antibodies forms an intra-heavy chain disulfide bond with a cysteine ​​residue of the second parent monospecific antibody molecule, and at the same time, the CH3 domain of the parent antibody is released and reformed by dissociation-association. The CH3 domain of the Fab arm can be engineered to favor heterodimerization over homodimerization. The resulting product is a bispecific antibody with two Fab arms or half molecules that each bind a different epitope, i.e., an epitope in EGFR and an epitope in c-Met. For example, bispecific antibodies of the invention can be generated using the techniques described in WO 2011 / 131746. For IgG1 antibodies, the mutation F405L in one heavy chain and K409R in the other heavy chain can be used. For IgG2 antibodies, wild type IgG2 and IgG2 antibodies with F405L and R409K substitutions may be used. For IgG4 antibodies, wild type IgG4 and IgG4 antibodies with F405L and R409K substitutions may be used. To generate a bispecific antibody, a first monospecific bivalent antibody and a second monospecific bivalent antibody are engineered to have the aforementioned mutations in the Fc region, and the antibodies are incubated together under sufficient reducing conditions to allow the cysteines in the hinge region to undergo disulfide bond isomerization, thereby generating the bispecific antibody by Fab arm exchange. Incubation conditions may optimally be returned to non-reducing conditions.Exemplary reducing agents that may be used are 2-mercaptoethylamine (2-MEA), dithiothreitol (DTT), dithioerythritol (DTE), glutathione, tris(2-carboxyethyl)phosphine (TCEP), L-cysteine, and β-mercaptoethanol. For example, incubation at a temperature of at least 20° C., in the presence of at least 25 mM 2-MEA or in the presence of at least 0.5 mM dithiothreitol, at a pH of 5-8, e.g., pH 7.0 or pH 7.4, for at least 90 minutes may be used.

[0232] Bispecific anti-EGFR / c-Met antibodies for use in the methods of the disclosure can also be generated using designs such as knobs-in-holes (Genentech), CrossMAb (Roche) and electrostatically-matched (Chugai, Amgen, NovoNordisk, Oncomed), LUZ-Y (Genentech), Strand Exchange Engineered Domain body (SEEDbody) (EMD Serono), and Biclonic (Merus).

[0233] In the "knobs-in-holes" approach (see, for example, WO 2006 / 028936), selected amino acids that form the interface of the CH3 domain of human IgG can be mutated at positions that affect CH3 domain interactions to promote heterodimer formation. Amino acids with small side chains (holes) are introduced into the heavy chain of an antibody that specifically binds to a first antigen, and amino acids with large side chains (knobs) are introduced into the heavy chain of an antibody that specifically binds to a second antigen. After co-expression of the two antibodies, heterodimers are formed as a result of the preferential interaction of heavy chains with "holes" and heavy chains with "knobs". Exemplary CH3 substitution pairs that form knobs and holes are T366Y / F405A, T366W / F405W, F405W / Y407A, T394W / Y407T, T394S / Y407A, T366W / T394S, F405W / T394S, and T366W / T366S_L368A_Y407V (expressed as modification position in the first CH3 domain of the first heavy chain / modification position in the second CH3 domain of the second heavy chain).

[0234] In addition to utilizing a "knobs-in-holes" approach to promote Fab arm exchange, CrossMAb technology utilizes CH1 / CL domain swapping in one of the half-arms to ensure correct light chain pairing of the resulting bispecific antibody (see, e.g., U.S. Pat. No. 8,242,247).

[0235] Other crossover techniques may be used to generate full length bispecific antibodies of the invention by swapping variable or constant, or both, domains in one or both arms between the heavy and light chains of the bispecific antibody or within the heavy chain, including, for example, VH-CH1 and VL-CL, VH and VL, CH3 and CL, and CH3 and CH1 as described in WO 2009 / 080254, WO 2009 / 080251, WO 2009 / 018386, and WO 2009 / 080252.

[0236] Other approaches, such as promoting heavy chain heterodimerization using electrostatic interactions by substituting positively charged residues on one CH3 surface and negatively charged residues on the second CH3 surface, may be used as described in U.S. Patent Application Publication No. 2010 / 0015133, U.S. Patent Application Publication No. 2009 / 0182127, U.S. Patent Application Publication No. 2010 / 028637, or U.S. Patent Application Publication No. 2011 / 0123532. In another approach, heterodimerization can be achieved by the following substitutions: L351Y_F405A_Y407V / T394W, T366I_K392M_T394W / F405A_Y407V, T366L_K392M_T394W / F405A_Y407V, L351Y ... A / T366A_K409F, L351Y_Y407A / T366V_K409F, Y407A / T366A_K409F, or T350V_L351Y_F405A_Y407V / T350V_T366L_K392L_T394W (represented as modification position in the first CH3 domain of the first heavy chain / modification position in the second CH3 domain of the second heavy chain).

[0237] SEEDbody technology may be utilized to generate bispecific antibodies of the invention. SEEDbodies have selected IgG residues in their constant domains substituted with IgA residues to promote heterodimerization, as described in US Patent Application Publication No. 20070287170.

[0238] Mutations are typically made at the DNA level, using standard methods, on molecules such as the constant domain of an antibody.

[0239] kit The present invention also provides kits that include any of one or more reagents for determining the presence or level of one or more biomarkers described herein. The kits can be used for therapeutic applications and as diagnostic kits.

[0240] The kit may include one or more other components, including packaging; instructions for use; other reagents, such as a label, a therapeutic agent, or an agent useful for chelating or otherwise coupling, an antibody to a label or therapeutic agent, or a radioprotective composition; a device or other materials for preparing the antibody for administration; a pharma- ceutically acceptable carrier, and a device or other materials for administration to a subject.

[0241] In some embodiments, the kits of the present disclosure include one or more reagents for determining the presence of any one or more mutations described herein, including but not limited to, mutations in tumor DNA from a subject with cancer, e.g., lung cancer. In some embodiments, the tumor DNA is circulating tumor DNA (ctDNA).

[0242] The kit can be used to detect the presence of a mutation. Non-limiting examples of mutations include PIK3CA E545K, PIK3CA E542K / V, PIK3CA H1047R, PIK3CA amplification, mutations in one or more genes from the RAS / RAF / MEK pathway as described herein, mutations in one or more genes from the WNT / b-catenin pathway as described herein, KRAS G12V / C / D / X (wherein X is any amino acid other than G, V, C, and D), KRAS amplification, BRAF V600E, BRAF amplification, CCND1 amplification, CCND2 amplification, CCNE1 amplification, CDK4 amplification, CDK6 amplification, HER2 amplification, HER2 oncogenic alterations, PTEN deletions, PTEN N48K, CDKN2A G101W, CDKN2B mutations, ALK fusions, FGFR3-TACC3 and other fusions (e.g., TPM3-NTRK1 fusions), RET fusions, BRAF fusions and other oncogenic fusion events, EGFR These are EGFR C797S, EGFR L792H, EGFR amplification, EGFR G796S, EGFR L718X (where X is any amino acid), EGFR E709K, EGFR G724S, MET amplification, and MET exon 14 skipping (METex14) mutations.

[0243] In some embodiments, the mutations in one or more genes from the RAS / RAF / MEK pathway include FGFR3 fusions, BRAF G469A, BRAF V600E, ERBB2 copy number alterations, ALK fusions, ERBB2 I767M, ERBB2 V777L, KRAS A18V, KRAS copy number alterations, KRAS G12X (wherein X is any amino acid), NRAS Q61R, PDGFRA copy number alterations, and RET fusions. In some embodiments, the KRAS G12X mutation is KRAS G12D, KRAS G12A, KRAS G12C, and KRAS G12V.

[0244] In some embodiments, the mutations in one or more genes from the WNT / b-catenin pathway include APC Q1469, APC R405, APC S713, CTNNB1 S33P, CTNNB1 S37C, CTNNB1 S37F, and CTNNB1 S45P.

[0245] Non-limiting examples of HER2 oncogenic alterations include HER2 Y772_A775 duplication, HER2 L755M / S / W, and HER2 S310F / Y. Non-limiting examples of PTEN deletions include PTEN I33del and PTEN I14del. Non-limiting examples of ALK fusions include SQSTM1-ALK fusion and EML4-ALK fusion. Non-limiting examples of RET fusions include CCDC6-RET fusion, KIF5B-RET fusion, and NCOA4-RET fusion. Non-limiting examples of BRAF fusions include those described by Ross et al. al., Int. J. Cancer: 138, 881-890 (2016), which is incorporated by reference in its entirety, e.g., KIAA1549-BRAF, MKRN1-BRAF, TRIM24-BRAF, AGAP3-BRAF, ZC3HAV1-BRAF, AKAP9-BRAF, CCDC6-BRAF, AGK-BRAF, EPS15-BRAF, NUP214-BRAF, ARMC10-BRAF, BTF3L4-BRAF, GHR-BRAF, ZNF767 -BRAF, CCDC91-BRAF, DYNC1I2-BRAF, ZKSCAN1-BRAF, GTF2I-BRAF, MZT1-BRAF, RAD18-BRAF, CUX1-BRAF, SLC12A7-BRAF, MYRIP-BRAF, SND1-BRAF, NUB1-BRAF, KLHL7-BRAF, TANK-BRAF, RBMS3-BRAF, STRN3-BRAF, STK35-BRAF, ETFA-BRAF, SVOPL-BRAF, and JHDM1D-BRAF. Other oncogenic fusion events include, but are not limited to, those described in Figure 1 of Gao et al., Cell Rep. 2018 April 03; 23(1): 227-238. e3. (incorporated herein by reference in its entirety).

[0246] In another aspect, provided herein is a diagnostic kit comprising (i) one or more reagents for determining the presence of one or more mutations in tumor DNA from a subject having cancer, and (ii) optionally packaging and / or instructions, wherein the one or more mutations are selected from a mutation in one or more genes from the RAS / RAF / MEK pathway and a mutation in PIK3CA. In some embodiments, the one or more genes from the RAS / RAF / MEK pathway are FGFR3, KRAS, BRAF, ERBB2, ALK, NRAS, PDGFRA, and / or RET. In some embodiments, the mutations in one or more genes from the RAS / RAF / MEK pathway include FGFR3 fusions, BRAF G469A, BRAF V600E, ERBB2 copy number changes, ALK fusions, ERBB2 I767M, ERBB2 V777L, KRAS A18V, KRAS copy number changes, KRAS G12X (X is any amino acid), NRAS Q61R, PDGFRA copy number changes, and RET fusions. In some embodiments, the KRAS G12X mutations are KRAS G12D, KRAS G12A, KRAS G12C, and KRAS G12V. In some embodiments, the mutations in PIK3CA include PIK3CA E545K.

[0247] In some embodiments of the above kit, the one or more mutations are further selected from mutations in one or more genes from the WNT / b-catenin pathway. In some embodiments, the one or more genes from the WNT / b-catenin pathway are APC and CTNNB1. In some embodiments, the mutations in one or more genes from the WNT / b-catenin pathway include APC Q1469, APC R405, APC S713, CTNNB1 S33P, CTNNB1 S37C, CTNNB1 S37F, and CTNNB1 S45P.

[0248] In some embodiments, the kit may be a diagnostic kit, the diagnostic kit comprising (i) one or more reagents for determining the presence of one or more mutations in tumor DNA (e.g., circulating tumor DNA (ctDNA)) from a subject having cancer, and (ii) optionally, packaging and / or instructions for use, wherein the one or more mutations are selected from the following two groups: (1) PIK3CA E545K, PIK3CA E542K / V, PIK3CA H1047R, PIK3CA amplification, KRAS G12V / C / D / X (wherein X is any amino acid other than G, V, C, and D), KRAS amplification, BRAF V600E, BRAF amplification, CCND1 amplification, CCND2 amplification, CCNE1 amplification, CDK4 amplification, CDK6 amplification, HER2 amplification, HER2 oncogenic alterations, PTEN deletion, PTEN N48K, CDKN2A amplification, HER2 oncogenic alterations ... G101W, CDKN2B mutations, ALK fusions, FGFR3-TACC3 and other fusions (e.g., TPM3-NTRK1 fusions), RET fusions, BRAF fusions and other oncogenic fusion events; and (2) selected from EGFR C797S, EGFR L792H, EGFR amplification, EGFR G796S, EGFR L718X (wherein X is any amino acid), EGFR E709K, EGFR G724S, MET amplification, and MET exon 14 skipping (METex14) mutations. In some embodiments, the ctDNA may be present in a biological sample isolated from the subject. The biological sample may be any of the biological samples of the present disclosure, such as, but not limited to, a blood sample or a plasma sample. In some embodiments, the tumor DNA may be present in a tumor sample isolated from the subject.

[0249] In some embodiments, the diagnostic kit may further comprise one or more reagents for purifying tumor DNA (e.g., ctDNA) from a biological sample from a subject. In some embodiments, the one or more reagents may be used in conjunction with a sequencing technology (e.g., next generation sequencing (NGS)) to determine one or more mutations disclosed herein.

[0250] In certain aspects, a diagnostic kit is provided that includes (i) one or more reagents for determining the expression level of EGFR and / or MET in a tumor sample from a subject having cancer, and (ii) optionally, packaging and / or instructions for use. In some embodiments, the one or more reagents may be used in conjunction with immunohistochemistry (IHC) to determine the expression level of EGFR and / or MET.

[0251] Exemplary embodiments 1. A method for determining whether a cancer in a subject is susceptible to treatment with a combination therapy comprising a bispecific anti-epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) bispecific antibody and an EGFR tyrosine kinase inhibitor (TKI), comprising: a) determining the presence of one or more mutations in tumor DNA obtained from the subject, wherein the one or more mutations are selected from a mutation in one or more genes from the RAS / RAF / MEK pathway and a mutation in PIK3CA; b) (i) identifying the cancer in the subject as susceptible to treatment with the combination therapy if tumor DNA from the subject does not have the mutation, or (ii) identifying the cancer in the subject as not susceptible to treatment with the combination therapy if tumor DNA from the subject has one or more of the mutations. 2. A method for treating cancer in a subject in need thereof, comprising: a) determining the presence of one or more mutations in tumor DNA obtained from the subject, wherein the one or more mutations are selected from a mutation in one or more genes from the RAS / RAF / MEK pathway and a mutation in PIK3CA; b) (i) administering to the subject a therapeutically effective amount of a combination therapy comprising a bispecific anti-epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) bispecific antibody and an EGFR tyrosine kinase inhibitor (TKI) if the tumor DNA from the subject does not have the mutations, or (ii) administering to the subject a cancer therapy that does not include the combination therapy used in (i) if the tumor DNA from the subject has one or more of the mutations. 3. The method of claim 1 or 2, wherein the one or more genes from the RAS / RAF / MEK pathway are FGFR3, KRAS, BRAF, ERBB2, ALK, NRAS, PDGFRA and / or RET. 4. The method of claim 3, wherein the mutations in one or more genes from the RAS / RAF / MEK pathway comprise FGFR3 fusions, BRAF G469A, BRAF V600E, ERBB2 copy number alterations, ALK fusions, ERBB2 I767M, ERBB2 V777L, KRAS A18V, KRAS copy number alterations, KRAS G12X (wherein X is any amino acid), NRAS Q61R, PDGFRA copy number alterations, and RET fusions. 5. The method of claim 4, wherein the KRAS G12X mutation is KRAS G12D, KRAS G12A, KRAS G12C, or KRAS G12V. 6. The method of any one of claims 1 to 5, wherein the mutation in PIK3CA comprises PIK3CA E545K. 7. The method of any one of claims 1 to 5, wherein the one or more mutations are further selected from mutations in one or more genes from the WNT / b-catenin pathway. 8. The method of claim 6, wherein the one or more genes from the WNT / b-catenin pathway are APC and CTNNB1. 9. The method of claim 7, wherein the mutations in one or more genes from the WNT / b-catenin pathway include APC Q1469, APC R405, APC S713, CTNNB1 S33P, CTNNB1 S37C, CTNNB1 S37F, and CTNNB1 S45P. 10. A method for determining whether a cancer in a subject is susceptible to treatment with a combination therapy comprising a bispecific anti-epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) bispecific antibody and an EGFR tyrosine kinase inhibitor (TKI), comprising: a) determining the presence of one or more mutations in tumor DNA obtained from the subject, the one or more mutations falling into one of the following two groups: (1) PIK3CA E545K, PIK3CA E542K / V, PIK3CA H1047R, PIK3CA amplification, KRAS G12V / C / D / X (X is any amino acid other than G, V, C, and D), KRAS amplification, BRAF V600E, BRAF amplification, CCND1 amplification, CCND2 amplification, CCNE1 amplification, CDK4 amplification, CDK6 amplification, HER2 amplification, HER2 oncogenic alteration, PTEN deletion, PTEN N48K, CDKN2A G101W, CDKN2B mutation, ALK fusion, FGFR3-TACC3 fusion, TPM3-NTRK1 fusion, RET fusion, BRAF fusion, and other oncogenic fusion events; (2) selected from EGFR C797S, EGFR L792H, EGFR amplification, EGFR G796S, EGFR L718X (wherein X is any amino acid), EGFR E709K, EGFR G724S, MET amplification, and MET exon 14 skipping (METex14) mutation; b) (i) identifying the cancer in the subject as susceptible to treatment with the combination therapy if tumor DNA from the subject either has no mutations from group (1), or has one or more mutations from group (1) and one or more mutations from group (2); or (ii) identifying the cancer in the subject as not susceptible to treatment with the combination therapy if tumor DNA from the subject has one or more mutations from group (1) and no mutations from group (2). 11. A method for treating cancer in a subject in need thereof, comprising: a) determining the presence of one or more mutations in tumor DNA obtained from the subject, the one or more mutations falling into one of the following two groups: (1) PIK3CA E545K, PIK3CA E542K / V, PIK3CA H1047R, PIK3CA amplification, KRAS G12V / C / D / X, KRAS amplification, BRAF V600E, BRAF amplification, CCND1 amplification, CCND2 amplification, CCNE1 amplification, CDK4 amplification, CDK6 amplification, HER2 amplification, HER2 oncogenic alterations, PTEN deletion, PTEN N48K, CDKN2A G101W, CDKN2B, ALK fusions, FGFR3-TACC3 and other fusions, RET fusions, BRAF fusions, and other oncogenic fusion events; (2) selected from EGFR C797S, EGFR L792H, EGFR amplification, EGFR G796S, EGFR L718X (wherein X is any amino acid), EGFR E709K, EGFR G724S, MET amplification, and MET exon 14 skipping (METex14) mutation; b) (i) administering to the subject a therapeutically effective amount of a combination therapy comprising a bispecific anti-epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) bispecific antibody and an EGFR tyrosine kinase inhibitor (TKI), if the tumor DNA from the subject either does not have a mutation from group (1) or has one or more mutations from group (1) and one or more mutations from group (2); or (ii) administering to the subject a cancer therapy that does not include the combination therapy used in (i), if the tumor DNA from the subject has one or more mutations from group (1) and does not have a mutation from group (2). 12. The method of embodiment 10 or 11, wherein the HER2 oncogenic alterations include HER2 Y772_A775 duplication, HER2 L755M / S / W and HER2 S310F / Y. 13. The method of any one of embodiments 10-12, wherein the PTEN deletion comprises PTEN I33del and PTEN I14del. 14. The method of any one of embodiments 10 to 13, wherein the ALK fusion comprises an SQSTM1-ALK fusion and an EML4-ALK fusion. 15. The method of any one of embodiments 10 to 14, wherein the RET fusion comprises a CCDC6-RET fusion, a KIF5B-RET fusion, and an NCOA4-RET fusion. 16. The method of any one of embodiments 1 to 15, wherein the cancer is lung cancer. 17. The method of embodiment 16, wherein the lung cancer is non-small cell lung cancer (NSCLC). 18. The method of any one of embodiments 1-17, wherein the cancer in the subject is resistant to treatment with an EGFR TKI that is not the same as the EGFR TKI used in the combination therapy. 19. The method of embodiment 18, wherein the EGFR TKI to which the cancer is resistant is selected from osimertinib, erlotinib, afatinib, rociletinib, olmutinib, and any combination thereof. 20. The method of embodiment 19, wherein the EGFR TKI to which the cancer is resistant is osimertinib. 21. The method of any one of embodiments 1-20, wherein the subject is chemotherapy naive. 22. The method of any one of embodiments 1 to 21, wherein the tumor DNA from the subject has at least one EGFR activating mutation. 23. The method of embodiment 22, wherein the EGFR activating mutation is selected from exon 19 deletion, L858R, and T790M. 24. The method of any one of embodiments 1 to 23, wherein the tumor DNA is circulating tumor DNA (ctDNA). 25. The method of embodiment 24, wherein the ctDNA is present in a biological sample isolated from the subject. 26. The method of embodiment 25, wherein the biological sample is a blood sample or a plasma sample. 27. The method of embodiment 25 or 26, wherein ctDNA is isolated from the biological sample prior to mutation identification. 28. The method of any one of embodiments 1 to 27, wherein the tumor DNA is present in a tumor sample isolated from the subject. 29. The method of embodiment 28, wherein tumor DNA is isolated from the tumor sample prior to mutation identification. 30. The method of any one of embodiments 1 to 29, wherein one or more mutations are determined by sequencing. 31. The method of embodiment 30, wherein the one or more mutations are determined using next generation sequencing (NGS). 32. The method of any one of embodiments 1 to 31, wherein the bispecific anti-EGFR / c-Met antibody comprises a first domain that specifically binds to EGFR and a second domain that specifically binds to c-Met, wherein the first domain comprises heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 1, HCDR2 of SEQ ID NO: 2, HCDR3 of SEQ ID NO: 3, light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 4, LCDR2 of SEQ ID NO: 5, and LCDR3 of SEQ ID NO: 6, and the second domain that binds to c-Met comprises HCDR1 of SEQ ID NO: 7, HCDR2 of SEQ ID NO: 8, HCDR3 of SEQ ID NO: 9, LCDR1 of SEQ ID NO: 10, LCDR2 of SEQ ID NO: 11, and LCDR3 of SEQ ID NO: 12. 33. The method of embodiment 32, wherein the first domain that specifically binds to EGFR comprises a heavy chain variable region (VH) of SEQ ID NO: 13 and a light chain variable region (VL) of SEQ ID NO: 14, and the second domain that specifically binds to c-Met comprises a VH of SEQ ID NO: 15 and a VL of SEQ ID NO: 16. 34. The method of embodiment 32 or 33, wherein the bispecific anti-EGFR / c-Met antibody is of the IgG1 isotype. 35. The method of any one of embodiments 1 to 34, wherein the bispecific anti-EGFR / c-Met antibody comprises a first heavy chain (HC1) of SEQ ID NO: 17, a first light chain (LC1) of SEQ ID NO: 18, a second heavy chain (HC2) of SEQ ID NO: 19, and a second light chain (LC2) of SEQ ID NO: 20. 36. The method of any one of embodiments 1 to 35, wherein the bispecific anti-EGFR / c-Met antibody comprises a biantennary glycan structure having a fucose content of about 1% to about 15%. 37. The method of any one of embodiments 1-36, wherein the bispecific anti-EGFR / c-Met antibody is administered intravenously to the subject. 38. The method of embodiment 37, wherein the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 140 mg to about 2240 mg. 39. The method of embodiment 38, wherein the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 700 mg, about 750 mg, about 800 mg, about 850 mg, 900 mg, 950 mg, 1000 mg, 1050 mg, 1100 mg, 1150 mg, 1200 mg, 1250 mg, 1300 mg, 1350 mg, 1400 mg, 1575 mg, 1600 mg, 2100 mg, or 2240 mg. 40. The method of embodiment 39, wherein the bispecific anti-EGFR / c-Met antibody is administered at a dose of 1050 mg if the subject has a body weight of less than 80 kg. 41. The method of embodiment 39, wherein the bispecific anti-EGFR / c-Met antibody is administered at a dose of 1400 mg if the subject has a body weight of 80 kg or more. 42. The method of any one of embodiments 1-36, wherein the bispecific anti-EGFR / c-Met antibody is administered to the subject subcutaneously or intradermally. 43. The method of embodiment 42, wherein the bispecific anti-EGFR / c-Met antibody is administered subcutaneously or intradermally at a dose sufficient to achieve a therapeutic effect in the subject. 44. The method of any one of embodiments 1-43, wherein the bispecific anti-EGFR / c-Met antibody is administered twice a week, once a week, once every two weeks, once every three weeks, or once every four weeks. 45. The method according to any one of embodiments 1 to 44, wherein the EGFR TKI administered in combination with the bispecific anti-EGFR / c-Met antibody is lazertinib. 46. ​​The method of any one of embodiments 1 to 45, wherein the EGFR TKI administered in combination with the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 20 to about 320 mg. 47. The method of any one of embodiments 1-46, wherein the EGFR TKI administered in combination with the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 240 mg. 48. The method of any one of embodiments 1-47, wherein the EGFR TKI administered in combination with the bispecific anti-EGFR / c-Met antibody is administered daily, every other day, twice weekly, or once weekly. 49. The method according to any one of embodiments 1 to 48, wherein the EGFR TKI administered in combination with the bispecific anti-EGFR / c-Met antibody is administered daily. 50. The method of any one of embodiments 1-49, wherein the EGFR TKI administered in combination with the bispecific anti-EGFR / c-Met antibody is administered orally. 51. The method according to any one of embodiments 2 to 9 and 11 to 50, wherein the cancer therapy not including the concomitant therapy used in (i) is a platinum-based chemotherapy. 52. The method of embodiment 51, wherein the platinum-based chemotherapy comprises carboplatin and / or cisplatin. 53. The method of any one of embodiments 1 to 52, comprising obtaining a biological sample from the subject prior to step (a), wherein the biological sample contains tumor DNA, and optionally purifying the tumor DNA from the biological sample. 54. A method for determining whether a cancer in a subject is susceptible to treatment with a combination therapy comprising a bispecific anti-epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) bispecific antibody and an EGFR tyrosine kinase inhibitor (TKI), comprising: a) determining the expression level of EGFR or MET in a tumor sample obtained from the subject using immunohistochemistry (IHC); b) determining a staining intensity score on a scale of 0 to 3+ based on the expression level of EGFR or MET determined in step (a); c) (i) identifying the cancer in the subject as susceptible to treatment with the combination therapy if the staining intensity score is 3+, or (ii) identifying the cancer in the subject as not susceptible to treatment with the combination therapy if the staining intensity score is less than 3+. 55. The method of claim 54, wherein step (c) comprises identifying the cancer in the subject as sensitive to treatment with the combination therapy if 25% or more of the cells in the tumor sample have a staining intensity score of 3+; or (ii) identifying the cancer in the subject as not sensitive to treatment with the combination therapy if less than 25% of the cells in the tumor sample have a staining intensity score of 3+. 56. A method for treating cancer in a subject in need thereof, comprising: a) determining the expression level of EGFR or MET in a tumor sample obtained from the subject using immunohistochemistry (IHC); b) determining a staining intensity score on a scale of 0 to 3+ based on the expression level of EGFR or MET determined in step (a); c) (i) administering to the subject a therapeutically effective amount of a combination therapy comprising a bispecific anti-epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) bispecific antibody and an EGFR tyrosine kinase inhibitor (TKI) if the staining intensity score is 3+; or (ii) not administering to the subject the combination therapy used in (i), or administering to the subject a cancer therapy that does not include the combination therapy used in (i), if the staining intensity score is less than 3+. 57. The method of claim 56, wherein in step (c), (i) if 25% or more of the cells in the tumor sample have a staining intensity score of 3+, a therapeutically effective amount of the combination therapy is administered to the subject, or (ii) if less than 25% of the cells in the tumor sample have a staining intensity score of 3+, then the combination therapy used in (i) is not administered to the subject, or a cancer therapy not including the combination therapy used in (i) is administered to the subject. 58. A method for determining whether a cancer in a subject is susceptible to treatment with a combination therapy comprising a bispecific anti-epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) bispecific antibody and an EGFR tyrosine kinase inhibitor (TKI), comprising: a) determining the expression levels of EGFR and MET in a tumor sample obtained from the subject using immunohistochemistry (IHC); b) calculating a combined H-score based on the expression levels of EGFR and MET determined in step (a); and c) (i) identifying the cancer in the subject as susceptible to treatment with the combination therapy if the combined H-score is 400 or greater, or (ii) identifying the cancer in the subject as not susceptible to treatment with the combination therapy if the combined H-score is less than 400. 59. A method for treating cancer in a subject in need thereof, comprising: a) determining the expression levels of EGFR and MET in a tumor sample obtained from the subject using immunohistochemistry (IHC); b) calculating a combined H-score based on the expression levels of EGFR and MET determined in step (a); and c) (i) administering to the subject a therapeutically effective amount of a combination therapy comprising a bispecific anti-epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) bispecific antibody and an EGFR tyrosine kinase inhibitor (TKI) if the combined H-score is 400 or greater; and (ii) not administering to the subject the combination therapy used in (i), or administering to the subject a cancer therapy that does not include the combination therapy used in (i), if the combined H-score is less than 400. 60. The method of any one of embodiments 54-59, wherein the cancer is lung cancer. 61. The method of embodiment 60, wherein the lung cancer is non-small cell lung cancer (NSCLC). 62. The method of any one of embodiments 54-61, wherein the cancer in the subject is resistant to treatment with an EGFR TKI that is not the same as the EGFR TKI used in the combination therapy. 63. The method of embodiment 62, wherein the EGFR TKI to which the cancer is resistant is selected from osimertinib, erlotinib, afatinib, rociletinib, olmutinib, and any combination thereof. 64. The method of embodiment 63, wherein the EGFR TKI to which the cancer is resistant is osimertinib. 65. The method of any one of embodiments 54-64, wherein the subject is chemotherapy naive. 66. The method of any one of embodiments 54 to 65, wherein the subject's tumor has at least one EGFR activating mutation. 67. The method of embodiment 66, wherein the EGFR activating mutation is selected from exon 19 deletion, L858R, and T790M. 68. The method of any one of embodiments 64 to 67, wherein the bispecific anti-EGFR / c-Met antibody comprises a first domain that specifically binds to EGFR and a second domain that specifically binds to c-Met, wherein the first domain comprises heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 1, HCDR2 of SEQ ID NO: 2, HCDR3 of SEQ ID NO: 3, light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 4, LCDR2 of SEQ ID NO: 5, and LCDR3 of SEQ ID NO: 6, and the second domain that binds to c-Met comprises HCDR1 of SEQ ID NO: 7, HCDR2 of SEQ ID NO: 8, HCDR3 of SEQ ID NO: 9, LCDR1 of SEQ ID NO: 10, LCDR2 of SEQ ID NO: 11, and LCDR3 of SEQ ID NO: 12. 69. The method of embodiment 68, wherein the first domain that specifically binds to EGFR comprises a heavy chain variable region (VH) of SEQ ID NO: 13 and a light chain variable region (VL) of SEQ ID NO: 14, and the second domain that specifically binds to c-Met comprises a VH of SEQ ID NO: 15 and a VL of SEQ ID NO: 16. 70. The method of embodiment 68 or 69, wherein the bispecific anti-EGFR / c-Met antibody is of the IgG1 isotype. 71. The method of any one of embodiments 68 to 70, wherein the bispecific anti-EGFR / c-Met antibody comprises a first heavy chain (HC1) of SEQ ID NO: 17, a first light chain (LC1) of SEQ ID NO: 18, a second heavy chain (HC2) of SEQ ID NO: 19, and a second light chain (LC2) of SEQ ID NO: 20. 72. The method of any one of embodiments 54 to 71, wherein the bispecific anti-EGFR / c-Met antibody comprises a biantennary glycan structure having a fucose content of about 1% to about 15%. 73. The method of any one of embodiments 54-72, wherein the bispecific anti-EGFR / c-Met antibody is administered intravenously to the subject. 74. The method of embodiment 73, wherein the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 140 mg to about 2240 mg. 75. The method of embodiment 74, wherein the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 700 mg, about 750 mg, about 800 mg, about 850 mg, 900 mg, 950 mg, 1000 mg, 1050 mg, 1100 mg, 1150 mg, 1200 mg, 1250 mg, 1300 mg, 1350 mg, 1400 mg, 1575 mg, 1600 mg, 2100 mg, or 2240 mg. 76. The method of embodiment 75, wherein the bispecific anti-EGFR / c-Met antibody is administered at a dose of 1050 mg if the subject has a body weight of less than 80 kg. 77. The method of embodiment 75, wherein the bispecific anti-EGFR / c-Met antibody is administered at a dose of 1400 mg if the subject has a body weight of 80 kg or more. 78. The method of any one of embodiments 54 to 72, wherein the bispecific anti-EGFR / c-Met antibody is administered to the subject subcutaneously or intradermally. 79. The method of embodiment 78, wherein the bispecific anti-EGFR / c-Met antibody is administered subcutaneously or intradermally at a dose sufficient to achieve a therapeutic effect in the subject. 80. The method of any one of embodiments 54-79, wherein the bispecific anti-EGFR / c-Met antibody is administered twice a week, once a week, once every two weeks, once every three weeks, or once every four weeks. 81. The method according to any one of embodiments 54 to 80, wherein the EGFR TKI administered in combination with the bispecific anti-EGFR / c-Met antibody is lazertinib. 82. The method according to any one of embodiments 54 to 81, wherein the EGFR TKI administered in combination with the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 20 to about 320 mg. 83. The method according to any one of embodiments 54-82, wherein the EGFR TKI administered in combination with the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 240 mg. 84. The method according to any one of embodiments 54 to 83, wherein the EGFR TKI administered in combination with the bispecific anti-EGFR / c-Met antibody is administered daily, every other day, twice weekly, or once weekly. 85. The method according to any one of embodiments 54-84, wherein the EGFR TKI administered in combination with the bispecific anti-EGFR / c-Met antibody is administered daily. 86. The method according to any one of embodiments 54 to 85, wherein the EGFR TKI administered in combination with the bispecific anti-EGFR / c-Met antibody is administered orally. 87. The method according to any one of embodiments 56-57 and 59-86, wherein the cancer therapy not including the concomitant therapy used in (i) is a platinum-based chemotherapy. 88. The method of embodiment 87, wherein the platinum-based chemotherapy comprises carboplatin and / or cisplatin. 89. The method of any one of embodiments 54 to 88, comprising obtaining a tumor sample from the subject prior to step (a). 90. A diagnostic kit comprising: (i) one or more reagents for determining the presence of one or more mutations in tumor DNA from a subject having cancer; and (ii) optionally packaging and / or instructions for use, wherein the one or more mutations are selected from a mutation in one or more genes from the RAS / RAF / MEK pathway and a mutation in PIK3CA. 91. The diagnostic kit of claim 90, wherein the one or more genes from the RAS / RAF / MEK pathway are FGFR3, KRAS, BRAF, ERBB2, ALK, NRAS, PDGFRA and / or RET. 92. The diagnostic kit of claim 91, wherein the mutations in one or more genes from the RAS / RAF / MEK pathway include FGFR3 fusions, BRAF G469A, BRAF V600E, ERBB2 copy number alterations, ALK fusions, ERBB2 I767M, ERBB2 V777L, KRAS A18V, KRAS copy number alterations, KRAS G12X (wherein X is any amino acid), NRAS Q61R, PDGFRA copy number alterations, and RET fusions. 93. The diagnostic kit of claim 92, wherein the KRAS G12X mutation is KRAS G12D, KRAS G12A, KRAS G12C, or KRAS G12V. 94. The diagnostic kit of any one of claims 90 to 93, wherein the mutation in PIK3CA comprises PIK3CA E545K. 95. The diagnostic kit of any one of claims 90 to 94, wherein the one or more mutations are further selected from mutations in one or more genes from the WNT / b-catenin pathway. 96. The diagnostic kit of claim 95, wherein the one or more genes from the WNT / b-catenin pathway are APC and CTNNB1. 97. The diagnostic kit of claim 96, wherein the mutations in one or more genes from the WNT / b-catenin pathway include APC Q1469, APC R405, APC S713, CTNNB1 S33P, CTNNB1 S37C, CTNNB1 S37F, and CTNNB1 S45P. 98. A diagnostic kit comprising: (i) one or more reagents for determining the presence of one or more mutations in tumor DNA from a subject with cancer; and (ii) optionally packaging and / or instructions for use, wherein the one or more mutations are classified into one of the following two groups: (1) PIK3CA E545K, PIK3CA E542K / V, PIK3CA H1047R, PIK3CA amplification, KRAS G12V / C / D / X (X is any amino acid other than G, V, C, and D), KRAS amplification, BRAF V600E, BRAF amplification, CCND1 amplification, CCND2 amplification, CCNE1 amplification, CDK4 amplification, CDK6 amplification, HER2 amplification, HER2 oncogenic alteration, PTEN deletion, PTEN N48K, CDKN2A G101W, CDKN2B mutation, ALK fusion, FGFR3-TACC3 fusion, TPM3-NTRK1 fusion, RET fusion, BRAF fusion, and other oncogenic fusion events; (2) A diagnostic kit selected from EGFR C797S, EGFR L792H, EGFR amplification, EGFR G796S, EGFR L718X (X is any amino acid), EGFR E709K, EGFR G724S, MET amplification, and MET exon 14 skipping (METex14) mutations. 99. The method of embodiment 98, wherein the HER2 oncogenic alterations include HER2 Y772_A775 duplication, HER2 L755M / S / W and HER2 S310F / Y. 100. The method of embodiment 98 or 99, wherein the PTEN deletion comprises PTEN I33del and PTEN I14del. 101. The method of embodiment 98 or 99, wherein the ALK fusion comprises an SQSTM1-ALK fusion and an EML4-ALK fusion. 102. The method of any one of embodiments 98 to 101, wherein the RET fusion comprises a CCDC6-RET fusion, a KIF5B-RET fusion, and an NCOA4-RET fusion. 103. A diagnostic kit according to any one of embodiments 90 to 102, wherein the tumor DNA is circulating tumor DNA (ctDNA). 104. The diagnostic kit of embodiment 103, wherein the ctDNA is present in a biological sample isolated from the subject. 105. The diagnostic kit according to embodiment 104, wherein the biological sample is a blood sample or a plasma sample. 106. A diagnostic kit according to any one of embodiments 90 to 102, wherein the tumor DNA is present in a tumor sample isolated from the subject. 107. A diagnostic kit according to any one of embodiments 103 to 106, further comprising one or more reagents for purifying tumor DNA from a biological sample derived from a subject. 108. A diagnostic kit according to any one of embodiments 90 to 107, wherein one or more reagents can be used in conjunction with sequencing techniques to determine one or more mutations. 109. A diagnostic kit according to any one of embodiments 90 to 108, wherein one or more reagents can be used in conjunction with next generation sequencing (NGS) to determine one or more mutations. 110. A diagnostic kit comprising: (i) one or more reagents for determining the expression levels of EGFR and / or MET in a tumor sample from a subject having cancer; and (ii) optionally, packaging and / or instructions for use. 111. The diagnostic kit of embodiment 110, wherein one or more reagents can be used in conjunction with immunohistochemistry (IHC) to determine the expression levels of EGFR and / or MET. EXAMPLES

[0252] The following examples are provided to further describe some of the embodiments disclosed herein and are intended to be illustrative and not limiting of the embodiments of the present disclosure.

[0253] Example 1. Amivantamab in combination with lazertinib for the treatment of osimertinib-relapsed, chemotherapy-naïve EGFR mutant (EGFRm) non-small cell lung cancer (NSCLC) and potential biomarkers for response This example investigated the preliminary efficacy of the combination of amivantamab (an epidermal growth factor receptor (EGFR) and mesenchymal-epithelial transition factor (MET) bispecific antibody) and lazertinib (a third-generation tyrosine kinase inhibitor (TKI)) in treatment-naïve and osimertinib (osi)-relapsed patients with EGFR mutant (EGFRm) non-small cell lung cancer (NSCLC). Briefly, pretreatment tumor biopsies and circulating tumor DNA (ctDNA) were collected prior to administration of amivantamab in combination lazertinib. Osimertinib-resistant mutations or amplifications in EGFR / MET identified by next-generation sequencing (NGS) in either ctDNA or tumor biopsies (biomarker positive [pos]) were evaluated for enrichment response. Immunohistochemistry (IHC) staining for EGFR and MET expression was also investigated as potential biomarkers for response.

[0254] A schematic diagram of the structures of amivantamab and lazertinib, as well as a detailed description of the mechanism of action (MOA) of amivantamab, is shown in Figure 1. The progression of acquired resistance to osimertinib in epidermal growth factor receptor mutant (EGFRm) non-small cell lung cancer (NSCLC) is illustrated in Figure 2. Specifically, primary mutations, e.g., EGFR driver mutations (exon 19 deletion + L858R), may co-occur with resistance mutations such as those that are EGFR-dependent (C797S) or MET-dependent (MET amplification), may involve other pathways (e.g., PIK3CA, RAS / RAF / MEK, fusion, cycle), may result from transformation, or are still unclear (approximately 40-50%), each contributing to osimertinib resistance. Ultimately, the complexity of osimertinib resistance likely arises from heterogeneous resistance patterns with the co-occurrence of multiple resistance mechanisms. Sequencing of single tumor lesions may not reveal heterogeneous patterns in resistance or co-occurring mutations, and in this sense, plasma next-generation sequencing (NGS) may be more useful. NGS of circulating tumor DNA (ctDNA) is the most frequently used method to characterize osimertinib resistance mechanisms due to the difficulty of obtaining tissue (Papadimitrakopoulou et al., Annals of Oncol 29: VIII741, 2018; Ramalingam et al., Annals of Oncol 29: VIII740, 2018).

[0255] According to the methods of the present disclosure, patients (N=45) with EGFR exon 19 deletion or L858R mutation NSCLC that progressed on osimertinib without chemotherapy intervention were enrolled in the combination cohort of the ongoing CHRYSALIS study (NCT02609776, Cohort E), as shown in Figure 3. A description of patient demographics and baseline disease characteristics is shown in Figure 4. Pretreatment tumor biopsies and ctDNA were collected prospectively and patients received a combination dose of 1050 / 1400 mg amivantamab + 240 mg lazertinib to evaluate safety and efficacy in the osimertinib relapse population. Response was assessed by the investigator according to RECIST v1.1. Osimertinib resistance mutations or amplifications in EGFR / MET identified by next generation sequencing (NGS) in either ctDNA or tumor biopsies (biomarker positive [pos]) were evaluated for enrichment response. Durable responses were observed with the combination of amivantamab + lazertinib with manageable safety (Figures 5A-5B). The sum of the diameters (SoD) of target lesions shown in Figure 5A were measured as described in EAEisenhauer et al., New response evaluation criteria in solid tumours: Revised RECIST guideline (version 1.1), European J of Cancer 45 (2009) 228-247. The safety profile was consistent with our previous experience with amivantamab + lazertinib (Cho et al., Ann Oncol 31:S813, 2020). The most common adverse events (AEs) were infusion-related reactions (IRR; 78%), rash (acneiform dermatitis 51% + rash 27%), and paronychia (49%), the majority of which were grade 1-2. Of treatment-related events, 16% were grade ≥3 AEs, 4% led to discontinuation, and 18% led to dose reductions.

[0256] Among the key findings of this example, of 45 osimertinib relapsed patients, 36% (95% confidence interval [CI], 22-51) had a confirmed response (1 complete response and 15 partial responses [PR]). At a median follow-up of 8.2 months (1.0-11.8), 20 of 45 patients (44%) continued treatment. Eleven of 16 patients (69%) had ongoing responses (2.6-9.6+ months) with median duration of response not reached (NR). Median progression-free survival (mPFS) was 4.9 months (95% CI, 3.7-8.3).

[0257] In total, 44 of 45 patients were evaluable by ctDNA and 29 of 45 were evaluable by tumor NGS. Genetic testing identified 17 biomarker-positive patients, of whom 8 (47%) responded (Figures 6A-6B). At a median follow-up of 8.2 months (1.0-11.8), 20 of 45 patients (44%) continue treatment. Eleven of 16 patients (69%) continue to respond (2.6-9.6+ months) with median duration of response not reached (NR). Median progression-free survival (mPFS) was 4.9 months (95% CI, 3.7-8.3). Figure 6A shows plots of best percentage change in tumor volume for the EGFR-based, MET-based, and EGFR+MET (EGFR+MET)-based resistant groups. FIG. 6B shows a summary chart of the genetic alterations determined for EGFR-based, MET-based, and additional resistance groups.

[0258] Of the remaining 28 patients, 8 (29%) responded (Figures 7A-7B). Of these 28 patients, 18 had an unknown mechanism of osimertinib resistance (8 PR) and 10 had an identified non-EGFR / MET resistance mechanism (did not respond). The mPFS (95% CI) for biomarker-positive and remaining patients was 6.7 months (3.4-NR) and 4.1 months (4-9-1.5), respectively. Figure 7A shows plots of the best percentage change in tumor volume for the unknown resistance mechanism and EGFR / MET-independent resistance groups. Figure 7B shows a summary chart of the subset of genetic alterations, e.g., mutations, determined for the EGFR / MET-independent group. By way of non-limiting example, exemplary EGFR / MET-independent genetic alterations may include PIK3CA E545K, PIK3CA E542K / V, PIK3CA H1047R, PIK3CA amplification, KRAS G12V / C / D / X, KRAS amplification, BRAF V600E, BRAF amplification, CCND1 amplification, CCND2 amplification, CCNE1 amplification, CDK4 amplification, CDK6 amplification, HER2 amplification, HER2 oncogenic alterations, PTEN deletion, PTEN N48K, CDKN2A G101W, CDKN2B, ALK fusions, FGFR3-TACC3 and other fusions, RET fusions, BRAF fusions, and other oncogenic fusion events. According to the stratification methods of the present invention, in the absence of such EGFR / MET-independent mutations, a patient may be a candidate for the combination treatment of amivantamab and lazertinib disclosed herein. As illustrated in Figures 7A-7B, patients who are determined to have a non-EGFR, non-MET (i.e., EGFR / MET-independent) resistance mechanism and who further lack EGFR-based resistance mutations, e.g., EGFR C797S, EGFR L792H, EGFR amplification, EGFR G796S, EGFR L718X (where X is any amino acid), EGFR E709K, and EGFR G724S, and / or MET-based resistance mutations, e.g., MET amplification, and MET exon 14 skipping (METex14) mutations (see, e.g., Figure 6B), are excluded from treatment with amivantamab in combination with lazertinib treatment.These data indicate that such patients have a low probability of responding to this combination and, given the current standard of care, would instead be treated with, for example, platinum-based chemotherapy.

[0259] Adequate tissue was available for 20 patients to perform IHC testing for EGFR and MET (Figure 8). Ten patient biopsies were immunopositive (IHC+) for EGFR / MET and showed a combined EGFR+MET (EGFR+MET) H-score ≥ 400. The remaining 10 patient biopsies were defined as IHC-. IHC+ patients had an overall response rate (ORR) of 90% (9 / 10 patients), a median duration of response (mDOR) of 9.7 months, a clinical benefit rate (CBR) of 100%, and a median progression-free survival (mPFS) of 12.5 months. Five responders in the IHC+ group had unknown genetic mechanisms. A higher representation of positive responders (PR) was observed in the IHC+ group and was associated with a greater percentage reduction in tumor volume. These data suggest that IHC showing high expression of MET and / or EGFR is a positive predictor of treatment response to amivantamab / lasertinib combination therapy.

[0260] This example demonstrated that treatment with a combination of amivantamab and lazertinib produced a response in 36% of chemotherapy-naive patients who progressed on osimertinib. Among these patients, genetic EGFR and MET-based biomarkers of resistance identified a subgroup of patients more likely to respond to amivantamab and lazertinib, but additional patients lacking the identified resistance markers also responded. An IHC-based approach may identify patients most likely to benefit from the combination regimen.

[0261] Example 2. Validation of biomarkers for response in an expansion cohort study The CHRYSALIS-2 Phase 1 / 1b expansion cohorts will generally be conducted according to the exemplary study design, as shown in Figure 9. Key patient inclusion criteria for Phase 1b expansion cohorts A-D will be applied as follows: Inclusion criteria for expansion cohort A are EGFR exon 19 deletion or L858R, progression after osimertinib (first / second line), and platinum-based chemotherapy as final treatment. Inclusion criteria for expansion cohort B are EGFR exon 20 insertion, previous standard of care (SOC) platinum-based chemotherapy, or alternatively, EGFR TKI, which may include investigational EGFR-TKIs targeting exon 20 insertions (e.g., mobocertinib and poziotinib) or immuno-oncology therapy (IO), and chemotherapy with 3 or fewer prior therapies as final treatment. Inclusion criteria for expansion cohort C are rare non-exon 20 insertion mutations (e.g., S768I, L861Q, G719X), treatment naïve or having already received one 1st / 2nd generation EGFR TKI as last line, and no more than 2 previous lines of therapy. Inclusion criteria for expansion cohort D are EGFR exon 19 deletion or L858R, post-osimertinib (first / second line) as last line, and suitable tumor biopsy for biomarker validation after progression on most recent systemic therapy or from first biopsy in the metastatic setting. Phase 1b expansion cohorts received lazertinib (240 mg) in combination with amivantamab (1050 / 1400 mg (1050 mg, body weight < 80 kg; 1400 mg, body weight ≥ 80 kg). For expansion cohort D, osimertinib resistance mutations or amplifications in EGFR / MET as exemplified in Example 1 will be verified by next generation sequencing (NGS) in either ctDNA or tumor biopsies (biomarker positive [pos]) and assessed for enrichment response. Immunohistochemistry (IHC) staining for EGFR and MET expression will be further validated as biomarkers for response.

[0262] Example 3. Biomarker strategy based on NGS analysis of baseline plasma ctDNA Materials and Methods To evaluate biomarker strategies to identify patients with increased or decreased probability of tumor response to treatment with the amivantamab and lazertinib combination in participants who previously tested positive for EGFR exon 19del or L858R mutant NSCLC who progressed on or after osimertinib (phase 1b expansion cohort D), ORR by RECIST v1.1 was assessed in populations dichotomized by next-generation sequencing (NGS) analysis of circulating tumor DNA (ctDNA) in baseline plasma. The RECIST v1.1 guideline was adapted from EAEisenhauer et al., New response evaluation criteria in solid tumours: Revised RECIST guideline (version 1.1), European J of Cancer. 45 (2009) 228-247, which is incorporated herein by reference in its entirety. These participants had previously tested positive for either EGFR exon 19del or L858R mutant NSCLC. Participants progressed on or after osimertinib, were enrolled without biomarker selection, and were required to submit plasma for ctDNA NGS analysis. Each patient underwent classification based on the mechanism of osimertinib resistance as determined by ctDNA NGS analysis. Patients were classified as NGS1 if ctDNA NGS analysis identified a pathogenic PIK3CA E545K mutation or a pathogenic alteration in the RAS / RAF / MEK pathway. Patients were classified as NGS2 if ctDNA NGS analysis identified a pathogenic PIK3CA E545K mutation or a pathogenic alteration in the RAS / RAF / MEK pathway or a pathogenic alteration in the WNT / b-catenin pathway. Patients were classified as NGS3 if they identified E545K mutations or pathogenic alterations in the RAS / RAF / MEK pathway or pathogenic alterations in the WNT / b-catenin pathway, or if ctDNA NGS failed to detect EGFR L858R mutations or EGFR exon 19 deletion mutations (possibly due to the sensitivity limits of the ctDNA assay).Progression-free survival (PFS) was also compared between each NGS group.

[0263] A complete list of mutations observed in patients treated with amivantamab and lazertinib is provided below.

[0264] [Table 1] * Mutations ending in "DELINSX" mean that the amino acids at the listed positions (inclusive) have been deleted and replaced by an insertion of residue "X." For example, "EGFR_L747_P753DELINSS" means that the amino acids at positions 747 to 753 (inclusive) have been deleted and replaced by an insertion of "S."

[0265] result

[0266] [Table 2]

[0267] Exclusion of NGS1, NGS2, or NGS3 positive patients, who were enriched for PR and uPR compared to the unselected population. When treating uPR as PR, the ORRs for the NGS1, NGS2, and NGS3 negative populations are 39.5% (95% CI: 28.4% to 51.4%), 42% (95% CI: 28.4% to 51.4%), and 45.8% (95% CI: 32.7% to 59.3%), respectively (see Table 2). Meanwhile, the ORRs for the NGS1, NGS2, and NGS3 positive populations are 4.3% (95% CI: 0.1% to 22.0%), 6.7% (95% CI: 0.8% to 22.1%), and 10% (95% CI: 2.8% to 23.7%), respectively (Table 2). The ORR for the unselected population of subjects with evaluable ctDNA NGS results is 31.3% (95% CI: 22.4% to 41.4%) (Table 3).

[0268] [Table 3] * Cohort E was described in Example 1. # Dependent = EGFR / MET dependent, i.e., EGFR C797S mutation or MET amplification. Dependent=EGFR / MET dependent, such as KRAS or PIK3CA. Δ Unknown = Does not fit into dependent or non-dependent categories.

[0269] In support of the ORR, waterfall plots demonstrate enrichment for the best change from baseline in target lesion size with at least a 30% reduction in all NGS-negative groups (Figures 10B, 11B, and 12B). At 3 months, all NGS-negative groups show better PFS survival compared to their respective positive groups (Figures 10A, 11A, and 12A).

[0270] Example 4. Biomarker strategy based on IHC analysis of baseline tumor biopsies Materials and Methods To evaluate biomarker strategies to identify patients with increased or decreased probability of tumor response upon treatment with amivantamab and lazertinib combination in participants who previously tested positive for EGFR exon 19del or L858R mutant NSCLC and progressed on or after osimertinib (Phase 1b expansion cohort D), ORR by RECIST v1.1 was evaluated in populations dichotomized by immunohistochemistry (IHC) analysis of EGFR and MET expression on baseline tumor biopsies. These participants previously tested positive for EGFR exon 19del or L858R mutant NSCLC and progressed on or after osimertinib. These patients were enrolled without biomarker selection and were required to submit tumor tissue for IHC analysis (EGFR and MET expression). Each patient underwent classification for each IHC assay. Patients were classified as EGFR positive (IHC1) if they had an EGFR 3+ IHC intensity score of 25% or more per cell, and similarly, patients were classified as MET positive (IHC2) if their biopsy sample had a MET 3+ IHC intensity score of 25% or more. Progression-free survival was also compared between each IHC group.

[0271] result

[0272] [Table 4]

[0273] For the analysis, partial responders (PRs) and unconfirmed partial responders (uPRs) were considered as "responders," while progressive disease, stable disease, and non-evaluable / unknown were considered as "non-responders." Both the IHC1-positive and IHC2-positive groups were enriched for partial responders (PRs) and unconfirmed partial responders (uPRs) compared with the unselected population. When treating uPRs as PRs, the ORRs for the IHC1- and IHC2-positive populations are 45.7% (95% exact CI: 28.8-63.4%) and 55.6% (95% exact CI: 30.8-78.5%), respectively. Meanwhile, the ORRs for the IHC1- and IHC2-negative populations are 0% (95% exact CI: 0-21.8%) and 18.8% (95% exact CI: 7.21-36.4%), respectively. The ORR for the unselected population of subjects with evaluable IHC results is 32% (95% exact CI: 19.5-46.7%). In support of the ORR, swim lane plots demonstrate enrichment for best change from baseline (defined by RECIST v1.1) in target lesion size with at least a 30% reduction in both IHC positive groups (Figures 13A-14B). The sum of total diameters (SoD) of target lesions was measured as described in EAEisenhauer et al., New response evaluation criteria in solid tumours: Revised RECIST guideline (version 1.1), European J of Cancer 45 (2009) 228-247. At 3 months, both IHC positive groups show better PFS survival compared to their respective negative groups (Table 5, Figures 15A-15B).

[0274] [Table 5] * * *

[0275] The present invention is not to be limited in scope by the specific embodiments and examples described herein. Indeed, various modifications of the invention in addition to those described herein will become apparent to those skilled in the art from the foregoing description. Such modifications are intended to be within the scope of the appended claims.

[0276] All patents, applications, publications, test methods, literature, and other materials cited herein are incorporated by reference in their entirety as if physically present herein.

Claims

1. 1. A method for determining whether a cancer in a subject is susceptible to treatment with a combination therapy comprising a bispecific anti-epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) bispecific antibody and an EGFR tyrosine kinase inhibitor (TKI), comprising: a) determining the presence of one or more mutations in tumor DNA obtained from the subject, wherein the one or more mutations are selected from a mutation in one or more genes from the RAS / RAF / MEK pathway and a mutation in PIK3CA; b) (i) identifying the cancer in the subject as susceptible to said treatment with the combination therapy if tumor DNA from the subject does not have said mutations, or (ii) identifying the cancer in the subject as not susceptible to said treatment with the combination therapy if tumor DNA from the subject has one or more of said mutations; A method comprising:

2. A method for testing whether a cancer in a subject is susceptible to treatment with a combination therapy comprising a bispecific anti-epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) bispecific antibody and an EGFR tyrosine kinase inhibitor (TKI), comprising: a) determining the presence of one or more mutations in tumor DNA obtained from the subject, wherein the one or more mutations are selected from a mutation in one or more genes from the RAS / RAF / MEK pathway and a mutation in PIK3CA; b) testing whether the cancer in the subject is sensitive to treatment with the combination therapy by comparing with a criterion that if the tumor DNA from the subject does not have the mutation, the cancer in the subject is sensitive to treatment with the combination therapy, and if the tumor DNA from the subject has the mutation, the cancer in the subject is not sensitive to treatment with the combination therapy; The method includes:

3. The following (A) to (B): (A) the one or more genes from the RAS / RAF / MEK pathway are FGFR3, KRAS, BRAF, ERBB2, ALK, NRAS, PDGFRA and / or RET, and optionally the mutations in one or more genes from the RAS / RAF / MEK pathway comprise FGFR3 fusion, BRAF G469A, BRAF V600E, ERBB2 copy number alteration, ALK fusion, ERBB2 I767M, ERBB2 V777L, KRAS A18V, KRAS copy number alteration, KRAS G12X (X is any amino acid), NRAS Q61R, PDGFRA copy number alteration, and RET fusion, e.g., the KRAS G12X mutation is selected from the group consisting of KRAS G12D, KRAS G12E, KRAS G12F, KRAS G12G, KRAS G12H ... KRAS G12A, KRAS G12C and KRAS G12V, (B) the mutation in PIK3CA comprises PIK3CA E545K; The method according to claim 1 or 2, wherein any one of the above is satisfied.

4. The method of claim 1 or 2, wherein the one or more mutations are further selected from mutations in one or more genes from the WNT / b-catenin pathway, optionally wherein the one or more genes from the WNT / b-catenin pathway are APC and CTNNB1, for example, the mutations in one or more genes from the WNT / b-catenin pathway include APC Q1469, APC R405, APC S713, CTNNB1 S33P, CTNNB1 S37C, CTNNB1 S37F, and CTNNB1 S45P.

5. 3. The method of claim 1 or 2, wherein the cancer is lung cancer, optionally wherein the lung cancer is non-small cell lung cancer (NSCLC).

6. 3. The method of claim 1 or 2, wherein the cancer in the subject is resistant to treatment with an EGFR TKI that is not the same as the EGFR TKI used in the combination therapy, optionally wherein the EGFR TKI to which the cancer is resistant is selected from osimertinib, erlotinib, afatinib, rociletinib, olmutinib, and any combination thereof, e.g., the EGFR TKI to which the cancer is resistant is osimertinib.

7. 3. The method of claim 1 or 2, wherein the subject is chemotherapy naive.

8. The following (A) to (C): (A) the tumor DNA from the subject has at least one EGFR activating mutation, optionally wherein the EGFR activating mutation is selected from exon 19 deletion and L858R; (B) The tumor DNA is circulating tumor DNA (ctDNA), and optionally the ctDNA is present in a biological sample isolated from the subject, for example, as follows: (1) The biological sample is a blood sample or a plasma sample. (2) ctDNA is isolated from the biological sample prior to mutation identification; Either of the following is satisfied: (C) the tumor DNA is present in a tumor sample isolated from the subject, and optionally the tumor DNA is isolated from the tumor sample prior to mutation identification. The method according to claim 1 or 2, wherein any one of the above is satisfied.

9. 3. The method of claim 1 or 2, wherein the one or more mutations are determined by sequencing, optionally wherein the one or more mutations are determined using next generation sequencing (NGS).

10. The following (A) to (B): (A) The bispecific anti-EGFR / c-Met antibody comprises a first domain that specifically binds EGFR and a second domain that specifically binds c-Met, wherein the first domain comprises heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 1, HCDR2 of SEQ ID NO: 2, HCDR3 of SEQ ID NO: 3, light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 4, LCDR2 of SEQ ID NO: 5, and LCDR3 of SEQ ID NO: 6; and the second domain that binds c-Met comprises HCDR1 of SEQ ID NO: 7, HCDR2 of SEQ ID NO: 8, HCDR3 of SEQ ID NO: 9, LCDR1 of SEQ ID NO: 10, LCDR2 of SEQ ID NO: 11, and LCDR3 of SEQ ID NO: 12, and optionally one of the following (1) to (2): (1) The first domain that specifically binds to EGFR comprises a heavy chain variable region (VH) of SEQ ID NO: 13 and a light chain variable region (VL) of SEQ ID NO: 14, and the second domain that specifically binds to c-Met comprises a VH of SEQ ID NO: 15 and a VL of SEQ ID NO: 16; (2) The bispecific anti-EGFR / c-Met antibody is of the IgG1 isotype. Either of the following is satisfied: (B) the bispecific anti-EGFR / c-Met antibody comprises a first heavy chain (HC1) of SEQ ID NO: 17, a first light chain (LC1) of SEQ ID NO: 18, a second heavy chain (HC2) of SEQ ID NO: 19, and a second light chain (LC2) of SEQ ID NO: 20; (C) the bispecific anti-EGFR / c-Met antibody comprises a biantennary glycan structure having a fucose content of about 1% to about 15%. The method according to claim 1 or 2, wherein any one of the above is satisfied.

11. The following (A) to (C) (A) The bispecific anti-EGFR / c-Met antibody is administered intravenously to the subject, optionally at a dose of about 140 mg to about 2240 mg of the bispecific anti-EGFR / c-Met antibody, for example, at a dose of about 700 mg, about 750 mg, about 800 mg, about 850 mg, 900 mg, 950 mg, 1000 mg, 1050 mg, 1100 mg, 1150 mg, 1200 mg, 1250 mg, 1300 mg, 1350 mg, 1400 mg, 1575 mg, 1600 mg, 2100 mg, or 2240 mg of the bispecific anti-EGFR / c-Met antibody, and further optionally at a dose of (1)-(2) below: (1) if the subject has a body weight less than 80 kg, the bispecific anti-EGFR / c-Met antibody is administered at a dose of 1050 mg; (2) if the subject has a body weight of 80 kg or greater, the bispecific anti-EGFR / c-Met antibody is administered at a dose of 1400 mg; Either of the following is satisfied: (B) the bispecific anti-EGFR / c-Met antibody is administered subcutaneously or intradermally to the subject, optionally wherein the bispecific anti-EGFR / c-Met antibody is administered subcutaneously or intradermally in a dose sufficient to achieve a therapeutic effect in the subject. (C) the bispecific anti-EGFR / c-Met antibody is administered twice a week, once a week, once every two weeks, once every three weeks, or once every four weeks. The method according to claim 1 or 2, wherein any one of the above is satisfied.

12. The following (A) to (F): (A) the EGFR TKI administered in combination with the bispecific anti-EGFR / c-Met antibody is lazertinib; (B) the EGFR TKI administered in combination with the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 20 to about 320 mg. (C) the EGFR TKI administered in combination with the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 240 mg. (D) the EGFR TKI administered in combination with the bispecific anti-EGFR / c-Met antibody is administered daily, every other day, twice weekly, or once weekly. (E) the EGFR TKI administered in combination with the bispecific anti-EGFR / c-Met antibody is administered daily. (F) the EGFR TKI administered in combination with the bispecific anti-EGFR / c-Met antibody is administered orally. The method according to claim 1 or 2, wherein any one of the above is satisfied.

13. 3. The method of claim 1 or 2, comprising obtaining a biological sample from the subject prior to step (a), wherein the biological sample comprises tumor DNA, and optionally comprising purifying the tumor DNA from the biological sample.

14. 1. A method for determining whether a cancer in a subject is susceptible to treatment with a combination therapy comprising a bispecific anti-epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) bispecific antibody and an EGFR tyrosine kinase inhibitor (TKI), comprising: a) determining the expression level of EGFR or MET in a tumor sample obtained from said subject using immunohistochemistry (IHC); b) determining a staining intensity score on a scale of 0 to 3+ based on the expression level of EGFR or MET determined in step (a); c) (i) identifying the cancer in the subject as susceptible to said treatment with the combination therapy if the staining intensity score is 3+, or (ii) identifying the cancer in the subject as not susceptible to said treatment with the combination therapy if the staining intensity score is less than 3+. A method comprising:

15. 15. The method of claim 14, wherein step (c) comprises (i) identifying the cancer in the subject as sensitive to said treatment with the combination therapy if the staining intensity score is 3+ in 25% or more of the cells of the tumor sample, or (ii) identifying the cancer in the subject as not sensitive to said treatment with the combination therapy if the staining intensity score is 3+ in less than 25% of the cells of the tumor sample.

16. A method for testing whether a cancer in a subject is susceptible to treatment with a combination therapy comprising a bispecific anti-epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) bispecific antibody and an EGFR tyrosine kinase inhibitor (TKI), comprising: a) determining the expression level of EGFR or MET in a tumor sample obtained from said subject using immunohistochemistry (IHC); b) determining a staining intensity score on a scale of 0 to 3+ based on the expression level of EGFR or MET determined in step (a); c) testing whether the cancer in the subject is sensitive to treatment with the combination therapy by comparing with a criterion that if the staining intensity score is 3+, the cancer in the subject is sensitive to treatment with the combination therapy, and if the staining intensity score is less than 3+, the cancer in the subject is not sensitive to treatment with the combination therapy; The method includes:

17. The method of any one of claims 14 to 16, wherein the cancer is lung cancer, optionally wherein the lung cancer is non-small cell lung cancer (NSCLC).

18. 17. The method of any one of claims 14 to 16, wherein the cancer in the subject is resistant to treatment with an EGFR TKI that is not the same as the EGFR TKI used in the combination therapy, optionally wherein the EGFR TKI to which the cancer is resistant is selected from osimertinib, erlotinib, afatinib, rociletinib, olmutinib, and any combination thereof, e.g., the EGFR TKI to which the cancer is resistant is osimertinib.

19. The method of any one of claims 14 to 16, wherein the subject is chemotherapy naive.

20. 17. The method of any one of claims 14 to 16, wherein the tumor of the subject has at least one EGFR activating mutation, optionally wherein the EGFR activating mutation is selected from exon 19 deletion and L858R.

21. The bispecific anti-EGFR / c-Met antibody comprises a first domain that specifically binds EGFR and a second domain that specifically binds c-Met, wherein the first domain comprises heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 1, HCDR2 of SEQ ID NO: 2, HCDR3 of SEQ ID NO: 3, light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 4, LCDR2 of SEQ ID NO: 5, and LCDR3 of SEQ ID NO: 6; and the second domain that binds c-Met comprises HCDR1 of SEQ ID NO: 7, HCDR2 of SEQ ID NO: 8, HCDR3 of SEQ ID NO: 9, LCDR1 of SEQ ID NO: 10, LCDR2 of SEQ ID NO: 11, and LCDR3 of SEQ ID NO: 12, and optionally one of the following (A) to (D): (A) the first domain that specifically binds to EGFR comprises a heavy chain variable region (VH) of SEQ ID NO: 13 and a light chain variable region (VL) of SEQ ID NO: 14, and the second domain that specifically binds to c-Met comprises a VH of SEQ ID NO: 15 and a VL of SEQ ID NO: 16; (B) the bispecific anti-EGFR / c-Met antibody is of the IgG1 isotype; (C) the bispecific anti-EGFR / c-Met antibody comprises a first heavy chain (HC1) of SEQ ID NO: 17, a first light chain (LC1) of SEQ ID NO: 18, a second heavy chain (HC2) of SEQ ID NO: 19, and a second light chain (LC2) of SEQ ID NO: 20; (D) the bispecific anti-EGFR / c-Met antibody comprises a biantennary glycan structure having a fucose content of about 1% to about 15%. The method according to any one of claims 14 to 16, wherein any one of the above is satisfied.

22. The following (A) to (C): (A) The bispecific anti-EGFR / c-Met antibody is administered intravenously to the subject, optionally at a dose of about 140 mg to about 2240 mg of the bispecific anti-EGFR / c-Met antibody, for example, at a dose of about 700 mg, about 750 mg, about 800 mg, about 850 mg, 900 mg, 950 mg, 1000 mg, 1050 mg, 1100 mg, 1150 mg, 1200 mg, 1250 mg, 1300 mg, 1350 mg, 1400 mg, 1575 mg, 1600 mg, 2100 mg, or 2240 mg of the bispecific anti-EGFR / c-Met antibody, and further optionally at a dose of (1)-(2) below: (1) if the subject has a body weight less than 80 kg, the bispecific anti-EGFR / c-Met antibody is administered at a dose of 1050 mg; (2) if the subject has a body weight of 80 kg or greater, the bispecific anti-EGFR / c-Met antibody is administered at a dose of 1400 mg; Either of the following is satisfied: (B) the bispecific anti-EGFR / c-Met antibody is administered subcutaneously or intradermally to the subject, optionally wherein the bispecific anti-EGFR / c-Met antibody is administered subcutaneously or intradermally in a dose sufficient to achieve a therapeutic effect in the subject. (C) the bispecific anti-EGFR / c-Met antibody is administered twice a week, once a week, once every two weeks, once every three weeks, or once every four weeks. The method according to any one of claims 14 to 16, wherein any one of the above is satisfied.

23. The following (A) to (F): (A) the EGFR TKI administered in combination with the bispecific anti-EGFR / c-Met antibody is lazertinib; (B) the EGFR TKI administered in combination with the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 20 to about 320 mg. (C) the EGFR TKI administered in combination with the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 240 mg. (D) the EGFR TKI administered in combination with the bispecific anti-EGFR / c-Met antibody is administered daily, every other day, twice weekly, or once weekly. (E) the EGFR TKI administered in combination with the bispecific anti-EGFR / c-Met antibody is administered daily. (F) the EGFR TKI administered in combination with the bispecific anti-EGFR / c-Met antibody is administered orally. The method according to any one of claims 14 to 16, wherein any one of the above is satisfied.

24. The method of any one of claims 14 to 16, comprising obtaining a tumor sample from the subject prior to step (a).

25. A diagnostic kit comprising: (i) one or more reagents for determining the presence of one or more mutations in tumor DNA from a subject having cancer; and (ii) optionally packaging and / or instructions for use, wherein the one or more mutations are selected from mutations in one or more genes from the RAS / RAF / MEK pathway and a mutation in PIK3CA.

26. The following (A) to (B): (A) the one or more genes from the RAS / RAF / MEK pathway are FGFR3, KRAS, BRAF, ERBB2, ALK, NRAS, PDGFRA and / or RET, and optionally the mutations in one or more genes from the RAS / RAF / MEK pathway comprise FGFR3 fusion, BRAF G469A, BRAF V600E, ERBB2 copy number alteration, ALK fusion, ERBB2 I767M, ERBB2 V777L, KRAS A18V, KRAS copy number alteration, KRAS G12X (X is any amino acid), NRAS Q61R, PDGFRA copy number alteration, and RET fusion, e.g., the KRAS G12X mutation is selected from the group consisting of KRAS G12D, KRAS G12E, KRAS G12F, KRAS G12G, KRAS G12H ... KRAS G12A, KRAS G12C and KRAS G12V, (B) the mutation in PIK3CA comprises PIK3CA E545K; The diagnostic kit according to claim 25, which satisfies any one of the above.

27. The diagnostic kit of claim 25, wherein the one or more mutations are further selected from mutations in one or more genes from the WNT / b-catenin pathway, optionally wherein the one or more genes from the WNT / b-catenin pathway are APC and CTNNB1, e.g., the mutations in one or more genes from the WNT / b-catenin pathway include APC Q1469, APC R405, APC S713, CTNNB1 S33P, CTNNB1 S37C, CTNNB1 S37F, and CTNNB1 S45P.

28. The following (A) to (B): (A) the tumor DNA is circulating tumor DNA (ctDNA), and optionally the ctDNA is present in a biological sample isolated from the subject, e.g., the biological sample is a blood sample or a plasma sample; (B) the tumor DNA is present in a tumor sample isolated from the subject. The diagnostic kit according to claim 25, which satisfies any one of the above.

29. 26. The diagnostic kit of claim 25, further comprising one or more reagents for purifying the tumor DNA from the biological sample from the subject.

30. The following (A) to (B): (A) the one or more reagents can be used in conjunction with sequencing techniques to determine the one or more mutations; (B) the one or more reagents can be used in conjunction with next generation sequencing (NGS) to determine the one or more mutations; The diagnostic kit according to any one of claims 25 to 29, which satisfies any one of the above.

31. A diagnostic kit comprising: (i) one or more reagents for determining the expression level of EGFR and / or MET in a tumor sample from a subject having cancer; and (ii) optionally, packaging and / or instructions for use.

32. 32. The diagnostic kit of claim 31, wherein the one or more reagents can be used in conjunction with immunohistochemistry (IHC) to determine the expression levels of EGFR and / or MET.

33. A pharmaceutical composition for use in the treatment of cancer, comprising a bispecific anti-epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) bispecific antibody and a pharma- ceutical acceptable carrier, It is used in combination with EGFR tyrosine kinase inhibitors (TKIs), the combination therapy is administered in the absence of one or more mutations in tumor DNA obtained from a subject in need of cancer treatment; The pharmaceutical composition, wherein the one or more mutations are selected from one or more genes from the RAS / RAF / MEK pathway and a mutation in PIK3CA.

34. Use of a bispecific anti-epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) bispecific antibody in the manufacture of a medicament for use in the treatment of cancer, comprising: It is used in combination with EGFR tyrosine kinase inhibitors (TKIs), the combination therapy is administered in the absence of one or more mutations in tumor DNA obtained from a subject in need of cancer treatment; The one or more mutations are selected from one or more genes from the RAS / RAF / MEK pathway and a mutation in PIK3CA.

Images