Methods of treating non-small cell lung cancer using mesenchymal epithelial transformation factor (MET) targeting agents

Abstract

The present disclosure provides methods for treating MET-associated non-small cell lung cancer or inhibiting the growth of MET-associated non-small cell lung cancer comprising selecting a subject having cancer and administering a therapeutically effective amount of a MET x MET bispecific antibody wherein each binding arm of the bispecific antibody interacts with a different MET epitope. In certain embodiments, the cancer contains an MET alteration, such as an exon 14 alteration in DNA or a deletion that results in exon 14 skipping, MET gene amplification, or MET protein overexpression.

Description

Technical Field

[0001] The present invention relates to a MET×MET bispecific antibody that specifically binds to hepatocyte growth factor receptor (c-Met or MET) and regulates MET signal transduction, and is used for treating non-small cell lung cancer (NSCLC). Sequence Listing

[0002] An official copy of the sequence listing in XML format, named 11305WO01_Sequence Listing ST.26, with a creation date of August 31, 2023, is submitted electronically through the EFS website at the same time as the specification, and the size of the sequence listing is about 32,768 bytes. The sequence listing is part of the specification, and its entire content is incorporated herein by reference. Background Art

[0003] Hepatocyte growth factor (HGF) (also known as scatter factor [SF]) is a heterodimeric paracrine growth factor that exerts its activity by interacting with the HGF receptor (HGFR). HGFR is the product of the c-Met oncogene and is also known as MET. MET is a receptor tyrosine kinase composed of a transmembrane β chain connected to an extracellular α chain by a disulfide bond. The binding of HGF to MET activates the kinase catalytic activity of MET, leading to phosphorylation of Tyr 1234 and Tyr 1235 of the β chain and subsequent activation of downstream signaling pathways.

[0004] It has been shown that MET and / or HGF overexpression, activation or amplification are involved in non-small cell lung cancer (NSCLC) (Sierra and Tsao, Ther. Adv. Med. Oncol., 3(1 Suppl): S21-S35, 2011). MET amplification is considered a key driver of NSCLC tumorigenesis. In addition, mutations that lead to MET exon 14 deletions are considered to be oncogenic drivers of NSCLC subtypes. The growth and survival of tumor cell lines with MET gene amplification are highly dependent on MET. Preclinical data indicate that MET signaling resists targeted therapy in NSCLC.

[0005] Both preclinical and recent clinical results show that tumors containing these genetic alterations respond to MET inhibitors, indicating that MET is a cancer driver. A variety of monovalent MET blocking antibodies are in clinical development for the treatment of a variety of cancers (see U.S. Patent Nos. 5,686,292, 5,646,036, 6,099,841, 7,476,724, 9,260,531, and 9,328,173; and U.S. Patent Application Publication Nos. 2014 / 0349310 and 2005 / 0233960). These antibodies include onartuzumab (MetMab) and emibetuzumab (Xiang et al., Clin. Cancer Res. 19 (18): 5068-78, 2013, and Rosen et al., Clin. Cancer Res., published on October 10, 2016, doi: 10.1158 / 1078-0432.CCR-16-1418). Some of these antibodies block ligand-dependent MET signaling, but are not as effective in blocking ligand-independent MET activation.

[0006] There remains a significant unmet medical need for improved anticancer drugs that effectively block both ligand-dependent and ligand-independent MET signaling. Summary of the invention

[0007] The following methods are provided herein: using bispecific antibodies and fragments thereof, which bind to human c-Met receptor protein (MET×MET) for targeting non-small cell lung cancer (NSCLC) cells with MET changes, such as exon 14 changes in DNA or deletions that lead to exon 14 skipping, MET gene amplification, or MET protein overexpression. Patients with NSCLC can be treated according to acceptable standards of care, but may eventually exhaust all approved treatments available for patients because the tumor becomes resistant to treatment. Resistance to MET treatments (e.g., chemotherapy, immune checkpoint inhibitors, or targeted therapy) can be inherent or acquired.

[0008] By identifying those patients with tumors containing MET alterations and treating those patients with a MET×MET bispecific antibody, the unexpected effects as described herein were achieved.

[0009] Thus, provided herein are methods of treating NSCLC, reducing NSCLC tumor growth, and / or causing regression of NSCLC in a subject having a tumor containing a MET alteration, the method comprising administering to the subject a dose of 250 to 2000 mg of a MET×MET bispecific antibody.

[0010] Also provided herein is a method for treating NSCLC or inhibiting the growth of NSCLC, the method comprising: (1) selecting a subject having a tumor containing a MET alteration; and (2) administering to the subject (a) about 250 mg, 500 mg, 750 mg, 1000 mg, 1500 mg or 2000 mg of a MET×MET bispecific antibody. In some aspects, the administration of step (2) is performed once every 3 weeks.

[0011] Also provided herein is a method for treating a tumor, the method comprising: (a) selecting a subject having NSCLC; (b) determining that the tumor exhibits a MET alteration selected from the following: an exon 14 alteration in DNA or a deletion resulting in exon 14 skipping, MET gene amplification, and / or Met protein overexpression, comprising (i) obtaining a tissue sample and / or a liquid sample from the subject; and (ii) assessing MET gene amplification in the tissue sample using fluorescent in situ hybridization of the tumor tissue or by next-generation sequencing of the tumor tissue and / or ctDNA, and / or assessing Met protein overexpression in the tissue sample using immunohistochemistry, and / or assessing exon 14 mutations in the liquid sample using ctDNA; and, if the tumor exhibits a MET alteration, (c) administering one or more doses of a MET×MET bispecific antibody to the subject in need thereof.

[0012] Also provided herein is a method for identifying a candidate for a Met×Met anti-tumor therapy, the method comprising: obtaining a tissue sample and / or a fluid sample from a subject suffering from NSCLC; and assessing the tissue sample and / or the fluid sample for MET alterations selected from exon 14 alterations in DNA or deletions resulting in exon 14 skipping, MET gene amplification, and / or Met protein overexpression, wherein the presence of at least one Met alteration in the tissue sample or the fluid sample identifies the subject as a candidate for the anti-tumor therapy, wherein the MET×MET anti-tumor therapy comprises a MET×MET bispecific antibody.

[0013] MET alterations provided herein include exon 14 alterations in the DNA, MET gene amplification, or MET protein overexpression.

[0014] In some aspects, the MET change is an exon 14 change in the DNA or a deletion that causes exon 14 skipping. Exon 14 changes in the DNA or deletions that cause exon 14 skipping include missense changes, deletions, splice site changes, and whole exon deletions that cause exon 14 skipping of the MET gene. In some aspects, the MET change is an exon 14 mutation. In some embodiments, exon 14 mutations may include, but are not limited to, D1010N, D1010fs*19, D1010Y, D1010H, or R1004P.

[0015] In some aspects, the MET alteration is a MET gene amplification. In certain aspects, the highly amplified MET gene has a MET gene copy number (GCN) ≥ 5 in the tissue and / or a ratio of MET to chromosome 7 centromere (MET / CEP7) ≥ 2 by FISH, or a METGCN ≥ 6 by next generation sequencing (NGS), or a MET fold change ≥ 2 in ctDNA.

[0016] In some aspects, MET changes are MET protein overexpression. MET protein overexpression can be assessed by MET immunohistochemistry (IHC) expression in tumor tissues higher than that in normal tissues. In some aspects, elevated MET protein expression is measured as IHC ≥ 2+ or H score> 150. In some aspects, highly overexpressed MET proteins are measured as IHC 3+ or H score ≥ 200.

[0017] In some aspects, MET alterations are identified using ctDNA from a blood sample obtained from the patient prior to treatment (ie, a liquid biopsy).

[0018] In some aspects, MET alterations are identified in a tissue sample, ie, a tumor biopsy, obtained from the patient prior to treatment.

[0019] In some aspects, the NSCLC tumor has an EGFR mutation. The mutation may include, but is not limited to, L858R, G719S, E709A, E746_A750del, and S752_I759del. In some aspects, the subject is selected to have NSCLC with one or more mutations in the EGFR gene.

[0020] In some embodiments, the subject has not received prior anti-MET cancer therapy. In some embodiments, the subject has not received prior anti-MET cancer therapy. Tyrosine kinase inhibitor (TKI). In other words, the subject has not received previous treatment with a TKI targeting MET. In some embodiments, the subject has received previous anticancer treatments including one or more of the following: TKI targeting MET, PD-1 inhibitors, EGFR inhibitors, PD-L1 inhibitors, surgery, radiotherapy, or chemotherapy. In some aspects, the previous anticancer treatment includes a TKI targeting MET. In some aspects, the previous anticancer treatment includes a PD-1 inhibitor or a PD-L1 inhibitor. In some aspects, the previous anticancer treatment includes an EGFR inhibitor. In some aspects, the subject is resistant or insufficiently responsive to the previous treatment, or relapses after the previous treatment.

[0021] Patients who have not received MET TKIs have not been previously treated with any MET TKIs; similarly, patients who have received MET TKIs are MET TKI-experienced. Patients who have PD-(L)1 experience have previously been treated with PD-1 inhibitors (e.g., but not limited to, pembrolizumab, nivolumab, cemiplimab, dostarlimab, and retifanlimab) or PD-L1 inhibitors (e.g., but not limited to, atezolizumab, avelumab, and durvalumab) and are considered to be PD-(L)1-experienced; similarly, patients who do not have PD-(L)1 experience (i.e., have not received PD-(L)1) have not been previously treated with PD-1 or PD-L1 inhibitors. Patients may be those who have been treated with EGFR inhibitors, i.e., have been treated with EGFR inhibitors. Exemplary EGFR inhibitors include, but are not limited to, erlotinib, afatinib, gefitinib, osimertinib, dacomitinib, cetuximab, panitumumab, dacomitinib, etc.

[0022] In some embodiments, subjects are further selected based on one or more of the following criteria: (i) Subjects were MET TKI-naive; (ii) subjects had histologically confirmed NSCLC; (iii) NSCLC showing MET exon 14 alterations or deletions leading to exon 14 skipping in DNA; (iv) NSCLC showed MET gene amplification; (v) NSCLC showed elevated MET protein expression (IHC ≥2+ or H score >150); (vi) NSCLC showed MET exon 14 alterations or deletions leading to exon 14 skipping in DNA and was treated with MET TKI; (vii) NSCLC showing MET exon 14 alterations or deletions leading to exon 14 skipping in DNA and are MET TKI naive; (viii) NSCLC showed high MET gene amplification (MET GCN ≥ 5 and / or MET / CEP7 ratio ≥ 2 by FISH in tissue, or MET GCN ≥ 6 by NGS, or MET fold change ≥ 2 in ctDNA) and had not received MET TKI; (ix) NSCLC showing high MET protein overexpression (IHC 3+ or H score ≥200) and not treated with METKI; and / or (x) NSCLC showed high MET gene amplification (MET GCN ≥5 and / or MET / CEP7 ratio ≥2 by FISH in tissue, or MET GCN ≥6 by NGS, or MET fold change ≥2 in ctDNA), high MET protein overexpression (IHC 3+ or H score ≥200), and had not received MET TKI.

[0023] In some embodiments, the cancer is non-squamous NSCLC. In some embodiments, the cancer is NSCLC squamous carcinoma. In some aspects, the NSCLC is metastatic, for example, the cancer has metastasized to the brain and / or liver. In certain aspects, the NSCLC is unresectable.

[0024] Provided herein are methods for monitoring the efficacy of treatment with a MET×MET bispecific antibody in a subject having NSCLC with a MET alteration, the method comprising: (i) obtaining a tissue sample and / or a fluid sample from the subject, and assessing somatic mutations in one or more genes in the tissue sample and / or the fluid sample, the somatic mutations being selected from: ●a. On-target MET receptor gene mutations that confer resistance to MET TKIs, which are present in MET Ex14Mut patients with prior TKI Exp and MET gene silencing (loss of function); b. TK driven receptor activation selected from alternatively or concurrently TK receptor and ligand gene amplification and TKR activating mutations; and c. Activating gene mutations in a pathway selected from the group consisting of the JAK2 / STAT3 pathway, the RAS / RAF / MEK / MAPK pathway, the PI3K / AKT / MTOR pathway, TP53 mutations, and cell cycle gene amplifications; (ii) administering a MET×MET bispecific antibody to the subject; and (iii) repeating steps (i) and (ii) during the treatment; wherein the acquisition of one or more gene mutations indicates resistance to treatment and / or indicates a poor prognosis.

[0025] In some aspects, the on-target MET receptor gene mutation is selected from MET Y1230C, MET D1228H, MET D1228N; and MET gene silencing (loss of function) is selected from somatic mutations of DNMT3A and TET2.

[0026] In some aspects, the TKR activating mutation is selected from the group consisting of: EGFR L858R, EGFR G719S, EGFR E709A, EGFR E746_A750del, and EGFR S752_I759del.

[0027] In some aspects, the JAK2 / STAT3 pathway mutation is JAK2 V617F; the RAS / RAF / MEK / MAPK pathway mutation is selected from the group consisting of KRAS G12A / V, GNAS R201H, MKRN-BRAF fusion, BRAF S602Y, RICTOR Amp, and MAP2K1 K57N; the PI3K / AKT / MTOR pathway mutation is selected from the group consisting of PIK3CA H1047L, PIK3CA E545K, PIK3CA E542K, PIK3CAN345K, IDH1 R132L, and MTOR E2338Q; the PI3K / AKT / MTOR pathway amplification is selected from the group consisting of AKT2 Amp and RICTOR Amp; the TP53 mutation is selected from the group consisting of TP53 R280T and TP53 R248Q; and the cell cycle gene amplification is selected from the group consisting of CDK4Amp, CDK6Amp, CCND1 Amp, and CCNE1 Amp.

[0028] In some aspects, the patient has confirmed MET amplification and MET overexpression, but gene amplification occurs in one or more of HGF, EPH, EGFR, BRAF, BCL2L1, PI3KCB, KRAS, AKT2, ATR, VEFGA, FGF, CCND, CCNE, CDK6, RAD21 and MYC. In some aspects, the patient has confirmed MET amplification and MET overexpression, but gene deletion occurs in one or more of CDKN2A, CDKN2B, MTAP and RBM10. In some aspects, the patient has confirmed MET amplification and MET overexpression, but one or more point mutations provided in Table 15 occur.

[0029] In some aspects, the patient has confirmed MET amplification (and non-MET overexpression) and can be EGFR mutant, but in EGFR, BRAF, PI3KC2G, KRAS, HGF, EPHA3, ERCC4, RICTOR, RAD21, LYN, MYC, MDM2, CDK4 / 6, FgF3 / 4 / 19, FGF10 and CCND1, one or more gene amplification occurs. In some aspects, the patient has confirmed MET amplification (and no MET overexpression), and can be EGFR mutant, but in CDKN2A, CDKN2B, MTAP, TEK and BCOR, one or more gene deletion occurs. In some aspects, the patient has confirmed MET amplification (and no MET overexpression), and can be EGFR mutant, but one or more point mutations provided in Table 16 occur.

[0030] In some aspects, the patient has confirmed MET exon 14 changes and has not received TKI, but gene amplification occurs in one or more of MDM2, EGFR, FGFR1, ERBB3, CDK4, GNA13, MYC, RPTOR, TERC, IKZF1, EZH2, SDHA, SOX, WHSC1L1 and ZNF703. In some aspects, the patient has confirmed MET exon 14 changes and has not received TKI, but gene deletion occurs in one or more of CDKN2A, CDKN3A and MTAP. In some aspects, the patient has confirmed MET exon 14 changes and has not received TKI, but one or more point mutations provided in Table 17 have occurred.

[0031] In some aspects, the patient has confirmed MET exon 14 changes and has undergone TKI, but gene amplification occurs in one or more of EGFR, RAF1, PI3KC2G, CDK4, CEBPA, CDKN1A, CARD11, MYC, RICTOR, VEGFA, CD22, DDR1, RAC1, NBN, FGF19, MDM2, NFKBIA, CCND1, INPP4B, PPARG, PMS2, GATA4, SDHA and RAD21. In some aspects, the patient has confirmed MET exon 14 changes and has undergone TKI, but gene deletion occurs in one or more of CDKN2A, CDKN3A and MTAP. In some aspects, the patient has confirmed MET exon 14 changes and has undergone TKI, but one or more point mutations provided in Table 18 have occurred.

[0032] Also provided herein is a method for treating NSCLC in a subject, the method comprising: (i) obtaining a liquid sample from the subject and determining MET amplification in ctDNA from the liquid sample, and (ii) administering a MET×MET bispecific antibody to the subject; wherein steps (i) and (ii) are repeated every three weeks, and wherein loss of MET amplification after repeating step (ii) indicates a durable response to treatment.

[0033] Provided herein is a method for determining a therapeutically effective amount of a MET×MET bispecific antibody, the method comprising: (i) administering a dose of the bispecific antibody to a patient in need thereof, and (ii) measuring soluble MET in a blood sample, wherein a maximum increase in soluble MET (sMET) indicates receptor occupancy saturation and a therapeutically effective amount of the bispecific antibody.

[0034] In some aspects, the subject is administered an amount of the MET×MET bispecific antibody of about 250 mg to about 5000 mg, or about 250 mg, 300 mg, 400 mg, 500 mg, 600 mg, 700 mg, 800 mg, 900 mg, 1000 mg, 1100 mg, 1200 mg, 1300 mg, 1400 mg, 1500 mg, 1600 mg, 1700 mg, 1800 mg, 1900 mg, 2000 mg, 2100 mg, 2200 mg, 2300 mg, 2400 mg, 2500 mg, 3000 mg, 3500 mg, 4000 mg, 4500 mg, or 5000 mg. In some aspects, the dose is about 250 mg of the MET×MET bispecific antibody. In some aspects, the dosage is about 500 mg MET×MET bispecific antibody. In some aspects, the dosage is about 1000 mg MET×MET bispecific antibody. In some aspects, the dosage is about 2000 mg MET×MET bispecific antibody. In some aspects, the dosage is 250 mg, 500 mg, 1000 mg, or 2000 mg MET×MET bispecific antibody.

[0035] In some aspects, the bispecific antibody is administered intravenously, subcutaneously or intraperitoneally. The bispecific antibody can be administered once every five days, once a week, once every two weeks, once every three weeks, once every four weeks, once a month, once every five weeks, once every six weeks, or once every two months. In some aspects, the bispecific antibody is administered once every three weeks. In some aspects, the bispecific antibody is administered three weeks after the previous dose.

[0036] In some aspects, treatment produces a therapeutic effect selected from the group consisting of tumor growth delay, metastasis reduction, tumor cell number reduction, tumor regression, survival improvement, partial response, and complete response. In some aspects, tumor growth is delayed by at least 10 days, or at least 20 days, or at least 30 days, or at least 40 days, or at least 50 days compared to an untreated subject. In some aspects, tumor growth is inhibited by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, or at least 60% compared to an untreated subject.

[0037] The MET×MET bispecific antibody comprises a first antigen-binding domain (D1) comprising three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) within a heavy chain variable region (HCVR) comprising an amino acid sequence at least 95% identical to the sequence of SEQ ID NO: 1, and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) within a light chain variable region (LCVR), wherein the heavy chain variable region (HCVR) comprises an amino acid sequence at least 95% identical to the sequence of SEQ ID NO: 9, and the light chain variable region (LCVR) comprises an amino acid sequence at least 95% identical to the sequence of SEQ ID NO: 9; and a second antigen-binding domain (D2) comprising three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) within a heavy chain variable region (HCVR) comprising an amino acid sequence at least 95% identical to the sequence of SEQ ID NO: 5, and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) within a light chain variable region (LCVR), wherein the heavy chain variable region (HCVR) comprises an amino acid sequence at least 95% identical to the sequence of SEQ ID NO: 5, and the light chain variable region (LCVR) comprises an amino acid sequence at least 95% identical to the sequence of SEQ ID NO: 6. The sequence of ID NO: 9 has an amino acid sequence that is at least 95% identical.

[0038] In some aspects, D1 specifically binds to a first epitope of human MET.

[0039] In some embodiments, D1 comprises the HCDR1 amino acid sequence shown in SEQ ID NO:2; the HCDR2 amino acid sequence shown in SEQ ID NO:3; the HCDR3 amino acid sequence shown in SEQ ID NO:4; the LCDR1 amino acid sequence shown in SEQ ID NO:10; the LCDR2 amino acid sequence shown in SEQ ID NO:11; and the LCDR3 amino acid sequence shown in SEQ ID NO:12.

[0040] In some embodiments, D1 comprises a HCVR comprising the amino acid sequence of SEQ ID NO:1; and a LCVR comprising the amino acid sequence of SEQ ID NO:9.

[0041] In some aspects, D2 specifically binds to a second epitope of human MET.

[0042] In some embodiments, D2 comprises the HCDR1 amino acid sequence shown in SEQ ID NO:6; the HCDR2 amino acid sequence shown in SEQ ID NO:7; the HCDR3 amino acid sequence shown in SEQ ID NO:8; the LCDR1 amino acid sequence shown in SEQ ID NO:10; the LCDR2 amino acid sequence shown in SEQ ID NO:11; and the LCDR3 amino acid sequence shown in SEQ ID NO:12.

[0043] In some embodiments, D2 comprises a HCVR comprising the amino acid sequence of SEQ ID NO:5; and a LCVR comprising the amino acid sequence of SEQ ID NO:9.

[0044] In some embodiments, the MET change is an exon 14 change in the DNA or a deletion that results in exon 14 skipping, MET gene amplification, or MET protein overexpression. In some aspects, the MET change is an exon 14 change in the DNA or a deletion that results in exon 14 skipping. In some aspects, the MET change is a MET gene amplification. In some aspects, the MET change is MET protein overexpression.

[0045] Other embodiments will become apparent by review of the ensuing detailed description. BRIEF DESCRIPTION OF THE DRAWINGS

[0046] The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.

[0047] Figure 1A and 1B Included is a bar graph depicting the relative growth of EBC-1 cells as a function of treatment with control antibody and MET×MET bispecific antibody at 0.1 μg / mL and 1 μg / mL.

[0048] Figure 2 Depicted are immunoblots of MET (and tubulin as a loading control) expression in Hs746T cells after treatment with a control antibody and a MET×MET bispecific antibody for 2, 6, and 18 hours.

[0049] Figure 3 Depicted are immunoblots of pMET, MET, pErk, and tubulin (for loading control) extracted from EBC-1 cells after treatment with a control antibody and a MET×MET bispecific antibody.

[0050] Figure 4 is a line graph depicting the change in EBC-1 tumor volume (in cubic millimeters) as a function of time (in days) after implantation of EBC-1 cells in animals treated with control antibody (solid squares ■), MET monovalent antibody (solid circles ●), or MET×MET bispecific antibody (solid diamonds ◆).

[0051] Figure 5is a bar graph depicting relative cell growth of NCI-H596 cells as a function of treatment with control antibody (C), MET×MET bispecific antibody (MM), MET×MET parental monospecific antibody 1 (M1), MET×MET parental monospecific antibody 2 (M2), a combination of parental antibodies 1 and 2 (M1M2), and the MET agonist hepatocyte growth factor (HGF).

[0052] Figure 6 Depicts a study flow chart of the clinical trial using REGN5093.

[0053] Figure 7 is a schematic diagram showing the study design for dose escalation and expansion cohorts.

[0054] Figure 8 Tumor responses of patients characterized by centrally confirmed MET alterations are depicted.

[0055] Fig. 9A and 9B Depicts the effect of dose escalation on Fig. 9A ) and multiple extended groups ( Fig. 9B ) Serum concentrations after the first dose of REGN5093 in . The elimination half-life estimated by non-compartmental analysis was 15 days over a 3-week dosing interval.

[0056] Fig.10 Depicted is the interaction of the MET×MET bispecific antibody with the MET extracellular domain. The figure also shows how MET alterations affect MET signaling and the enhancement of ligand-independent activation.

[0057] Fig.11 Methods for confirming MET alterations in ctDNA and tissues, as well as exemplary commercial products that can be used for these methods are depicted. (CEP7, chromosome 7 centromere; ELISA, enzyme-linked immunosorbent assay; FFPE, formalin-fixed paraffin-embedded; FISH, fluorescent in situ hybridization; FMI, Foundation Medicine Inc.; GCN, gene copy number; HGF, hepatocyte growth factor; IHC, immunohistochemistry; MET, mesenchymal epithelial transition; sMET, soluble MET; TSO, Trusight Oncology.)

[0058] Fig.12The number of patients enrolled in the dose escalation study and the number and characteristics of patients enrolled in the expansion cohort are illustrated. (aNSCLC, advanced non-small cell lung cancer; GCN, gene copy number; FIH, first-in-human; IHC, immunohistochemistry; IV, intravenous; MET, mesenchymal epithelial transition; PK, pharmacokinetics; Q3W, every 3 weeks; Q6W, every 6 weeks; TKI, tyrosine kinase inhibitor.)

[0059] Fig.13 Response data for the 2000 mg dose escalation study and expansion cohorts at the 2000 mg dose are provided. The overall response rate was highest in the cohort of patients with exon 14 skipping mutations and in which the patients had not received prior MET TKIs, and in the cohort of patients with both MET gene amplification and MET protein overexpression and in which the patients had not received prior MET TKIs.

[0060] Fig.14 Tumor responses with confirmed MET alterations are shown. MET overexpression-IHC: ≥75% tumor cells w / 3+ membrane staining; MET amplification-FISH: GCN≥5 or MET / CEP7>2 (tissue); or NGS: GCN≥6 (tissue); or ≥2.2-fold (ctDNA); MET exon 14 mutation-NGS, in tissue or ctDNA. Responses to REGN5093 were observed in aNSCLC patients who had not received MET TKI (2L+) and who had MET exon 14 mutations or MET amplification and overexpression regardless of EGFR mutation status. For patients whose tumor tissues were analyzed for MET amplification using both platforms, the MET GCN by NGS was significantly higher than the MET GCN by FISH (p=0.0015). (aNSCLC, advanced non-small-cell lung cancer; CEP7, chromosome 7 centromere; FISH, fluorescence in situ hybridization; GCN, gene copy number; IHC, immunohistochemistry; MET, mesenchymal-epithelial transition; NA, not available or to be determined; NGS, next-generation sequencing; PD, progressive disease; PR, partial response; SD, stable disease; TKI, tyrosine kinase inhibitor.)

[0061] Fig.15Responses in subgroups with specific MET alterations (regardless of the presence of other MET alterations) relative to the total population are provided. Results are based on small sample sizes and require prospective validation. (CR, complete response; DCR, disease control rate; MET, mesenchymal epithelial transition; ORR, objective response rate; PR, partial response; SD, stable disease; TKI, tyrosine kinase inhibitor.)

[0062] Fig.16 Figure 2 is a graph of baseline demographic and clinical characteristics of the bypass resistance mutation study population. The median age of the study population was 66 years, 53.8% were male, 70.5% were Asian, and patients had received a median of 2.5 prior lines of therapy. (ECOG, Eastern Cooperative Oncology Group; EGFR, epidermal growth factor receptor; PS, performance status.)

[0063] Fig.17 Depicts the differences in somatic variants detected in tissue and ctDNA. Tumor profiling at baseline in 2L+aNSCLC identified somatic variants with known functional significance. The Venn diagram shows the total number of baseline somatic variants identified by NGS using the FMI-Dx (tissue; n = 51 patients) and FMI-L (ctDNA; n = 38 patients) groups (324 genes); genes are grouped by type of alteration and clinical sample type (ctDNA, tissue, or both). CNVs (including gene amplification and deletion) are more easily detected in tissues, while more NSVs (single nucleotide variants, splicing variants, and gene fusions) are detected in ctDNA. Therefore, bypass gene detection of NSVs and CNVs in ctDNA can, in some embodiments, supplement tissue results to provide a more comprehensive tumor profiling of Met×Met resistance mechanisms. (2L, second-line; aNSCLC, advanced non-small-cell lung cancer; CNV, copy number variation; FMI, Foundation Medicine, Inc.; NGS, next-generation sequencing; NSV, non-synonymous variation; SNV, single nucleotide variation.)

[0064] Fig.18Unbiased clustering of baseline somatic variants with confirmed MET alterations is illustrated. Several genes of known functional significance were detected and clustered by cohort assignment, centrally confirming MET alterations and EGFR status. (Amp, amplification; EGFR, epidermal growth factor receptor; Ex14, exon 14; FMI, Foundation Medicine; IHC, immunohistochemistry; MET, N, no; NA, not available; mesenchymal-epithelial transition; O / E, overexpression; SNV, single nucleotide variant; TKI, tyrosine kinase inhibitor; Y, yes.)

[0065] Fig.19 Unbiased clustering of baseline somatic variants with MET alterations and clinical response is illustrated. Several genes with known functional significance were detected and clustered by cohort assignment, confirming MET alterations, EGFR status, and clinical response.

[0066] Fig. 20 We describe a classification and examples of baseline somatic mutations identified by tumor profiling. These mutations were identified in nonresponders and can cause MET bypass resistance mechanisms and potentially impact clinical response to REGN5093, even in the presence of MET oncogenic drivers. (Amp, amplification; EGFR, epidermal growth factor receptor; Ex14, exon 14; GOF, gain of function; LOF, loss of function; TKI, tyrosine kinase inhibitor).

[0067] Fig.21 The total REGN5093 and sMET concentrations during the study treatment period, as well as the individual concentrations of sMET over time at the best overall response, are graphically represented. The total concentration of REGN5093 was several times higher than the total sMET concentration in serum, indicating that saturation of receptor occupancy was achieved with the 2000 mg Q3W dosing regimen. sMET and cHGF levels increased after dosing, indicating target engagement, but neither baseline nor post-treatment levels of sMET and cHGF were significantly correlated with response (data not shown). (cHGF, circulating hepatocyte growth factor; Q3W, every 3 weeks; sMET, soluble mesenchymal epithelial transition)

[0068] Fig. 22 The lack of significant association between baseline concentrations of sMET or cHGF and clinical response is illustrated. cHGF, circulating hepatocyte growth factor; PD, progressive disease; PR, partial response; SD, stable disease; sMET, soluble mesenchymal-epithelial transition. DETAILED DESCRIPTION

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

[0070] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as those generally understood by those of ordinary skill in the art to which the present invention belongs. When used with reference to a specific listed value, the term "about" as used herein means that the value may differ from the listed value by no more than 1%. For example, the expression "about 100" as used herein includes 99 and 101 and all values ​​therebetween (e.g., 99.1, 99.2, 99.3, 99.4, etc., including 100).

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

[0072] As used herein, the expressions "MET", "c-Met" and the like refer to a human transmembrane receptor tyrosine kinase comprising: (1) the amino acid sequence set forth in SEQ ID NO:13, and / or the amino acid sequence set forth in NCBI Accession No. NM_001127500.2, which represents an unprocessed proprotein of isoform "a", (2) the amino acid sequence set forth in SEQ ID NO:14, and / or the amino acid sequence set forth in NCBI Accession No. NM_000236.2, which represents an unprocessed proprotein of isoform "b", (3) the amino acid sequence set forth in SEQ ID NO:15, and / or the amino acid sequence set forth in NCBI Accession No. NM_001311330.1, which represents an unprocessed proprotein of isoform "c", and / or (3) a cytoplasmic α subunit (SEQ ID NO:16) and a transmembrane β subunit (isoforms a, b and c are SEQ ID NOs:17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 67, 68, 70, 71, 72, 73, 74, 75, 76 NO: 17, 18 or 19). The expression "MET" includes both monomeric MET molecules and multimeric MET molecules. The expression "monomeric human MET" used herein means a MET protein or a portion thereof that does not contain or have any multimerization domain, and that exists as a single MET molecule under normal conditions without direct physical connection to another MET molecule. An exemplary monomeric MET molecule is a molecule referred to herein as "hMET.mmh", which comprises the amino acid sequence of SEQ ID NO: 20 (SEQ ID NO: 152 from Example 3 of U.S. Patent No. 11,142,578). The expression "dimeric human MET" used herein means a construct comprising two MET molecules connected to each other by a linker, a covalent bond, a non-covalent bond, or by a multimerization domain (e.g., an antibody Fc domain). An exemplary dimeric MET molecule is the molecule referred to herein as "hMET.mFc," which comprises the amino acid sequence of SEQ ID NO: 21 (SEQ ID NO: 153 from US Pat. No. 11,142,578 - Example 3).

[0073] Unless explicitly indicated as being from a non-human species, all references to proteins, polypeptides and protein fragments herein are intended to refer to the human versions of the corresponding proteins, polypeptides or protein fragments. Thus, unless indicated as being from a non-human species, e.g., "mouse MET", "monkey MET", etc., the expression "MET" means human MET.

[0074] The expression "cell surface expressed MET" used herein means one or more MET proteins or their extracellular domains expressed on the cell surface in vitro or in vivo such that at least a portion of the MET protein is exposed on the extracellular side of the cell membrane and accessible to the antigen binding portion of the antibody. "Cell surface expressed MET" may include or consist of a MET protein expressed on the surface of a cell that normally expresses a MET protein. Alternatively, "cell surface expressed MET" may include or consist of a MET protein expressed on the surface of a cell that normally does not express human MET on its surface but has been artificially modified to express MET on its surface.

[0075] Measures of MET alterations in NSCLC include: MET exon 14 mutation, an oncogenic driver for NSCLC with recent TKI approval in 1L (first-line); MET gene amplification: the major resistance mechanism to EGFR TKI therapy in 2L (second-line) + NSCLC; and MET protein overexpression, which has been reported to enhance but not select for treatment response to MET TKIs in NSCLC. Treatments for NSCLC cancer

[0076] Lung cancer is one of the most commonly diagnosed cancers and is the leading cause of cancer-related mortality worldwide (Siegel et al., CA Cancer J Clin, 66(1):7-30, 2016). Non-small cell lung cancer (NSCLC) accounts for 80% to 85% of all lung cancers and is composed of several histopathological subtypes, the most common of which are adenocarcinoma (40% to 60%) and squamous cell carcinoma (30%) (Dela Cruz et al., Clin Chest Med, 32(4):605-44, 2011). Most patients with NSCLC are found to have advanced cancer at the time of diagnosis.

[0077] Anti-PD-1 and anti-PD-L1 therapy has changed the standard of care for many patients with NSCLC (Topalian et al., NEJM, 366(26):3443-54, 2012). However, emerging data suggest that patients with MET-driven NSCLC may not experience equivalent benefits with agents targeting the PD-1 / PD-L1 axis, even if their tumors highly express PD-L1 or exhibit high tumor mutation burden (TMB) (Sabari et al., Ann Oncol, 29(10):2085-91, 2018). This is consistent with data generated in lung cancers containing EGFR mutations or ALK rearrangements (Garassino et al., Lancet Oncol, 19(4):521-36, 2018) (Lee et al., JAMA Oncol, 4(2):210-16 2018) (Peters et al., J Clin Oncol, 35(24):2781-9, 2017), and suggests that monotherapy with anti-PD-1 or anti-PD-L1 may not be the preferred treatment for patients with MET-driven disease. Therefore, there remains a large unmet need for treatments that improve response rates and survival in patients with MET-altered NSCLC.

[0078] The inventors unexpectedly discovered that certain MET×MET bispecific antibodies are extremely useful for treating and / or inhibiting NSCLC or slowing down NSCLC metastasis associated with MET alterations, such as exon 14 alterations in DNA or deletions leading to exon 14 skipping, MET gene amplification, or MET protein overexpression. REGN5093 is an exemplary human bispecific antibody that binds to two epitopes on the MET receptor, resulting in blockade of ligand-dependent and ligand-independent signaling with potential activity in lung cancer. The examples below illustrate that tumor response to treatment with MET×MET bispecific antibodies is enhanced by identifying patients with these MET alterations.

[0079] Thus, useful according to the methods described herein are MET×MET bispecific antibodies comprising a first antigen binding domain (also referred to herein as “D1”) that specifically binds to a first epitope of human MET and a second antigen binding domain (also referred to herein as “D2”) that specifically binds to a second epitope of human MET. The simultaneous binding of the bispecific antibody to two separate MET epitopes results in effective ligand blocking and minimal activation of MET signaling. Such MET×MET bispecific antibodies are described in U.S. Publication No. 2018 / 0134794, which is incorporated herein by reference in its entirety.

[0080] The MET×MET bispecific antibody is particularly useful for treating, preventing and / or ameliorating NSCLC associated with or mediated by MET expression, signaling or activity, or can be treated by blocking the interaction between MET and HGF, or otherwise inhibiting MET activity and / or signaling, and / or promoting receptor internalization and / or reducing the number of cell surface receptors. The MET×MET bispecific antibody is particularly useful for treating and / or ameliorating NSCLC in subjects with tumors containing MET alterations, such as exon 14 alterations in DNA or deletions that result in exon 14 skipping, MET gene amplification, or MET protein overexpression. The MET×MET bispecific antibody is particularly useful for preventing NSCLC recurrence or metastasis in subjects with tumors containing MET alterations, such as exon 14 alterations in DNA or deletions that result in exon 14 skipping, MET gene amplification, or MET protein overexpression.

[0081] For example, the MET×MET bispecific antibodies of the present disclosure can be used to treat tumors expressing (or overexpressing) MET, for example, NSCLC with MET changes. Illustratively, MET changes can be exon 14 changes in DNA or deletions that cause exon 14 skipping, MET gene amplification, or MET protein overexpression. In some aspects, MET changes are exon 14 changes in DNA or deletions that cause exon 14 skipping. In some aspects, MET changes are MET gene amplification. In some aspects, MET changes are MET protein overexpression.

[0082] In some embodiments, the subject has non-squamous NSCLC. In some embodiments, the subject has NSCLC squamous carcinoma. In some aspects, the NSCLC has metastasized. In some aspects, the subject has NSCLC that has metastasized to the brain. In some aspects, the subject has NSCLC that has metastasized to the liver. In some embodiments, the NSCLC is unresectable. Treatment includes reducing NSCLC tumor growth and / or causing regression of NSCLC in the subject.

[0083] Illustrative methods of using MET×MET bispecific antibodies are provided throughout this disclosure and are described in detail below.

[0084] A method of treating NSCLC in a subject may comprise administering to a subject in need thereof a therapeutic composition comprising a MET×MET bispecific antibody (e.g., a MET×MET bispecific antibody comprising the D1 and D2 components shown in Table 1 herein, or an anti-MET antibody selected from onatuzumab, imatinib, telisotuzumab, SAIT301, ARGX-111, Sym015, HuMax-cMet, and CE-355621).

[0085] A method of treating NSCLC, reducing the growth of a NSCLC tumor, inhibiting or slowing invasion and / or metastasis, and / or causing regression of NSCLC in a subject having a tumor containing a MET alteration may comprise administering to a subject in need thereof a bispecific antibody comprising: a first antigen binding domain (D1), and a second antigen binding domain (D2); wherein D1 specifically binds to a first epitope of human MET, and wherein D2 specifically binds to a second epitope of human MET.

[0086] A method for treating a subject with a NSCLC tumor containing a MET change may include administering a MET×MET bispecific antibody to the subject, the MET×MET bispecific antibody comprising: a first antigen binding domain (D1), and a second antigen binding domain (D2); wherein D1 specifically binds to a first epitope of human MET; and wherein D2 specifically binds to a second epitope of human MET. In some aspects, the MET change is an exon 14 change in DNA or a deletion that causes exon 14 skipping. In some aspects, the subject is a patient who has not received a tyrosine kinase inhibitor (MET-TKI) targeting MET. In some aspects, the MET change is MET gene amplification. In some aspects, the MET change is MET protein expression.

[0087] The method of treating NSCLC, reducing NSCLC tumor growth, and / or causing regression of NSCLC in a subject having a tumor containing a MET alteration may comprise administering to the subject a dose of 250 to 2000 mg of a MET×MET bispecific antibody.

[0088] The method of treating or inhibiting the growth of NSCLC may include: (1) selecting a subject having a tumor containing a MET alteration; and (2) administering to the subject (a) about 250 mg, 500 mg, 750 mg, 1000 mg, 1500 mg or 2000 mg of a MET×MET bispecific antibody. In some aspects, the administration of step (2) is performed once every 3 weeks.

[0089] Another method of treating a tumor comprises: (a) selecting a subject having NSCLC; (b) determining that the tumor exhibits a MET alteration selected from the following: an exon 14 alteration in DNA or a deletion resulting in exon 14 skipping, MET gene amplification, and / or Met protein overexpression, comprising (i) obtaining a tissue sample and / or a liquid sample from the subject, and (ii) assessing the tissue sample for MET gene amplification using fluorescent in situ hybridization of the tumor tissue or by next-generation sequencing of the tumor tissue and / or ctDNA, and / or assessing the tissue sample for Met protein overexpression using immunohistochemistry, and / or assessing the liquid sample for exon 14 mutations using ctDNA; and, if the tumor exhibits a MET alteration, (c) administering one or more doses of a MET×MET bispecific antibody to the subject in need thereof.

[0090] At least from the patient's perspective, there is a need to identify patients or patient populations that may be successfully treated with a MET×MET antibody. Accordingly, the inventors conceived a method for identifying a candidate for a MET×MET anti-tumor therapy, the method comprising obtaining a tissue sample and / or a fluid sample from a subject with NSCLC; and assessing the tissue sample and / or the fluid sample for a MET alteration selected from an exon 14 alteration in DNA or a deletion resulting in exon 14 skipping, a MET gene amplification, and / or Met protein overexpression, wherein the presence of at least one Met alteration in the tissue sample or fluid sample identifies the subject as a candidate for an anti-tumor therapy, wherein the MET×MET anti-tumor therapy comprises a MET×MET bispecific antibody.

[0091] An additional method for treating NSCLC in a subject may comprise: (i) obtaining a liquid sample from the subject and determining MET amplification in ctDNA from the liquid sample, and (ii) administering a MET×MET bispecific antibody to the subject; wherein steps (i) and (ii) are repeated every three weeks, and wherein loss of MET amplification after repeating step (ii) indicates a durable response to treatment.

[0092] In some aspects, the subject according to any of the methods provided herein has one or more of the following: (i) histologically confirmed NSCLC; (ii) MET exon 14 gene mutation; (iii) MET gene amplification; (iv) elevated MET protein expression (IHC ≥2+ or H score>150); (v) MET exon 14 gene mutation and MET TKI-treated; (vi) MET exon 14 gene mutation and MET TKI-naive; (vii) MET gene highly amplified (METGCN ≥5 and / or MET / CEP7 ratio ≥2 by FISH in tissue, or MET GCN ≥6 by NGS, or MET fold change in ctDNA ≥2) and MET TKI-naive; (viii) MET protein highly overexpressed (IHC 3+ or H score ≥200) and MET TKI-naive; (ix) MET gene highly amplified (MET FISH in tissue GCN ≥5 and / or MET / CEP7 ratio ≥2, or MET GCN ≥6 by NGS, or MET fold change ≥2 in ctDNA), highly overexpressed MET protein (IHC 3+ or H score ≥200), and not received MET TKI.

[0093] In some aspects, the subject is selected as having one or more of the following: exon 14 alterations in DNA or deletions resulting in exon 14 skipping with prior MET TKI experience; exon 14 alterations in DNA or deletions resulting in exon 14 skipping (not having received MET TKI) with PD-(L)1 experience; exon 14 alterations in DNA or deletions resulting in exon 14 skipping (not having received MET TKI) with prior EGFR inhibitor experience and no PD-(L1) experience; MET gene amplification and / or MET protein overexpression (not having received MET TKI) and no PD-(L1) experience; MET gene amplification and / or MET protein overexpression (not having received MET TKI) with PD-(L1) experience; or MET gene amplification and / or MET protein overexpression (not having received MET TKI) with prior EGFR inhibitor experience and no PD-(L1) experience. Subjects can also be selected based on having a tumor that has one or more mutations in the EGFR gene.

[0094] In some embodiments, the subject has not received previous anticancer therapy. In some embodiments, the subject has not received a tyrosine kinase inhibitor (TKI) targeting MET. In other words, the subject has not received treatment with a previous TKI. In some embodiments, the subject has received an anticancer therapy previously including one or more of the following: TKI, PD-1 inhibitor, PD-L1 inhibitor, surgery, radiotherapy, or chemotherapy. In some aspects, the previous anticancer therapy includes TKI. In some aspects, the subject is resistant or insufficiently responsive to the previous treatment, or relapses after the previous treatment.

[0095] In the context of the treatment methods described herein, the MET×MET bispecific antibody can be administered as a monotherapy (ie, as the sole therapeutic agent) or in combination with one or more additional therapeutic agents.

[0096] As tumor cells develop bypass resistance mechanisms, responsiveness to treatment with MET×MET bispecific antibodies may change over time. Resistance to MET treatment can be acquired in heavily pre-treated NSCLC patients in response to previous treatment, such as patients treated with chemotherapy, immune checkpoint inhibitors, and / or EGFR inhibitors. If the tumor has been shown to be resistant to current treatment, it is desirable to provide alternative treatments as early as possible. Therefore, provided herein is a method for monitoring the efficacy of treatment with MET×MET bispecific antibodies in subjects with NSCLC having MET changes. The method comprises: (i) obtaining a tissue sample and / or a fluid sample from a subject, and assessing somatic mutations in one or more genes selected from the following in the tissue sample and / or fluid sample: ●a. On-target MET receptor gene mutations that confer resistance to MET TKIs, which are present in MET Ex14 Mut patients with prior TKIExp and MET gene silencing (loss of function); b. TK driven receptor activation selected from TK receptor and ligand gene amplification and TKR activating mutations; and c. Activating gene mutations in a pathway selected from the group consisting of the JAK2 / STAT3 pathway, the RAS / RAF / MEK / MAPK pathway, the PI3K / AKT / MTOR pathway, TP53 mutations, and cell cycle gene amplifications; (ii) administering a MET×MET bispecific antibody to the subject; and (iii) repeating steps (i) and (ii) during the treatment; wherein the acquisition of one or more gene mutations indicates resistance to treatment and / or indicates a poor prognosis.

[0097] In some aspects, the on-target MET receptor gene mutation is selected from MET Y1230C, MET D1228H, MET D1228N; and MET gene silencing (loss of function) is selected from somatic mutations of DNMT3A and TET2.

[0098] In some aspects, the TKR activating mutation is selected from the group consisting of: EGFR L858R, EGFR G719S, EGFR E709A, EGFR E746_A750del, and EGFR S752_I759del.

[0099] In some aspects, the JAK2 / STAT3 pathway mutation is JAK2 V617F; the RAS / RAF / MEK / MAPK pathway mutation is selected from the group consisting of KRAS G12A / V, GNAS R201H, MKRN-BRAF fusion, BRAF S602Y, RICTOR Amp, and MAP2K1 K57N; the PI3K / AKT / MTOR pathway mutation is selected from the group consisting of PIK3CA H1047L, PIK3CA E545K, PIK3CA E542K, PIK3CAN345K, IDH1 R132L, and MTOR E2338Q; the PI3K / AKT / MTOR pathway amplification is selected from the group consisting of AKT2 Amp and RICTOR Amp; the TP53 mutation is selected from the group consisting of TP53 R280T and TP53 R248Q; and the cell cycle gene amplification is selected from the group consisting of CDK4 Amp, CDK6 Amp, CCND1 Amp, and CCNE1 Amp.

[0100] Various aspects of the MET×MET bispecific antibodies are provided in the following paragraphs, but are described in more detail elsewhere herein.

[0101] In some aspects, D1 and D2 do not compete with each other in binding to human MET. In some aspects, the first epitope of human MET comprises amino acids 192 to 204 of SEQ ID NO: 22. In some aspects, the second epitope of human MET comprises amino acids 305 to 315 and 421 to 455 of SEQ ID NO: 22. In some aspects, the first epitope of human MET comprises amino acids 192 to 204 of SEQ ID NO: 22; and the second epitope of human MET comprises amino acids 305 to 315 and 421 to 455 of SEQ ID NO: 22.

[0102] In some embodiments, D1 comprises three heavy chain complementary determining regions (HCDR1, HCDR2, and HCDR3) within a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 1, and three light chain complementary determining regions (LCDR1, LCDR2, and LCDR3) within a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 9. In some embodiments, D2 comprises three heavy chain complementary determining regions (HCDR1, HCDR2, and HCDR3) within a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 5, and three light chain complementary determining regions (LCDR1, LCDR2, and LCDR3) within a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 9.

[0103] In some embodiments, the bispecific antibody comprises CDRs within the D1-HCVR amino acid sequence of SEQ ID NO:1 and CDRs within the D2-HCVR amino acid sequence of SEQ ID NO:5.

[0104] In some aspects, the bispecific antibody D1 comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) within a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 1, or an amino acid sequence having at least 95% identity thereto, and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) within a light chain variable region (LCVR), wherein the heavy chain variable region (HCVR) comprises the amino acid sequence of SEQ ID NO: 1, or an amino acid sequence having at least 95% identity thereto, and the light chain variable region (LCVR) comprises the amino acid sequence of SEQ ID NO: 9, or an amino acid sequence having at least 95% identity thereto.

[0105] In some aspects, D1 HCDR1 comprises the amino acid sequence of SEQ ID NO:2; HCDR2 comprises the amino acid sequence of SEQ ID NO:3; HCDR3 comprises the amino acid sequence of SEQ ID NO:4; LCDR1 comprises the amino acid sequence of SEQ ID NO:10; LCDR2 comprises the amino acid sequence of SEQ ID NO:11; and LCDR3 comprises the amino acid sequence of SEQ ID NO:12.

[0106] In some aspects, the bispecific antibody D1 comprises a HCVR comprising the amino acid sequence of SEQ ID NO: 1, or an amino acid sequence that is at least 95% identical thereto, and a LCVR comprising the amino acid sequence of SEQ ID NO: 9, or an amino acid sequence that is at least 95% identical thereto.

[0107] In some aspects, the bispecific antibody D1 comprises a HCVR comprising the amino acid sequence of SEQ ID NO:1 and a LCVR comprising the amino acid sequence of SEQ ID NO:9.

[0108] In some aspects, the bispecific antibody D2 comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) within a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO:5, or an amino acid sequence having at least 95% identity thereto, and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) within a light chain variable region (LCVR), wherein the heavy chain variable region (HCVR) comprises the amino acid sequence of SEQ ID NO:5, or an amino acid sequence having at least 95% identity thereto, and the light chain variable region (LCVR) comprises the amino acid sequence of SEQ ID NO:9, or an amino acid sequence having at least 95% identity thereto.

[0109] In some aspects, the bispecific antibody D2 HCDR1 comprises the amino acid sequence of SEQ ID NO:6; HCDR2 comprises the amino acid sequence of SEQ ID NO:7; HCDR3 comprises the amino acid sequence of SEQ ID NO:8; LCDR1 comprises the amino acid sequence of SEQ ID NO:10; LCDR2 comprises the amino acid sequence of SEQ ID NO:11; and LCDR3 comprises the amino acid sequence of SEQ ID NO:12.

[0110] In some aspects, the bispecific antibody D2 comprises a HCVR comprising the amino acid sequence of SEQ ID NO:5, or an amino acid sequence that is at least 95% identical thereto, and a LCVR comprising the amino acid sequence of SEQ ID NO:9, or an amino acid sequence that is at least 95% identical thereto.

[0111] In some aspects, the bispecific antibody D2 comprises a HCVR comprising the amino acid sequence of SEQ ID NO:5 and a LCVR comprising the amino acid sequence of SEQ ID NO:9. Biological characteristics of the antibodies provided herein

[0112] Useful according to the methods provided herein are bispecific antibodies and antigen-binding fragments thereof that inhibit NSCLC cell proliferation, inhibit NSCLC cell invasion, induce NSCLC cell apoptosis and / or reduce NSCLC cell viability. Bispecific antibodies and antigen-binding fragments thereof bind to the MET receptor and prevent interaction with HGF.

[0113] Also useful according to the methods provided herein are bispecific antibodies and antigen-binding fragments thereof that bind to monomeric human MET with high affinity. D An anti-MET×MET antibody that binds to monomeric human MET (e.g., hMET.mmh) at less than about 230 nM, wherein the K D As measured, for example, by surface plasmon resonance at 25° C. or 37° C. using the assay format defined in Example 3 of U.S. Patent No. 11,142,578 or a substantially similar assay. According to certain embodiments, the anti-MET antibodies can be used according to the methods provided herein at 37° C. with a K of less than about 230 nM, less than about 200 nM, less than about 150 nM, less than about 100 nM, less than about 50 nM, less than about 25 nM, less than about 20 nM, less than about 10 nM, less than about 8 nM, less than about 6 nM, less than about 5 nM, less than about 4 nM, or less than about 3 nM. D Binding to monomeric human MET, the K D As measured, for example, by surface plasmon resonance using the assay format defined in Example 3 of US Patent No. 11,142,578, or a substantially similar assay.

[0114] Such bispecific antibodies and antigen-binding fragments thereof bind to monomeric human MET (e.g., hMET.mmh) with a dissociation half-life (t1 / 2) of greater than about 1 minute, as measured by surface plasmon resonance at 25° C. or 37° C. using, for example, the assay format defined in Example 3 of U.S. Patent No. 11,142,578, or a substantially similar assay. Provided are bispecific antibodies that bind to monomeric human MET at 37° C. with a t1 / 2 of greater than about 1 minute, greater than about 2 minutes, greater than about 4 minutes, greater than about 6 minutes, greater than about 8 minutes, greater than about 10 minutes, greater than about 12 minutes, greater than about 14 minutes, greater than about 16 minutes, greater than about 18 minutes, or greater than about 20 minutes, or longer, as measured by surface plasmon resonance using, for example, the assay format defined in Example 3 of U.S. Patent No. 11,142,578, or a substantially similar assay.

[0115] Such bispecific antibodies and antigen-binding fragments thereof bind to dimeric human MET with high affinity (e.g., hMET.mFc). For example, a bispecific antibody binds with a K of less than about 3 nM. D Binds to dimeric human MET, the K DAs measured, for example, by surface plasmon resonance at 25° C. or 37° C. using the assay format defined in Example 3 of U.S. Patent No. 11,142,578 or a substantially similar assay. According to certain embodiments, anti-MET antibodies are provided that have a K of less than about 3 nM, less than about 2 nM, less than about 1 nM, less than about 0.9 nM, less than about 0.8 nM, less than about 0.7 nM, less than about 0.6 nM, less than about 0.5 nM, less than about 0.4 nM, less than about 0.3 nM, or less than about 0.25 nM at 37° C. D Binds to dimeric human MET, the K D As measured, for example, by surface plasmon resonance using the assay format defined in Example 3 of US Patent No. 11,142,578, or a substantially similar assay.

[0116] Such bispecific antigen-binding molecules and antigen-binding fragments thereof can bind to dimeric human MET (e.g., hMET.mFc) with a dissociation half-life (t1 / 2) of greater than about 4 minutes, as measured by surface plasmon resonance at 25°C or 37°C, for example, using the assay format defined in U.S. Patent No. 11,142,578_Example 3 or a substantially similar assay. According to certain embodiments, an anti-MET antibody is provided that binds to dimeric human MET at 37° C. with a t1 / 2 of greater than about 4 minutes, greater than about 5 minutes, greater than about 10 minutes, greater than about 20 minutes, greater than about 30 minutes, greater than about 40 minutes, greater than about 50 minutes, greater than about 60 minutes, greater than about 70 minutes, greater than about 80 minutes, greater than about 90 minutes, greater than about 100 minutes, greater than about 105 minutes, or longer, as measured by surface plasmon resonance using, for example, the assay format defined in Embodiment 3 of U.S. Patent No. 11,142,578 or a substantially similar assay.

[0117] Also useful according to the methods provided herein are MET×MET bispecific antigen binding proteins that bind to dimeric human MET (e.g., hMET.mFc) with a dissociation half-life (t1 / 2) of greater than about 10 minutes, as measured by surface plasmon resonance at 25°C or 37°C using, for example, the assay format defined in Example 6 of U.S. Patent No. 11,142,578 or a substantially similar assay. According to certain embodiments, a MET×MET bispecific antigen binding protein is provided that binds to dimeric human MET at 37° C. with a t1 / 2 of greater than about 10 minutes, greater than about 20 minutes, greater than about 30 minutes, greater than about 40 minutes, greater than about 50 minutes, greater than about 60 minutes, greater than about 70 minutes, greater than about 80 minutes, greater than about 90 minutes, greater than about 100 minutes, greater than about 200 minutes, greater than about 300 minutes, greater than about 400 minutes, greater than about 500 minutes, greater than about 600 minutes, greater than about 700 minutes, greater than about 800 minutes, greater than about 900 minutes, greater than about 1000 minutes, greater than about 1100 minutes, or longer, as measured, for example, by surface plasmon resonance using the assay format defined in Example 6 of U.S. Patent No. 11,142,578 or a substantially similar assay.

[0118] Also useful according to the methods provided herein are MET×MET bispecific antibodies that block the interaction between HGF and MET (e.g., in an in vitro ligand binding assay). According to certain embodiments provided herein, the MET×MET bispecific antigen binding protein blocks the binding of HGF to cells expressing human MET and induces minimal or no MET activation in the absence of HGF signaling. For example, the present disclosure provides a MET×MET bispecific antigen binding protein that exhibits a degree of MET agonist activity in a cell-based MET activity reporter assay that is less than 50%, less than 40%, less than 30%, less than 20%, less than 10%, less than 5%, less than 3%, less than 2%, or less than 1% of the MET agonist activity observed in an equivalent activity reporter assay using a monospecific antibody comprising only D1 or D2.

[0119] Bispecific antibodies available according to the present disclosure may have one or more of the aforementioned biological characteristics, or any combination thereof. The biological characteristics of the antibodies listed above are not intended to be exhaustive. Other biological characteristics of the antibodies provided herein will be apparent to those of ordinary skill in the art by reviewing the present disclosure including the working examples herein. Epitope mapping and related technologies

[0120] Useful according to the methods provided herein are MET×MET bispecific antibodies that bind to a human MET epitope comprising amino acids 192 to 204, amino acids 305 to 315, and / or amino acids 421 to 455 of SEQ ID NO:22. Preparation of human antibodies

[0121] The MET×MET bispecific antibodies useful according to the methods provided herein can be fully human antibodies. Methods for producing monoclonal antibodies, including fully human monoclonal antibodies, are known in the art. Any such known methods can be used in the context of the present disclosure to prepare human antibodies that specifically bind to human MET.

[0122] For example, using VELOCIMMUNE TM The method is based on the invention of the invention, or any other similar known method for producing fully human monoclonal antibodies, first isolating a high-affinity chimeric antibody against MET having a human variable region and a mouse constant region. As in the experimental section below, the antibodies are characterized and selected for desired characteristics including affinity, ligand blocking activity, selectivity, epitope, etc. If necessary, the mouse constant region is replaced with a desired human constant region, such as wild-type or modified IgG1 or IgG4, to produce a fully human anti-MET antibody. Although the selected constant region may vary depending on the specific use, the high-affinity antigen binding and target-specific characteristics are present in the variable region. In some cases, fully human anti-MET antibodies are isolated directly from antigen-positive B cells. Bioequivalents

[0123] The methods described herein can use MET×MET bispecific antibodies and antibody fragments thereof that contain proteins having an amino acid sequence different from that of the antibody but retaining the ability to bind to human MET. Such variant molecules and antibody fragments contain one or more amino acid additions, deletions or substitutions compared to the parent sequence, but exhibit biological activities that are substantially equivalent to the biological activities of the molecules. Similarly, the DNA sequences encoding anti-MET×MET antibodies of the present disclosure include sequences that contain one or more nucleotide additions, deletions or substitutions compared to the disclosed sequences, but that encode anti-MET×MET antibodies or antibody fragments that are substantially biologically equivalent to the anti-MET antibodies or antibody fragments of the present disclosure. Some examples of such variant amino acids and DNA sequences are discussed above.

[0124] If, for example, two antigen binding proteins or antibodies are pharmaceutical equivalents or pharmaceutical substitutes that do not show significant differences in their absorption rate and extent when administered in single or multiple doses at the same molar dose under similar experimental conditions, they are considered bioequivalent. If some antibodies are equivalent in their extent of absorption but different in their absorption rate, these antibodies will be considered equivalents or pharmaceutical substitutes and may still be considered bioequivalent because such differences in absorption rate are intentional and reflected in the labeling, are not necessary to achieve effective in vivo drug concentrations (e.g., long-term use), and are considered medically insignificant for the specific drug product under study.

[0125] In one embodiment, two antigen binding proteins are bioequivalent if there are no clinically meaningful differences in their safety, purity, and potency.

[0126] In one embodiment, two antigen binding proteins are bioequivalent if a subject can switch between the reference product and the biological product one or more times without an increased risk of expected adverse effects (including clinically significant changes in immunogenicity, or reduced effectiveness) compared to continued treatment without such a switch.

[0127] In one embodiment, two antigen binding proteins are bioequivalent if they both act by one or more common mechanisms of action for one or more conditions of use to the extent such mechanisms are known.

[0128] Bioequivalence can be demonstrated by both in vivo and in vitro methods. Bioequivalence measurements include, for example: (a) in vivo tests in humans or other mammals, in which the concentration of the antibody or its metabolites is measured as a function of time in blood, plasma, serum or other biological fluids; (b) in vitro tests that correlate with and are reasonably predictive of in vivo human bioavailability data; (c) in vivo tests in humans or other mammals, in which the appropriate acute pharmacological effect of the antibody (or its target) is measured as a function of time; and (d) in well-controlled clinical trials that establish the safety, efficacy, or bioavailability or bioequivalence of the antibody.

[0129] Bioequivalent variants of the anti-MET antibodies provided herein can be constructed by, for example, making various substitutions of residues or sequences or deleting terminal or internal residues or sequences that are not required for biological activity. For example, cysteine ​​residues that are not required for biological activity can be deleted or replaced with other amino acids to prevent the formation of unnecessary or incorrect intramolecular disulfide bridges during renaturation. In other cases, bioequivalent antibodies may include anti-MET antibody variants that include amino acid changes that alter the glycosylation characteristics of the antibody, such as mutations that eliminate or remove glycosylation. Species selectivity and cross-species reactivity

[0130] According to certain embodiments, the present disclosure provides methods of using anti-MET antibodies (and antibodies comprising anti-MET antigen binding domains) that bind to human MET but not to MET from other species. Anti-MET antibodies (and antibodies comprising anti-MET antigen binding domains) that bind to human MET and MET from one or more non-human species are also available. For example, anti-MET×MET antibodies and antibodies can bind to human MET and may bind or not bind to one or more of mouse, rat, guinea pig, hamster, gerbil, pig, cat, dog, rabbit, goat, sheep, cattle, horse, camel, cynomolgus, marmoset, rhesus monkey, or chimpanzee MET, as appropriate. According to certain exemplary embodiments, anti-MET×MET antibodies and antibodies that specifically bind to human MET and cynomolgus (e.g., cynomolgus macaque (Macaca fascicularis)) MET are provided. Other anti-MET×MET antibodies and antibodies bind to human MET but not to cynomolgus monkey MET, or bind only weakly to cynomolgus monkey MET. Therapeutic formulations and administration

[0131] Useful herein are pharmaceutical compositions comprising the MET×MET bispecific antibodies of the present invention. The pharmaceutical compositions can be formulated with suitable carriers, excipients, and other agents to provide improved transfer, delivery, tolerability, and the like.

[0132] In some aspects, a pharmaceutical composition comprising a MET×MET bispecific antibody is formulated for administration to a subject for the treatment of lung cancer, and in particular, the treatment of NSCLC.

[0133] Provided herein are methods in which a MET×MET bispecific antibody administered to a subject is contained within a pharmaceutical formulation. The pharmaceutical formulation may comprise a MET×MET bispecific antibody and at least one inactive ingredient, such as, for example, a pharmaceutically acceptable carrier. Other agents may be incorporated into the pharmaceutical composition to provide improved transfer, delivery, tolerability, and the like. The term "pharmaceutically acceptable" means approved by a regulatory agency of a federal or state government or listed in the U.S. Pharmacopeia or other recognized pharmacopeia for use in animals and more specifically in humans. The term "carrier" refers to a diluent, adjuvant, excipient, or carrier administered with the antibody. Many suitable formulations can be found in formularies known to all pharmaceutical chemists: Remington's Pharmaceutical Sciences (15th ed., Mack Publishing Company, Easton, Pa., 1975), specifically Chapter 87 of Blaug, Seymour. These formulations include, for example, powders, pastes, ointments, jellies, waxes, oils, lipids, vesicles containing lipids (cationic or anionic) (e.g., LIPOFECTIN.TM.), DNA conjugates, anhydrous absorption pastes, oil-in-water and water-in-oil emulsions, emulsions carbowax (polyethylene glycols of various molecular weights), semisolid gels, and semisolid mixtures containing carbowax. Any of the foregoing mixtures may be suitable in the context of the methods of the present disclosure, provided that the anti-MET antibody or MET×MET bispecific antibody is not inactivated by the formulation and that the formulation is physiologically compatible and tolerable under the route of administration. See also Powell et al. PDA (1998) J Pharm Sci Technol. 52:238-311 and references therein for additional information on excipients and carriers known to pharmaceutical chemists.

[0134] Pharmaceutical preparations that can be used for injection in the context of the present disclosure can be prepared by dissolving, suspending or emulsifying the anti-MET antibody or the MET×MET bispecific antibody in a sterile aqueous medium or an oily medium conventionally used for injection. As aqueous media for injection, there are, for example, physiological saline, isotonic solutions containing glucose and other adjuvants, which can be used in combination with appropriate solubilizers such as alcohols (e.g., ethanol), polyols (e.g., propylene glycol, polyethylene glycol), nonionic surfactants [e.g., polysorbate 80, HCO-50 (polyoxyethylene (50 mol) adduct of hydrogenated castor oil)], etc. As oily media, for example, sesame oil, soybean oil, etc. can be used, which can be used in combination with solubilizers such as benzyl benzoate, benzyl alcohol, etc. If necessary, the injection thus prepared can be filled into a suitable ampoule. Administration regimen

[0135] According to certain embodiments, multiple doses of an anti-MET antibody or MET×MET bispecific antibody (or a pharmaceutical composition comprising a combination of an anti-MET antibody or MET×MET bispecific antibody and any additional therapeutically active agent mentioned herein) may be administered to a subject over a defined time course. The method according to this aspect comprises sequentially administering multiple doses of an anti-MET antibody or MET×MET bispecific antibody provided herein to a subject. “Sequential administration” as used herein means that each dose of the antibody is administered to a subject at different time points (e.g., on different days) separated by a predetermined interval (e.g., hours, days, weeks, or months). The present disclosure includes methods comprising sequentially administering a single initial dose of an anti-MET antibody or MET×MET bispecific antigen binding molecule to a subject, followed by one or more second doses of an anti-MET antibody or MET×MET bispecific antigen binding molecule, and optionally followed by one or more third doses of an anti-MET antibody or MET×MET bispecific antibody.

[0136] The terms "initial dose", "second dose" and "third dose" refer to the temporal order of administration of an anti-MET antibody or a MET×MET bispecific antibody. Thus, the "initial dose" is the dose administered at the start of the treatment regimen (also referred to as a "baseline dose"); the "second dose" is the dose administered after the initial dose; and the "third dose" is the dose administered after the second dose. The initial dose, the second dose, and the third dose may all contain the same amount of anti-MET antibody or MET×MET bispecific antigen binding molecule, but may generally differ from each other in terms of the frequency of administration. However, in certain embodiments, during the course of treatment, the amount of antibody contained in the initial dose, the second dose, and / or the third dose differs from each other (e.g., appropriately adjusted upward or downward). In certain embodiments, two or more (e.g., 2, 3, 4, or 5) doses are administered at the start of the treatment regimen as a "loading dose", followed by subsequent doses (e.g., "maintenance doses") based on lower frequency administration.

[0137] Anti-MET antibodies or MET×MET bispecific antibodies can be administered according to any regimen that provides a therapeutic effect. In some aspects, the bispecific antibody is administered at the following dosing frequency: about four times a week, twice a week, once a week, once every two weeks, once every three weeks, once every four weeks, once every five weeks, once every six weeks, once every eight weeks, once every twelve weeks, or less, as long as a therapeutic response is achieved. In some aspects, the bispecific antibody is administered once a week, once every two weeks, once every three weeks, or once a month. In some aspects, the bispecific antibody is administered once every three weeks.

[0138] In some aspects, the bispecific antibody is administered one week, two weeks, three weeks, or four weeks after the previous dose. In some aspects, the bispecific antibody is administered three weeks after the previous dose.

[0139] According to certain embodiments of the present disclosure, multiple doses of bispecific antibodies can be administered to an object during a limited time course. The method according to this aspect of the present disclosure includes sequentially administering more than one dose of bispecific antibodies to an object. "Sequential administration" as used herein means that at different time points (e.g., hours, days, weeks or months) separated by a predetermined interval, each dose of antibody is administered to an object. The present disclosure includes such a method, which includes sequentially administering a single initial dose of bispecific antigen binding molecules to an object, followed by administration of one or more second doses of bispecific antigen binding molecules, and optionally subsequently administering one or more third doses of bispecific antibodies.

[0140] According to certain embodiments of the present disclosure, multiple doses of a bispecific antibody may be administered to a subject once every 3 or 6 weeks for months or years.

[0141] The terms "initial dose", "second dose" and "third dose" refer to the temporal order of administration. Thus, the "initial dose" is the dose administered at the start of the treatment regimen (also referred to as the "baseline dose"); the "second dose" is the dose administered after the initial dose; and the "third dose" is the dose administered after the second dose. The initial dose, the second dose, and the third dose may all contain the same amount of antibody (anti-MET×MET bispecific antigen binding molecule). However, in certain embodiments, during the course of treatment, the amounts contained in the initial dose, the second dose, and / or the third dose are different from each other (e.g., appropriately adjusted upward or downward). In certain embodiments, one or more (e.g., 1, 2, 3, 4, or 5) doses are administered at the start of the treatment regimen as a "loading dose", followed by subsequent doses (e.g., "maintenance doses") based on lower frequency administration. For example, a bispecific antibody can be administered to a subject with NSCLC with a loading dose of about 1000 to 3000 mg, followed by one or more maintenance doses of about 500 mg, 1000 mg, or 2000 mg.

[0142] In an exemplary embodiment of the present disclosure, each second and / or third dose is 1 / 2 to 14 (e.g., 1 / 2, 1, 1 1 / 2 , 2, 2 1 / 2 ,3,3 1 / 2 , 4, 4 1 / 2, 5, 5 1 / 2 ,6,6 1 / 2 ,7,7 1 / 2 ,8,8 1 / 2 , 9, 9 1 / 2 , 10, 10 1 / 2 , 11, 11 1 / 2 , 12, 12 1 / 2 , 13, 13 1 / 2 , 14, 14 1 / 2 As used herein, the phrase "immediately preceding dose" means the dose of the bispecific antibody administered to the patient immediately prior to administration of the next dose in the sequence without an intervening dose in the order of multiple administrations.

[0143] According to some aspects of the method, the bispecific antibody of the second and / or third dose of any number may be administered to the subject. For example, in certain embodiments, only a single second dose is administered to the subject. In other embodiments, two or more (e.g., 2, 3, 4, 5, 6, 7, 8 or more) second doses are administered to the subject. Similarly, in certain embodiments, only a single third dose is administered to the subject. In other embodiments, two or more (e.g., 2, 3, 4, 5, 6, 7, 8 or more) third doses are administered to the subject.

[0144] In embodiments involving multiple second doses, each second dose can be administered at the same frequency as other second doses. For example, each second dose can be administered to a subject 1 to 2 weeks immediately after the previous dose. Similarly, in embodiments involving multiple third doses, each third dose can be administered to a subject at the same frequency as other third doses. For example, each third dose can be administered to a subject 2, 3 or 4 weeks immediately after the previous dose. Alternatively, the frequency of the second dose and / or third dose administered to a subject can vary during the course of the treatment regimen. Depending on the needs of individual subjects after clinical examination, physicians can also adjust the frequency of use during the course of treatment.

[0145] In certain embodiments, at the beginning of the treatment regimen, one or more doses of the bispecific antibody are administered on a higher frequency basis (twice a week, once a week, or once every two weeks) as an "induction dose," followed by subsequent doses ("consolidation doses" or "maintenance doses") that are administered on a lower frequency basis (e.g., once every 4 to 12 weeks). dose

[0146] The amount of the MET×MET bispecific antibody administered to a subject according to the methods of the present disclosure is generally a therapeutically effective amount. As used herein, the phrase "therapeutically effective amount" means an amount of the bispecific antibody that results in one or more of the following or has one or more of the following therapeutic effects: (a) reducing the severity or duration of cancer symptoms compared to untreated subjects; (b) inhibiting tumor growth, or increasing tumor necrosis, tumor shrinkage and / or tumor disappearance; (c) delaying tumor growth and occurrence; (d) inhibiting or delaying or preventing tumor metastasis; (e) preventing tumor growth recurrence; (f) improving the survival of subjects with cancer; and / or (g) reducing the use or need for conventional anti-cancer treatments (e.g., reducing or eliminating the use of chemotherapeutic agents or cytotoxic agents).

[0147] In some cases, a therapeutically effective amount can be about 250 mg to about 8000 mg, for example, about 250 mg, about 300 mg, about 350 mg, about 400 mg, about 450 mg, about 500 mg, about 550 mg, about 600 mg, about 700 mg, about 800 mg, about 900 mg, about 1000 mg, about 1050 mg, about 1100 mg, about 1500 mg, about 1600 mg, about 1700 mg, about 2000 mg, about 2050 mg, about 2100 mg, about 2200 mg, about 2500 mg, about 2700 mg, about 2800 mg, about 2900 mg, about 3000 mg, about 3200 mg, about 4000 mg, about 5000 mg, about 6000 mg, about 7000 mg or about 8000 mg of a MET×MET bispecific antibody.

[0148] In some aspects, the bispecific antibody can be administered at a dosage of about 250 mg to about 5000 mg, or about 250 mg, 300 mg, 400 mg, 500 mg, 600 mg, 700 mg, 750 mg, 800 mg, 900 mg, 1000 mg, 1100 mg, 1200 mg, 1300 mg, 1400 mg, 1500 mg, 1600 mg, 1700 mg, 1800 mg, 1900 mg, 2000 mg, 2100 mg, 2200 mg, 2300 mg, 2400 mg, 2500 mg, 3000 mg, 3500 mg, 4000 mg, 4500 mg or 5000 mg. In some aspects, the dosage is about 500 mg MET×MET bispecific antibody. In some aspects, the dosage is about 1000 mg MET×MET bispecific antibody. In some aspects, the bispecific antibody is administered at a dose of about 2000 mg. In some aspects, the MET×MET bispecific antibody can be administered at a dose of 250 mg, 500 mg, 1000 mg, or 2000 mg.

[0149] The bispecific antibody can be administered intravenously, subcutaneously, or intraperitoneally. In some aspects, the bispecific antibody is administered by intravenous infusion. Example

[0150] The following examples are presented to provide a complete disclosure and description of how to prepare and use the methods and compositions provided herein to those of ordinary skill in the art, and are not intended to limit the scope of what the inventors regard as their invention. Efforts have been made to ensure accuracy with respect to the numbers used (e.g., amounts, temperatures, etc.), but some experimental errors and deviations should be considered. Unless otherwise indicated, parts are parts by weight, molecular weights are average molecular weights, temperatures are in degrees Celsius, and pressures are at or near atmospheric pressure. Example 1. Construction of a bispecific antibody having two different antigen binding domains specific for different epitopes of MET

[0151] Examples 1 to 3 of U.S. Pat. No. 11,142,578, the entire contents of which are incorporated herein by reference, describe the construction of bispecific antibodies that comprise two different antigen-binding domains derived from two bivalent monospecific anti-MET antibodies (the "D1" arm derived from the exemplary anti-MET antibody H4H13306P2 and the "D2" arm derived from the exemplary anti-MET antibody H4H13312P2), and thus bind to separate epitopes on the extracellular domain of MET:

[0152] Binding epitope of anti-Met antibody H4H13312P2: AA 192 to 204 in SEQ ID NO:22: VRRLKETKDGFMF (SEQ ID NO:23).

[0153] Binding epitope of anti-Met antibody H4H13306P2: AA 305 to 315 in SEQ ID NO:22: LARQIGASLND (SEQ ID NO:24) and AA 421 to 455 in SEQ ID NO:22: FIKGDLTIANLGTSEGRFMQVVVSRSGPSTPHVNF (SEQ ID NO:25).

[0154] The two antigen binding domains (D1 and D2) comprise a common light chain variable region. The components of bispecific antibodies useful according to the methods provided herein are summarized in Table 1. Table 1: Summary of MET×MET bispecific antibody components

[0155] The binding kinetic parameters of H4H14639D to monomeric Met protein (hMET.mmh) are shown in Table 2. Table 2: Biacore binding affinity of H4H14639D at 37°C

[0156] H4H14639D exhibited a significantly lower off-rate than each of its parental antibodies H4H13306P2 and H4H13312P2. See Table 3. Table 3: Biacore binding affinity of bispecific anti-MET mAbs and monospecific parents at 37°C

[0157] As described in Example 7 of US Patent No. 11,142,578, MET×MET bispecific antibodies block HGF signaling and exhibit low MET agonist activity. Example 2. Anti-Met antibodies inhibit the growth of Met-amplified cells

[0158] The blocking activity of the MET×MET bispecific antibody (i.e., H4H14639D) was evaluated in the non-small cell lung cancer (NSCLC) cell line EBC-1, which exhibits an amplified Met gene and overexpresses MET (Lutterbach et al., Cancer Res. 67(5):2081-2088, 2007). The complete growth medium for EBC-1 cells contains MEM Earle salts, 10% fetal bovine serum (FBS), penicillin / streptomycin / glutamine, and non-essential amino acids for MEM. According to the SRE-luciferase readout, H4H14369D showed the greatest percentage inhibition of MET activity. In the current experiment, 3.0×10 3 EBC-1 cells were seeded in complete growth medium in the presence of H4H14639D at concentrations ranging from 15 pM to 100 nM. Cells were cultured under 5% CO 2 The cells were then fixed in 4% formaldehyde and stained with 3 μg / ml Hoechst 33342 to mark the nucleus. Images were acquired on a Micro XL (Molecular Devices, Sunnyvale, CA) and analyzed by Image analysis software (Molecular Devices, Sunnyvale, CA) was used to determine nuclear counts. Background nuclear counts of cells treated with 40 nM digitonin were subtracted from all wells, and viability was expressed as a percentage of the untreated control. The 10-point response curve (GRAPHPAD ) The IC50 value was determined using a four-parameter logistic equation in . As summarized in Table 4 below, the MET×MET bispecific antibody H4H14639D inhibited the growth of EBC-1 cells by 37 and had an IC50 of 0.82 nM. Table 4: Anti-Met bispecific antibodies block EBC-1

[0159] The effect of the MET×MET bispecific antibody on the growth of EBC-1 cells was evaluated. 2,500 EBC-1 cells were seeded in a 96-well plate and cultured in Dulbecco's medium supplemented with 10% FBS. The cells were treated with 0.1 μg / mL or 1 μg / mL of a control antibody or the MET×MET bispecific antibody and then incubated at 37°C under 5% CO 2 After 5 days, The indicator dye was measured in an M3 plate reader (Molecular Devices, LLC, Sunnyvale, CA). Relative cell growth was determined by reduction to its highly fluorescent form. The results are shown in Table 5 and Figure 1. The MET×MET bispecific antibody (H4H14639D) significantly reduced the relative cell growth of EBC-1 cells compared to the control antibody (Figure 1).

[0160] Several anti-MET antibodies (both bivalent monospecific and MET×MET bivalent) are potent inhibitors of SRE-Luc activation and inhibit the growth of MET-amplified and MET-overexpressing cell lines. Table 5: Anti-Met bispecific antibodies block EBC-1 cell growth Relative cell growth (n=3) Standard Deviation Comparison 1.000 0.045 0.1μg / mL H4H14639D 0.397 0.032 1μg / mL H4H14639D 0.462 0.028 Example 3. MET×MET bispecific antibody induces modest and transient MET pathway activity in NCI-H596 NSCLC cells

[0161] The effect of the MET×MET bispecific antibody on the MET pathway in human lung adenosquamous carcinoma cells was evaluated in vitro.

[0162] 250,000 NCI-H596 cells were seeded in 12-well plates and cultured in RPMI medium supplemented with 10% FBS. Cells were treated with 50 ng / ml of hepatocyte growth factor (HGF) or 10 μg / ml of the MET×MET bispecific antibody H4H14639D in duplicate. The cells were then incubated at 37°C in 5% CO 2 Incubate in 5% 4% PBS. After 0, 2, 6 or 18 hours, cell lysates were prepared, protein content was normalized and immunoblot analysis was performed. MET phosphorylation and ERK phosphorylation were quantified using ImageJ image processing program (T.Collins, BioTechniques 43:S25-S30, 2007). Phosphorylation levels were normalized to tubulin loading controls and expressed as fold changes relative to control treatments. Results are summarized in Table 6. Table 6: Phosphorylation of MET and ERK Processing (hours) Phospho-MET (mean ± SD) Phospho-ERK (mean ± SD) Control (hFc) (18) 1.0±0.5 1.0±0.3 HGF(2) 202.3±38.7 16.7±1.6 HGF(6) 38.9±4.9 12.4±3.9 HGF(18) 59.2±24.4 12.4±0.9 H4H14639D(2) 69.7±7.0 2.2±0.9 H4H14639D(6) 9.9±7.4 0.3±0.4 H4H14639D(18) 1.4±0.1 0.1±0.1

[0163] HGF treatment of NCI-H596 cells induced strong activation of MET and ERK, which peaked at 2 hours and persisted after 18 hours. Treatment with the H4H14636D bispecific antibody detected modest phosphorylation of MET and ERK, which returned to baseline levels at 18 hours or 6 hours, respectively. Example 4. MET×MET bispecific antibody induces MET degradation more potently than monospecific antibody in NCI-H596 lung cancer cells

[0164] The effects of the MET×MET bispecific antibody and the parental bivalent monospecific anti-MET antibody on the expression levels of hepatocyte growth factor receptor (HGFR or MET) on human lung adenosquamous carcinoma cells were evaluated. 250,000 NCI-H596 human lung adenosquamous carcinoma cells were seeded in 12-well plates and cultured in RPMI medium supplemented with 10% FBS. The cells were treated with: (1) 5 μg / mL of hFc control molecule, (2) 5 μg / mL of parental bivalent monospecific anti-MET antibody H4H13306P2, (3) 5 μg / mL of the parent bivalent monospecific anti-MET antibody H4H13312P2, (4) 2.5 μg / mL of H4H13306P2 and 2.5 μg / mL of H4H13312P2 in combination, or (5) 5 μg / mL of the MET×MET bispecific antibody H4H14639D. The cells were then incubated at 37°C in 5% CO 2 After 18 hours, cell lysates were prepared, protein content was normalized and immunoblot analysis was performed. MET expression was quantified using ImageJ image processing program (T. Collins, BioTechniques 43: S25-S30, 2007). The results are summarized in Table 7. Table 7: Relative levels of MET protein molecular Relative MET level Control (hFc) 1±0.03 H4H13306P2 0.50±0.01 H4H13312P2 0.35±0.04 H4H13306P2+H4H13312P2 0.61±0.04 H4H14639D 0.24±0.01

[0165] NCI-H596 (MET exon 14 skipping mutation) lung cancer cells were also treated with control or MET×MET bispecific antibody (10 μg / ml) for 2, 6, or 18 hours. MET expression was determined by immunoblotting ( Figure 2 ), which shows MET degradation induced by the MET×MET bispecific antibody with increasing treatment time.

[0166] In NCI-H596 lung cancer cells, the bispecific antibody H4H14636D induced MET degradation more potently than its parent conventional antibody. Example 5. In EBC-1 cells, MET×MET bispecific antibodies more potently induce MET degradation, inhibit pathway activity, and suppress tumor growth than monospecific antibodies

[0167] As described above, MET-amplified human lung squamous cell carcinoma EBC-1 cells (Lutterbach et al., "Lung cancer cell lines harboring MET gene amplification are dependent on Met for growth and survival," Cancer Res. 2007 Mar 1; 67(5): 2081-8) were treated with control antibody or 10 μg / ml of MET×MET bispecific antibody for 18 hours. MET expression and MET pathway activation confirmed by pMET and pErk expression were determined by immunoblotting with the indicated antibodies. Figure 3 Shown in.

[0168] Treatment of EBC-1 cells containing MET gene amplification with MET×MET bispecific antibody induced MET degradation more potently than treatment with control antibody. Treatment of EBC-1 cells with MET×MET bispecific antibody inhibited downstream effectors of the MET pathway.

[0169] In another experiment, 5 million EBC-1 cells were implanted subcutaneously into the flank of CB-17 SCID mice. Once tumors reached a volume of approximately 150 mm 3 , the mice were randomly divided into 6 groups and treated twice a week with 25 mg / kg of control antibody or 25 mg / kg of MET×MET bispecific antibody H4H14639D. Tumor growth was monitored 30 days after implantation, and the tumor volume (mm) of each experimental group was measured over time. 3 ). The results are shown in Table 8 and Figure 4 , which showed that the MET×MET bispecific antibody significantly inhibited the growth of EBC-1 tumors. Table 8: Relative EBC-1 Tumor Growth Example 6. MET×MET bispecific antibody does not induce growth of NCI-H596 lung cancer cells in vitro

[0170] The effect of MET×MET bispecific antibody on the growth of human non-small cell lung cancer (NSCLC) cells (NCI-H596) was evaluated in vitro. 10,000 NCI-H596 lung adenosquamous carcinoma cells (Nair et al., J. Nat'l. Cancer Inst. 86 (5): 378-383, 1994) were seeded on a 0.66% agar layer in a 96-well plate, the 0.66% agar layer in a medium supplemented with 1% fetal bovine serum (FBS). The cells were cultured in RPMI 1640 medium supplemented with 1% FBS and 0.3% agarose. Cells were treated with (1) 5 μg / ml of a single parental bivalent monospecific anti-MET antibody (H4H13306P2 or H4H13312P2), (2) a combination of two parental bivalent monospecific anti-MET antibodies (H4H13306P2 and H4H13312P2) at 2.5 μg / ml each, (3) 5 μg / ml of a bispecific antibody containing one binding arm from H4H13306P2 and the other binding arm from H4H13312P2 (H4H14639D), or (4) 100 ng / mL of hepatocyte growth factor (HGF). Cells were then incubated at 37°C in the presence of 5% CO. 2 After two weeks, The indicator dye ALAMAR was measured in an M3 plate reader (Molecular Devices, Sunnyvale, CA). Relative cell growth was determined by reducing the 5-mercaptoethanol to its highly fluorescent form (Thermo Fischer Scientific, Waltham, MA). Increased fluorescence correlates with cell growth. Table 9 and Figure 5 Relative NCI-H596 cell growth for each antibody treatment normalized to control (untreated) NCI-H596 cell growth is depicted. Treatment of NCI-H596 lung cancer cells with HGF resulted in a potent induction of growth in soft agar. MET×MET( Figure 5 The bispecific antibody H4H14639D in M1 (MM) did not significantly alter growth. A modest induction of cell growth was observed with each parental bivalent monospecific antibody H4H13306P2 (M1) or H4H13312P2 (M2) alone or in combination (H4H13306P2 and H4H13312P2) (M1M2). Table 9: Normalized NCI-H596 cell growth Example 7. Clinical trial of Met×Met bispecific antibody in non-small cell lung cancer background:

[0171] Lung cancer is one of the most commonly diagnosed cancers and is the leading cause of cancer-related mortality worldwide (Siegel et al., CA Cancer J Clin, 66(1):7-30, 2016). Non-small cell lung cancer (NSCLC) accounts for 80% to 85% of all lung cancers and is composed of several histopathological subtypes, the most common of which are adenocarcinoma (40% to 60%) and squamous cell carcinoma (30%) (Dela Cruz et al., Clin Chest Med, 32(4):605-44, 2011). Most patients with NSCLC are found to have advanced cancer at the time of diagnosis.

[0172] First-line treatment of advanced NSCLC is guided by the presence of molecular alterations. Patients can receive targeted small molecule tyrosine kinase inhibitors (TKIs) with or without maintenance therapy (Besse et al., Ann Oncol, 25(8):1475-84, 2014) (Ettinger et al., J Natl Compr Canc Netw 15(4):504-35, 2017) (Reckett al., Ann Oncol, 25Suppl 3:27-39, 2014), immune checkpoint inhibitor antibodies that inhibit the PD-1 receptor or PD-1 ligand (PD-L1), or platinum-based doublet chemotherapy regimens (Besse et al., Ann Oncol, 25(8):1475-84, 2014) (Ettinger et al., J Natl Compr Canc Netw 15(4):504-35, 2017) (Reckett al., Ann Oncol, 25Suppl 3:27-39, 2014). In advanced NSCLC, long-term survival remains an unmet need. The overall survival (OS) of PD-1 / PD-L1 inhibitors is about 12 months, with some studies exceeding 2 years in patients treated with TKIs (Camidge et al., J Clin Oncol, 32: abstract 8001, 2014).

[0173] Mesenchymal epithelial transition factor (MET) is a single-pass transmembrane tyrosine kinase receptor for hepatocyte growth factor (HGF). It is expressed in normal tissues such as liver, breast and fat, and is upregulated in several cancers. In NSCLC tumors and gastric tumors, high levels of MET expression can occur by increased protein expression or by gene amplification, and are associated with negative outcomes in patients (Catenacci et al., Cancer, 123 (6): 1061-70, 2017) (Topalian et al., NEJM, 366 (26): 2443-54, 2012) (Zhang et al., Hum Pathol, 72: 59-65, 2018). MET mutations that cause exon 14 deletions promote prolonged ligand-dependent signaling, resulting in receptor stability and increased tumorigenicity (Kong-Beltran et al., Cancer Res, 66 (1): 283-9, 2006).

[0174] It has been reported that approximately 3% of NSCLCs contain MET exon 14 alterations (Cancer Genome Atlas Research Network, Nature, 511(7511):543-50, 2014) (Schrock et al., J Thorac Oncol, 11(9):1493-1502, 2016). In addition, hereditary amplification of the MET gene has been reported in approximately 3% of NSCLCs, and elevated MET protein expression has been reported in 25% of NSCLCs (Bubendorf et al., Lung Cancer, 111: 143-9, 2017)(Cappuzzo et al., J Clin Oncol, 27(10): 1667-74, 2009)(Fang et al., Oncotarget, 9(16): 12959-70, 2018)(Reis et al., Clin Lung Cancer, 19(4): e441-63, 2018)(Sterlacci et al., Virchows Arch, 471(1): 49-55, 2017). Increased MET gene amplification and expression are resistance mechanisms to epidermal growth factor receptor (EGFR) targeted therapies, and up to 25% of tumors resistant to third-generation TKIs may contain elevated MET through these mechanisms (Bean et al., PNAS, 104(52):20932-7, 2007)(Catenacci et al., Cancer, 123(6):1061-70, 2017)(Go et al., J Thorac Oncol, 5(3):283-9, 2010)(Le et al., JAMA Oncol, 4(2):210-6, 2018)(Zhang et al., Hum Pathol, 72:59-65, 2018).

[0175] Tumors with MET amplification or exon 14 deletion are responsive to MET TKIs (crizotinib [Package insert], Pfizer Pharmaceutical Company, New York, NY, 2017)(Angevin et al, Eur J Cancer, 87:131-9, 2017)(Camidge et al., J Clin Oncol, 32:abstract 8001, 2014)(Camidge et al., Nat Rev Clin Oncol, 16(6):341-55, 2019)(Paiket al., Cancer Discov, 5(8):842-9, 2015).

[0176] REGN5093 is a human bispecific antibody (bsAb) that binds to two different epitopes of MET with nanomolar affinity, blocks HGF binding to MET, and induces internalization and degradation of MET without inducing MET-driven biological responses. In preclinical studies, REGN5093 showed dose-dependent antitumor activity in immunodeficient mouse models of MET-driven cancers, including both exon 14 alteration models and MET amplification models. Unmet needs in lung cancer

[0177] The first-line treatment of advanced NSCLC is guided by the presence of molecular changes. Small molecule TKIs are usually used to target patients with tumors showing sensitizing mutations in EGFR, anaplastic lymphoma kinase (anaplastic lymphoma kinase, ALK) or c-ros oncogene 1 receptor tyrosine kinase (ROS1) fusions (Besse et al., Ann Oncol, 25 (8): 1475-84, 2014) (Ettinger et al., J Natl Compr Canc Netw 15 (4): 504-35, 2017) (Reck et al., Ann Oncol, 25 Suppl 3: 27-39, 2014). Patients without any of these activating mutations can receive immune checkpoint inhibitor antibodies that inhibit PD-1 receptors or PD-L1 with or without chemotherapy. In addition to these targeted systemic and immunotherapy approaches, advanced NSCLC is treated with platinum-based dual chemotherapy regimens with or without maintenance therapy (Besse et al., Ann Oncol, 25(8): 1475-84, 2014) (Ettinger et al., J Natl Compr Canc Netw 15(4): 504-35, 2017) (Reck et al., Ann Oncol, 25 Suppl 3: 27-39, 2014). In advanced NSCLC, long-term survival remains an unmet need. The overall survival of PD-1 / PD-L1 inhibitors is about 12 months, with some studies in patients treated with TKIs exceeding 2 years (Camidge et al., Nat Rev Clin Oncol, 16(6): 341-55, 2019).

[0178] Anti-PD-1 and anti-PD-L1 therapy has changed the standard of care for many patients with NSCLC (Topalian et al., NEJM, 366(26):2443-54, 2012). However, emerging data suggest that patients with MET-driven NSCLC may not experience equivalent benefit from agents targeting the PD-1 / PD-L1 axis, even for tumors that express high PD-L1 or exhibit high tumor mutational burden (TMB) (Sabari et al., Ann Oncol, 29(10):2085-91, 2018). This is consistent with data generated in lung cancers containing EGFR mutations or ALK rearrangements (Garassino et al., Lancet Oncol, 19(4):521-36, 2018) (Lee et al., JAMA Oncol, 4(2):210-16, 2018) (Peters et al., J Clin Oncol, 35(24):2781-89, 2017) and suggests that monotherapy with anti-PD-1 or anti-PD-L1 may not be the preferred treatment for patients with MET-driven disease. Therefore, there remains a large unmet need for treatments that improve response rates and survival in patients with MET-altered NSCLC. Purpose

[0179] The primary objective of the dose escalation (Phase 1) portion of this study is to evaluate the safety, tolerability, and pharmacokinetics (PK) of REGN5093 to determine the maximum tolerated dose (MTD) and / or define the recommended phase 2 dose (RP2D) of REGN5093 (in patients with MET-altered NSCLC). The secondary objective of the dose escalation (Phase 1) portion of this study is to evaluate the preliminary anti-tumor activity of REGN5093 as measured by objective response rate (ORR) according to the response evaluation criteria in solid tumors (RECIST 1.1).

[0180] The primary objective of the dose expansion (Phase 2) portion of this study is to evaluate the preliminary antitumor activity of REGN5093 as measured by ORR according to RECIST 1.1. The secondary objectives of the dose escalation (Phase 2) portion of this study are to evaluate the safety and tolerability of REGN5093 in each expansion cohort, as well as to evaluate the concentration and PK of REGN5093 in serum.

[0181] Secondary objectives of both parts of this study were to assess immunogenicity as measured by anti-drug antibody (ADA) to REGN5093, as well as to evaluate other measures of preliminary anti-tumor activity.

[0182] The exploratory objectives of the two parts of this study are to evaluate the relationship between the efficacy of REGN5093 and baseline MET alterations / mutations or amplification / expression and / or prior MET TKI treatment across cohorts, to assess pharmacodynamic changes in putative serum or plasma biomarkers, and to evaluate the impact of tumor mutation profiles in tissue and circulating tumor DNA (ctDNA amplification phase only) at baseline and after treatment on efficacy. Target Group

[0183] Adult patients aged ≥18 years (or the legal age of adults permitted to participate in clinical research according to country-specific regulations).

[0184] Dose escalation : Patients with advanced NSCLC, disease showing a previously documented presence of MET alterations in any of the following: exon 14 gene mutation, MET gene amplification, or elevated MET protein expression. In the dose escalation phase of this study, patients will be enrolled based on the documentation of any MET alteration defined by any of the above criteria, regardless of prior experience with a TKI targeting MET.

[0185] Dose expansion Patients with advanced NSCLC demonstrating MET-altered disease were assigned to cohorts based on the presence of the following prior documentation: disease with MET exon 14 mutations and according to prior MET-targeting TKI experience (Cohort 1A experienced MET TKI and Cohort 1B no prior MET TKI), high MET gene amplification (Cohort 2A no prior MET TKI), high MET protein overexpression (Cohort 2B no prior MET TKI), high MET gene amplification + high MET protein overexpression (Cohort 2C no prior MET TKI).

[0186] A documented MET status based on at least 1 test is sufficient to qualify a patient for the relevant cohort; testing in more than 1 category is not required. Thus, cohorts 2A and 2B may include patients with unknown overexpression or gene amplification status, respectively.

[0187] The dose expansion phase of this study is designed to further explore the safety and biological activity of REGN5093 at the RP2D. Patients will receive REGN5093 at the RP2D administered by IV over 30 minutes.

[0188] Patients will be recruited into separate cohorts based on previously documented MET-altered disease and based on prior experience with TKIs targeting MET. These cohorts are designed to provide relatively homogeneous patient populations based on different cutoff values ​​for 3 different types of biomarkers (MET exon 14 alterations, MET amplification, and MET protein overexpression).

[0189] The expanded groups are as follows (see Table 10): ●Expansion cohort 1A (NSCLC with MET exon 14 alterations; MET TKI-experienced) ●Expanded cohort 1B (NSCLC with MET exon 14 alterations; no prior MET TKI) ●Expansion cohort 2A (highly amplified NSCLC, MET GCN ≥5 and / or MET / CEP7 ratio ≥2 by FISH in tissue, or MET GCN ≥6 by NGS, or MET fold change ≥2 in ctDNA; no prior MET TKI) ●Expansion cohort 2B (high MET protein expression, IHC 3+ or H score ≥200; no prior MET TKI) ●Expanded cohort 2C (highly amplified NSCLC, MET GCN ≥5 and / or MET / CEP7 ratio ≥2 by FISH in tissue, or MET GCN ≥6 by NGS, or MET fold change ≥2 in ctDNA; and high MET protein overexpression, IHC 3+ or H score ≥200; no prior MET TKI) *If <40% tumor content in the biopsy is observed, the FISH or ctDNA local results must be within the specified range. Table 10: Criteria for assigning patients to the expansion cohort according to documented MET change status ●CEP7: The centromere of chromosome 7 ctDNA: circulating tumor DNA ●GCN: gene copy number IHC: Immunohistochemistry MET: mesenchymal epithelial transition TKI: tyrosine kinase inhibitor *Unless Group 1B is full **If <40% tumor content in the biopsy is observed, the FISH or ctDNA local results must be within the specified range.

[0190] Patients are not required to have prior testing for all 3 types of MET-altered disease to qualify for the relevant expansion cohort. For dose expansion, in cases where patients have not received prior MET-TKI and have more than one type of test data for MET-altered disease indicating that they may qualify for more than one cohort, they will be assigned to the expansion cohort in the following order of priority (if space is available): ●First priority group: MET exon 14 alterations = groups 1A and 1B ● Second priority (MET TKI-naive patients without MET exon 14 alterations, or patients with exon 14 alterations only after Cohort 1B is filled): MET gene amplification + protein expression will be considered for assignment to Cohorts 2A, 2B, and 2C.

[0191] The expansion cohort will have a Simon 2-stage design. Enrollment in the expansion cohort will be paused after the required number of patients in Phase 1 are enrolled in the cohort. Depending on the number of responders observed in Phase 1 and the corresponding Phase 1 criteria, the cohort will be stopped or expanded. Inclusion criteria

[0192] Patients must meet the following criteria to be eligible for inclusion in this study: ● Histologically confirmed advanced stage NSCLC for which no standard treatment options provide clinical benefit. Advanced is defined as unresectable or metastatic disease. Patients must have exhausted all available approved treatments for that patient. ●Have archived tumor tissue available unless discussed with the medical monitor. ●The existence of the following has been previously recorded: -For the dose escalation cohort: MET exon 14 gene mutation and / or MET gene amplification (by any local CLIA laboratory MET amplification call), and / or elevated MET protein expression (IHC ≥2+ or H score >150) - For dose expansion cohorts 1A and 1B: MET exon 14 mutation; MET TKI-experienced or prior MET TKI-naive, respectively - For dose expansion cohort 2A: MET gene highly amplified (MET GCN ≥5 and / or MET / CEP7 ratio ≥2 by FISH in tissue, or MET GCN ≥6 by NGS, or MET fold change ≥2 in ctDNA); no prior MET TKI -For dose expansion cohort 2B: MET protein is highly overexpressed (IHC 3+ or H score ≥200); no prior MET TKI) -For dose expansion cohort 2C: MET gene highly amplified (MET GCN ≥5 and / or MET / CEP7 ratio ≥2 by FISH in tissue, or MET GCN ≥6 by NGS, or MET fold change ≥2 in ctDNA) and MET protein highly overexpressed (IHC 3+ or H score ≥200); no prior MET TKI) *If the tumor content in the biopsy is observed to be <40%, the FISH or ctDNA local results must be within the specified range Willingness to provide tumor tissue from a newly obtained biopsy. A newly obtained biopsy is required at screening unless medically contraindicated and discussed with the medical monitor. For patients in the expansion cohort, the biopsy should be taken from a previously non-irradiated tumor site and not the only measurable target lesion. ● For the expansion cohort only: At least 1 lesion was measurable by RECIST 1.1. Tumor lesions in previously irradiated areas were considered measurable if such lesions showed progression after irradiation. Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1 Adequate organ and bone marrow function as documented by: Hemoglobin ≥9.0 g / dL Absolute neutrophil count ≥1.5 x 10 9 / L Platelet count ≥75 × 10 9 / L Serum creatinine ≤1.5 × ULN or estimated glomerular filtration rate (GFR) ≥30 mL / minute / 1.73 m 2 ●Adequate liver function: Total bilirubin ≤1.5 × ULN (≤3 × ULN if liver involvement) AST ≤2.5 × ULN (≤5 × ULN if liver involvement) ALT ≤2.5 × ULN (≤5 × ULN if liver involvement) Alkaline phosphatase ≤2.5 × ULN (≤5 × ULN if liver or bone involvement) Note: (a) Regardless of the above criteria, patients with tumor involvement of the liver will be excluded if the AST level is ≥3×ULN or the ALT level is ≥3×ULN, and the bilirubin level is ≥2×ULN. (b) Patients with Gilbert's syndrome do not need to meet the total bilirubin requirement, provided that their total bilirubin is not elevated above their historical level. Gilbert's syndrome must be appropriately documented as a past medical history. ●Adult patients aged ≥18 years (or the legal age of adults permitted to participate in clinical research according to country-specific regulations). ● Willing and able to comply with clinic visit and study-related procedures and requirements ●Must be willing and able to provide informed consent as specified by health department and institutional guidelines ● Provide an informed consent signed by the research patient or a legally acceptable representative. Exclusion criteria

[0193] Patients who meet any of the following criteria will be excluded from this study: Have been treated with approved systemic therapy or have participated in a study with any investigational agent or investigational device according to the following schedule: - For small molecule cytotoxins or other agents unlikely to interact with study drug: within 2 weeks or 5 half-lives (whichever is shorter) of prior therapy and at least 7 days from the first dose of study treatment. - Exceptions: Patients who have been treated with investigational immuno-PET agents or are enrolled in studies involving treatment with investigational immuno-PET agents are not excluded. ●Has not recovered from any acute toxic effect from prior therapy (ie, Grade ≤1 or baseline), except: - Laboratory changes as described in the inclusion criteria, and - Patients with grade ≤2 neuropathy For immune-related AEs affecting any organ system within 2 months prior to enrollment, an improvement trajectory of the irAE (to ≤ Grade 1 or baseline at enrollment) must be documented, and for those toxic effects that remain at Grade 1, 2 stability assessments at least 4 weeks apart must be documented. Note: AEs mediated by endocrine immunity controlled by hormonal or other non-immunosuppressive therapy that did not resolve prior to enrollment were allowed ●Radiation therapy or major surgery within 14 days of first dose of study drug or failure to recover from an AE (ie, Grade ≤1 or baseline), except for laboratory changes as described in the inclusion criteria and patients with Grade ≤2 neuropathy ● For expansion cohorts only: Prior treatment with biologic therapy targeting MET (function-blocking antibodies or ADCs). In addition, for expansion cohorts 1B, 2A, 2B, and 2C, prior treatment with any agent targeting MET, including small molecule tyrosine kinase inhibitors such as crizotinib, capmatinib, tepotinib ●For expanded cohort only: Another malignancy with the following exceptions: - Non-melanoma skin cancer who has undergone potentially curative treatment, or - Cervical cancer in situ or - Any other tumor that has been treated and the patient is considered to be in complete remission for at least 2 years prior to enrollment and does not require additional treatment during the study period ●Untreated or active primary brain tumor, CNS metastasis, leptomeningeal disease, or spinal cord compression - Exceptions: Patients with previously treated CNS metastases or spinal cord compression may participate provided that: - No signs of progression for at least 2 weeks prior to the first dose of study treatment and any neurological symptoms have returned to baseline Encephalitis, meningitis, organic brain disease (eg, Parkinson's disease), or uncontrolled seizures in the year before the first dose of study treatment ●Uncontrolled infection with human immunodeficiency virus, hepatitis B, or hepatitis C infection; or diagnosis of immunodeficiency Note: (a) Patients with known HIV infection whose infection is controlled (undetectable viral load [HIV RNA PCR] and CD4 count greater than 350, either spontaneously or on a stable antiviral regimen) are permitted. Patients whose HIV infection is controlled will be monitored according to local standards. (b) Patients with known hepatitis B (HepBsAg+) whose infection is controlled (serum HBV DNA PCR is below the limit of detection and who are receiving antiviral treatment for hepatitis B) are permitted. Patients whose infection is controlled must undergo regular monitoring of HBV DNA. Patients must maintain antiviral treatment for at least 6 months after the last dose of the investigational study drug. (c) Patients with known hepatitis C virus antibody positive (HCV Ab+) whose infection was controlled (no detectable HCV RNA by PCR, either spontaneously or in response to a previous successful course of anti-HCV therapy) were allowed. Any infection requiring hospitalization or IV anti-infective therapy within 2 weeks prior to the first dose of study treatment. ● Placeholder for deleted exclusion criteria ● Placeholder for deleted exclusion criteria ● Mental or substance abuse disorders known to interfere with participation in research requirements ●Any medical condition, comorbidity, physical examination findings, or metabolic dysfunction or clinical laboratory abnormality that, in the opinion of the investigator, makes the patient unsuitable for participation in a clinical trial due to a high safety risk and / or the potential to affect the interpretation of the study results Women with a positive serum hCG pregnancy test at the screening / baseline visit. Women who are breastfeeding are also excluded ●Women of childbearing potential* or men who do not wish to use highly effective contraception before the initial dose / first treatment, during the study, and for at least 6 months after the last dose. Highly effective contraceptive measures include: - Stable use of combined (estrogen and progestin) hormonal contraception (oral, intravaginal, transdermal) or progestin-only hormonal contraception (oral, injectable, implantable) associated with ovulation suppression for the initiation of 2 or more menstrual cycles prior to screening -Intrauterine device (IUD); intrauterine hormone-releasing system (IUS) - Bilateral tubal ligation - A vasectomy partner (provided the vasectomy male partner is the only sexual partner of the study participant and the partner has been medically evaluated as a successful procedure) - and / or sexual abstinence *Women of childbearing potential are defined as fertile women after menarche until they are postmenopausal, unless permanently sterilized. Permanent sterilization methods include hysterectomy, bilateral salpingectomy, and bilateral oophorectomy. The postmenopausal state is defined as the absence of menstruation for 12 consecutive months without other medical reasons. High follicle stimulating hormone (FSH) levels in the postmenopausal range can be used to confirm the postmenopausal state in women who are not using hormonal contraception or hormone replacement therapy. However, in the absence of 12 months of amenorrhea, FSH measurements alone are insufficient to confirm the occurrence of the postmenopausal state. The above definition is based on the Clinical Trial Facilitation Group (CTFG) guidelines. Pregnancy testing and contraception are not required for women with a history of hysterectomy or tubal ligation. Sexual abstinence is considered to be an effective approach only if it is defined as abstention from heterosexual intercourse during the entire risk period associated with the study drug. The reliability of sexual abstinence needs to be evaluated based on the duration of the clinical trial and the patient's preferences and usual lifestyle. Periodic abstinence (calendar method, symptom-temperature method, postovulation method), withdrawal method (excretion with ejaculation), spermicide-only method, and lactational amenorrhea method (LAM) are unacceptable contraceptive methods. Female and male condoms should not be used together. Methods - Study Design

[0194] Patients with documentation of at least 1 of the 3 major categories of MET alterations were included in this first-in-human (FIH) open-label study: exon 14 skipping mutations, MET gene amplification (MET GCN ≥5 and / or MET / CEP7 ratio ≥2 by FISH in tissue, or MET GCN ≥6 by NGS, or MET fold change ≥2 in ctDNA), and MET protein overexpression (immunohistochemistry 3+ or H score ≥200). Patients were not required to have prior disease testing for all 3 categories of MET alterations.

[0195] Figure 6 The study flow from the initial screening period to post-treatment follow-up is depicted. Description of Study Cohorts and Dose Escalation

[0196] Patients underwent a screening procedure to determine eligibility prior to the first administration of REGN5093 and within 28 days after signing the informed consent form (ICF). Dose escalation

[0197] In the dose escalation phase, a series of 3 DLs of REGN5093 were studied: 500, 1000 and 2000 mg, administered IV every three weeks (Q3W) by 30-minute infusion (see Table 11). Table 11: Dose escalation 1 If DL1 is not tolerated, 1DL-1 can be used 2 DL1a and DL2a are alternative dose levels to be studied if the immediately higher dose is not tolerated and the immediately lower dose shows insufficient clinical and PK signals. A modified 3+3 dose escalation design ("4+3") will be used (Le Tourneau et al., J Natl Cancer Inst, 101(10):708-20, 2009). Dose escalation will be performed until the MTD is reached or an expansion dose (RP2D) is selected based on safety / tolerability and sufficient response evidence. If the initial or subsequent levels are considered intolerant, a dose reduction (DL-1) cohort and an intermediate cohort may be recruited to study the intermediate dose. In order to further evaluate safety and collect biological information, the sponsor may decide (in consultation with the investigator) to recruit up to 6 additional patients at any DL that is considered tolerable. A schematic diagram of the study design is in Figure 7 Shown in.

[0198] The dose limiting toxicity (DLT) evaluation period will be 21 days, starting on Day 1 of Cycle 1. Although at least 3 patients at each DL are required to be evaluable for DLT, in order to maximize the efficiency of Phase 1 dose escalation while maintaining patient safety, 4 patients will be enrolled at each DL to prevent patients from discontinuing before DLT evaluation can be performed. The tolerability rules are as follows:

[0199] Tolerability of the DL was considered achieved if all potentially DLT-evaluable patients completed the 21-day DLT period without DLT (0 of 3 patients or 0 of 4 patients).

[0200] NOTE: If 3 patients complete the DLT period without experiencing a DLT, but there is a fourth patient in the DLT evaluation period, the DL is considered to be tolerable only if the fourth patient completes the DLT evaluation period or discontinues treatment before a DLT evaluation can be performed.

[0201] If there is 1 DLT in 3 or 4 patients evaluable for DLT, 4 or 3 more patients will be enrolled, respectively, for a total of 7 patients. If there is 1 DLT in 6 or 7 patients, the dose is considered to be tolerable. If there are 2 or more DLTs in 2 to 7 evaluable patients, the MTD is reached.

[0202] At the highest DL tolerated, an additional 3 to 4 patients may be enrolled to evaluate further safety, for a total of 6 to 10 DLT-evaluable patients. The dose is considered acceptable if there are 0 to 1 DLT in 6 to 8 patients, or up to 2 DLTs in 9 to 10 patients.

[0203] Once all of the initial 3 or 4 patients enrolled in a cohort have been observed for at least 21 days and have completed safety assessments on Day 22 (Cycle 2 Day 1), and the data have been reviewed at the Dose Escalation Review Meeting, they will be escalated to the next dose cohort.

[0204] After the required number of patients have been enrolled in a given dose cohort, enrollment will be paused for DLT evaluation (although screening of the next dose cohort may begin before the current dose is confirmed to be safe). The dose escalation review meeting will be chaired by a designated member of the sponsor's clinical team (usually the medical or study lead) and will be attended by at least the sponsor's medical / study lead and the Global Patient Safety Lead; other individuals, including the investigator, may also be included. Dose cohorts are stopped, expanded, or escalated according to the dose escalation criteria. Dose-limiting toxicity

[0205] Dose-limiting toxicity (DLT) is any toxicity that prevents escalation to a higher dose as specified in the protocol. The DLT observation period for determining the safety of dose escalation is defined as 21 days starting on Day 1 of Cycle 1, which is intended to monitor the safety and tolerability of the first dose of REGN5093. In order to be evaluated for DLT, patients must have: - Has received at least 1 dose of study drug and has been monitored for at least 21 days after the first administration of study drug. - or is subjected to a DLT (defined below) prior to completion of the DLT Period.

[0206] For patients who experience an AE, the duration of the DLT observation period may be longer, and the duration of the AE must be evaluated to determine whether the event is a DLT.

[0207] If an event occurred during the DLT observation period, the event was counted as a DLT for the cohort involved, regardless of whether the patient remained on study treatment and / or continued to participate in study procedures. Dose-limiting toxicity definition

[0208] DLT is generally defined as any of the following treatment-emergent toxicities, excluding toxicities clearly related to disease progression or intercurrent illness. The grade of these toxicities is defined according to CTCAE version 5.0. Hematological toxicity: - Grade 4 neutropenia lasting >7 days -Grade 4 thrombocytopenia - Grade 4 Anemia - Grade 3 thrombocytopenia with bleeding - Grade 3 or higher febrile neutropenia (fever ≥38.5°C with an absolute neutrophil count [ANC] <1.0 × 10 9 / L) or ≥ grade 3 neutropenia with documented infection Non-hematologic toxicity: - Non-hematologic Grade ≥ 3 toxicities, except: a. Baldness b. Grade 3 nausea, vomiting, or diarrhea unless persistent (>72 hours duration) after initiation of supportive care measures as prescribed by the treating physician c. Clinically insignificant laboratory abnormalities - Clinically significant Grade ≥ 3 laboratory values ​​that required medical intervention or resulted in hospitalization - Abnormal liver function test consistent with Hy's law (Temple, J Allergy Clin Immunol, 117(2):391-97, 2006) or ALT or AST>3×ULN and bilirubin>2×ULN.

[0209] The frequency, onset, and severity of toxicities, as well as the success of standard medical management and dosing interruptions / delays, were analyzed to determine whether a given toxicity should be considered a DLT for dose escalation purposes.

[0210] Generally, due to the limited clinical experience with the new biomolecule REGN5093, any AE will be treated as unexpected.

[0211] Treatment-emergent adverse events that appear to meet the definition of a DLT will be discussed between the Sponsor and the Investigator. The final decision on whether a TEAE meets the definition of a DLT will be based on a careful review of all relevant data and consensus between the Medical / Study Lead and the designated Safety Lead from Global Patient Safety. You can also consult the researchers.

[0212] Regardless of whether the patient remains on study treatment and / or continues to participate in study procedures, if an event meeting DLT criteria occurs during the DLT observation period, the event is counted as a DLT for the cohort involved. Maximum tolerated dose

[0213] The MTD was defined as the DL immediately below the level at which dosing was stopped due to the occurrence of 2 or more DLTs in up to 7 evaluable patients. If the study was not stopped due to the occurrence of DLTs, the MTD was considered undetermined.

[0214] If the MTD is not reached, an RP2D may be selected for further evaluation based on clinical, PK, and / or biomarker data indicating that a pharmacologically active dose has been reached in combination with available safety information.

[0215] REGN5093 is provided as a lyophilisate in sterile single-use vials. During the trial, it is planned to introduce a new presentation form: sterile solution. Each vial contains REGN5093 at a concentration of 25 mg / ml. Both presentation forms will include labeled vials in labeled cartons.

[0216] Instructions for dose preparation are provided in the pharmacy leaflet.

[0217] No premedication is required prior to administration of REGN5093. REGN5093 is administered by IV infusion over 30 minutes Q3W. Prohibited drugs

[0218] According to the dosing regimen specified in this study, patients were not allowed to receive any standard or investigational agents for tumor treatment except REGN5093 while participating in this study.

[0219] No systemic treatment for cancer was allowed during this study period.

[0220] Patients were not allowed to receive live vaccines during this study.

[0221] Radiation therapy was not allowed during the study period, with the following exceptions: Palliative treatment of lesions (e.g., radiation) for local tumor control was allowed after communication with the Sponsor. Palliative radiation therapy for pain management at the site of bone disease or brain lesions was allowed after discussion with the Sponsor (as long as the lesion was not subsequently evaluated for treatment response).

[0222] The investigator may, at his or her discretion, administer any other medication that is deemed necessary for the patient's welfare and is not expected to interfere with the evaluation of the study drug. Permitted drugs

[0223] Gonadotropin-releasing hormone agonist therapy may be continued and is not contraindicated. Hormone replacement therapy is permitted. Inhaled, topical, ophthalmic, or intranasal steroids are permitted. Treatment of bone metastases (bisphosphonates, denosumab) and systemic corticosteroids are permitted. The use of high doses of steroids for extended periods of time requires discussion with the medical monitor. Security

[0224] The safety and tolerability of REGN5093 were monitored by clinical assessment of AEs, physical examination (complete and limited), repeated measurements of vital signs (temperature, blood pressure, pulse, and respiration), 12-lead electrocardiogram (ECG), and laboratory evaluations (including standard hematology, chemistry, and urinalysis). Vital signs, including temperature, sitting blood pressure, pulse, and respiration, were collected at pre-dose time points.

[0225] AE is any unfortunate medical occurrence in a patient taking a study drug that may or may not have a causal relationship with the study drug. Therefore, AE is any unfavorable and unexpected sign (including abnormal laboratory findings), symptom or disease associated with the duration of study drug use, whether or not considered to be related to the study drug (ICHE2A Guidelines, Clinical Safety Data Management: Definitions and Standards for Rapid Reporting, October 1994).

[0226] A SAE is any unfortunate medical occurrence at any dose: ● Resulting in death - includes all deaths, even those that appear to be completely unrelated to the study drug (e.g., a car accident in which the patient was a passenger). ● Is life-threatening - in the investigator's opinion, the patient is at immediate risk of death at the time of the event. It does not include AEs that occur in a more serious form and may lead to death. ● Requires hospitalization or prolongation of existing hospitalization. Hospitalization is defined as admission to a hospital or emergency room for longer than 24 hours. Prolongation of existing hospitalization is defined as a hospital stay longer than initially expected for the event or due to the occurrence of a new AE as determined by the investigator or treating physician. ● Resulting in permanent or severe disability / incapacity (significant disruption of the individual's ability to carry out normal life functions) ●Congenital malformation / birth defect ●A major medical event. A major medical event may not be immediately life-threatening or result in death or hospitalization but may endanger the patient or may require intervention to prevent one of the other serious outcomes listed above (e.g., intensive treatment in the emergency department or at home for allergic bronchospasm; hematologic cachexia or seizures that do not result in hospitalization; or development of drug dependence or abuse).

[0227] Hospitalization or death solely due to manifestations consistent with the typical progression of the underlying malignancy was not considered a SAE. Effectiveness

[0228] Radiographic tumor response was used to determine the overall response for each patient as defined by RECIST 1.1 (Eisenhauer et al., Eur J Caner, 45(2):228-47, 2009). Radiographic disease assessment informed the following calculations: ORR, defined as the percentage of patients with a complete response (CR) or partial response (PR) ●DOR, defined as the time from the first response of CR or PR to the first radiographic progression or death due to any cause in patients with a confirmed CR or PR. In the absence of radiographic progression or death before the analysis cutoff date or the date of initiation of further anticancer treatment, the DOR will be censored at the date of the last valid response assessment that did not show progression, performed before the analysis cutoff date or the start of further anticancer treatment, whichever is earlier. DCR, defined as the percentage of patients with a BOR of CR, PR, or stable disease (SD) PFS was defined as the time from the first administration of study treatment to the first radiographic progression or death from any cause. The same censoring rules as for DOR were used. ●OS was defined as the time from the first administration of study treatment to death due to any cause. For patients who did not die, OS was censored at the last date the patient was known to be alive. OS was assessed based on investigator-reported survival data.

[0229] Diagnostic quality CT with contrast and enhanced MRI Contrast-enhanced MRI is the preferred imaging modality for assessing radiographic tumor response. In patients in whom contrast agents are strictly contraindicated, plain CT of the chest and plain MRI of the body other than the chest are sufficient. The chest, abdomen, and pelvis must be imaged, as well as any other known or suspected sites of disease. If more than one imaging modality is used for screening, the most accurate imaging modality according to RECIST 1.1 (Appendix 1) should be used when recording data. The same imaging modality and technique used for screening should be used for all subsequent evaluations.

[0230] For screening, contrast-enhanced brain MRI or CT should be performed in patients with a known history of treatment for brain metastases. (If contraindicated, non-enhanced MRI was performed).

[0231] Additional sites of known disease should be imaged during screening.

[0232] Diagnostic quality (≤5 mm slices) enhanced CT scans of the chest and abdomen and any other known disease sites (e.g., neck) will be performed at screening, on day 1 of the 2nd and 3rd treatment cycles, every three months after the last visit, and at any time when disease progression is suspected. The scan will include a description of the tumor location, and up to 5 of the largest overt disease masses (no more than 2 per organ) should be selected as target lesions according to RECIST 1.1 and measured by the longest diameter (if non-nodal lesions) and short axis (for nodal lesions). All lesions should be evaluated and recorded. If a CT scan is not feasible, an MRI scan may be performed.

[0233] For each patient, the same measurements and the same technique must be used to evaluate each lesion throughout the study. If a patient inadvertently misses a scheduled tumor evaluation or a technical error prevents an evaluation, the patient can continue treatment until the next regularly scheduled evaluation unless there are signs of clinical progression. If disease progression is suspected at any time during the treatment period based on clinical or laboratory findings (and before the next regularly scheduled evaluation), an unscheduled tumor evaluation should be performed. Procedures and Assessment

[0234] Antitumor activity was assessed by computed tomography (CT) or magnetic resonance imaging (MRI). The safety and tolerability of REGN5093 were monitored by clinical assessment of AEs, physical examination (complete and limited), repeated measurements of vital signs (temperature, blood pressure, pulse, and respiration), 12-lead electrocardiogram (ECG), and laboratory evaluations (including standard hematology, chemistry, and urinalysis).

[0235] Blood was collected to assess the PK and concentration of REGN5093 in serum and immunogenicity (ADA) in serum; and for additional biomarker assessments. Additional biomarkers were measured in serum or plasma. Exploratory predictive and pharmacodynamic biomarkers associated with REGN5093 treatment exposure, clinical activity, or underlying disease were studied using samples from collected serum, plasma, archived tumor tissue, and tumor biopsy tissue in the study, tumor DNA (including circulating tumor DNA), and tumor RNA samples. Study End Points

[0236] The primary endpoints of the dose-escalation (Phase 1) portion of this study are: Safety, measured by the incidence and severity of treatment-emergent adverse events (TEAEs), adverse events of special interest (AESIs), serious adverse events (SAEs), and laboratory abnormalities of grade ≥3 during the treatment period and up to 90 days after the last dose Tolerability, measured by the incidence of dose-limiting toxicities (DLTs) of REGN5093 from the first dose to the end of the DLT observation period - REGN5093 concentrations in serum over time.

[0237] The primary endpoint of the dose-expansion (Phase 2) portion of this study was ORR according to RECIST 1.1, which was defined as the percentage of patients with a confirmed CR or PR according to RECIST 1.1 criteria for best overall response (BOR).

[0238] The secondary endpoint of the dose-escalation portion of this study was ORR according to RECIST 1.1.

[0239] Secondary endpoints of the dose expansion portion of this study are: Safety, measured by the incidence and severity of TEAEs, AESI / SAEs, and laboratory abnormalities of grade 3 or higher ●PK and concentration of REGN5093 in serum over time.

[0240] The secondary endpoints for both periods of this study were: ●Duration of response (DOR) according to RECIST 1.1. ●Disease control rate (DCR) according to RECIST 1.1. ●Progression free survival (PFS) according to RECIST 1.1. OS • Immunogenicity as measured by ADA of REGN5093.

[0241] The exploratory endpoints for the two parts of this study are: Response to REGN5093 by type (and extent) of baseline MET change and prior experience with MET-targeting TKIs ●Response to REGN5093 by baseline tumor mutation status. Example 8. Safety, Tolerability, and Efficacy of REGN5093 in Patients with Advanced NSCLC with MET Alterations

[0242] REGN5093 had therapeutic benefit in patients with MET-altered NSCLC and showed promising efficacy signals with a tolerable safety profile.

[0243] REGN5093 is being studied as a monotherapy and is administered intravenously every 3 weeks in a dose escalation cohort (Phase 1), followed by an expansion phase (Phase 2). For each patient, the study consists of a 28-day screening phase followed by a 3-week cycle of REGN5093 monotherapy. 2000 mg of REGN5093 is the recommended Phase 2 dose; previous dose levels were 500 mg and 1000 mg.

[0244] Tumor measurements were performed at baseline and every 6 weeks until disease progression, withdrawal of consent, death, or initiation of another anticancer treatment.

[0245] Tumor tissue (archive and research tumor biopsies) is obtained and used for retrospective analysis of MET alterations (and additional biomarker analysis if tissue permits). A newly obtained biopsy is also required at the time of screening unless it is considered unsafe.

[0246] Sixty-nine patients received REGN5093 in both dose escalation and dose expansion periods. Patient characteristics were consistent with the heavily treated population, with a median of 2.5 prior lines of treatment (range: 1 to 8). Most patients had an ECOGPS of 1 (80%). Most patients had non-squamous histology (93.6%), and EGFR mutations were present in 37.2% of patients (Table 12). The mean age of the study population was 66 years, 53.8% were male, and 70.5% were Asian. Safety Data

[0247] No dose-limiting toxicity (DLT) was observed. REGN5093 showed a similar safety profile in the dose escalation and dose expansion phases, especially only 6 (9%) patients had grade 1 / 2 peripheral edema (Table 13). At the time of data cutoff, nine (13%) patients were still on treatment and 60 (87%) patients had discontinued treatment. The main reasons for discontinuation of treatment were: 52 (75%) patients had disease progression; five (7%) patients had patient decisions; and only three (4%) patients had adverse events. Tumor response

[0248] Partial responses (assessed by the investigator) were observed in the subgroup of patients with exon 14 alterations in DNA or deletions leading to exon 14 skipping who had not received prior MET TKI treatment and in patients with MET gene amplification and / or MET protein overexpression (Tables 14 and Figure 8 ).

[0249] ORR in patients with centrally confirmed MET alterations: · 33% (3 / 9): MET exon 14 mutation in tumor tissue or by NGS in ctDNA (naive of MET TKI) · 25% (5 / 20): MET gene amplification (GCN ≥ 5 by FISH or NGS in tumor tissue) · 23% (5 / 22): MET protein overexpression (IHC 3+ in ≥ 50% of tumor cells) · 36% (5 / 14): MET protein overexpression (IHC 3+ in ≥ 75% of tumor cells) · 50% (4 / 8): MET protein overexpression (IHC 3+ in ≥ 90% of tumor cells). Pharmacokinetics

[0250] Serum exposure of REGN5093 was linear and dose proportional over the dose range of 500 mg to 2000 mg Q3W intravenous (IV), and serum concentrations at 2000 mg Q3W IV were similar in the dose escalation cohort and multiple expansion cohorts (see Figure 9). The elimination half-life estimated by non-compartmental analysis was 15 days over the 3-week dosing interval. in conclusion

[0251] In this heavily treated patient population with advanced NSCLC with MET alterations, REGN5093 monotherapy demonstrated an acceptable safety profile. No DLTs were observed. Eighty-six percent of patients experienced TEAEs of any grade. Twenty-six percent of patients experienced TEAEs of ≥ Grade 3. Three (4%) patients discontinued treatment due to TEAEs. REGN5093 exposure in serum was shown to increase in a dose-dependent manner. REGN5093 monotherapy demonstrated preliminary efficacy signals in patients with MET exon 14 alterations in DNA or deletions leading to exon 14 skipping, as well as in patients with MET gene amplification and / or MET protein overexpression. Tumor responses were enhanced with centrally validated biomarker selection. Example 9: Predictive Biomarkers of Response to REGN5093

[0252] In some cases, when treating MET-altered advanced non-small cell lung cancer (aNSCLC), patient selection may be guided by the use of predictive biomarkers of response to REGN5093. REGN5093 monotherapy is being studied in patients with MET-altered aNSCLC. Tumor measurements are performed at baseline and Q6W until progression, withdrawal of consent, death, or initiation of another anticancer therapy.

[0253] Provided herein are selection criteria that can be used to improve the percentage of responders when treating aNSCLC. In some aspects, the identification of MET actionable mutations can be used for patient selection for MET targeted therapy in late stage (2L+, i.e., patients who have received two or more previous treatments) NSCLC. MET exon 14 skipping / deletion is an oncogenic driver of 1L NSCLC, leading to the loss of c-CbI binding sites, which weakens receptor degradation and ultimately leads to increased MET signaling. In 2L+NSCLC, MET gene amplification is a resistance mechanism to EGFR tyrosine kinase inhibitor (TKI) treatment. MET protein overexpression enhances but does not select for treatment response to MET TKI, especially in NSCLC populations driven by TKI resistance to MET amplification ( Fig.10 ).

[0254] Both tumor biopsies and liquid biopsies can be used to identify and confirm MET alterations. Exon 14 alterations can be determined by genetic sequencing of tumor biopsies or ctDNA obtained from blood samples. Similarly, MET gene amplification can be assessed by genetic sequencing or fluorescence in situ hybridization [FISH; GCN or MET: chromosome 7 centromere (CEP7) ratio] of tumor biopsies, or sequencing of ctDNA obtained from blood samples. MET protein expression can be assessed in tumor biopsies by immunohistochemistry using, for example, a specific c-MET antibody to stain for total MET protein ( Fig.11 ).

[0255] In some cases, assessing ctDNA supplements tissue profiling and overcomes the limitations of biopsy collection and analysis. For example, some tumors are located in areas where it is difficult or impossible to obtain tissue biopsies. The ctDNA assessment captures all active "drivers" from all tumor sources in the body (i.e., both primary and metastatic tumors) and allows the identification of mutations that are not present in a single tissue biopsy to overcome spatial heterogeneity. As tumors evolve over time (either inherently or in response to one or both of the treatments), temporal heterogeneity can be overcome by assessing tissue and ctDNA samples obtained simultaneously, thereby obtaining higher consistency and accurate coverage.

[0256] MET amplification status was assessed using tissue and / or ctDNA by FISH and NGS, for example, using gene copy number (GCN) thresholds in tissue by fluorescence in situ hybridization (FISH) (GCN ≥ 5) or NGS (GCN ≥ 6) and / or ≥ 2.2× in ctDNA by NGS to assess the efficacy of additional MET inhibitors.

[0257] Using multiple next-generation sequencing (NGS) panels on tissue and ctDNA ( CD× (tissue-based 324 genomes) and LiquidCD× (a blood-based 324 gene panel) is an exemplary gene panel, but other gene panels are contemplated as being available herein) to centrally confirm MET exon 14 (i.e., assayed by one central laboratory to control for any variability in sample collection and assay implementation). Exemplary exon 14 alterations include, but are not limited to, D1010N, D1010fs*19, D1010Y, D1010H, or R1004P mutations and exon 14 skipping.

[0258] Fig.12 A study design is provided. Multiple cohorts include patients with confirmed MET alterations as determined by a given patient's medical record. Fig.12 As shown in , patient recruitment during the trial extension phase was based on documented MET alterations (MET exon 14 alterations in DNA or deletions leading to exon 14 skipping, MET gene amplification, and / or MET protein overexpression) and association with clinical response. Fig.13 As can be seen in the 65 patients who received a 2000 mg dose of REGN5093, 9 patients had a partial response. The distribution of responders according to the group is as follows: Group 1A (experienced MET Ex 14TKI): 0; Group 1B (not received MET Ex 14TKI: 4; Group 2A (MET amplification): 0; Group 2B (MET overexpression): 1; Group 2C (MET amplification and overexpression): 3; DL3 (dose level 3, MET amplification and OE): 1.

[0259] like Fig.14 As shown in Figure 2, regardless of EGFR mutation status, tumor responses were observed in MET TKI-naive patients with centrally confirmed (1) MET exon 14 (4 / 15) or 27% or (2) MET amplification + OE (5 / 14) 36%. Fig.15 As shown in , by centralized analysis, the overall response rate (%ORR) of the subgroup with specific MET alterations was higher than that of the total population. Example 10: Bypass resistance mutations at baseline and clinical response to REGN5093

[0260] Resistance to MET therapy can be intrinsic or acquired in response to prior therapy in the pre-treated 2L+ NSCLC patient population. The number of prior therapies received by patients ranged from one to eight, with a median of 2.5 ( Fig.16 ). Previous treatments for patients in this study included chemotherapy, immune checkpoint inhibitors (ICIs), and EGFR inhibitors. Patients who initially responded to treatment in some cases acquired additional mutations that promoted disease progression after treatment.

[0261] Bypass gene detection of nonsynonymous variants, such as single nucleotide variants (SNVs), insertions and deletions (Indels), frameshifts, nonsense mutations, splice variants or gene fusions, and copy number variants (CNVs), such as gene amplifications or deletions (in ctDNA), complements tissue results and provides a more comprehensive tumor profile of MET×MET resistance mechanisms. The different sensitivities of the FMI NGS panel are based on the variant allele frequency (VAF) of each of the 324 genes present in each platform. Fig.17 Detection of nonsynonymous variants and CNVs and overlap of detection in ctDNA and tissue are illustrated. NSV detection in ctDNA was 39%; in tissue, 33%; and in both, 28%. CNV detection in ctDNA was 9%; in tissue, 84%; and in both, 6%.

[0262] Fig.18 Unbiased co-clustering of baseline somatic mutations detected in both ctDNA and tumor tissue by cohort assignment, EGFR status, and centrally confirmed MET alterations is illustrated. Fig.19 Shown as Fig.18 Several baseline somatic mutations based on the clustered dataset were identified in non-responders that could lead to MET bypass resistance mechanisms and potentially affect clinical response to REGN5093, even in the presence of MET oncogenic drivers.

[0263] Fig. 20A classification and examples of bypass alterations detected in study patients with centrally confirmed MET oncogenic drivers who did not respond to REGN5093 are provided. On-target receptor mutations include MET TKI resistance mutations (i.e., MET Y1230C, MET D1228H, MET D1228N), MET fusions / rearrangements (MET:MET gene rearrangements), and MET gene silencing (loss of function) (i.e., DNMT3A, TET2). Resistance mechanisms involving selective or concurrent tyrosine kinase-driven receptor activation include tyrosine kinase receptor (TKR) and TK ligand gene amplifications (e.g., FGFR1-4 Amp, FGF14 Amp, NTRK1-3 Amp, MERTK Amp, ERBB2 / 3 Amp, and VEGFA Amp), TKR-activating mutations (EGFR L858R, G719S, E709A, E746_A750del, S752_I759del), and oncogenic fusions and rearrangements (MKRN-BRAF fusion). Resistance mechanisms involving activation of downstream proliferation / survival / anti-apoptotic pathways, including the JAK2 / STAT3 pathway (e.g., JAK2 V617F; SFK amp); RAS / RAF / MEK / MAPK pathway (e.g., KRAS G12A / V and G12D / V; GNAS R201H, MKRN-BRAF fusion, BRAF S602Y; RICTORAmp; MAP2K1 K57N; RAS mut / gain; RAF mut; ERK-MAPK amp); PI3K / AKT / MTOR pathway (e.g., PI3KCA H1047L, E545K, E542K, N345K; IDH1 R132L, MTOR E2338Q, AKT2 Amp, RICTOR Amp; PI3K mut; PTEN loss); TP53 mutations (e.g., TP53 R280T, TP53 R248Q) and cell cycle mutations (e.g., CDK4 Amp, CDK6 Amp, CCND1 Amp, CCNE1 Amp).

[0264] Gene amplifications identified in patients with confirmed Met amplification and overexpression who did not respond to RENG5093 included the following: HGF, EPH, EGFR, BRAF, BCL2L1, PI3KCB, KRAS, AKT2, ATR, VEFGA, FGF, CCND, CCNE, CDK6, RAD21, and MYC. The following gene deletions were also identified: CDKN2A, CDKN2B, MTAP, and RBM10. The MET bypass resistance mechanisms identified in these patients are provided in Table 15. Table 15: MET bypass resistance mechanisms - MET overexpression and MET amplification

[0265] Gene amplification was identified in patients with confirmed MET amplification (not MET overexpression) mainly EGFR mutants that were unresponsive to REGN5093: EGFR, BRAF, PI3KC2G, KRAS, HGF, EPHA3, ERCC4, RICTOR, RAD21, LYN, MYC, MDM2, CDK 4 / 6, FgF3 / 4 / 19, FGF10 and CCND1. Gene deletions in these patients include: CDKN2A, CDKN2B, MTAP, TEK and BCOR. MET bypass resistance mechanisms identified in these patients are provided in Table 16. Table 16: MET bypass resistance mechanisms - MET amplification and EGFR mutations

[0266] Gene amplifications identified in patients with MET exon 14 alterations who had not received TKI but were non-responders to REGN5093 included: MDM2, EGFR, FGFR1, ERBB3, CDK4, GNA13, MYC, RPTOR, TERC, IKZF1, EZH2, SDHA, SOX, WHSC1L1, and ZNF703. Gene deletions identified in these patients included CDKN2A, CDKN3A, and MTAP. MET bypass resistance mechanisms identified in these patients are provided in Table 17. Table 17: MET bypass resistance mechanisms - MET Ex 14, TKI naive

[0267] Gene amplifications identified in patients with MET exon 14 changes who have undergone TKI but are non-responders to REGN5093 include: EGFR, RAF1, PI3KC2G, CDK4, CEBPA, CDKN1A, CARD11, MYC, RICTOR, VEGFA, CD22, DDR1, RAC1, NBN, FGF19, MDM2, NFKBIA, CCND1, INPP4B, PPARG, PMS2, GATA4, SDHA and RAD21. Gene deletions identified in these patients include CDKN2A, CDKN3A and MTAP. MET bypass resistance mechanisms identified in these patients are provided in Table 18. Table 18: MET bypass resistance mechanisms - MET Ex 14, TKI-experienced Example 11: Circulating Biomarkers of Target Engagement

[0268] Soluble MET (sMET) is an extracellular domain fragment of MET that yields the receptor in tumor tissue free of protease cleavage. Total sMET and HGF were measured by ELISA.

[0269] The total concentration of REGN5093 in serum was several times higher than the total sMET concentration, indicating that receptor occupancy saturation was achieved at the dose-expanded 2000 mg Q3W dose regimen ( Fig.21 ).

[0270] HGF is a ligand for the MET receptor, and when REGN5093 binds to the MET receptor in tumors, the ligand is displaced and increased (in circulation). Both circulating HGF (cHGF) and total sMET levels increased after dosing, indicating target engagement, but neither baseline nor post-treatment levels of sMET or cHGF were significantly associated with clinical response ( Fig. 22 ). in conclusion

[0271] Of the 36 pts who received the 2000 mg dose, 6 had partial responses (5 with prior anti-PD-(L)1 therapy). These responses occurred in: 2 / 5 pts with exon 14 skipping mutations who had not received prior MET tyrosine kinase inhibitor (TKI) therapy (Cohort 1B); 0 / 10 pts with exon 14 skipping mutations who had been previously treated with TKIs (Cohort 1A); and 4 / 21 pts with MET gene amplification, protein overexpression, or both who had not received prior MET TKI therapy (Cohorts 2A to C).

[0272] REGN5093 monotherapy induced tumor responses in patients with MET-altered aNSCLC. Heterogeneity in response rates was observed between subgroups of MET alterations in MET TKI-naive patients, with a response rate of 4 / 15 (27%) in pts with MET Ex14 mutations and 5 / 16 (36%) in pts with MET amplification and overexpression, but this was based on a small sample size.

[0273] In clinical nonresponders, certain baseline somatic mutations co-occurred with MET alterations, which served as potential bypass resistance mechanisms and influenced clinical responses to REGN5093 monotherapy.

[0274] Total concentrations of REGN5093 in serum were several-fold higher than total sMET concentrations, supporting the selection of a 2000 mg Q3W dosing regimen. Total sMET and cHGF levels increased after dosing, indicating target engagement of REGN5093, but neither baseline nor post-treatment changes in total sMET and cHGF concentrations correlated with response. Table 19: List of informal sequences

Claims

1. A method of treating non-small cell lung cancer (NSCLC), reducing NSCLC tumor growth, and / or causing NSCLC regression in a subject having a tumor containing a MET alteration, the method comprising administering to the subject a dose of about 250 to 2000 mg of a bispecific antibody, the bispecific antibody comprising: a first antigen binding domain (D1) comprising three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) within a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 1 and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) within a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 9; and a second antigen binding domain (D2) comprising three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) within a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO:5 and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) within a light chain variable region (LCVR), wherein the heavy chain variable region (HCVR) comprises the amino acid sequence of SEQ ID NO:5 and the light chain variable region (LCVR) comprises the amino acid sequence of SEQ ID NO:9; wherein D1 specifically binds to the first epitope of human MET; and Among them, D2 specifically binds to the second epitope of human MET.

2. The method of claim 1, wherein the subject is further selected to have one or more of the following criteria: (i) Not received MET TKI; (ii) histologically confirmed NSCLC; (iii) MET exon 14 alterations in DNA or deletions leading to exon 14 skipping; (iv) MET gene amplification; (v) elevated MET protein expression (IHC ≥ 2+ or H score > 150); (vi) MET exon 14 alterations in DNA or deletions leading to exon 14 skipping and treatment with MET TKI; (vii) MET exon 14 alterations in DNA or deletions leading to exon 14 skipping and no prior MET TKI treatment; (viii) highly amplified MET gene (MET GCN ≥ 5 and / or MET / CEP7 ratio ≥ 2 by FISH in tissue, or MET GCN ≥ 6 by NGS, or MET fold change ≥ 2 in ctDNA) and not receiving MET TKI; (ix) MET protein is highly overexpressed (IHC 3+ or H score ≥200) and has not received MET TKI; and / or (x) MET gene is highly amplified (MET GCN ≥ 5 and / or MET / CEP7 ratio ≥ 2 by FISH in tissue or MET GCN ≥ 6 by NGS, or MET fold change ≥ 2 in ctDNA), MET protein is highly overexpressed (IHC 3+ or H score ≥ 200), and has not received MET TKI.

3. The method of claim 1, wherein the MET alteration is an exon 14 alteration in DNA, MET gene amplification, or MET protein overexpression.

4. The method of claim 1, wherein the MET alteration is an exon 14 alteration or a deletion resulting in exon 14 skipping in the DNA.

5. The method of claim 1, wherein the MET alteration is an exon 14 mutation.

6. The method of claim 5, wherein the exon 14 mutation is D1010N, D1010fs*19, D1010Y, D1010H, or R1004P.

7. The method of claim 1, wherein the MET alteration is MET gene amplification.

8. The method of claim 1, wherein the overexpression of MET protein means that the expression of MET protein in tumor tissue is higher than that in normal tissue.

9. The method of claim 1, wherein the MET alteration is identified using ctDNA from a blood sample obtained from the patient prior to treatment.

10. The method of claim 1, wherein the MET alteration is identified in a tissue sample obtained from the patient prior to treatment.

11. The method of any one of claims 1 to 10, wherein the subject is MET tyrosine kinase inhibitor (TKI) naive.

12. The method of any one of claims 1 to 10, wherein the subject has received a prior anti-cancer therapy comprising one or more of the following: a PD-1 inhibitor, an EGFR inhibitor, a PD-L1 inhibitor, surgery, radiation therapy, or chemotherapy.

13. The method of claim 11, wherein the prior anti-cancer therapy comprises a PD-1 inhibitor or a PD-L1 inhibitor.

14. The method of claim 11, wherein the prior anti-cancer therapy comprises an EGFR inhibitor.

15. The method of claim 11, wherein the subject is resistant or inadequately responsive to a previous treatment, or has relapsed following a previous treatment.

16. The method of any one of claims 1 to 10, wherein the subject has not received prior anti-cancer therapy.

17. The method of any one of claims 1 to 16, wherein the tumor has an EGFR mutation.

18. The method of claim 17, wherein the EGFR mutation is selected from L858R, G719S, E709A, E746_A750del and S752_I759del.

19. The method of any one of claims 1 to 18, wherein the tumor is non-squamous NSCLC.

20. The method of any one of claims 1 to 18, wherein the tumor is squamous NSCLC.

21. The method of any one of claims 1 to 18, wherein the NSCLC is metastatic.

22. The method of claim 21, wherein the NSCLC has metastasized to the brain.

23. The method of claim 21, wherein the NSCLC has metastasized to the liver.

24. The method of any one of claims 1 to 23, wherein the NSCLC is unresectable.

25. The method of any one of claims 1 to 24, wherein D1 comprises the HCDR1 amino acid sequence shown in SEQ ID NO:2; the HCDR2 amino acid sequence shown in SEQ ID NO:3; the HCDR3 amino acid sequence shown in SEQ ID NO:4; the LCDR1 amino acid sequence shown in SEQ ID NO:10; the LCDR2 amino acid sequence shown in SEQ ID NO:11; and the LCDR3 amino acid sequence shown in SEQ ID NO:

12.

26. The method of any one of claims 1 to 25, wherein D1 comprises a HCVR comprising the amino acid sequence of SEQ ID NO: 1 and a LCVR comprising the amino acid sequence of SEQ ID NO:

9.

27. The method of any one of claims 1 to 26, wherein D2 comprises the HCDR1 amino acid sequence shown in SEQ ID NO:6; the HCDR2 amino acid sequence shown in SEQ ID NO:7; the HCDR3 amino acid sequence shown in SEQ ID NO:8; the LCDR1 amino acid sequence shown in SEQ ID NO:10; the LCDR2 amino acid sequence shown in SEQ ID NO:11; and the LCDR3 amino acid sequence shown in SEQ ID NO:

12.

28. The method of any one of claims 1 to 27, wherein D2 comprises a HCVR comprising the amino acid sequence of SEQ ID NO:5 and a LCVR comprising the amino acid sequence of SEQ ID NO:

9.

29. The method of any one of claims 1 to 28, wherein the bispecific antibody is administered at a dose of about 250 mg, 500 mg, 750 mg, 1000 mg, 1500 mg, or 2000 mg.

30. The method of any one of claims 1 to 29, wherein the bispecific antibody is administered at a dose of 500 mg.

31. The method of any one of claims 1 to 29, wherein the bispecific antibody is administered at a dose of 1000 mg.

32. The method of any one of claims 1 to 29, wherein the bispecific antibody is administered at a dose of 2000 mg.

33. The method of any one of claims 1 to 32, wherein the bispecific antibody is administered intravenously, subcutaneously, or intraperitoneally.

34. The method of any one of claims 1 to 33, wherein the bispecific antibody is administered once every three weeks.

35. The method of any one of claims 1 to 33, wherein the bispecific antibody is administered 3 weeks after the immediately preceding dose.

36. The method of any one of claims 1 to 35, wherein the treatment produces a therapeutic effect selected from the group consisting of tumor growth delay, reduction in tumor cell number, reduction in metastasis, tumor regression, improved survival, partial response, and complete response.

37. The method of claim 36, wherein tumor growth is delayed by at least 10 days compared to untreated subjects.

38. The method of claim 36, wherein tumor growth is inhibited by at least 50% compared to untreated subjects.

39. A method for treating or inhibiting the growth of NSCLC, the method comprising: (1) Selecting subjects with tumors containing MET alterations; as well as (2) administering to the subject (a) a bispecific antibody at a dose of about 250 mg, 500 mg, 750 mg, 1000 mg, 1500 mg, or 2000 mg, the bispecific antibody comprising: a first antigen binding domain (D1) comprising three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) within a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 1 and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) within a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 9; and a second antigen binding domain (D2) comprising three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) within a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO:5 and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) within a light chain variable region (LCVR), wherein the heavy chain variable region (HCVR) comprises the amino acid sequence of SEQ ID NO:5 and the light chain variable region (LCVR) comprises the amino acid sequence of SEQ ID NO:9; wherein D1 specifically binds to the first epitope of human MET; and Among them, D2 specifically binds to the second epitope of human MET.

40. The method of claim 39, wherein the administration of step (2) is performed once every 3 weeks.

41. The method of claim 39, wherein the subject is further selected to have one or more of the following criteria: (i) Not received MET TKI; (ii) histologically confirmed NSCLC; (iii) MET exon 14 alterations in DNA or deletions leading to exon 14 skipping; (iv) MET gene amplification; (v) elevated MET protein expression (IHC ≥ 2+ or H score > 150); (vi) MET exon 14 alterations in DNA or deletions leading to exon 14 skipping and treatment with MET TKI; (vii) MET exon 14 alterations in DNA or deletions leading to exon 14 skipping and no prior MET TKI treatment; (viii) highly amplified MET gene (MET GCN ≥ 5 and / or MET / CEP7 ratio ≥ 2 by FISH in tissue, or MET GCN ≥ 6 by NGS, or MET fold change ≥ 2 in ctDNA) and not receiving MET TKI; (ix) MET protein is highly overexpressed (IHC 3+ or H score ≥200) and has not received MET TKI; and / or (x) The MET gene is highly amplified (MET GCN ≥ 5 and / or MET / CEP7 ratio ≥ 2 by FISH in tissue, or MET GCN ≥ 6 by NGS, or MET fold change ≥ 2 in ctDNA), the MET protein is highly overexpressed (IHC 3+ or H score ≥ 200), and the patient has not received MET TKI.

42. The method of claim 39, wherein the MET alteration is an exon 14 alteration in DNA, MET gene amplification, or MET protein overexpression.

43. The method of claim 39, wherein the MET alteration is an exon 14 alteration or a deletion resulting in exon 14 skipping in the DNA.

44. The method of claim 39, wherein the MET alteration is an exon 14 mutation.

45. The method of claim 44, wherein the exon 14 mutation is D1010N, D1010fs*19, D1010Y, D1010H, or R1004P.

46. ​​The method of claim 39, wherein the MET alteration is MET gene amplification.

47. The method of claim 39, wherein the overexpression of MET protein means that the expression of MET protein in tumor tissue is higher than that in normal tissue.

48. The method of claim 39, wherein the MET alteration is identified using ctDNA from a blood sample obtained from the patient prior to treatment.

49. The method of claim 39, wherein the MET alteration is identified in a tissue sample obtained from the patient prior to treatment.

50. The method of claim 39, wherein the subject is further selected as having NSCLC with an EGFR mutation.

51. The method of claim 39, wherein the subject is further selected as having non-squamous NSCLC.

52. The method of claim 39, wherein the subject is further selected as having squamous NSCLC.

53. The method of any one of claims 39 to 52, wherein the NSCLC is metastatic.

54. The method of claim 53, wherein the NSCLC has metastasized to the brain.

55. The method of claim 53, wherein the NSCLC has metastasized to the liver.

56. The method of any one of claims 39 to 55, wherein the NSCLC is unresectable.

57. A method for treating a tumor, the method comprising: (a) selecting a subject suffering from NSCLC; (b) determining that the tumor exhibits a MET alteration selected from an exon 14 alteration in DNA or a deletion resulting in exon 14 skipping, MET gene amplification, and / or Met protein overexpression, comprising (i) obtaining a tissue sample and / or a liquid sample from the subject; and (ii) assessing the tissue sample for MET gene amplification using fluorescence in situ hybridization of tumor tissue or by next generation sequencing of tumor tissue and / or ctDNA, and / or assessing the tissue sample for Met protein overexpression using immunohistochemistry, and / or assessing the liquid sample for exon 14 mutations using ctDNA; and if the tumor exhibits MET alterations, then (c) administering one or more doses of the MET×MET bispecific antibody to a subject in need thereof, Wherein the Met×Met anti-tumor therapy comprises a bispecific antibody, wherein the bispecific antibody comprises: a first antigen binding domain (D1) comprising three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) within a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 1 and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) within a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 9; and a second antigen binding domain (D2) comprising three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) within a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO:5 and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) within a light chain variable region (LCVR), wherein the heavy chain variable region (HCVR) comprises the amino acid sequence of SEQ ID NO:5 and the light chain variable region (LCVR) comprises the amino acid sequence of SEQ ID NO:9; wherein D1 specifically binds to the first epitope of human MET; and Among them, D2 specifically binds to the second epitope of human MET.

58. A method for identifying a candidate for Met×Met anti-tumor therapy, the method comprising obtaining a tissue sample and / or a fluid sample from a subject with NSCLC; and assessing the tissue sample and / or the fluid sample for MET alterations selected from exon 14 alterations or deletions leading to exon 14 skipping in DNA, MET gene amplification, and / or Met protein overexpression, wherein the presence of at least one Met alteration in the tissue sample or fluid sample identifies the subject as a candidate for anti-tumor therapy; and wherein the Met×Met anti-tumor therapy comprises a bispecific antibody comprising a first antigen binding domain (D1), the first antigen binding domain (D1) comprising three heavy chain complementarity determining regions (HCDR1, HCDR2 and HCDR3) within a heavy chain variable region (HCVR) and three light chain complementarity determining regions (LCDR1, LCDR2 and LCDR3) within a light chain variable region (LCVR), the heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 1, the light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 9; and a second antigen binding domain (D2), comprising three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) within a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO:5, and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) within a light chain variable region (LCVR), wherein the heavy chain variable region (HCVR) comprises the amino acid sequence of SEQ ID NO:5, and the light chain variable region (LCVR) comprises the amino acid sequence of SEQ ID NO:9; wherein D1 specifically binds to the first epitope of human MET; and Among them, D2 specifically binds to the second epitope of human MET.

59. The method of claim 58, wherein the MET alteration is an exon 14 alteration or a deletion resulting in exon 14 skipping in the DNA.

60. The method of claim 58, wherein the MET alteration is an exon 14 mutation.

61. The method of claim 60, wherein the exon 14 mutation is D1010N, D1010fs*19, D1010Y, D1010H, or R1004P.

62. The method of claim 58, wherein the MET alteration is MET gene amplification.

63. The method of claim 58, wherein the overexpression of MET protein means that the expression of MET protein in tumor tissue is higher than that in normal tissue.

64. The method of claim 58, wherein the MET alteration is identified using ctDNA from a blood sample obtained from the patient prior to treatment.

65. The method of claim 58, wherein the MET alteration is identified in a tissue sample obtained from the patient prior to treatment.

66. A method for monitoring the efficacy of a MET×MET bispecific antibody in a subject having NSCLC with a MET alteration, the method comprising: (i) obtaining a tissue sample and / or a fluid sample from the subject, and assessing somatic mutations in one or more genes in the tissue sample and / or the fluid sample, wherein the somatic mutations are selected from: a. On-target MET receptor gene mutations conferring resistance to MET TKIs, present in MET Ex14 Mut patients with prior TKI Exp and MET gene silencing (loss of function); b. TK driven receptor activation selected from TK receptor and ligand gene amplification and TKR activating mutations; as well as c. Activating gene mutations in a pathway selected from the group consisting of the JAK2 / STAT3 pathway, the RAS / RAF / MEK / MAPK pathway, the PI3K / AKT / MTOR pathway, TP53 mutations, and cell cycle gene amplifications; (ii) administering a MET×MET bispecific antibody to the subject; (iii) repeating steps (i) and (ii) during treatment; wherein acquisition of a mutation in one or more of the genes indicates resistance to treatment and / or indicates a poor prognosis.

67. The method of claim 66, wherein: a. The on-target MET receptor gene mutation is selected from: MET Y1230C, MET D1228H, MET D1228N; and the MET gene silencing (loss of function) is selected from somatic mutations of DNMT3A and TET2; b. The TKR activating mutation is selected from: EGFR L858R, EGFR G719S, EGFR E709A, EGFR E746_A750del and EGFR S752_I759del; and c. The JAK2 / STAT3 pathway mutation is JAK2 V617F; the RAS / RAF / MEK / MAPK pathway mutation is selected from: KRAS G12A / V, GNAS R201H, MKRN-BRAF fusion, BRAF S602Y, RICTOR Amp and MAP2K1 K57N; the PI3K / AKT / MTOR pathway mutation is selected from: PIK3CAH1047L, PIK3CAE545K, PIK3CAE542K, PIK3CAN345K, IDH1 R132L and MTOR E2338Q; the PI3K / AKT / MTOR pathway amplification is selected from: AKT2 Amp and RICTOR Amp; the TP53 mutation is selected from TP53 R280T and TP53 R248Q; and the cell cycle gene amplification is selected from: CDK4 Amp, CDK6 Amp, CCND1 Amp and CCNE1 Amp.

68. A method for treating NSCLC in a subject, the method comprising: (i) obtaining a liquid sample from the subject and determining MET amplification in ctDNA from the liquid sample, and (ii) administering a MET×MET bispecific antibody to the subject; wherein steps (i) and (ii) are repeated every three weeks, and wherein loss of MET amplification after repeating step (ii) indicates a durable response to treatment.

69. The method of claim 68, wherein the bispecific antibody is administered intravenously, subcutaneously, or intraperitoneally.

70. The method of claim 68, wherein the bispecific antibody is administered once every three weeks. E 71. The method of E 68, wherein the bispecific antibody is administered 3 weeks after the immediately preceding dose.

72. A method for determining a therapeutically effective amount of a MET×MET bispecific antigen binding molecule, the method comprising: (i) administering a dose of the bispecific antibody to a patient in need thereof, and (ii) measuring soluble MET in a blood sample, wherein a maximal increase in soluble MET (sMET) indicates receptor occupancy saturation and a therapeutically effective amount of the bispecific antibody.

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