Methods for treating non-small cell lung cancer using mesenchymal-epithelial transition factor (MET)-targeted drugs

Abstract

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

Description

[Technical Field]

[0001] The present invention relates to a METxMET bispecific antibody that specifically binds to hepatocyte growth factor receptor (c-Met or MET) and modulates MET signaling for use in the treatment of non-small cell lung cancer (NSCLC).

[0002] Sequence Listing An official copy of the Sequence Listing is being submitted electronically via EFS-Web contemporaneously herewith as an XML-formatted Sequence Listing entitled 11305WO01_Sequence Listing ST.26, with a creation date of August 31, 2023, and approximately 32,768 bytes in size. This Sequence Listing is a part of the present specification and is incorporated herein by reference in its entirety. [Background technology]

[0003] Hepatocyte growth factor (HGF) (also known as scatter factor [SF]) is a heterodimeric paracrine growth factor that exerts its activity through interaction with the HGF receptor (HGFR). HGFR is the product of the c-Met oncogene, also known as MET. MET is a receptor tyrosine kinase composed of a transmembrane beta chain linked via a disulfide bridge to an extracellular alpha chain. Binding of HGF to MET activates the kinase catalytic activity of MET, leading to phosphorylation of Tyr 1234 and Tyr 1235 of the beta chain and subsequent activation of downstream signaling pathways.

[0004] Overexpression, activation, or amplification of MET and / or HGF has been shown to be associated with non-small cell lung cancer (NSCLC) (Non-Patent Document 1). MET amplification is believed to be a key driver of oncogenesis in NSCLC. In addition, mutations resulting in MET exon 14 deletion have been described as an oncogenic driver in a subset of NSCLC. Tumor cell lines with MET gene amplification are highly dependent on MET for proliferation and survival. Preclinical data have demonstrated the involvement of MET signaling in resistance to targeted therapy in NSCLC.

[0005] Both preclinical and recent clinical results have shown that tumors with these genetic abnormalities respond to MET inhibitors, thereby identifying MET as a cancer driver. A variety of monovalent MET-blocking antibodies are in clinical development for the treatment of various cancers (see Patent Document 1; Patent Document 2; Patent Document 3; Patent Document 4; Patent Document 5 and Patent Document 6; and Patent Documents 7 and 8). These antibodies include onartuzumab (MetMab) and emibetuzumab (Non-Patent Document 2 and Non-Patent Document 3). Some of these antibodies block ligand-dependent MET signaling but are less effective at blocking ligand-independent MET activation. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] U.S. Patent No. 5,686,292 [Patent Document 2] U.S. Patent No. 5,646,036 [Patent Document 3] U.S. Patent No. 6,099,841 [Patent Document 4] U.S. Patent No. 7,476,724 [Patent Document 5] U.S. Patent No. 9,260,531 [Patent Document 6] U.S. Patent No. 9,328,173 [Patent Document 7] U.S. Patent Application Publication No. 2014 / 0349310 [Patent Document 8] U.S. Patent Application Publication No. 2005 / 0233960 [Non-patent literature]

[0007] [Non-Patent Document 1] Sierra and Tsao, Ther. Adv. Med. Oncol., 3(Suppl 1):S21-S35, 2011 [Non-patent document 2] Xiang et al., Clin. Cancer Res. 19(18):5068-78, 2013 [Non-patent document 3] Rosen et al., Clin. Cancer Res., published October 10, 2016, doi:10.1158 / 1078-0432.CCR-16-1418 Summary of the Invention [Problem to be solved by the invention]

[0008] There remains a significant unmet medical need for improved anticancer drugs that potently block both ligand-dependent and ligand-independent MET signaling. [Means for solving the problem]

[0009] Provided herein are methods using bispecific antibodies and fragments thereof that bind to the human c-Met receptor protein (MET x MET) to target non-small cell lung cancer (NSCLC) cells with MET abnormalities, such as DNA deletions resulting in exon 14 aberrations or skipping of exon 14, MET gene amplification, or overexpression of the MET protein. While patients with NSCLC can be treated according to accepted standard therapies, tumors develop resistance to treatment, and eventually, patients may have exhausted all approved and available therapies appropriate for them. Resistance to MET therapy can be intrinsic or acquired, for example, chemotherapy, immune checkpoint inhibitors, or targeted therapies.

[0010] By identifying such patients whose tumors have a MET aberration and treating them with a METxMET bispecific antibody, surprising benefits were achieved, as described herein.

[0011] Accordingly, provided herein is a method of treating NSCLC, reducing NSCLC tumor growth, and / or causing regression of NSCLC in a subject afflicted with a tumor that has a MET aberration, comprising administering to the subject a 250-2000 mg dose of a METxMET bispecific antibody.

[0012] Provided herein are methods of treating or inhibiting the growth of NSCLC, comprising: Further provided are methods comprising: (1) selecting a subject having a tumor with a MET aberration; and (2) administering to subject (a) a dose of about 250 mg, 500 mg, 750 mg, 1000 mg, 1500 mg, or 2000 mg of a METxMET bispecific antibody. In some embodiments, the administering in step (2) occurs once every three weeks.

[0013] Provided herein is a method for treating a tumor, comprising: (a) Selecting subjects with NSCLC; (b) determining that the tumor exhibits a MET abnormality selected from the group consisting of an exon 14 abnormality or deletion in DNA resulting in exon 14 skipping, MET gene amplification, and / or Met protein overexpression; (i) obtaining a tissue sample and / or a body fluid sample from a subject; and (ii) evaluating tissue samples for MET gene amplification using fluorescence in situ hybridization in tumor tissue or by next-generation sequencing in tumor tissue and / or ctDNA and / or evaluating tissue samples for Met protein overexpression using immunohistochemistry and / or evaluating body fluid samples for exon 14 mutations using ctDNA. including; and if the tumor exhibits MET abnormalities, (c) administering one or more doses of the MET×MET bispecific antibody to a subject in need thereof. Still further provided is a method comprising:

[0014] Provided herein is a method for identifying candidates for MetxMet anti-tumor therapy, comprising: Obtaining a tissue and / or fluid sample from a subject with NSCLC; and evaluating the tissue and / or fluid samples for MET abnormalities selected from the group consisting of exon 14 abnormalities or deletions resulting in exon 14 skipping in DNA, MET gene amplification, and / or Met protein overexpression. wherein the presence of at least one Met abnormality in the tissue or fluid sample identifies the subject as a candidate for anti-tumor therapy, wherein the METxMET anti-tumor therapy comprises a METxMET bispecific antibody.

[0015] MET abnormalities provided herein include abnormalities of exon 14 in DNA, amplification of the MET gene, or overexpression of the MET protein.

[0016] In some embodiments, the MET abnormality is an abnormality in exon 14 in DNA or a deletion that results in the skipping of exon 14. The abnormality in exon 14 in DNA or a deletion that results in the skipping of exon 14 includes missense abnormalities, deletions, splice site changes, and deletion of the entire exon that results in the skipping of exon 14 of the MET gene. In some embodiments, the MET abnormality is a mutation in exon 14. In some embodiments, mutations in exon 14 include, but are not limited to, D1010N, D1010fs ... * 19, D1010Y, D1010H or R1004P.

[0017] In some embodiments, the MET abnormality is MET gene amplification. In some embodiments, a highly amplified MET gene has a MET gene copy number (GCN) of ≧5 by FISH and / or a MET to chromosome 7 centromere (MET / CEP7) ratio of ≧2 in tissue, or a MET GCN of ≧6 by next generation sequencing (NGS), or a MET fold change of ≧2 in ctDNA.

[0018] In some embodiments, the MET abnormality is overexpression of MET protein. Overexpression of MET protein can be evaluated by MET immunohistochemistry (IHC) in tumor tissue, where the expression is higher than that in normal tissue. In some embodiments, elevated expression of MET protein is evaluated by IHC ≧2+ or H score >150. In some embodiments, highly overexpressed MET protein is evaluated by IHC 3+ or H score ≧200.

[0019] In some embodiments, MET abnormalities are identified using ctDNA from a blood sample obtained from the patient prior to treatment, i.e., a body fluid biopsy.

[0020] In some embodiments, the MET abnormality is identified in a tissue sample obtained from the patient prior to treatment, i.e., a tumor biopsy.

[0021] In some embodiments, NSCLC tumor has EGFR mutation.These mutations can include but are not limited to L858R, G719S, E709A, E746_A750del and S752_I759del.In some embodiments, the subject is selected as having NSCLC with one or more mutations in EGFR gene.

[0022] In some embodiments, the subject has not received a prior anti-MET cancer therapy. In some embodiments, the subject is tyrosine kinase inhibitor (TKI) naive. In other words, the subject has not received a prior treatment with a MET-targeting TKI. In some embodiments, the subject has received a prior anti-cancer therapy comprising one or more of a MET-targeting TKI, a PD-1 inhibitor, an EGFR inhibitor, a PD-L1 inhibitor, surgery, radiation therapy, or chemotherapy. In some aspects, the prior anti-cancer therapy comprises a MET-targeting TKI. In some aspects, the prior anti-cancer therapy comprises a PD-1 inhibitor or a PD-L1 inhibitor. In some aspects, the prior anti-cancer therapy comprises an EGFR inhibitor. In some aspects, the subject is resistant or has not adequately responded to the prior therapy, or has relapsed after the prior therapy.

[0023] A MET TKI-naive patient has not received any prior treatment with a MET TKI; similarly, a patient who has received treatment with a MET TKI is MET TKI-experienced. A PD-(L)1-experienced patient has received prior treatment with a PD-1 inhibitor (such as, but not limited to, pembrolizumab, nivolumab, cemiplimab, dostarlimab, and retifanlimab) or a PD-L1 inhibitor (such as, but not limited to, atezolizumab, avelumab, and durvalumab) and is considered PD-(L)1-experienced; similarly, a PD-(L)1-naive patient (i.e., PD-(L)1-naive) has not received prior treatment with a PD-1 inhibitor or PD-L1 inhibitor. A patient may be EGFR inhibitor-experienced, i.e., has received treatment with an EGFR inhibitor. Exemplary EGFR inhibitors include, but are not limited to, erlotinib, afatinib, gefitinib, osimertinib, dacomitinib, cetuximab, panitumumab, dacomitinib, and the like.

[0024] In some embodiments, the subject is further selected according to one or more of the following criteria: (i) Subjects were MET TKI-naive; (ii) subjects have histologically confirmed NSCLC; (iii) NSCLC presents with MET-exon 14 abnormalities in DNA or deletions resulting in exon 14 skipping; (iv) NSCLC presents with MET gene amplification; (v) NSCLC presents elevated expression of MET protein (IHC ≥2+ or H score >150); (vi) NSCLC presenting with MET exon 14 abnormality or deletion resulting in exon 14 skipping in DNA and prior MET TKI treatment; (vii) NSCLC presents with abnormalities of MET exon 14 in DNA or deletions resulting in exon 14 skipping and untreated MET TKI; (viii) NSCLC presents with a highly amplified MET gene (MET GCN ≥5 and / or MET / CEP7 ratio ≥2 in tissue by FISH or MET GCN ≥6 by NGS or MET fold change ≥2 in ctDNA) and MET TKI-naïve; (ix) NSCLC presenting with highly overexpressed MET protein (IHC 3+ or H-score ≥200) and MET TKI-naive; and / or (x) NSCLC presents with highly amplified MET gene (MET GCN ≥5 and / or MET / CEP7 ratio ≥2 in tissue by FISH or MET GCN ≥6 by NGS or MET fold change ≥2 in ctDNA), highly overexpressed MET protein (IHC 3+ or H-score ≥200), and MET TKI-naïve.

[0025] In some embodiments, cancer is non-squamous NSCLC.In some embodiments, cancer is NSCLC squamous cell carcinoma.In some aspects, NSCLC is metastatic, for example, cancer is metastasized to brain and / or liver.In some aspects, NSCLC is unresectable.

[0026] Provided herein is a method for monitoring the efficacy of treatment with a METxMET bispecific antibody in a subject with NSCLC with a MET aberration, comprising: (i) obtaining a tissue and / or fluid sample from a subject; and a. On-target MET receptor gene mutations and MET gene silencing (loss of function) conferring resistance to MET TKIs found in MET Ex14 Mut patients with prior TKI Exp; b. TK driver receptor activation selected from alternative or parallel TK receptor and ligand gene amplifications and TKR activating mutations; and c. Activating gene mutations in pathways selected from the group consisting of the JAK2 / STAT3 pathway, the RAS / RAF / MEK / MAPK pathway, and the PI3K / AKT / MTOR pathway, TP53 mutations, and cell cycle gene amplification: evaluating the tissue and / or fluid sample for somatic mutations in one or more genes selected from the group consisting of: (ii) administering a MET×MET bispecific antibody to a subject; and (iii) repeating steps (i) and (ii) over the course of treatment. Includes; The acquisition of mutations in one or more genes is indicative of resistance to therapy and / or a poor prognosis; A method is provided.

[0027] In some embodiments, the on-target MET receptor genetic mutation is selected from the group consisting of MET Y1230C, MET D1228H, MET D1228N; and the MET gene silencing (loss of function) is selected from somatic mutations in DNMT3A and TET2.

[0028] In some embodiments, the TKR activating mutation is selected from the group consisting of EGFR L858R, EGFR G719S, EGFR E709A, EGFR E746_A750del, and EGFR S752_I759del.

[0029] In some embodiments, 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, PIK3CA N345K, 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.

[0030] In some embodiments, patients have confirmed MET amplification and MET overexpression but have developed gene amplification in one or more of HGF, EPH, EGFR, BRAF, BCL2L1, PI3KCB, KRAS, AKT2, ATR, VEFGA, FGF, CCND, CCNE, CDK6, RAD21, and MYC. In some embodiments, patients have confirmed MET amplification and MET overexpression but have developed gene deletions in one or more of CDKN2A, CDKN2B, MTAP, and RBM10. In some embodiments, patients have confirmed MET amplification and MET overexpression but have developed one or more point mutations provided in Table 15.

[0031] In some embodiments, patients have confirmed MET amplification (but not MET overexpression) and may be EGFR mutated, but have developed gene amplifications in one or more of EGFR, BRAF, PI3KC2G, KRAS, HGF, EPHA3, ERCC4, RICTOR, RAD21, LYN, MYC, MDM2, CDK 4 / 6, FgF3 / 4 / 19, FGF10, and CCND1. In some embodiments, patients have confirmed MET amplification (but not MET overexpression) and may be EGFR mutated, but have developed gene deletions in one or more of CDKN2A, CDKN2B, MTAP, TEK, and BCOR. In some embodiments, patients have confirmed MET amplification (but not MET overexpression) and may be EGFR mutated, but have developed one or more of the point mutations provided in Table 16.

[0032] In some embodiments, patients have a confirmed MET exon 14 abnormality and are TKI-naïve, but have developed gene amplifications in one or more of MDM2, EGFR, FGFR1, ERBB3, CDK4, GNA13, MYC, RPTOR, TERC, IKZF1, EZH2, SDHA, SOX, WHSC1L1, and ZNF703. In some embodiments, patients have a confirmed MET exon 14 abnormality and are TKI-naïve, but have developed gene deletions in one or more of CDKN2A, CDKN3A, and MTAP. In some embodiments, patients have a confirmed MET exon 14 abnormality and are TKI-naïve, but have developed one or more point mutations provided in Table 17.

[0033] In some embodiments, patients have a confirmed MET exon 14 abnormality and are TKI-experienced, but have developed gene amplifications 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 embodiments, patients have a confirmed MET exon 14 abnormality and are TKI-experienced, but have developed gene deletions in one or more of CDKN2A, CDKN3A, and MTAP. In some embodiments, patients have a confirmed MET exon 14 abnormality and are TKI-experienced, but have developed one or more of the point mutations provided in Table 18.

[0034] Also provided herein is a method for treating NSCLC in a subject, comprising: (i) obtaining a body fluid sample from the subject and determining MET amplification in ctDNA from the body fluid sample, and (ii) administering a METxMET bispecific antibody to the subject; steps (i) and (ii) are repeated once every three weeks, and loss of MET amplification after step (ii) is repeated is an indication of a sustained response to the treatment.

[0035] Provided herein are methods for determining a therapeutically effective amount of a METxMET bispecific antibody, comprising: (i) administering a dosage 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 saturation of receptor occupancy and a therapeutically effective amount of the bispecific antibody.

[0036] In some embodiments, a subject is administered a METxMET bispecific antibody in an amount of about 250 mg to about 5000 mg, i.e., a dose of about 250 mg to about 5000 mg, or a dose of 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 embodiments, the dose is about 250 mg of METxMET bispecific antibody. In some embodiments, the dose is about 500 mg of the METxMET bispecific antibody. In some embodiments, the dose is about 1000 mg of the METxMET bispecific antibody. In some embodiments, the dose is about 2000 mg of the METxMET bispecific antibody. In some embodiments, the dose is 250 mg, 500 mg, 1000 mg, or 2000 mg of the METxMET bispecific antibody.

[0037] In some embodiments, the bispecific antibody is administered intravenously, subcutaneously, or intraperitoneally.The bispecific antibody can be administered once every 5 days, once a week, once every 2 weeks, once every 3 weeks, once every 4 weeks, once a month, once every 5 weeks, once every 6 weeks, or once every 2 months.In some embodiments, the bispecific antibody is administered once every 3 weeks.In some embodiments, the bispecific antibody is administered 3 weeks after the previous dose.

[0038] In some embodiments, the treatment produces a therapeutic effect selected from the group consisting of delayed tumor growth, reduced metastasis, reduced tumor cell count, tumor regression, improved survival, partial response, and complete response. In some embodiments, 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 untreated subjects. In some embodiments, 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 untreated subjects.

[0039] The METxMET bispecific antibody comprises a first antigen-binding domain (D1) comprising three heavy chain complementarity determining regions (HCDR1, HCDR2 and HCDR3) within the heavy chain variable region (HCVR) comprising an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 1 and three light chain complementarity determining regions (LCDR1, LCDR2 and LCDR3) within the light chain variable region (LCVR) comprising an amino acid sequence that is 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 the heavy chain variable region (HCVR) comprising an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO:5, and three light chain complementarity-determining regions (LCDR1, LCDR2, and LCDR3) within the light chain variable region (LCVR) comprising an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO:9.

[0040] In some embodiments, D1 specifically binds to the first epitope of human MET.

[0041] In some embodiments, D1 comprises an HCDR1 amino acid sequence set forth in SEQ ID NO: 2; an HCDR2 amino acid sequence set forth in SEQ ID NO: 3; an HCDR3 amino acid sequence set forth in SEQ ID NO: 4; an LCDR1 amino acid sequence set forth in SEQ ID NO: 10; an LCDR2 amino acid sequence set forth in SEQ ID NO: 11; and an LCDR3 amino acid sequence set forth in SEQ ID NO: 12.

[0042] In some embodiments, D1 comprises an HCVR comprising the amino acid sequence of SEQ ID NO:1; and an LCVR comprising the amino acid sequence of SEQ ID NO:9.

[0043] In some embodiments, D2 specifically binds to a second epitope of human MET.

[0044] In some embodiments, D2 comprises the HCDR1 amino acid sequence set forth in SEQ ID NO: 6; the HCDR2 amino acid sequence set forth in SEQ ID NO: 7; the HCDR3 amino acid sequence set forth in SEQ ID NO: 8; the LCDR1 amino acid sequence set forth in SEQ ID NO: 10; the LCDR2 amino acid sequence set forth in SEQ ID NO: 11; and the LCDR3 amino acid sequence set forth in SEQ ID NO: 12.

[0045] In some embodiments, D2 comprises an HCVR comprising the amino acid sequence of SEQ ID NO:5; and an LCVR comprising the amino acid sequence of SEQ ID NO:9.

[0046] In some embodiments, the MET abnormality is an abnormality of exon 14 in DNA or a deletion resulting in skipping of exon 14, an amplification of the MET gene, or overexpression of the MET protein. In some aspects, the MET abnormality is an abnormality of exon 14 in DNA or a deletion resulting in skipping of exon 14. In some aspects, the MET abnormality is an amplification of the MET gene. In some aspects, the MET abnormality is overexpression of the MET protein.

[0047] Other embodiments will be apparent from consideration of the detailed description that follows.

[0048] 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. [Brief explanation of the drawings]

[0049] [Figure 1]1A and 1B include bar graphs illustrating the relative proliferation of EBC-1 cells as a function of treatment with a control antibody and a MET×MET bispecific antibody at 0.1 μg / mL and 1 μg / mL. [Figure 2] FIG. 1 depicts 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. [Figure 3] FIG. 1 depicts immunoblots of pMET, MET, pErk, and tubulin (for loading control) extracted from EBC-1 cells after treatment with control antibody and MET×MET bispecific antibody. [Figure 4] This is a line graph illustrating the change in EBC-1 tumor volume in cubic millimeters as a function of time in days after transplantation of EBC-1 cells in animals treated with a control antibody (black square ■), a MET monovalent antibody (black circle ●), or a MET×MET bispecific antibody (black diamond ◆). [Figure 5] FIG. 1 is a bar graph illustrating relative cell proliferation of NCI-H596 cells as a function of treatment with a control antibody (C), a METxMET bispecific antibody (MM), a METxMET parental monospecific antibody 1 (M1), a METxMET parental monospecific antibody 2 (M2), a combination of parental antibodies 1 and 2 (M1M2), and the MET agonist hepatocyte growth factor (HGF). [Figure 6] FIG. 1 illustrates a study flow diagram for a clinical trial using REGN5093. [Figure 7] FIG. 1 is a schematic depiction of the study design showing dose escalation and expansion cohorts. [Figure 8] FIG. 1 illustrates tumor response in patients characterized by centrally confirmed MET aberrations. [Figure 9]9A and 9B illustrate serum concentrations after the first dose of REGN5093 in the dose-escalation cohort, FIG. 9A, and in various expansion cohorts, FIG. 9B. The elimination half-life estimated by noncompartmental analysis over the 3-week dose interval is 15 days. [Figure 10]

[0023] Figure 1 illustrates the interaction of a METxMET bispecific antibody with the MET ectodomain. This figure also illustrates how MET aberrations impact elevated and ligand-independent activation of MET signaling. [Figure 11] FIG. 1 illustrates methods used to identify MET aberrations in ctDNA and tissues, as well as exemplary commercially available products that can be used in such methods. (CEP7, chromosome 7 centromere; ELISA, enzyme-linked immunosorbent assay; FFPE, formalin-fixed paraffin-embedded; FISH, fluorescence 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.) [Figure 12] Figure illustrating the number of patients enrolled in the dose-escalation study, as well as the demographics and number of patients enrolled in the expansion cohort. (aNSCLC, advanced non-small cell lung cancer; GCN, gene copy number; FIH, first-in-human; IHC, immunohistochemistry; IV, intravenous; MET, mesenchymal-to-epithelial transition; PK, pharmacokinetics; Q3W, every 3 weeks; Q6W, every 6 weeks; TKI, tyrosine kinase inhibitor.) [Figure 13] Figure 1 provides response data at the 2000 mg dose for the dose escalation study—2000 mg and expansion cohorts. The overall response rate was highest for the cohort with patients with exon 14 skipping mutations and MET TKI-naïve patients, and for the cohort with patients with both MET gene amplification and MET protein overexpression and MET TKI-naïve patients. [Figure 14]Figure 1 shows tumor response by confirmed MET aberration. MET overexpression - IHC: ≥75% tumor cells with membrane staining 3+; 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. Response to REGN5093 was observed in patients with MET TKI-naive (2L+) aNSCLC with MET exon 14 mutation or MET amplification and overexpression, regardless of EGFR mutation status. For patients whose tumor tissue was analyzed for MET amplification using both platforms, MET GCN was significantly higher by NGS than 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 applicable or undetermined; NGS, next-generation sequencing; PD, progressive disease; PR, partial response; SD, stable disease; TKI, tyrosine kinase inhibitor.) [Figure 15] Figure providing response of specific MET abnormality subgroups (regardless of the presence of other MET abnormalities) versus response of the overall population. Results are based on small sample sizes and require prospective validation. (CR, complete response; DCR, disease control rate; MET, mesenchymal-to-epithelial transition; ORR, objective response rate; PR, partial response; SD, stable disease; TKI, tyrosine kinase inhibitor.) [Figure 16] Figure 1. Chart of baseline demographic and clinical characteristics of the bypass resistance mutation study population. The study population had a median age of 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.) [Figure 17]This figure illustrates the difference in the detection of somatic variants in tissues versus ctDNA. Baseline tumor profiling of 2L+ aNSCLC identified somatic variants with known functional significance. The Venn diagram shows the total number of baseline somatic variants (324 genes) identified by NGS using the FMI-Dx (tissue; n=51 patients) and FMI-L (ctDNA; n=38 patients) panels; genes are grouped by the type of abnormality and the type of clinical sample (ctDNA, tissue, or both). CNVs, including gene amplifications and deletions, were more easily detected in tissues, while more NSVs (single-nucleotide variations, splice variants, and gene fusions) were detected in ctDNA. Therefore, in some embodiments, detecting bypass genes of NSVs and CNVs in ctDNA can complement tissue results and provide a more comprehensive tumor profiling of MetxMet resistance mechanisms. (2L, second-line therapy; 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 variant.) [Figure 18] Figure 1 illustrates unbiased clustering of baseline somatic variants with confirmed MET aberrations. Several genes of known functional importance were detected and clustered by cohort allocation, centrally confirmed MET aberrations, and EGFR status. (Amp, amplification; EGFR, epidermal growth factor receptor; Ex14, exon 14; FMI, Foundation Medicine, Inc.; IHC, immunohistochemistry; MET; N, no; NA, not applicable; mesenchymal-to-epithelial transition; O / E, overexpression; SNV, single nucleotide variant; TKI, tyrosine kinase inhibitor; Y, yes.) [Figure 19]Figure 1 illustrates unbiased clustering of baseline somatic variants with MET aberrations and clinical response. Several genes of known functional importance were detected and clustered by cohort assignment, confirmed MET aberrations, EGFR status, and clinical response. [Figure 20] Figure 1 illustrates classification and examples of baseline somatic mutations identified by tumor profiling. These mutations were identified in non-responders but may confer MET bypass resistance mechanisms and influence 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.) [Figure 21] Figure 1 graphically depicts total REGN5093 and sMET concentrations during study treatment, as well as individual concentrations of sMET over time according to best overall response. Total REGN5093 concentrations were several-fold higher than total sMET concentrations in serum, suggesting that saturation of receptor occupancy was achieved with the 2000 mg Q3W dosing regimen. While elevated levels of sMET and cHGF after dosing suggest target capture, neither baseline nor posttreatment levels of sMET and cHGF were significantly associated with response (data not shown). (cHGF, circulating hepatocyte growth factor; Q3W, every 3 weeks; sMET, soluble mesenchymal-epithelial transition) [Figure 22] FIG. 1 illustrates the lack of a significant association between baseline concentrations of sMET or cHGF and clinical response. cHGF, circulating hepatocyte growth factor; PD, progressive disease; PR, partial response; SD, stable disease; sMET, soluble mesenchymal-epithelial transition. DETAILED DESCRIPTION OF THE INVENTION

[0050] Before describing the present invention, it is to be understood that this invention is not limited to the particular methods and experimental conditions described, as such methods and conditions may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope of the present invention will be limited only by the appended claims.

[0051] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as that commonly understood by those skilled in the art to which this invention belongs.As used herein, the term "about" when used in relation to a specific stated numerical value means that this value can vary by 1% or less from the stated value.For example, as used herein, the expression "about 100" includes 99 and 101 and all values ​​therebetween (for example, including 100, 99.1, 99.2, 99.3, 99.4, etc.).

[0052] Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, the preferred methods and materials are now described. All patents, patent applications, and non-patent publications mentioned herein are incorporated by reference in their entirety.

[0053] MET protein The terms "MET," "c-Met," and the like, as used herein, refer to human transmembrane receptor tyrosine kinases, including: (1) an amino acid sequence having the amino acid sequence set forth in SEQ ID NO: 13 and / or set forth in NCBI Accession No. NM_001127500.2, which represents the unprocessed preproprotein of isoform "a"; (2) an amino acid sequence having the amino acid sequence set forth in SEQ ID NO: 14 and / or set forth in NCBI Accession No. NM_000236.2, which represents the unprocessed preproprotein of isoform "b"; (3) an amino acid sequence having the amino acid sequence set forth in SEQ ID NO: 15 and / or set forth in NCBI Accession No. NM_001311330.1, which represents the unprocessed preproprotein of isoform "c", and / or (3) A mature protein comprising a cytoplasmic alpha subunit (SEQ ID NO: 16) and a transmembrane beta subunit (SEQ ID NO: 17, 18, or 19 for isoforms a, b, and c, respectively) shared by all three isoforms. The term "MET" includes both monomeric and multimeric MET molecules. As used herein, the term "monomeric human MET" refers to a MET protein or a portion thereof that does not contain or possess any multimerization domain and exists under normal conditions as a single MET molecule without direct physical connection to another MET molecule. An exemplary monomeric MET molecule is the molecule referred to herein as "hMET.mmh," which comprises the amino acid sequence of SEQ ID NO: 20 (SEQ ID NO: 152, Example 3 of U.S. Pat. No. 11,142,578). As used herein, the term "dimeric human MET" refers to a construct comprising two MET molecules connected to each other through a linker, a covalent bond, a noncovalent bond, or a multimerization domain, such as 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 according to U.S. Pat. No. 11,142,578, Example 3).

[0054] All references herein to proteins, polypeptides, and protein fragments are intended to refer to the human version of the respective protein, polypeptide, or protein fragment, unless expressly specified as being from a non-human species. Thus, the term "MET" refers to human MET, unless specified as being from a non-human species, e.g., "mouse MET," "simian MET," etc.

[0055] As used herein, the phrase "cell surface-expressed MET" refers to one or more MET proteins, or extracellular domains thereof, that are expressed on the surface of a cell 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 is accessible to the antigen-binding portion of an antibody. "Cell surface-expressed MET" can include or consist of a MET protein expressed on the surface of a cell that normally expresses MET protein. Alternatively, "cell surface-expressed MET" can include or consist of a MET protein expressed on the surface of a cell that does not normally express human MET on its surface but has been artificially engineered to express MET on its surface.

[0056] Measures of MET aberrations in NSCLC include MET exon 14 mutations; oncogenic drivers in NSCLC with recent TKI approval in 1L (first-line); MET gene amplification; a 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 therapeutic response to MET TKIs in NSCLC.

[0057] Therapeutic methods for treating NSCLC cancer Lung cancer is one of the most commonly diagnosed cancers and the leading cause of cancer-related deaths worldwide (Siegel et al., CA Cancer J Clin, 66(1):7-30, 2016). Non-small cell lung cancer (NSCLC) accounts for 80%-85% of all lung cancers and is composed of several histopathological subtypes, the most common of which are adenocarcinoma (40%-60%) and squamous cell carcinoma (30%) (Dela Cruz et al., Clin Chest Med, 32(4):605-44, 2011). The majority of patients with NSCLC are found to have advanced disease at the time of diagnosis.

[0058] Anti-PD-1 and anti-PD-L1 therapy has transformed 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, even those with tumors that highly express PD-L1 or exhibit high tumor mutation burden (TMB), may not receive the same benefit from agents targeting the PD-1 / PD-L1 axis (Sabari et al., Ann Oncol, 29(10):2085-91, 2018). This is consistent with data generated in lung cancers with 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), indicating that anti-PD-1 or anti-PD-L1 monotherapy may not be the preferred treatment for patients with MET-driven disease. Thus, there remains a substantial unmet need for therapies that improve response rates and survival for patients with MET-abnormal NSCLC.

[0059] The present inventors have surprisingly discovered that certain METxMET bispecific antibodies are highly suitable for treating and / or inhibiting NSCLC associated with MET abnormalities, such as DNA deletions resulting in exon 14 aberrations or exon 14 skipping, MET gene amplification, or MET protein overexpression, or for alleviating NSCLC metastasis. 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 METxMET bispecific antibodies can be enhanced by identifying patients with such MET abnormalities.

[0060] Thus, useful in accordance with the methods described herein are METxMET 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 blockade with minimal activation of MET signaling. Such METxMET bispecific antibodies are described in U.S. Publication No. 2018 / 0134794, which is incorporated herein by reference in its entirety.

[0061] METxMET bispecific antibodies are useful, inter alia, for the treatment, prevention, and / or amelioration of NSCLC associated with or mediated by MET expression, signaling, or activity, or NSCLC treatable 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. METxMET bispecific antibodies are useful, inter alia, for the treatment and / or amelioration of NSCLC in subjects suffering from tumors with MET abnormalities, such as DNA abnormalities of exon 14 or deletions resulting in exon 14 skipping, MET gene amplification, or MET protein overexpression. METxMET bispecific antibodies are useful, inter alia, for the prevention of NSCLC recurrence or metastasis in subjects suffering from tumors with MET abnormalities, such as DNA abnormalities of exon 14 or deletions resulting in exon 14 skipping, MET gene amplification, or MET protein overexpression.

[0062] For example, the METxMET bispecific antibody of the present disclosure is useful for treating tumors that express (or overexpress) MET, such as NSCLCs with MET abnormalities. Illustratively, the MET abnormality can be an abnormality of exon 14 in DNA or a deletion that results in exon 14 skipping, an amplification of the MET gene, or overexpression of the MET protein. In some embodiments, the MET abnormality is an abnormality of exon 14 in DNA or a deletion that results in exon 14 skipping. In some embodiments, the MET abnormality is an amplification of the MET gene. In some embodiments, the MET abnormality is overexpression of the MET protein.

[0063] In some embodiments, the subject suffers from non-squamous NSCLC. In some embodiments, the subject suffers from NSCLC squamous cell carcinoma. In some aspects, the NSCLC is metastatic. 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 the growth of NSCLC tumors in the subject and / or causing regression of NSCLC.

[0064] Exemplary methods of using METxMET bispecific antibodies are provided throughout this disclosure and detailed below.

[0065] A method of treating NSCLC in a subject can include administering to a subject in need thereof a therapeutic composition comprising a METxMET bispecific antibody (e.g., a METxMET bispecific antibody comprising a D1 component and a D2 component as described in Table 1 herein, or an anti-MET antibody selected from the group consisting of onartuzumab, emibetuzumab, telisotuzumab, SAIT301, ARGX-111, Sym015, HuMax-cMet, and CE-355621).

[0066] A method of treating NSCLC, reducing NSCLC tumor growth, inhibiting or attenuating invasion and / or metastasis, and / or causing regression of NSCLC in a subject having a tumor with a MET aberration can 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 D2 specifically binds to a second epitope of human MET.

[0067] A method for treating a subject suffering from an NSCLC tumor with a MET aberration can include administering to the subject a METxMET bispecific antibody, the METxMET bispecific antibody comprising: a first antigen-binding domain (D1); and a second antigen-binding domain (D2); D1 specifically binds to a first epitope of human MET; and D2 specifically binds to a second epitope of human MET. In some embodiments, the MET aberration is an aberration of exon 14 in DNA or a deletion resulting in exon 14 skipping. In some embodiments, the subject is naive to a MET-targeted tyrosine kinase inhibitor (MET-TKI). In some embodiments, the MET aberration is MET gene amplification. In some embodiments, the MET aberration is overexpression of MET protein.

[0068] A method for treating NSCLC, reducing NSCLC tumor growth, and / or causing regression of NSCLC in a subject afflicted with a tumor having a MET aberration can include administering to the subject a 250-2000 mg dose of a METxMET bispecific antibody.

[0069] A method for treating or inhibiting the growth of NSCLC can include: (1) selecting a subject having a tumor with a MET aberration; and (2) administering to the subject (a) a dose of about 250 mg, 500 mg, 750 mg, 1000 mg, 1500 mg, or 2000 mg of a METxMET bispecific antibody. In some embodiments, the administering in step (2) occurs once every three weeks.

[0070] Further methods for treating tumors include: (a) Selecting subjects with NSCLC; (b) determining that the tumor exhibits a MET abnormality selected from the group consisting of an exon 14 abnormality or deletion in DNA resulting in exon 14 skipping, MET gene amplification, and / or Met protein overexpression; (i) obtaining a tissue sample and / or a body fluid sample from a subject; and (ii) evaluating tissue samples for MET gene amplification using fluorescence in situ hybridization in tumor tissue or by next-generation sequencing in tumor tissue and / or ctDNA and / or evaluating tissue samples for Met protein overexpression using immunohistochemistry and / or evaluating body fluid samples for exon 14 mutations using ctDNA. including; and if the tumor exhibits MET abnormalities, (c) administering one or more doses of a METxMET bispecific antibody to a subject in need thereof; Includes.

[0071] Identifying patients or patient populations that may be successfully treated with a METxMET antibody is desirable, at least from the patient's perspective. Accordingly, the inventors have provided a method for identifying candidates for METxMET anti-tumor therapy, comprising: obtaining a tissue and / or fluid sample from a subject with NSCLC; and evaluating the tissue and / or fluid sample for a MET abnormality selected from the group consisting of an exon 14 abnormality or a deletion resulting in exon 14 skipping in DNA, an amplification of the MET gene, and / or overexpression of the Met protein; The presence of at least one Met abnormality in the tissue or fluid sample identifies the subject as a candidate for an anti-tumor therapy, wherein the METxMET anti-tumor therapy comprises a METxMET bispecific antibody. The method was devised.

[0072] A further method for treating NSCLC in a subject can include: (i) obtaining a body fluid sample from the subject and determining MET amplification in ctDNA from the body fluid sample, and (ii) administering a METxMET bispecific antibody to the subject; steps (i) and (ii) are repeated once every three weeks, and loss of MET amplification after step (ii) is repeated is an indication of a sustained response to the treatment.

[0073] In some embodiments, a subject according to any one of the methods provided herein comprises: (i) Histologically confirmed NSCLC; (ii) MET exon 14 mutation; (iii) MET gene amplification; (iv) elevated MET protein expression (IHC ≥2+ or H score >150); (v) MET exon 14 mutation and MET TKI-experienced; (vi) MET exon 14 mutation and MET TKI-naive; (vii) highly amplified MET gene (MET GCN ≥5 and / or MET / CEP7 ratio ≥2 in tissue by FISH or MET GCN ≥6 by NGS or MET fold change ≥2 in ctDNA) and MET TKI-naive; (viii) highly overexpressed MET protein (IHC 3+ or H score ≥ 200) and MET TKI-naïve; (ix) highly amplified MET gene (MET GCN ≥ 5 and / or MET / CEP7 ratio ≥ 2 in tissue by FISH or MET GCN ≥ 6 by NGS or MET fold change ≥ 2 in ctDNA), highly overexpressed MET protein (IHC 3+ or H score ≥ 200) and MET TKI-naïve.

[0074] In some embodiments, subjects are selected as having one or more of the following: prior MET TKI experience and an exon 14 aberration in DNA or a deletion resulting in exon 14 skipping; PD-(L)1 experience and an exon 14 aberration in DNA or a deletion resulting in exon 14 skipping (MET TKI-naive); prior EGFR inhibitor experience and PD-(L1)-naive and an exon 14 aberration in DNA or a deletion resulting in exon 14 skipping (MET TKI-naive); MET gene amplification and / or MET protein overexpression (MET TKI-naive) and PD-(L1)-naive; PD-(L)1 experience and MET gene amplification and / or MET protein overexpression (MET TKI-naive); or prior EGFR inhibitor experience and PD-(L)1-naive and MET gene amplification and / or MET protein overexpression (MET TKI-naive). Subjects can also be selected based on having a tumor with one or more mutations in the EGFR gene.

[0075] In some embodiments, the subject has not received a previous anti-cancer therapy. In some embodiments, the subject is MET-targeted tyrosine kinase inhibitor (TKI)-naive. In other words, the subject has not received a previous treatment with a TKI. In some embodiments, the subject has received a previous anti-cancer therapy including one or more of a TKI, a PD-1 inhibitor, a PD-L1 inhibitor, surgery, radiation therapy, or chemotherapy. In some aspects, the previous anti-cancer therapy includes a TKI. In some aspects, the subject is resistant or does not respond adequately to the previous therapy, or has relapsed after the previous therapy.

[0076] In the context of the methods of treatment described herein, the METxMET bispecific antibody can be administered as a monotherapy (i.e., as the only therapeutic agent) or in combination with one or more additional therapeutic agents.

[0077] Responsiveness to METxMET bispecific antibody treatment may vary over time as tumor cells develop bypass resistance mechanisms. Resistance to MET therapy may be acquired in response to prior therapy in heavily pretreated NSCLC patients, for example, in patients treated with chemotherapy, immune checkpoint inhibitors, and / or EGFR inhibitors. If a tumor proves resistant to the current therapy, it is desirable to provide an alternative therapy as soon as possible. Therefore, a method for monitoring the efficacy of METxMET bispecific antibody treatment in subjects with NSCLC with MET abnormalities is provided herein. Here's how: (i) obtaining a tissue and / or fluid sample from a subject; and a. On-target MET receptor gene mutations and MET gene silencing (loss of function) conferring resistance to MET TKIs found in MET Ex14 Mut patients with prior TKI Exp; b. TK driver receptor activation selected from TK receptor and ligand gene amplifications and TKR-activating mutations; and c. Activating gene mutations in pathways selected from the group consisting of the JAK2 / STAT3 pathway, the RAS / RAF / MEK / MAPK pathway, and the PI3K / AKT / MTOR pathway, TP53 mutations, and cell cycle gene amplification: evaluating the tissue and / or fluid sample for somatic mutations in one or more genes selected from the group consisting of: (ii) administering a MET×MET bispecific antibody to a subject; and (iii) repeating steps (i) and (ii) over the course of treatment. Includes; The acquisition of mutations in one or more genes is indicative of resistance to therapy and / or a poor prognosis.

[0078] In some embodiments, the on-target MET receptor genetic mutation is selected from the group consisting of MET Y1230C, MET D1228H, MET D1228N; and the MET gene silencing (loss of function) is selected from somatic mutations in DNMT3A and TET2.

[0079] In some embodiments, the TKR activating mutation is selected from the group consisting of EGFR L858R, EGFR G719S, EGFR E709A, EGFR E746_A750del, and EGFR S752_I759del.

[0080] In some embodiments, 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, PIK3CA N345K, 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.

[0081] Various embodiments of METxMET bispecific antibodies are provided in the following paragraphs and are described in more detail elsewhere herein.

[0082] In some embodiments, D1 and D2 do not compete with each other for binding to human MET. In some embodiments, the first epitope of human MET comprises amino acids 192-204 of SEQ ID NO: 22. In some embodiments, the second epitope of human MET comprises amino acids 305-315 and 421-455 of SEQ ID NO: 22. In some embodiments, the first epitope of human MET comprises amino acids 192-204 of SEQ ID NO: 22; and the second epitope of human MET comprises amino acids 305-315 and 421-455 of SEQ ID NO: 22.

[0083] In some embodiments, D1 comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) within the 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 the light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 9. In some embodiments, D2 comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) within the 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 the light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 9.

[0084] In some embodiments, the bispecific antibody comprises the CDRs within the D1-HCVR amino acid sequence of SEQ ID NO:1 and the CDRs within the D2-HCVR amino acid sequence of SEQ ID NO:5.

[0085] In some embodiments, D1 of the bispecific antibody comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) within the heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 1, or an amino acid sequence at least 95% identical thereto, and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) within the light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 9, or an amino acid sequence at least 95% identical thereto.

[0086] In some embodiments, 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.

[0087] In some embodiments, D1 of the bispecific antibody comprises an HCVR comprising the amino acid sequence of SEQ ID NO:1, or an amino acid sequence that is at least 95% identical thereto; and an LCVR comprising the amino acid sequence of SEQ ID NO:9, or an amino acid sequence that is at least 95% identical thereto.

[0088] In some embodiments, D1 of the bispecific antibody comprises an HCVR comprising the amino acid sequence of SEQ ID NO:1; and an LCVR comprising the amino acid sequence of SEQ ID NO:9.

[0089] In some embodiments, D2 of the bispecific antibody comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) within the heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 5, or an amino acid sequence at least 95% identical thereto, and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) within the light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 9, or an amino acid sequence at least 95% identical thereto.

[0090] In some embodiments, the D2 HCDR1 of the bispecific antibody 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.

[0091] In some embodiments, D2 of the bispecific antibody comprises an HCVR comprising the amino acid sequence of SEQ ID NO:5, or an amino acid sequence that is at least 95% identical thereto; and an LCVR comprising the amino acid sequence of SEQ ID NO:9, or an amino acid sequence that is at least 95% identical thereto.

[0092] In some embodiments, D2 of the bispecific antibody comprises an HCVR comprising the amino acid sequence of SEQ ID NO:5; and an LCVR comprising the amino acid sequence of SEQ ID NO:9.

[0093] Biological Properties of the Antibodies Provided herein Useful in accordance with the methods provided herein are bispecific antibodies and antigen-binding fragments thereof that inhibit the proliferation, inhibit invasion, induce apoptosis, and / or reduce the viability of NSCLC cells. The bispecific antibodies and antigen-binding fragments thereof bind to the MET receptor and prevent its interaction with HGF.

[0094] 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. For example, the present disclosure provides antibodies and antigen-binding fragments thereof that have a K of less than about 230 nM 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. Pat. No. 11,142,578 or a substantially similar assay. D According to certain embodiments, anti-METxMET antibodies useful in accordance with the methods provided herein have 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, as measured by surface plasmon resonance, e.g., using the assay format defined in Example 3 of U.S. Pat. No. 11,142,578 or a substantially similar assay. D and binds to monomeric human MET at 37°C.

[0095] Such bispecific antibodies and antigen-binding fragments thereof have a dissociation half-life (t ) 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. Pat. No. 11,142,578 or a substantially similar assay. 1 Such bispecific antibodies bind to monomeric human MET (e.g., hMET.mmh) at a t 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, e.g., using the assay format defined in Example 3 of U.S. Pat. No. 11,142,578 or a substantially similar assay. 1 / 2 and binds to monomeric human MET at 37°C.

[0096] Such bispecific antibodies and antigen-binding fragments thereof bind to dimeric human MET (e.g., hMET.mFc) with high affinity. For example, the bispecific antibodies may have a K of less than about 3 nM 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. Pat. No. 11,142,578 or a substantially similar assay. D 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, as measured by surface plasmon resonance, e.g., using the assay format defined in Example 3 of U.S. Pat. No. 11,142,578 or a substantially similar assay. D It binds to dimeric human MET at 37°C.

[0097] Such bispecific antigen binding molecules and antigen-binding fragments thereof have a dissociation half-life (t) of greater than about 4 minutes 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. Pat. No. 11,142,578 or a substantially similar assay. 1 / 2) to dimeric human MET (e.g., hMET.mFc). According to certain embodiments, anti-MET antibodies are provided that have a t 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, e.g., using the assay format defined in Example 3 of U.S. Pat. No. 11,142,578 or a substantially similar assay. 1 It binds to dimeric human MET at 37°C with γ-glucan at 1 / 2.

[0098] Also useful in accordance with the methods provided herein are compounds that have a dissociation half-life (t ) 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. Pat. No. 11,142,578 or a substantially similar assay. 1

[0013] According to certain embodiments, a METxMET bispecific antigen-binding protein is provided that binds to dimeric human MET (e.g., hMET.mFc) at a t 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 by surface plasmon resonance, e.g., using the assay format defined in Example 6 of U.S. Pat. No. 11,142,578 or a substantially similar assay. 1 It binds to dimeric human MET at 37°C with γ-glucan at 1 / 2.

[0099] Also useful in accordance with the methods provided herein are METxMET bispecific antibodies that block the interaction between HGF and MET, for example, in in vitro ligand binding assays. According to certain embodiments provided herein, METxMET bispecific antigen-binding proteins block HGF binding to cells expressing human MET and induce minimal or no MET activation in the absence of HGF signaling. For example, the present disclosure provides METxMET bispecific antigen-binding proteins that exhibit some MET agonist activity in a cell-based MET activity reporter assay, where the MET agonist activity is less than 50%, 40%, 30%, 20%, 10%, 5%, 3%, 2%, or 1% of the MET agonist activity observed in a comparable activity reporter assay using a monospecific antibody comprising D1 or D2 alone.

[0100] Bispecific antibodies useful according to the present disclosure can possess one or more of the above biological properties, or any combination thereof. The foregoing list of antibody biological properties is not intended to be exhaustive. Other biological properties of the antibodies provided herein will be apparent to those of skill in the art from a review of the present disclosure, including the working examples herein.

[0101] Epitope mapping and related techniques Useful in accordance with the methods provided herein are METxMET bispecific antibodies that bind to a human MET epitope comprising amino acids 192-204, amino acids 305-315, and / or amino acids 421-455 of SEQ ID NO:22.

[0102] Human antibody production The METxMET bispecific antibodies useful according to the methods provided herein can be fully human antibodies. Methods for generating monoclonal antibodies, including fully human monoclonal antibodies, are known in the art. Any such known method can be used in the context of the present disclosure to generate human antibodies that specifically bind to human MET.

[0103] For example, using VELOCIMMUNE™ technology or any other similar known method for generating fully human monoclonal antibodies, high-affinity chimeric antibodies against MET, having human variable regions and mouse constant regions, are first isolated. As in the experimental section below, the antibodies are characterized and selected for desired properties, including affinity, ligand-blocking activity, selectivity, epitope, etc. If necessary, the mouse constant region is replaced with a desired human constant region, e.g., wild-type or modified IgG1 or IgG4, to generate a fully human anti-MET antibody. While the constant region selected may vary depending on the particular application, the properties of high-affinity antigen binding and target specificity reside in the variable region. In certain cases, fully human anti-MET antibodies are isolated directly from antigen-positive B cells.

[0104] biological equivalent The methods described herein can utilize METxMET bispecific antibodies and antibody fragments thereof, including proteins with amino acid sequences that differ from those of the described antibodies but retain the ability to bind to human MET. Such variant molecules and antibody fragments contain one or more additions, deletions, or substitutions of amino acids compared to the parent sequences, but exhibit biological activity that is essentially equivalent to that of the described molecules. Similarly, DNA sequences encoding the anti-METxMET antibodies of the present disclosure include sequences that contain one or more additions, deletions, or substitutions of nucleotides compared to the disclosed sequences, but encode anti-METxMET antibodies or antibody fragments that are essentially biologically equivalent to the anti-MET antibodies or antibody fragments of the present disclosure. Examples of amino acid and DNA sequences for such variants are discussed above.

[0105] Two antigen-binding proteins or antibodies are considered to be bioequivalent if, for example, they are pharmaceutical equivalents or pharmaceutical substitutes that do not show significant differences in the rate and extent of absorption when administered at the same molar dose under similar experimental conditions, either in single or multiple doses. Some antibodies would be considered equivalents or pharmaceutical substitutes if: Drugs that are equivalent in their extent of absorption but not in their rate of absorption may still be considered bioequivalent because such differences in absorption rate are intended, reflected in labeling, are not important for achieving effective body drug concentrations, for example, during chronic use, and are considered medically insignificant for the particular drug being tested.

[0106] In one embodiment, two antigen binding proteins are bioequivalent if there are no clinically meaningful differences in the safety, purity, and potency of the two.

[0107] In one embodiment, two antigen binding proteins are bioequivalent if a patient can be switched between the reference product and the biologic product one or more times without an expected increased risk of adverse effects, including clinically significant changes in immunogenicity or reduced efficacy, compared to continuous therapy without switching between the reference product and the biologic product.

[0108] In one embodiment, two antigen binding proteins are bioequivalent if they both act through one or more common mechanisms of action for one or more conditions of use, to the extent that such mechanisms are known.

[0109] Bioequivalence can be demonstrated by in vivo and in vitro methods. Measurements of bioequivalence include, for example, (a) in vivo tests in humans or other mammals in which the concentration of an antibody or its metabolites is measured as a function of time in blood, plasma, serum, or other biological fluids; (b) in vitro tests that have previously correlated with and are reasonably predictive of human in vivo bioavailability data; (c) in vivo testing in humans or other mammals that measures the relevant acute pharmacological effects of the antibody (or its target) as a function of time; and (d) in well-controlled clinical trials that establish the safety, efficacy, or bioavailability or bioequivalence of the antibody.

[0110] Biologically equivalent variants of the anti-MET antibodies provided herein can be constructed, for example, by making various substitutions of residues or sequences or by deleting terminal or internal residues or sequences that are not required for biological activity. For example, cysteine ​​residues that are not essential for biological activity can be deleted or replaced with other amino acids to prevent the formation of unwanted or incorrect intramolecular disulfide bridges during renaturation. In other contexts, biologically equivalent antibodies can include anti-MET antibody variants that contain amino acid changes that alter the glycosylation characteristics of the antibody, e.g., mutations that eliminate or remove glycosylation.

[0111] Species selectivity and species cross-reactivity 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. Also useful are anti-MET antibodies (and antibodies comprising anti-MET antigen-binding domains) that bind to human MET and to MET from one or more non-human species. For example, anti-METxMET antibodies and antibodies can bind to human MET and, accordingly, may or may not bind to MET from one or more of the following species: mouse, rat, guinea pig, hamster, gerbil, pig, cat, dog, rabbit, goat, sheep, cow, horse, camel, macaque, marmoset, rhesus monkey, or chimpanzee. According to certain exemplary embodiments, anti-METxMET antibodies and antibodies that specifically bind to human MET and macaque (e.g., cynomolgus monkey (Macaca fascicularis)) MET are provided. Other anti-METxMET antibodies and antibodies bind to human MET but do not bind at all or only weakly to macaque MET.

[0112] Therapeutic Formulations and Administration Useful herein are pharmaceutical compositions comprising the METxMET bispecific antibodies of the invention, which can be formulated with suitable carriers, excipients, and other agents that improve mobility, delivery, tolerability, etc.

[0113] In some embodiments, a pharmaceutical composition comprising a METxMET bispecific antibody is formulated for administration to a subject to treat lung cancer, and specifically to treat NSCLC.

[0114] Provided herein are methods in which the METxMET bispecific antibody administered to a subject is contained within a pharmaceutical formulation. The pharmaceutical formulation can include the METxMET bispecific antibody along with at least one inactive ingredient, such as a pharmaceutically acceptable carrier. Other agents can be incorporated into the pharmaceutical composition to improve transport, delivery, tolerability, etc. The term "pharmaceutically acceptable" means approved by a federal or state regulatory agency or listed in the United States Pharmacopeia or other generally recognized pharmacopeia for use in animals, and more specifically, in humans. The term "carrier" refers to a diluent, adjuvant, excipient, or vehicle with which the antibody is administered. Many suitable formulations can be found in the formulary known to all pharmaceutical chemists: Remington's Pharmaceutical Sciences (15th ed., Mack Publishing Company, Easton, Pa., 1975), particularly Chapter 87 by Blaug and Seymour therein. These formulations include, for example, powders, pastes, ointments, jellies, waxes, oils, lipids, lipid (cationic or anionic)-containing vesicles (e.g., LIPOFECTIN™), DNA conjugates, anhydrous absorbent pastes, oil-in-water and water-in-oil emulsions, emulsion carbowax (polyethylene glycol of various molecular weights), semi-solid gels, and carbowax-containing semi-solid mixtures. Any of the above mixtures may be suitable in the context of the disclosed methods, provided that the anti-MET antibody or anti-METxMET bispecific antibody is not inactivated by the formulation and that the formulation is physiologically compatible with and tolerable by the route of administration. See also Powell et al. PDA (1998) J Pharm Sci Technol. 52:238-311 and citations therein for additional information related to excipients and carriers familiar to pharmaceutical chemists.

[0115] In the context of the present disclosure, pharmaceutical formulations useful for administration by injection can be prepared by dissolving, suspending, or emulsifying an anti-MET antibody or anti-MET x MET bispecific antibody in a sterile aqueous or oily medium conventionally used for injections. Aqueous media for injections include, for example, saline, isotonic solutions containing glucose and other auxiliary agents, and the like, which can be used in combination with appropriate solubilizers such as alcohols (e.g., ethanol), polyhydric alcohols (e.g., propylene glycol, polyethylene glycol), nonionic surfactants (e.g., polysorbate 80, HCO-50 (polyoxyethylene (50 mol) adduct of hydrogenated castor oil)), and the like. Oily media include, for example, sesame oil, soybean oil, and the like, which can be used in combination with solubilizers such as benzyl benzoate, benzyl alcohol, and the like. The injections prepared in this manner can be filled into appropriate ampoules, if desired.

[0116] Dosing regimen According to certain embodiments, multiple doses of an anti-MET antibody or anti-METxMET bispecific antibody (or a pharmaceutical composition comprising an anti-MET antibody or anti-METxMET bispecific antibody in combination with any of the additional therapeutically active agents mentioned herein) can be administered to a subject over a defined time course. A method according to this aspect comprises sequentially administering to a subject multiple doses of an anti-MET antibody or anti-METxMET bispecific antibody provided herein. As used herein, "sequentially administering" means that each dose of antibody is administered to a subject at different times, separated by a predetermined interval (e.g., hours, days, weeks, or months), e.g., on different days. The present disclosure includes methods comprising sequentially administering to a subject a single initial dose of an anti-MET antibody or METxMET bispecific antigen-binding molecule, followed by one or more secondary doses of the anti-MET antibody or METxMET bispecific antigen-binding molecule, optionally followed by one or more tertiary doses of the anti-MET antibody or anti-METxMET bispecific antibody.

[0117] The terms "initial dose," "secondary dose," and "tertiary dose" refer to the time sequence of administration of an anti-MET antibody or anti-METxMET bispecific antibody. Thus, an "initial dose" is a dose administered at the beginning of a treatment regimen (also referred to as a "baseline dose"); a "secondary dose" is a dose administered after the initial dose; and a "tertiary dose" is a dose administered after the secondary dose. The initial, secondary, and tertiary doses can all contain the same amount of anti-MET antibody or METxMET bispecific antigen-binding molecule, but may differ from one another overall in terms of frequency of administration. However, in certain embodiments, the amount of antibody contained in the initial, secondary, and / or tertiary doses differs from one another (e.g., adjusted up or down, as appropriate) during the course of treatment. In certain embodiments, two or more (e.g., 2, 3, 4, or 5) doses are administered at the beginning of a treatment regimen as a "loading dose," followed by subsequent doses (e.g., "maintenance doses") administered on a less frequent basis.

[0118] Anti-MET antibodies or anti-METxMET bispecific antibodies can be administered according to any regimen that provides a therapeutic effect. In some embodiments, the bispecific antibody is administered at a dosing frequency of about 4 times per week, twice per week, once per week, once every 2 weeks, once every 3 weeks, once every 4 weeks, once every 5 weeks, once every 6 weeks, once every 8 weeks, once every 12 weeks, or less frequently, as long as a therapeutic response is achieved. In some embodiments, the bispecific antibody is administered once per week, once every 2 weeks, once every 3 weeks, or once per month. In some embodiments, the bispecific antibody is administered once every 3 weeks.

[0119] In some embodiments, the bispecific antibody is administered 1 week, 2 weeks, 3 weeks, or 4 weeks after the immediately preceding dose, hi some embodiments, the bispecific antibody is administered 3 weeks after the immediately preceding dose.

[0120] According to certain embodiments of the present disclosure, multiple doses of a bispecific antibody can be administered to a subject over a defined time course. Methods according to this aspect of the disclosure include sequentially administering two or more doses of a bispecific antibody to a subject. As used herein, "sequentially administering" means that each dose of antibody is administered to a subject at different times, 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 to a subject a single initial dose of a bispecific antigen-binding molecule, followed by one or more secondary doses of the bispecific antigen-binding molecule, optionally followed by one or more tertiary doses of a bispecific antibody.

[0121] According to certain embodiments of the present disclosure, multiple doses of the bispecific antibody can be administered to a subject once every three weeks or once every six weeks over the course of several months or years.

[0122] The terms "initial dose," "secondary dose," and "tertiary dose" refer to a time sequence of administration. Thus, an "initial dose" is a dose administered at the beginning of a treatment regimen (also referred to as a "baseline dose"); a "secondary dose" is a dose administered after the initial dose; and a "tertiary dose" is a dose administered after the secondary dose. The initial, secondary, and tertiary doses can all contain the same amount of antibody (anti-METxMET bispecific antigen-binding molecule). However, in certain embodiments, the amount contained in the initial, secondary, and / or tertiary doses varies from one another (e.g., adjusted up or down, as appropriate) during the course of treatment. In certain embodiments, one or more doses (e.g., 1, 2, 3, 4, or 5) are administered at the beginning of a treatment regimen as a "loading dose," followed by subsequent doses (e.g., "maintenance doses") administered on a less frequent basis. For example, the bispecific antibody can be administered to a subject with NSCLC at 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.

[0123] In an exemplary embodiment of the present disclosure, each secondary and / or tertiary dose is 1 / 2~14 (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 The phrase "immediately preceding dose" as used herein means the dose of a bispecific antibody administered to a patient prior to the administration of the very next dose in a multiple dose series, provided there are no intervening doses in the series.

[0124] The methods provided in some embodiments can include administering any number of secondary and / or tertiary doses of a bispecific antibody to a subject. For example, in certain embodiments, only a single secondary dose is administered to a subject. In other embodiments, two or more (e.g., 2, 3, 4, 5, 6, 7, 8, or more) secondary doses are administered to a subject. Similarly, in certain embodiments, only a single tertiary dose is administered to a subject. In other embodiments, two or more (e.g., 2, 3, 4, 5, 6, 7, 8, or more) tertiary doses are administered to a subject.

[0125] In embodiments involving multiple secondary doses, each secondary dose can be administered with the same frequency as the other secondary doses. For example, each secondary dose can be administered to a subject 1-2 weeks after the immediately preceding dose. Similarly, in embodiments involving multiple tertiary doses, each tertiary dose can be administered with the same frequency as the other tertiary doses. For example, each tertiary dose can be administered to a subject 2, 3, or 4 weeks after the immediately preceding dose. Alternatively, the frequency with which the secondary and / or tertiary doses are administered to a subject can vary over the course of a treatment regimen. The administration frequency can also be adjusted by a physician during the course of treatment depending on the needs of an individual subject following clinical testing.

[0126] In certain embodiments, one or more doses of the bispecific antibody are administered on a more frequent basis (e.g., twice a week, once a week, or once every two weeks) at the beginning of the treatment regimen as an "induction dose," followed by subsequent doses ("consolidation doses" or "maintenance doses") administered on a less frequent basis (e.g., once every 4-12 weeks).

[0127] Dosage The amount of METxMET 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 bispecific antibody that results in or has a therapeutic effect of one or more of the following, compared to a subject not receiving treatment: (a) a reduction in the severity or duration of cancer symptoms; (b) inhibition of tumor growth or increased tumor necrosis, tumor regression, and / or tumor disappearance; (c) delay in tumor growth and / or development; (d) inhibition, slowing, or cessation of tumor metastasis; (e) prevention of recurrence of tumor growth; (f) increased survival of a subject with cancer; and / or (g) a reduction in the use of or need for conventional anti-cancer therapy (e.g., reduction or elimination of the use of chemotherapeutic or cytotoxic agents).

[0128] In some examples, the therapeutically effective amount can be from about 250 mg to about 8000 mg, e.g., 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 METxMET bispecific antibody.

[0129] In some embodiments, the bispecific antibody can be administered at a dose of about 250 mg to about 5000 mg, or at a dose of 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 embodiments, the dose is about 500 mg of METxMET bispecific antibody. In some embodiments, the dose is about 1000 mg of METxMET bispecific antibody. In some embodiments, the bispecific antibody is administered at a dose of about 2000 mg. In some embodiments, the METxMET bispecific antibody can be administered at a dose of 250 mg, 500 mg, 1000 mg, or 2000 mg.

[0130] Bispecific antibodies can be administered intravenously, subcutaneously, or intraperitoneally. In some aspects, the bispecific antibody is administered by intravenous infusion. [Example]

[0131] The following examples are presented so as to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the methods and compositions provided herein, 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 numbers used (e.g., amounts, temperatures, etc.), but some experimental error and deviation should be accounted for. Unless otherwise indicated, parts are parts by weight, molecular weight is average molecular weight, temperature is in degrees Celsius, and pressure is at or near atmospheric. [Example]

[0132] Construction of a bispecific antibody with two distinct antigen-binding domains specific for different epitopes of MET Examples 1 to 3 of U.S. Pat. No. 11,142,578 (incorporated herein by reference in its entirety) describe the construction of a bispecific antibody that is derived from two bivalent monospecific anti-MET antibodies (the "D1" arm derived from exemplary anti-MET antibody H4H13306P2 and the "D2" arm derived from exemplary anti-MET antibody H4H13312P2), and thus contains two distinct antigen-binding domains that bind to separate epitopes on the MET extracellular domain:

[0133] Binding epitope of anti-Met antibody H4H13312P2: AA 192-204: VRRLKETKDGFMF (SEQ ID NO: 23) of SEQ ID NO: 22.

[0134] Binding epitope of anti-Met antibody H4H13306P2: AA 305-315: LARQIGASLND (SEQ ID NO: 24) of SEQ ID NO: 22 and AA 421-455: FIKGDLTIANLGTSEGRFMQVVVSRSGPSTPHVNF (SEQ ID NO: 25) of SEQ ID NO: 22.

[0135] Both antigen-binding domains (D1 and D2) contain a common light chain variable region. The components of bispecific antibodies useful according to the methods provided herein are summarized in Table 1.

[0136] [Table 1]

[0137] The binding kinetic parameters of H4H14639D to the monomeric Met protein (hMET.mmh) are shown in Table 2.

[0138] [Table 2]

[0139] H4H14639D exhibits a significantly lower dissociation rate than each of its parent antibodies, H4H13306P2 and H4H13312P2. See Table 3.

[0140] [Table 3]

[0141] As described in Example 7 of US Pat. No. 11,142,578, a METxMET bispecific antibody blocks HGF signaling and exhibits low MET agonist activity. [Example]

[0142] Anti-Met antibodies inhibit the proliferation of Met-amplified cells The blocking activity of a MET x MET bispecific antibody (i.e., H4H14639D) was evaluated in the non-small cell lung cancer (NSCLC) cell line EBC-1, which displays an amplified Met gene and overexpresses MET (Lutterbach et al., Cancer Res. 67(5):2081-2088, 2007). Complete growth medium for EBC-1 cells contained MEM Earle's salts, 10% fetal bovine serum (FBS), penicillin / streptomycin / glutamine, and non-essential amino acids for MEM. H4H14369D displayed the greatest percent inhibition of MET activity as readout by SRE-luciferase. In this experiment, 3.0 x 10 3EBC-1 cells were seeded in complete growth medium in the presence of H4H14639D at concentrations ranging from 15 pM to 100 nM. Cells were incubated at 37°C in 5% CO for 3 days. Cells were then fixed with 4% formaldehyde and stained with Hoechst 33342 at 3 μg / ml to label nuclei. Images were acquired with an IMAGEXPRESS® Micro XL (Molecular Devices, Sunnyvale, CA), and nuclei counts were determined via METAXPRESS® Image Analysis software (Molecular Devices, Sunnyvale, CA). Background nuclei counts from cells treated with 40 nM digitonin were subtracted from all wells, and viability was expressed as a percentage of the untreated control. IC50 values ​​were determined from a four-parameter logistic equation over a 10-point response curve (GRAPHPAD PRISM®). As summarized below in Table 4, the METxMET bispecific antibody H4H14639D inhibited EBC-1 cell proliferation by 37 with an IC50 of 0.82 nM.

[0143] [Table 4]

[0144] The effect of the METxMET bispecific antibody on EBC-1 cell proliferation was evaluated. 2,500 EBC-1 cells were seeded into 96-well plates and cultured in Dulbecco's medium supplemented with 10% FBS. Cells were treated with a control antibody or the METxMET bispecific antibody at 0.1 μg / mL or 1 μg / mL, followed by incubation at 37°C with 5% CO2. After 5 days, relative cell proliferation was determined by measuring the reduction of the indicator dye ALAMARBLUE® to its highly fluorescent form using a SPECTRAMAX® M3 plate reader (Molecular Devices, LLC, Sunnyvale, CA). The results are shown in Table 5 and Figure 1. The METxMET bispecific antibody (H4H14639D) significantly reduced the relative cell proliferation of EBC-1 cells compared with the control antibody (Figure 1).

[0145] Several anti-MET antibodies, both bivalent monospecific antibodies and MET × MET bivalent antibodies, are potent inhibitors of SRE-Luc activation and inhibit the growth of Met-amplified and MET-overexpressing cell lines.

[0146] [Table 5] [Example]

[0147] METxMET bispecific antibody induces moderate and transient MET pathway activity in NCI-H596 NSCLC cells The effect of the MET × MET bispecific antibody on the MET pathway in human lung adenosquamous carcinoma cells was evaluated in vitro.

[0148] NCI-H596 cells (250,000 cells) were seeded in 12-well plates and cultured in RPMI medium supplemented with 10% FBS. Cells were treated in duplicate with hepatocyte growth factor (HGF) at 50 ng / ml or the MET x MET bispecific antibody H4H14639D at 10 μg / ml. Subsequently, cells were incubated at 37°C in 5% CO2. After 0, 2, 6, or 18 hours, cell lysates were prepared, normalized for protein content, and immunoblot analysis was performed. MET and ERK phosphorylation were quantified using the ImageJ image processing program (T. Collins, BioTechniques 43:S25-S30, 2007). Phosphorylation levels were normalized to tubulin loading control and are expressed as fold change compared to control treatment. The results are summarized in Table 6.

[0149] [Table 6]

[0150] HGF treatment of NCI-H596 cells induced robust activation of MET and ERK, which peaked at 2 h and persisted for 18 h. Moderate MET and ERK phosphorylation was detected with H4H14636D bispecific antibody treatment, and these phosphorylations returned to baseline levels by 18 h or 6 h, respectively. [Example]

[0151] MET×MET bispecific antibodies induce MET degradation more potently than monospecific antibodies in NCI-H596 lung cancer cells The effects of the METxMET 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. NCI-H596 human lung adenosquamous carcinoma cells (250,000 cells) were seeded in 12-well plates and cultured in RPMI medium supplemented with 10% FBS. Cells were treated with (1) 5 μg / ml of hFc control molecule, (2) 5 μg / ml of the parental bivalent monospecific anti-MET antibody H4H13306P2, (3) 5 μg / ml of the parental bivalent monospecific anti-MET antibody H4H13312P2, (4) a combination of 2.5 μg / ml of H4H13306P2 and 2.5 μg / ml of H4H13312P2, or (5) 5 μg / ml of the METxMET bispecific antibody H4H14639D. The cells were then incubated at 37°C with 5% CO2. After 18 hours, cell lysates were prepared, protein content was normalized, and immunoblot analysis was performed. MET expression was quantified using the ImageJ image processing program (T. Collins, BioTechniques 43:S25-S30, 2007). The results are summarized in Table 7.

[0152] [Table 7]

[0153] NCI-H596 (MET exon 14 skipping mutation) lung cancer cells were also treated with control or METxMET bispecific antibody at 10 μg / ml for 2, 6, or 18 hours. MET expression was determined by immunoblotting (Fig. 2), demonstrating METxMET bispecific antibody-induced degradation of MET with increasing treatment time.

[0154] The bispecific antibody H4H14636D induces MET degradation more potently than its parent antibody in NCI-H596 lung cancer cells. [Example]

[0155] METxMET bispecific antibodies induce MET degradation, inhibit pathway activity, and inhibit tumor growth more potently than monospecific antibodies in EBC-1 cells EBC-1 cells, a MET-amplified human lung squamous cell carcinoma (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 10 μg / ml of control antibody or MET x MET bispecific antibody for 18 hours, as described above. MET expression and MET pathway activation, confirmed by pMET and pErk expression, were determined by immunoblotting with the indicated antibodies. The immunoblots are shown in Figure 3.

[0156] Treatment of EBC-1 cells harboring MET gene amplification with the METxMET bispecific antibody induced more potent degradation of MET than treatment with a control antibody, and treatment of EBC-1 cells with the METxMET bispecific antibody inhibited downstream effectors of the MET pathway.

[0157] In another experiment, 5 million EBC-1 cells were implanted subcutaneously into the flanks of CB-17 SCID mice. Tumor volumes reached approximately 150 mm. 3Once tumor volume reached 1000 mg / kg, mice were randomized into six groups and treated twice weekly with control antibody at 25 mg / kg or METxMET bispecific antibody H4H14639D at 25 mg / kg. Tumor growth was monitored for 30 days post-implantation, and tumor volume (mm) was determined for each experimental group over time. 3 ) were measured. The results are shown in Table 8 and illustrated in Figure 4, and demonstrate that the METxMET bispecific antibody significantly inhibits the growth of EBC-1 tumors.

[0158] [Table 8] [Example]

[0159] MET×MET bispecific antibody does not induce proliferation of NCI-H596 lung cancer cells in vitro The effect of the MET x MET bispecific antibody on the proliferation of human non-small cell lung cancer (NSCLC) cells (NCI-H596) was evaluated in vitro. Ten thousand NCI-H596 lung adenosquamous carcinoma cells (Nai et al., J. Nat'l. Cancer Inst. 86(5):378-383, 1994) were seeded in 96-well plates on a 0.66% agar layer in medium supplemented with 1% fetal bovine serum (FBS). Cells were cultured in RPMI 1640 medium supplemented with 1% FBS containing 0.3% agarose. Cells were treated with: (1) an individual parental bivalent monospecific anti-MET antibody (H4H13306P2 or H4H13312P2) at 5 μg / ml, (2) a combination of two parental bivalent monospecific anti-MET antibodies (H4H13306P2 and H4H13312P2) at 2.5 μg / ml each, (3) a bispecific antibody (H4H14639D) containing one binding arm derived from H4H13306P2 and the other from H4H13312P2 at 5 μg / ml, or (4) hepatocyte growth factor (HGF) at 100 ng / ml. Cells were then incubated at 37°C with 5% CO. After two weeks, relative cell proliferation was determined by measuring the reduction of the indicator dye ALAMARBLUE® (Thermo Fischer Scientific, Waltham, MA) to its highly fluorescent form on a SPECTRAMAX® M3 plate reader (Molecular Devices, Sunnyvale, CA). Increased fluorescence correlates with cell proliferation. Table 9 and Figure 5 illustrate the relative NCI-H596 cell proliferation for each antibody treatment, normalized to control (untreated) NCI-H596 cell proliferation. Treatment of NCI-H596 lung cancer cells with HGF resulted in a strong induction of proliferation in soft agar. The MET×MET (MM in Figure 5) bispecific antibody H4H14639D did not significantly alter proliferation compared to treated control cells. A moderate induction of cell proliferation was observed with each parental bivalent monospecific antibody H4H13306P2 (M1) or H4H13312P2 (M2) individually or in combination (H4H13306P2 and H4H13312P2) (M1M2).

[0160] [Table 9] [Example]

[0161] Clinical trial of MetxMet bispecific antibody in non-small cell lung cancer background: Lung cancer is one of the most commonly diagnosed cancers and the leading cause of cancer-related deaths worldwide (Siegel et al., CA Cancer J Clin, 66(1):7-30, 2016). Non-small cell lung cancer (NSCLC) accounts for 80%-85% of all lung cancers and is composed of several histopathological subtypes, the most common of which are adenocarcinoma (40%-60%) and squamous cell carcinoma (30%) (Dela Cruz et al., Clin Chest Med, 32(4):605-44, 2011). The majority of patients with NSCLC are found to have advanced disease at the time of diagnosis.

[0162] First-line treatment of advanced NSCLC is guided by the presence of molecular abnormalities. Patients can receive targeted small molecule tyrosine kinase inhibitors (TKIs) (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), immune checkpoint inhibitor antibodies that block 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) (Reck et al., Ann Oncol, 25 Suppl 3:27-39, 2014), with or without maintenance therapy. Long-term survival remains an unmet need in advanced NSCLC. Overall survival (OS) is approximately 12 months for PD-1 / PD-L1 inhibitors, but exceeds 2 years in some studies in TKI-treated patients (Camidge et al., J Clin Oncol, 32:Abstract 8001, 2014).

[0163] 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 adipose tissue and is upregulated in several cancers. High levels of MET expression can occur through elevated protein expression or gene amplification in NSCLC and gastric tumors and are associated with negative patient outcomes (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). Mutations in MET resulting in exon 14 deletions promote prolonged ligand-dependent signaling, resulting in receptor stabilization and enhanced oncogenic potential (Kong-Beltran et al., Cancer Res, 66(1):283-9, 2006).

[0164] Approximately 3% of NSCLCs have been reported to contain MET exon 14 abnormalities (Cancer Genome Atlas Research Network, Nature, 511(7511):543-50, 2014) (Schrock et al., J Thorac Oncol, 11(9):1493-1502, 2016). Furthermore, gene 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). Amplification and increased expression of the MET gene are mechanisms of resistance to epidermal growth factor receptor (EGFR)-targeted therapy, and up to 25% of tumors resistant to third-generation TKIs may have 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).

[0165] Tumors with MET amplification or exon 14 deletion respond 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) (Paik, Cancer Discov, 5(8):842-9, 2015).

[0166] REGN5093 is a human bispecific antibody (bsAb) that binds to two distinct epitopes on MET with nanomolar affinity, blocks HGF binding to MET, and induces MET internalization and degradation without inducing MET-driven biological responses. In preclinical studies, REGN5093 demonstrated dose-dependent antitumor activity in immunocompromised mouse models of MET-driven cancer, including both exon 14 aberration and MET amplification models.

[0167] Unmet needs in lung cancer First-line treatment for advanced NSCLC is guided by the presence of molecular abnormalities. Patients with tumors exhibiting sensitive mutations in EGFR, anaplastic lymphoma kinase (ALK), or c-ros oncogene 1 receptor tyrosine kinase (ROS1) fusions are often targeted using small-molecule TKIs (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 lacking any of these activating mutations can receive immune checkpoint inhibitor antibodies that block the PD-1 receptor or PD-L1, with or without chemotherapy. Beyond these targeted systemic and immunotherapeutic approaches, advanced NSCLC is treated with platinum-based doublet 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). Long-term survival remains an unmet need in advanced NSCLC. Overall survival is generally 12 months for PD-1 / PD-L1 inhibitors, but some studies in TKI-treated patients have exceeded 2 years (Camidge et al., Nat Rev Clin Oncol, 16(6):341-55, 2019).

[0168] Anti-PD-1 and anti-PD-L1 therapy has transformed 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, even those with tumors that highly express PD-L1 or exhibit high tumor mutation burden (TMB), may not receive the same benefit from agents targeting the PD-1 / PD-L1 axis (Sabari et al., Ann Oncol, 29(10):2085-91, 2018). This is consistent with data generated in lung cancers with 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), indicating that anti-PD-1 or anti-PD-L1 monotherapy may not be the preferred treatment for patients with MET-driven disease. Thus, there remains a substantial unmet need for therapies that improve response rates and survival for patients with MET-abnormal NSCLC.

[0169] the purpose The primary objective of the dose-escalation (Phase I) portion of the 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 II dose (RP2D) of REGN5093 in patients with MET-altered NSCLC. A secondary objective of the dose-escalation (Phase I) portion of the study is to evaluate the preliminary antitumor activity of REGN5093 as assessed by objective response rate (ORR) according to Response Evaluation Criteria in Solid Tumors (RECIST 1.1).

[0170] The primary objective of the dose-expansion (Phase II) part of this study is to evaluate the preliminary antitumor activity of REGN5093 as assessed by ORR per RECIST 1.1. Secondary objectives of the dose-expansion (Phase II) part of this study are to evaluate the safety and tolerability of REGN5093 in each expansion cohort, as well as to evaluate the PK and concentrations of REGN5093 in serum.

[0171] Secondary objectives of both parts of this study are to evaluate the immunogenicity of REGN5093, as assessed by anti-drug antibodies (ADA), and other measures of preliminary anti-tumor activity.

[0172] Exploratory objectives of both parts of the study include assessing the relationship between REGN5093 efficacy and baseline MET aberrations / mutations or amplifications / expression and / or prior MET TKI treatment across the entire cohort; evaluating pharmacodynamic changes in putative serum or plasma biomarkers; and assessing the impact on efficacy of baseline and post-treatment tumor mutation spectrum in tissue and circulating tumor DNA (ctDNA expansion phase only).

[0173] Target population Adult patients ≥ 18 years of age (or legal age of adulthood to consent to participate in a clinical trial according to country-specific regulations).

[0174] Dose Escalation: Patients with advanced NSCLC presenting with previously documented MET aberration disease with 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 any documented MET aberration defined by any of the above criteria, regardless of prior experience with a MET-targeted TKI.

[0175] Dose expansion: Patients with advanced NSCLC presenting with MET-altered disease will be assigned to cohorts based on previously documented presence of: MET exon 14 mutant disease and prior MET-targeted TKI experience (Cohort 1A with MET TKI experience, 1B without prior MET TKI), high MET gene amplification (Cohort 2A without prior MET TKI), high MET protein overexpression (Cohort 2B without prior MET TKI), or both high MET gene amplification and high MET protein overexpression (Cohort 2C without prior MET TKI).

[0176] Documented MET status based on at least one test is sufficient to qualify a patient for the relevant cohort; testing in more than one category is not required. Thus, cohorts 2A and 2B may include patients with unknown overexpression or gene amplification status, respectively.

[0177] A dose-expansion phase of the study is planned to further explore the safety and biological activity of REGN5093 at the RP2D. Patients will receive the RP2D of REGN5093 administered IV via a 30-minute infusion.

[0178] Patients will be enrolled into separate cohorts according to their previously documented MET abnormalities and prior MET-targeted TKI experience. The cohorts will be designed to generate a relatively homogeneous patient population according to different cutoffs for three different types of biomarkers: MET exon 14 abnormalities, MET amplification, and MET protein overexpression.

[0179] The expansion cohorts are as follows (see also Table 10): Expansion cohort 1A (MET exon 14 aberrant NSCLC; MET TKI-experienced) Expansion cohort 1B (NSCLC with MET exon 14 abnormalities; no prior MET TKI) Expansion Cohort 2A (NSCLC with highly amplified MET GCN ≥5 and / or MET / CEP7 ratio ≥2 in tissue by FISH or MET GCN ≥6 by NGS or MET fold change ≥2 in ctDNA; no prior MET TKI) Expansion Cohort 2B (high MET protein expression as determined by IHC 3+ or H-score ≥200; no prior MET TKI) Expansion Cohort 2C (NSCLC with highly amplified MET GCN ≥5 and / or MET / CEP7 ratio ≥2 in tissue by FISH or MET GCN ≥6 by NGS or high MET protein overexpression in ctDNA by MET fold change ≥2 and IHC 3+ or H-score ≥200; no prior MET TKI) * If biopsy shows <40% tumor content, FISH or ctDNA results from local laboratories must be within the specified range

[0180] [Table 10]

[0181] Patients do not need to have been previously tested for all three categories of MET abnormality to be eligible for the relevant expansion cohort. For dose expansion, patients will be assigned to expansion cohorts (if there are open slots) in the following priority order if they are MET-TKI naive and have data from testing for more than one type of MET abnormality that indicates they could have been eligible for more than one cohort: Priority cohort: MET exon 14 abnormalities = Cohorts 1A and 1B Second priority (MET TKI-naive patients without MET exon 14 abnormalities or patients with exon 14 abnormalities only after Cohort 1B is filled): MET gene amplification + protein expression will lead to consideration for allocation to Cohorts 2A, 2B, and 2C.

[0182] The expansion cohort will have a Simon two-stage design. Enrollment in the expansion cohort will cease after the number of patients required for Stage 1 has been enrolled in the cohort. The cohort will be stopped or expanded pending the number of responses observed in Stage 1 and the corresponding Stage 1 criteria.

[0183] Inclusion criteria Patients must meet the following criteria to be eligible for inclusion in the study: Histologically confirmed NSCLC, advanced stage with no standard treatment options that may provide clinical benefit. Progressive is defined as unresectable or metastatic disease. Patients must have exhausted all approved and available therapies appropriate for the patient. Archived tumor tissue is available unless otherwise discussed with the medical monitor. Historical records of the following entities: - For the dose escalation cohort: MET-exon 14 mutation and / or MET gene amplification (MET amplification call by any local CLIA laboratory) 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 and no prior MET TKI, respectively - For dose expansion cohort 2A: Highly amplified MET gene (MET GCN ≥5 and / or MET / CEP7 ratio ≥2 in tissue by FISH or MET GCN ≥6 by NGS or MET fold change ≥2 in ctDNA); no prior MET TKI - For dose expansion cohort 2B: highly overexpressed MET protein (IHC 3+ or H-score ≥200; no prior MET TKI) - For dose expansion cohort 2C: Highly amplified MET gene (MET GCN ≥5 and / or MET / CEP7 ratio ≥2 in tissue by FISH or MET GCN ≥6 by NGS or MET fold change ≥2 in ctDNA) and highly overexpressed MET protein (IHC 3+ or H-score ≥200); no prior MET TKI * If biopsy shows <40% tumor content, FISH or ctDNA results from local laboratories must be within the specified range Willingness to donate tumor tissue from a newly obtained biopsy. A newly obtained biopsy at screening is required unless medically contraindicated and in consultation with the medical monitor. For patients in the expansion cohort, the biopsy must be taken from a tumor site that has not been previously irradiated and is not the only measurable target lesion. For expansion cohort only: At least one lesion measurable by RECIST 1.1. Tumor lesions in previously irradiated areas are considered measurable if there is documented progression in such lesions after radiation. Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1 Adequate organ and bone marrow function documented by: Hemoglobin ≧9.0g / dL Absolute neutrophil count ≥ 1.5 × 10 9 / L Platelet count ≥ 75 × 10 9 / L Serum creatinine ≤1.5× ULN or estimated glomerular filtration rate (GFR) ≥30 mL / min / 1.73 m 2 Proper liver function: Total bilirubin ≤ 1.5 × ULN (≤ 3 × ULN if tumor infiltrates the liver) AST ≤ 2.5 × ULN (≤ 5 × ULN if tumor invades the liver) ALT ≤ 2.5 × ULN (≤ 5 × ULN if tumor invades the liver) Alkaline phosphatase ≤2.5x ULN (≤5x ULN if tumor invades liver or bone) Special note: (a) Patients with tumor hepatic infiltration will be excluded regardless of the above criteria if their AST level is ≥3× ULN or their ALT level is ≥3× ULN and their bilirubin level is ≥2× ULN. (b) Patients with Gilbert syndrome do not need to meet the total bilirubin requirement if their total bilirubin is not greater than their preexisting level. Gilbert syndrome is adequately documented in their past medical history. Adult patients ≥ 18 years of age (or the legal age of adulthood to consent to participate in a clinical trial according to country-specific regulations). · Willingness and ability to comply with all medical and study-related procedures and requirements. · Must be willing and able to provide informed consent as specified by health authorities and institutional guidelines Provide signed informed consent from the study patient or legal representative

[0184] Exclusion criteria Patients who meet any of the following criteria will be excluded from the study: Have been treated with approved systemic therapy or participated in any study of an investigational drug or device according to the following timeframes: - For small molecule cytotoxins or other agents unlikely to interact with the investigational drug: at least 7 days after the first dose of investigational therapy and within 2 weeks or 5 half-lives of the preceding treatment, whichever is shorter. - Exception: Patients who have undergone or are participating in a study involving treatment with an investigational immunoPET reagent are not excluded. Have not yet recovered (i.e., grade ≤ 1 or baseline) from any acute toxicity attributable to prior therapy, except for the following: - Laboratory changes as described in the inclusion criteria, and - Patients with grade ≤2 neuropathy For immune-mediated AEs affecting any organ system within 2 months prior to enrollment, there must be documentation of the trajectory of improvement of this irAE (Grade ≤ 1 by time of enrollment or to baseline) and of those toxicities that remain Grade 1, with documentation of two stable assessments at least 4 weeks apart. Note: Endocrine-immune mediated AEs controlled with hormone therapy or other non-immunosuppressive therapies that have not resolved prior to enrollment are acceptable. - Have undergone radiation therapy or major surgery within 14 days of the first dose of study drug, or have not recovered from an AE (i.e., grade ≤ 1 or baseline), except for patients with laboratory changes described in the inclusion criteria and neuropathy grade ≤ 2 For expansion cohorts only: Prior treatment with a MET-targeted biopharmaceutical therapy (function-blocking antibody or ADC). Additionally, for expansion cohorts 1B, 2A, 2B, and 2C, prior treatment with any MET-targeted agent, including small molecule tyrosine kinase inhibitors, e.g., crizotinib, capmatinib, and tepotinib. For expansion cohort only: Other malignancies, with the following exceptions: - Non-melanoma skin cancer that has been treated with potentially curative therapy, or - Cervical intraepithelial neoplasia or - Other tumors that have been previously treated, patients must be in complete remission for at least 2 years prior to enrollment and will not require further therapy during the study Untreated or active primary brain tumor, CNS metastases, leptomeningeal disease, or spinal cord compression - Exception: Patients previously treated for central nervous system metastases or spinal cord compression may participate if: - No evidence of progression for at least 2 weeks prior to the first dose of study treatment, with any neurological symptoms returning to baseline Encephalitis, meningitis, organic brain disease (e.g., Parkinson's disease), or uncontrolled seizures within 1 year prior to the first dose of investigational therapy Uncontrolled infection with human immunodeficiency virus, hepatitis B, or hepatitis C infection; or a diagnosis of immunodeficiency Special note: (a) Patients with known HIV infection who have controlled their infection (either naturally or on a stable antiviral regimen, with an undetectable viral load [HIV RNA PCR] and CD4 count >350) are eligible. For patients with controlled HIV infection, monitoring will be performed according to local agency standards. (b) Patients with known hepatitis B (HepBsAg+) who have controlled infection (serum hepatitis B virus DNA PCR below the limit of detection and receiving hepatitis B antiviral therapy) are eligible. Controlled patients will undergo regular HBV DNA monitoring. Patients will remain on antiviral therapy for at least 6 months beyond the last dose of investigational drug. (c) Patients with known hepatitis C virus antibody positivity (HCV Ab+) who have controlled the infection (HCV RNA undetectable by PCR, either spontaneously or in response to a prior successful course of anti-HCV therapy) are eligible. Any infection requiring hospitalization or treatment with IV anti-infectives within 2 weeks prior to the first dose of investigational therapy. Placeholder for removed exclusion criteria Placeholder for removed exclusion criteria - Known psychiatric or substance abuse disorder that would interfere with participation in this study Any medical condition, comorbidity, physical examination findings, metabolic dysfunction, or laboratory abnormality that, in the opinion of the investigator, makes the patient unsuitable for participation in the study because it poses a high safety risk and / or may affect the interpretation of the results of the study. Women with a positive serum hCG pregnancy test at the screening / baseline visit. Lactating women will also be excluded. Women of childbearing potential who are unwilling to use highly effective contraception before the start of the initial dose / initial treatment, during the study, and for at least 6 months after the final dose * or men. Highly effective contraceptive measures include: - Stable use of combined (estrogen and progestogen-containing) hormonal contraceptives (oral, vaginal, transdermal) or progestogen-only hormonal contraceptives (oral, injectable, implantable) associated with ovulation inhibition, initiated for at least two menstrual cycles prior to screening - Intrauterine Device (IUD); Intrauterine Hormone-Releasing System (IUS) - Bilateral tubal obstruction - A partner who has had a vasectomy (provided that the partner of the male who has had a vasectomy is the study participant's only sexual partner and has undergone a medical evaluation of the surgical success of the procedure) - and / or sexual abstinence †,‡ * A woman of childbearing potential is defined as a woman who is capable of becoming pregnant from menarche until menopause, excluding those who are permanently sterile. Permanent methods of contraception include hysterectomy, bilateral salpingectomy, bilateral tubal ligation, and bilateral oophorectomy. Postmenopausal status is defined as 12 months of amenorrhea without other medical causes. In women not using hormonal contraceptives or hormone replacement therapy, a high level of follicle-stimulating hormone (FSH) within the postmenopausal range may be used as confirmation of menopausal status. However, in the absence of 12 months of amenorrhea, a single FSH measurement is insufficient to determine the occurrence of postmenopausal status. The above definition conforms to the guidance of the Clinical Trial Facilitation Group (CTFG). Pregnancy testing and contraception are not required for women with a documented hysterectomy or tubal ligation. † Sexual abstinence is considered highly effective only if it is defined as abstinence from heterosexual intercourse for the entire duration of the risks associated with the investigational drug. The reliability of sexual abstinence should be assessed in relation to the duration of the trial and the patient's preferred usual lifestyle. ‡Periodic abstinence methods (calendar, symptom-thermal, post-ovulation), pull-out (abortive intercourse), spermicide alone, and lactational amenorrhea (LAM) are not acceptable methods of contraception. Do not use female and male condoms together.

[0185] Methods - Research Design Patients with documented MET abnormalities in at least one of three major categories will be 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 in tissue by FISH or MET GCN ≥6 by NGS or MET fold change ≥2 in ctDNA), and MET protein overexpression (immunohistochemistry 3+ or H-score ≥200). Patients are not required to have previously undergone testing for all three categories of MET abnormalities.

[0186] Figure 6 illustrates the flow of this study from the initial screening period to post-treatment follow-up.

[0187] Description of study cohorts and dose escalation Patients underwent a screening procedure to determine eligibility after signing an informed consent form (ICF) within 28 days prior to their first dose of REGN5093.

[0188] Dose escalation The dose escalation phase investigated a range of three DLs of REGN5093: 500, 1000, and 2000 mg administered IV via a 30-minute infusion once every three weeks (Q3W) (see Table 11).

[0189] [Table 11]

[0190] A modified 3+3 dose escalation design ("4+3") will be utilized (Le Tourneau et al., J Natl Cancer Inst, 101(10):708-20, 2009). Dose escalation will continue until the MTD is reached or a dose is selected for expansion based on sufficient evidence of safety / tolerability and response (RP2D). Dose-decreasing (DL-1) and intermediate cohorts may be enrolled to explore intermediate doses if the initial or subsequent levels, respectively, are deemed to be intolerable. At the sponsor's discretion (in consultation with the investigator), up to six additional patients may be enrolled at any DL deemed tolerable to further evaluate safety and gather biopharmaceutical information. A schematic diagram of the study design is shown in Figure 7.

[0191] The dose-limiting toxicity (DLT) evaluation period will be 21 days, starting on Day 1 of Cycle 1. A minimum of three patients per DL must be evaluable for DLT, but to maximize the efficiency of Phase 1 dose escalation while maintaining patient safety, four patients per DL will be enrolled in case a patient discontinues before being evaluable for DLT. Tolerability rules are as follows:

[0192] DL tolerability will be considered achieved if all potential DLT-evaluable patients complete the 21-day DLT period without DLT (0 of 3 patients or 0 of 4 patients).

[0193] Of note: If three patients complete the DLT period without experiencing a DLT, but there is a fourth patient in the DLT evaluation period, DL tolerability will be considered achieved only if the fourth patient discontinues therapy before completing the DLT evaluation period or being evaluable for DLT.

[0194] If there is one DLT in either three or four DLT-evaluable patients, then enroll four or three more patients, respectively, for a total of seven patients. If there is one DLT in six or seven patients, the dose is considered tolerable. If there are two or more DLTs in two to seven evaluable patients, the MTD is reached.

[0195] At the highest tolerated DL, an additional 3-4 patients may be enrolled to further evaluate safety, for a total of 6-10 DLT-evaluable patients. A dose will be considered tolerable if there are 0-1 DLT in 6-8 patients or up to 2 DLTs in 9-10 patients.

[0196] All of the first three or four patients enrolled in a cohort will be observed for at least 21 days, complete safety assessments on Day 22 (Day 1 of Cycle 2), and escalation to the next dose cohort will occur once the data have been reviewed at a Dose Escalation Review meeting.

[0197] After the required number of patients have been enrolled in a given dose cohort, enrollment is paused for DLT assessment (although screening of the next dose cohort may begin before the current dose is confirmed to be safe). A dose escalation review meeting is led by a designated member of the sponsor's clinical team (typically either the Medical Director or Investigator) and is attended by at least the sponsor's Medical Director / Investigator and Global Patient Safety Lead; other individuals, including the investigator, may be included. Dose cohorts are stopped, expanded, or escalated according to the dose escalation criteria.

[0198] Dose-limiting toxicity A dose-limiting toxicity (DLT) is any toxicity that may prevent progression to a higher dose as specified in this protocol. The DLT observation period for assessing the safety of dose escalation is defined as 21 days, beginning on Day 1 of Cycle 1, to monitor the safety and tolerability of the initial dose of REGN5093. To be evaluable for DLT, patients: - Received at least one dose of study drug and been monitored for at least 21 days following the first dose of study drug, or experiences a DLT (as defined below) prior to the completion of the DLT Period; It is necessary to do so.

[0199] The duration of the DLT observation period may be longer for patients experiencing an AE whose duration needs to be assessed to determine whether the event was a DLT.

[0200] If such an event occurs during the DLT observation period, regardless of whether the patient remains on study treatment and / or continues to participate in study procedures, the event will be counted as a DLT for the involved cohort.

[0201] Definition of dose-limiting toxicity DLT is generally defined as any of the following treatment-emergent toxicities, excluding those 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 febrile neutropenia (fever ≥38.5°C and absolute neutrophil count [ANC] <1.0 x 10 9 / L) or documented infection with grade ≥3 neutropenia Non-hematological toxicity: - Non-hematologic grade ≥ 3 toxicity, excluding: Alopecia b. Grade 3 nausea, vomiting, or diarrhea unless persistent (>72 hours duration) after initiation of supportive care measures prescribed by the treating physician C. Clinically insignificant laboratory abnormalities - Clinically significant grade ≥ 3 laboratory values ​​requiring medical intervention or leading to hospitalization Liver dysfunction consistent with Hy's rule (Temple, J Allergy Clin Immunol, 117(2):391-97, 2006) or ALT or AST >3x ULN and bilirubin >2x ULN

[0202] The frequency, time to onset, and severity of toxicities, as well as the success of standard medical management and dose interruptions / delays, will be analyzed to determine whether a given toxicity should be considered a DLT for dose escalation purposes.

[0203] In general, given the limited clinical experience with the novel biopharmaceutical molecule REGN5093, any AEs will be treated as unexpected.

[0204] Treatment-emergent adverse events that appear to meet the definition of a DLT will be discussed between the sponsor and the investigator. The final determination of whether a TEAE meets the DLT definition will be based on careful review of all relevant data and consensus between the Medical / Study Director and the designated Safety Leader from Global Patient Safety. Consultation with the investigator may be requested.

[0205] If an event that meets the DLT criteria occurs during the DLT observation period, regardless of whether the patient remains on study treatment and / or continues to participate in study procedures, the event will be counted as a DLT for the involved cohort.

[0206] maximum capacity The MTD is defined as the DL level immediately below which treatment is discontinued due to the occurrence of two or more DLTs among up to seven evaluable patients. If the study is not discontinued due to the occurrence of DLTs, the MTD is considered not to have been determined.

[0207] If the MTD is not reached, the RP2D for further evaluation may be selected based on clinical, PK, and / or biomarker data demonstrating that a pharmacologically active dose has been reached, in conjunction with available safety information.

[0208] REGN5093 is supplied as a lyophilized product in sterile, single-use vials. A new presentation, a sterile solution, is planned to be introduced during the trial. Each vial contains REGN5093 at a concentration of 25 mg / ml. Both presentations contain labeled vials in labeled boxes.

[0209] Instructions for dose preparation will be provided in the Investigational Product Administration Procedures.

[0210] No premedication is required prior to administration of REGN5093. REGN5093 is administered by IV infusion over 30 minutes Q3W.

[0211] Contraindicated drugs While participating in this study, patients must not receive any standard or investigational agents for the treatment of their tumors other than REGN5093, according to the study's specific dosing regimen.

[0212] No systemic cancer treatment is permitted during the study period.

[0213] Patients should not receive live vaccines during the study period.

[0214] Radiation therapy will not be permitted during the study, with the following exceptions: After consultation with the sponsor, local palliative treatment (e.g., radiation) will be permitted for local control of tumor. After consultation with the sponsor, palliative radiation therapy for pain control at the site of bone disease or brain lesions will be permitted (unless the lesions are followed for treatment response evaluation).

[0215] Any other medications deemed necessary for the patient's welfare and expected not to interfere with the evaluation of the investigational drug may be given at the investigator's discretion.

[0216] Permitted drugs 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. Long-term use of high-dose steroids requires consultation with a medical monitor.

[0217] safety The safety and tolerability of REGN5093 will be monitored by clinical evaluation of AEs, physical examination (complete and limited), repeated measurement 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, will be collected pre-dose.

[0218] An AE is any untoward medical occurrence in a patient administered an investigational drug, which may or may not have a causal relationship to the investigational drug. Thus, an AE is any untoward and unintended sign (including abnormal clinical laboratory findings), symptom, or disease temporally associated with the use of an investigational drug, whether or not considered related to the investigational drug (ICH E2A Guideline: Clinical Safety Data Management: Definitions and Standards for Expedited Reporting, October 1994).

[0219] An SAE is any untoward medical occurrence of any of the following at any dose: Fatal - all deaths, including those considered completely unrelated to the study drug (e.g., motor vehicle accidents in which the patient was a passenger). Life-threatening - Any AE that, in the investigator's opinion, poses an immediate risk of death to the patient at the time of the event. This does not include AEs that may have caused death if they had occurred in a more severe form. Requiring 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 originally expected for the event, or a hospital stay that is prolonged due to the occurrence of a new AE as determined by the investigator or treating physician. Permanent or serious impairment / disability (substantial destruction of the ability to carry out normal life functions) · Congenital anomalies / birth defects Significant Medical Event. A significant medical event is not immediately life-threatening or likely to result in death or hospitalization, but may endanger the patient or require intervention to prevent one of the other serious outcomes listed above (e.g., intensive care in the emergency room or at home for allergic bronchospasm; hematopoietic dysfunction or seizures that do not result in hospitalization; or the development of drug addiction or drug abuse).

[0220] Hospitalization or death due solely to symptomatic episodes consistent with the typical progression of the underlying malignancy is not considered an SAE.

[0221] efficacy Radiographic tumor response is 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 informs the calculation of: ORR: defined as the percentage of patients with a complete response (CR) or partial response (PR) DOR: For patients with confirmed CR or PR, defined as the time from the first CR or PR to the first radiographic progression or death from any cause. In the absence of radiographic progression or death before the analysis cutoff date or the start of further anti-cancer treatment, DOR will be censored at the date of the last valid response assessment without progression performed before the analysis cutoff date or the start of further anti-cancer treatment, whichever occurs first. DCR: defined as the percentage of patients with CR, PR, or stable disease (SD) PFS: Defined as the time from first study treatment administration to first radiographic progression or death from any cause, using the same censoring rules as DOR. OS: Defined as the time from the first administration of study treatment to death from any cause. For patients who do not die, OS will be censored at the last date the patient is known to be alive. OS will be assessed based on investigator-reported survival data.

[0222] Diagnostic-quality CT with contrast and contrast-enhanced MRI are the preferred imaging modalities for assessing radiographic tumor response. In patients with strict contraindications to contrast, a non-contrast CT scan of the chest and a non-contrast MRI scan of the rest of the body are sufficient. Image the chest, abdomen, and pelvis, along with other known or suspected sites of disease. If more than one imaging modality is used at screening, use the most accurate imaging modality that adheres to RECIST 1.1 (Appendix 1) when recording data. The same imaging modality and techniques used at screening should be used for all subsequent evaluations.

[0223] At screening, patients with known previously treated brain metastases should undergo brain MRI or CT with contrast (unless contraindicated, otherwise MRI without contrast).

[0224] Additional sites of known disease should also be imaged at screening.

[0225] Diagnostic-quality (≦5 mm slices) CT scans with contrast of the chest and abdomen and other sites of known disease (e.g., neck) will be performed at screening, on Day 1 of treatment cycles 2 and 3, every 3 months after the final visit, and whenever disease progression is suspected. Scans will include a description of tumor location, with up to five largest predominant disease masses (no more than two per organ) selected as target lesions and assessed by longest diameter for non-lymph node disease and short axis for nodal disease according to RECIST 1.1. All lesions will be assessed and recorded. If a CT scan is not feasible, an MRI scan may be performed.

[0226] For each patient, the same measurement method and the same technique will be used to evaluate each lesion throughout the study. If a patient inadvertently misses a routine tumor assessment, or if a technical error prevents the assessment, the patient may continue treatment until the next scheduled assessment unless there are signs of clinical progression. At any time during the treatment phase, unscheduled tumor assessments will be performed if there is suspicion of disease progression based on clinical or laboratory findings (and before the next scheduled assessment).

[0227] Procedures and Evaluation Antitumor activity will be assessed by computed tomography (CT) or magnetic resonance imaging (MRI). Safety and tolerability of REGN5093 will be monitored by clinical evaluation of AEs, physical examination (complete and limited), repeated measurement of vital signs (temperature, blood pressure, pulse, and respiration), 12-lead electrocardiogram (ECG), and laboratory evaluations including standard hematology, chemistry, and urinalysis.

[0228] Blood will be collected to assess serum REGN5093 PK and concentration, serum immunogenicity (ADA), and for further biomarker evaluation. Additional biomarkers will be measured in serum or plasma. Exploratory predictive and pharmacodynamic biomarkers related to REGN5093 treatment exposure, clinically active disease, or underlying disease will be investigated using collected serum, plasma, samples from archived tumor tissue and tumor biopsy tissue during the study, tumor DNA (including circulating tumor DNA), and tumor RNA samples.

[0229] Test evaluation items The primary endpoints of the dose-escalation (Phase I) portion of the study are: Safety, as assessed by the incidence and severity of treatment-emergent adverse events (TEAEs), adverse events of special interest (AESIs), serious adverse events (SAEs), and grade ≥3 laboratory abnormalities during treatment and up to 90 days after the last dose. Tolerability, as assessed by the incidence of dose-limiting toxicities (DLTs) from the first dose of REGN5093 to the end of the DLT observation period Serum REGN5093 concentrations over time

[0230] The primary endpoint of the dose-expansion (Phase II) part of the study was ORR per RECIST 1.1, defined as the percentage of patients with a best overall response (BOR) of confirmed CR or PR according to RECIST 1.1 criteria.

[0231] Secondary endpoints for the dose escalation portion of the study are: ORR per RECIST 1.1.

[0232] Secondary endpoints of the dose-expansion portion of the study include: Safety, as assessed by the incidence and severity of TEAEs, AESIs / SAEs, and grade ≥ 3 laboratory abnormalities. PK and concentration of REGN5093 in serum over time

[0233] Secondary endpoints for both phases of the study include: 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. Operating System Immunogenicity of REGN5093 as assessed by ADA.

[0234] Exploratory endpoints for both parts of the study include: Response to REGN5093 by type (and degree) of baseline MET aberration and prior MET-targeted TKI experience Response to REGN5093 by baseline tumor mutation status [Example]

[0235] Safety, tolerability, and efficacy of REGN5093 in patients with MET-altered advanced NSCLC REGN5093 has therapeutic benefit in patients with MET-altered NSCLC and has demonstrated promising efficacy signals with a tolerable safety profile.

[0236] REGN5093 was investigated as monotherapy, administered intravenously once every three weeks in dose-escalation cohorts (Phase I), followed by an expansion phase (Phase II). For each patient, the study consisted of a screening period of up to 28 days, followed by three-week cycles of REGN5093 monotherapy. REGN5093 at 2000 mg was the recommended dose for Phase II, with preceding dose levels of 500 mg and 1000 mg.

[0237] Tumor measurements were performed at baseline and every 6 weeks until disease progression, withdrawal of consent, death, or initiation of alternative anticancer treatment.

[0238] Tumor tissue (archive and on-study tumor biopsies) was collected and utilized for retrospective analysis of MET aberrations (and further biomarker analysis if tissue was available), as were fresh biopsies obtained at screening unless deemed unsafe.

[0239] Sixty-nine patients received REGN5093 in both the dose-escalation and dose-expansion phases. Patient demographics were consistent with a heavily pretreated population, with a median of 2.5 (range, 1-8) prior lines of therapy. Most patients had ECOG PS 1 (80%). Most patients had non-squamous histology (93.6%), and EGFR mutations were present in 37.2% of patients (Table 12). The study population had a median age of 66 years, 53.8% were male, and 70.5% were Asian.

[0240] [Table 12]

[0241] Safety Data No dose-limiting toxicities (DLTs) were observed. REGN5093 demonstrated a similar safety profile in the dose-escalation and dose-expansion phases, notably, only 6 (9%) patients experienced grade 1 / 2 peripheral edema (Table 13). At the time of data cutoff, 9 (13%) patients were still on treatment, and 60 (87%) patients had discontinued treatment. The primary reasons for treatment discontinuation were: disease progression in 52 (75%) patients; patient discretion in 5 (7%) patients; and adverse events in only 3 (4%) patients.

[0242] [Table 13]

[0243] Tumor response Partial responses (by investigator assessment) were observed in a subset of patients naive to MET TKI therapy who had exon 14 aberrations in DNA or deletions resulting in exon 14 skipping, as well as in patients with MET gene amplification and / or MET protein overexpression (Table 14 and Figure 8).

[0244] [Table 14]

[0245] ORR among patients with centrally confirmed MET aberrations: 33% (3 / 9) with MET exon 14 mutations in tumor tissue or ctDNA by NGS (MET TKI-naive); 25% (5 / 20) with MET gene amplification (GCN ≥5 in tumor tissue by FISH or NGS); 23% (5 / 22) with MET protein overexpression (IHC 3+ in ≥50% of tumor cells); 36% (5 / 14) with MET protein overexpression (IHC 3+ in ≥75% of tumor cells); and 50% (4 / 8) with MET protein overexpression (IHC 3+ in ≥90% of tumor cells).

[0246] Pharmacokinetics Serum REGN5093 exposure appeared linear and dose-proportional over the 500 mg to 2000 mg Q3W intravenous (IV) dose range, with serum concentrations at 2000 mg Q3W IV being similar in the dose-escalation cohorts and across expansion cohorts (see Figure 9). The elimination half-life estimated by noncompartmental analysis over a 3-week dose interval was 15 days.

[0247] conclusion Among this population of heavily treated patients with MET-altered advanced NSCLC, REGN5093 monotherapy demonstrated an acceptable safety profile. No DLTs were observed. 86% of patients experienced TEAEs of any grade. 26% experienced TEAEs of grade ≥3. Three (4%) patients discontinued treatment due to TEAEs. Serum REGN5093 exposure appeared to increase in a dose-dependent manner. REGN5093 monotherapy demonstrated preliminary efficacy signals among patients with MET exon 14 mutations or deletions resulting in exon 14 skipping in DNA, as well as patients with MET gene amplification and / or MET protein overexpression. Tumor responses were enhanced with centrally confirmed biomarker selection. [Example]

[0248] Predictive biomarkers of response to REGN5093 In some cases, when treating MET-altered advanced non-small cell lung cancer (aNSCLC), patient selection can be guided by the use of predictive biomarkers of response to REGN5093. REGN5093 monotherapy was investigated in patients with MET-altered aNSCLC. Tumor measurements were performed at baseline and Q6W until progression, withdrawal of consent, death, or initiation of alternative anticancer treatment.

[0249] Provided herein are selection criteria that can be used to improve the percentage of responders when treating aNSCLC. In some embodiments, identification of MET actionable mutations can be used to select patients for MET-targeted therapy in advanced (2L+, i.e., patients who have received two or more prior therapies) NSCLC. MET exon 14 skipping / deletion is an oncogenic driver in 1L NSCLC, causing loss of the c-Cbl binding site, which impairs receptor degradation and ultimately leads to increased MET signaling. MET gene amplification is a resistance mechanism to EGFR tyrosine kinase inhibitor (TKI) therapy in 2L+ NSCLC. MET protein overexpression enhances, but does not select for, therapeutic response to MET TKIs, particularly in the MET-amplified TKI resistance-driven NSCLC population (Figure 10).

[0250] Both tumor and fluid biopsies can be used to identify and confirm MET abnormalities. Exon 14 abnormalities can be determined via gene sequencing of ctDNA obtained from tumor biopsies or blood samples. Similarly, MET gene amplification can be assessed via gene sequencing or fluorescence in situ hybridization (FISH; GCN or MET: chromosome 7 centromere (CEP7) ratio) of tumor biopsies or via sequencing of ctDNA obtained from blood samples. MET protein expression can be assessed in tumor biopsies, for example, by immunohistochemistry using a specific c-MET antibody that stains total MET protein (Figure 11).

[0251] In some cases, assessing ctDNA complements tissue profiling and overcomes the limitations of biopsy collection and analysis. For example, some tumors are located in areas where obtaining tissue biopsies can be difficult or impossible. ctDNA assessment captures all active "drivers" from all tumor sources within the body, both primary and metastatic, allowing for the identification of mutations not present in a single tissue biopsy, thereby overcoming spatial heterogeneity. Because tumors progress over time, either intrinsically or in response to therapy, or both, assessing simultaneously obtained tissue and ctDNA samples provides greater concordance and accurate coverage, thereby overcoming temporal heterogeneity.

[0252] MET amplification status was assessed using tissue and / or ctDNA by FISH and NGS, for example, using the following gene copy number (GCN) thresholds: a GCN threshold in tissue by either fluorescence in situ hybridization (FISH) (GCN ≧5) or NGS (GCN ≧6), and / or a GCN threshold in ctDNA of ≧2.2× by NGS assay, to evaluate the efficacy of other MET inhibitors.

[0253] MET exon 14 was confirmed centrally (i.e., assayed by one central laboratory to control for any variability arising from sample collection and assay performance) using various next-generation sequencing (NGS) panels (FoundationOne® CDx (tissue-based 324-gene panel) and FoundationOne® LiquidCDx (blood-based 324-gene panel) are exemplary gene panels, although other panels are contemplated as useful herein) on tissue and ctDNA. Exemplary exon 14 abnormalities include, but are not limited to, D1010N, D1010fs ... * 19, D1010Y, D1010H or R1004P mutations and exon 14 skipping.

[0254] Figure 12 provides the study design. The various cohorts included patients with confirmed MET abnormalities, as determined by a given patient's medical records. As shown in Figure 12, patient enrollment during the expansion phase of the trial was based on documented MET abnormalities (MET exon 14 abnormalities or deletions resulting in exon 14 skipping in DNA, MET gene amplification, and / or MET protein overexpression) and correlated with clinical response. As can be seen in Figure 13, of the 65 patients who received the 2000 mg dose of REGN5093, 9 patients had a partial response. The distribution of responders by cohort was as follows: Cohort 1A (MET Ex 14 TKI-experienced): 0 patients; Cohort 1B (MET Ex 14 TKI-naive): 4 patients; Cohort 2A (MET amplification): 0 patients; Cohort 2B (MET overexpression): 1 patient; Cohort 2C (MET amplification and overexpression): 3 patients; DL3 (dose level 3, MET Amp and OE): 1 patient.

[0255] Tumor responses were observed in MET TKI-naive patients with centrally confirmed (1) MET exon 14 (4 / 15) or 27% or (2) MET Amp+OE (5 / 14) 36%, regardless of EGFR mutation status, as shown in Figure 14. Overall response rates (%ORR) were higher in specific MET aberration subgroups compared to the overall population by central analysis, as shown in Figure 15. [Example]

[0256] Baseline bypass resistance mutations and clinical response to REGN5093 Resistance to MET therapy can be intrinsic or acquired in response to prior therapy in a patient population with pretreated 2L+ NSCLC. Patients received a range of prior therapies from 1 to 8, with a median of 2.5 (Figure 16). Prior therapies received by patients in this study included chemotherapy, immune checkpoint inhibitors (ICIs), and EGFR inhibitors. Patients who initially responded to therapy, in some cases, acquired other mutations that promoted disease progression upon treatment.

[0257] Nonsynonymous variations such as single nucleotide variations (SNVs), insertion deletions (indels), frameshifts, nonsense mutations, splice variants, or gene fusions, and bypass gene detection of copy number variations (CNVs) such as gene amplifications or deletions in ctDNA complemented the tissue results and provided a more comprehensive tumor profiling of METxMET resistance mechanisms. The sensitivity variation for detection by the FMI NGS panel depended on the variant allele frequency (VAF) for each of the 324 genes present on each platform. Figure 17 illustrates the overlap between nonsynonymous variation and CNV detection and detection by ctDNA and tissue. NSV detection by ctDNA was 39%, compared with 33% for tissue and 28% for both. CNV detection by ctDNA was 9%, compared with 84% for tissue and 6% for both.

[0258] Figure 18 illustrates unbiased co-clustering of baseline somatic mutations detected in both ctDNA and tumor tissue by cohort assignment, EGFR status, and centrally confirmed MET aberrations. Figure 19 shows another unbiased set of clusterings, as in Figure 18, but with clinical response. Several baseline somatic mutations were identified in non-responders based on the clustering dataset; such mutations may confer MET bypass resistance mechanisms, potentially impacting clinical response to REGN5093 even in the presence of MET oncogenic drivers.

[0259] Figure 20 provides classification and examples of bypass aberrations detected in study patients with centrally confirmed MET oncogenic drivers who did not respond to REGN5093. 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 alternative or parallel activation of tyrosine kinase driver receptors include gene amplification of tyrosine kinase receptors (TKRs) and TK ligands (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 include the JAK2 / STAT3 pathway (e.g., JAK2 V617F; SFKs amp); RAS / RAF / MEK / MAPK pathway (e.g., KRAS G12A / V and G12D / V; GNAS R201H, MKRN-BRAF fusion, BRAF S602Y; RICTOR Amp; 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 deletion); TP53 mutations (e.g., TP53 R280T, TP53 R248Q) and cell cycle mutations (e.g., CDK4 Amp, CDK6 Amp, CCND1 Amp, CCNE1 Amp).

[0260] Gene amplifications identified in patients with confirmed Met amplification and overexpression who did not respond to RENG5093 included: HGF, EPH, EGFR, BRAF, BCL2L1, PI3KCB, KRAS, AKT2, ATR, VEFGA, FGF, CCND, CCNE, CDK6, RAD21, and MYC. Gene deletions: CDKN2A, CDKN2B, MTAP, and RBM10 were also identified. The MET bypass resistance mechanisms identified in these patients are provided in Table 15.

[0261] [Table 15]

[0262] Gene amplifications identified in patients with confirmed MET amplification (but not MET overexpression) who did not respond to REGN5093 were mostly EGFR mutants: 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 included: CDKN2A, CDKN2B, MTAP, TEK, and BCOR. The MET bypass resistance mechanisms identified in these patients are provided in Table 16.

[0263] [Table 16]

[0264] Gene amplifications identified in TKI-naive patients with MET exon 14 abnormalities who 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. The MET bypass resistance mechanisms identified in these patients are provided in Table 17.

[0265] [Table 17]

[0266] Gene amplifications identified in TKI-experienced patients with MET exon 14 aberrations who were non-responders to REGN5093 included: 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 included CDKN2A, CDKN3A, and MTAP. The MET bypass resistance mechanisms identified in these patients are provided in Table 18.

[0267] [Table 18] [Example]

[0268] Circulating biomarkers of target capture Soluble MET (sMET) is an extracellular domain fragment of MET derived from the receptor in tumor tissues cleaved by proteases. Total sMET and HGF were measured by ELISA.

[0269] Total concentrations of REGN5093 were several-fold higher than total sMET concentrations in serum, suggesting that saturation of receptor occupancy was achieved with the 2000 mg Q3W dose regimen at dose escalation (Figure 21).

[0270] HGF is a ligand for the MET receptor, which is displaced and increased in the circulation when REGN5093 binds to the MET receptor in tumors. Although circulating HGF (cHGF) and total sMET levels both increased after dosing, suggesting target capture, neither baseline nor post-treatment levels of sMET and cHGF were significantly associated with clinical response (Figure 22).

[0271] conclusion Of 36 pts receiving the 2000 mg dose, 6 had partial responses (5 had prior anti-PD-(L)1 therapy). These responses occurred in 2 / 5 pts with exon 14 skipping mutations who were MET tyrosine kinase inhibitor (TKI)-naïve (Cohort 1B); 0 / 10 pts with exon 14 skipping mutations who had previously been treated with a TKI (Cohort 1A); and 4 / 21 pts with MET TKI-naïve MET amplification, protein overexpression, or both (Cohorts 2A-C).

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

[0273] Certain baseline somatic mutations that co-occur with MET aberrations in clinical non-responders act as potential bypass resistance mechanisms and influence clinical response to REGN5093 monotherapy.

[0274] Total REGN5093 concentrations were several-fold higher than total sMET concentrations in serum, supporting the choice of the 2000 mg Q3W dosing regimen. Total sMET and cHGF levels increased after dosing, suggesting target capture by REGN5093, but neither baseline nor posttreatment changes in total sMET or cHGF concentrations were associated with response.

[0275] [Table 19-1] [Table 19-2] [Table 19-3] [Table 19-4] [Table 19-5] [Table 19-6]

Claims

1. A bispecific antibody for use in a method to treat non-small cell lung cancer (NSCLC) in subjects with tumors exhibiting MET abnormalities, to reduce the growth of NSCLC tumors, and / or to induce regression of NSCLC, The method includes administering a dose of approximately 250 to 2000 mg of the bispecific antibody to the target population. The bispecific antibody is A first antigen-binding domain (D1) comprising three heavy chain complementarity-determining regions (CDRs) (HCDR1, HCDR2, and HCDR3) contained within a heavy chain variable region (HCVR) and three light chain complementarity-determining regions (LCDR1, LCDR2, and LCDR3) contained within a light chain variable region (LCVR), wherein HCDR1 comprises the amino acid sequence of SEQ ID NO: 2, HCD2 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 AAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 12; A second antigen-binding domain (D2) comprising three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) contained within HCVR and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained within LCVR, wherein HCDR1 comprises the amino acid sequence of SEQ ID NO: 6, HCD2 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 AAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO:

12. Includes, D1 specifically binds to the first epitope of human MET; and D2 specifically binds to the second epitope of human MET. Bispecific antibodies.

2. The criteria for eligibility are as follows: (i) MET tyrosine kinase inhibitor (TKI) untreated; (ii) Histologically confirmed NSCLC; (iii) Deletion resulting from an abnormality in MET-exon 14 or skipping of exon 14 in DNA; (iv) Amplification of the MET gene; (v) Increased MET protein expression (IHC ≥ 2+ or H score > 150); (vi) Abnormalities in MET exon 14 in DNA, deletions resulting in exon 14 skipping, and a history of MET TKI; (vii) Abnormalities or deletions resulting in exon 14 skipping in MET exon 14 in DNA and MET TKI untreated; (viiii) Highly amplified MET gene (MET gene copy number (GCN) ≥ 5 and / or MET to chromosome 7 centromere (MET / CEP7) ratio ≥ 2 in tissue by FISH, or MET GCN ≥ 6 or MET change ratio ≥ 2 in ctDNA by next-generation sequencing (NGS)) and MET TKI untreated; (ix) Highly overexpressed MET protein (IHC 3+ or H score ≥200) and MET TKI untreated; and (x) 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 change ratio ≥ 2 in ctDNA), highly overexpressed MET protein (IHC 3+ or H score ≥ 200), and MET TKI untreated; A bispecific antibody for use according to claim 1, having one or more of the following:

3. The MET abnormality is an abnormality of exon 14 in DNA, amplification of the MET gene, or overexpression of the MET protein, a bispecific antibody for use according to claim 1.

4. A bispecific antibody for use according to claim 1, wherein a MET abnormality is a deletion resulting in an abnormality or skipping of exon 14 in DNA.

5. A bispecific antibody for use according to claim 1, wherein the MET abnormality is a mutation in exon 14.

6. The mutations in exon 14 are D1010N and D1010fs * 19. A bispecific antibody for use according to claim 5, which is D1010Y, D1010H, or R1004P.

7. A bispecific antibody for use according to claim 1, wherein the MET abnormality is amplification of the MET gene.

8. The bispecific antibody for use according to claim 3, wherein the overexpression of MET protein is higher expression of MET protein in tumor tissue than in normal tissue.

9. A bispecific antibody for use according to claim 1, wherein MET abnormalities are identified using ctDNA derived from a blood sample obtained from the patient before treatment.

10. A bispecific antibody for use according to claim 1, wherein MET abnormalities are identified in tissue samples obtained from the patient before treatment.

11. A bispecific antibody for use according to any one of claims 1 to 10, wherein the subject is MET tyrosine kinase inhibitor (TKI) untreated.

12. The subject is a bispecific antibody for use according to any one of claims 1 to 10, which is used by a person who has received prior anticancer therapy comprising one or more of the following: PD-1 inhibitors, EGFR inhibitors, PD-L1 inhibitors, surgery, radiotherapy, and chemotherapy.

13. A bispecific antibody for use according to claim 12, wherein the preceding anticancer therapy comprises a PD-1 inhibitor or a PD-L1 inhibitor.

14. A bispecific antibody for use according to claim 12, wherein the preceding anticancer therapy comprises an EGFR inhibitor.

15. The subject is a bispecific antibody for use according to claim 12, which is resistant to or has not adequately responded to prior therapy, or has relapsed after prior therapy.

16. The subject is a bispecific antibody for use according to any one of claims 1 to 10, which has not received prior anticancer therapy.

17. A bispecific antibody for use according to any one of claims 1 to 10, wherein the tumor has an EGFR mutation.

18. The bispecific antibody for use according to claim 17, wherein the EGFR mutation is selected from the group consisting of L858R, G719S, E709A, E746_A750del, and S752_I759del.

19. The tumor is non-squamous NSCLC, a bispecific antibody for use according to any one of claims 1 to 10.

20. A bispecific antibody for use according to any one of claims 1 to 10, wherein the tumor is squamous cell NSCLC.

21. A bispecific antibody for use according to any one of claims 1 to 10, wherein NSCLC is transferable.

22. A bispecific antibody for use according to claim 21, wherein NSCLC has metastasized to the brain.

23. A bispecific antibody for use according to claim 21, wherein NSCLC has metastasized to the liver.

24. A bispecific antibody for use according to any one of claims 1 to 10, wherein NSCLC is unresectable.

25. A bispecific antibody for use according to any one of claims 1 to 10, wherein D1 comprises an HCVR containing the amino acid sequence of SEQ ID NO: 1 and an LCVR containing the amino acid sequence of SEQ ID NO: 9, and D2 comprises an HCVR containing the amino acid sequence of SEQ ID NO: 5 and an LCVR containing the amino acid sequence of SEQ ID NO:

9.

26. The bispecific antibody for use according to any one of claims 1 to 10, wherein the bispecific antibody is administered in doses of approximately 250 mg, 500 mg, 750 mg, 1000 mg, 1500 mg, or 2000 mg.

27. A bispecific antibody for use according to any one of claims 1 to 10, wherein the bispecific antibody is administered in a dose of 2000 mg.

28. A bispecific antibody for use according to any one of claims 1 to 10, wherein the bispecific antibody is administered intravenously, subcutaneously, or intraperitoneally.

29. A bispecific antibody for use according to any one of claims 1 to 10, wherein the bispecific antibody is administered once every three weeks.

30. A bispecific antibody for use according to any one of claims 1 to 10, wherein the bispecific antibody is administered three weeks after the immediately preceding dose.

31. A bispecific antibody for use according to any one of claims 1 to 10, wherein the treatment produces a therapeutic effect selected from the group consisting of delayed tumor growth, reduced tumor cell count, reduced metastasis, tumor regression, improved survival, partial response, and complete response.

32. A bispecific antibody for use according to claim 31, wherein tumor growth is delayed by at least 10 days compared to untreated subjects.

33. A bispecific antibody for use according to claim 31, wherein tumor growth is inhibited by at least 50% compared to untreated subjects.

34. A bispecific antibody for use in methods to treat or inhibit the growth of NSCLC, The method is: (1) Select subjects who have tumors with MET abnormalities; and (2) Administer bispecific antibodies in doses of approximately 250 mg, 500 mg, 750 mg, 1000 mg, 1500 mg, or 2000 mg. Includes, The bispecific antibody is A first antigen-binding domain (D1) comprising three heavy chain complementarity-determining regions (CDRs) (HCDR1, HCDR2, and HCDR3) contained within a heavy chain variable region (HCVR) and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained within a light chain variable region (LCVR), wherein HCDR1 comprises the amino acid sequence of SEQ ID NO: 2, HCD2 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 AAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 12; A second antigen-binding domain (D2) comprising three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) contained within HCVR and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained within LCVR, wherein HCDR1 comprises the amino acid sequence of SEQ ID NO: 6, HCD2 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 AAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO:

12. Includes, D1 specifically binds to the first epitope of human MET; and D2 specifically binds to the second epitope of human MET. Bispecific antibodies.

35. The bispecific antibody for use according to claim 34, administered in a dose of approximately 2000 mg.

36. The bispecific antibody for use according to claim 34, wherein the bispecific antibody is administered once every three weeks.

37. The criteria for eligibility are as follows: (i) MET tyrosine kinase inhibitor (TKI) untreated; (ii) Histologically confirmed NSCLC; (iii) Deletion resulting from an abnormality in MET-exon 14 or skipping of exon 14 in DNA; (iv) Amplification of the MET gene; (v) Increased MET protein expression (IHC ≥ 2+ or H score > 150); (vi) Abnormalities in MET exon 14 in DNA, deletions resulting in exon 14 skipping, and a history of MET TKI; (vii) Abnormalities or deletions resulting in exon 14 skipping in MET exon 14 in DNA and MET TKI untreated; (viiii) Highly amplified MET gene (MET gene copy number (GCN) ≥ 5 and / or MET to chromosome 7 centromere (MET / CEP7) ratio ≥ 2 in tissue by FISH, or MET GCN ≥ 6 or MET change ratio ≥ 2 in ctDNA by next-generation sequencing (NGS)) and MET TKI untreated; (ix) Highly overexpressed MET protein (IHC 3+ or H score ≥200) and MET TKI untreated; and (x) 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 change ratio ≥ 2 in ctDNA), highly overexpressed MET protein (IHC 3+ or H score ≥ 200), and MET TKI untreated; A bispecific antibody for use according to claim 34, having one or more of the following:

38. The MET abnormality is an abnormality in exon 14 of DNA, amplification of the MET gene, or overexpression of the MET protein, according to the bispecific antibody for use according to claim 34.

39. A bispecific antibody for use according to claim 34, wherein a MET abnormality is a deletion resulting in an abnormality or skipping of exon 14 in DNA.

40. A bispecific antibody for use according to claim 34, wherein the MET abnormality is a mutation in exon 14.

41. The mutations in exon 14 are D1010N and D1010fs * 19. A bispecific antibody for use according to claim 40, which is D1010Y, D1010H, or R1004P.

42. A bispecific antibody for use according to claim 34, wherein the MET abnormality is amplification of the MET gene.

43. The bispecific antibody for use according to claim 38, wherein overexpression of MET protein is higher expression of MET protein in tumor tissue than in normal tissue.

44. A bispecific antibody for use according to claim 34, wherein MET abnormalities are identified using ctDNA derived from a blood sample obtained from the patient before treatment.

45. A bispecific antibody for use according to claim 34, wherein MET abnormalities are identified in tissue samples obtained from the patient before treatment.

46. The bispecific antibody for use according to claim 34, further selected as having NSCLC with EGER mutations.

47. The bispecific antibody for use according to claim 34, further selected as having non-squamous NSCLCs.

48. The bispecific antibody for use according to claim 34, further selected as having squamous epithelial NSCLCs.

49. A bispecific antibody for use according to any one of claims 34 to 48, wherein NSCLC is transferable.

50. A bispecific antibody for use according to claim 49, wherein NSCLC has metastasized to the brain.

51. A bispecific antibody for use according to claim 49, wherein NSCLC has metastasized to the liver.

52. A bispecific antibody for use according to any one of claims 34 to 48, wherein NSCLC is unresectable.

53. A method for identifying candidates for MET x MET antitumor therapy, the method is: Obtaining tissue samples and / or liquid samples from subjects having NSCLC; and Evaluate tissue and / or liquid samples for MET abnormalities selected from the group consisting of abnormalities or deletions resulting in exon 14 skipping in DNA, amplification of the MET gene, and overexpression of the MET protein. Includes, The presence of at least one MET abnormality in a tissue or fluid sample identifies the subject as a candidate for antitumor therapy; MET x MET antitumor therapy includes a bispecific antibody, and this bispecific antibody is A first antigen-binding domain (D1) comprising three heavy chain complementarity-determining regions (HCDR1, HCDR2, and HCDR3) contained within a heavy chain variable region (CDR) (HCVR) and three light chain complementarity-determining regions (LCDR1, LCDR2, and LCDR3) contained within a light chain CDR (LCVR), wherein HCDR1 comprises the amino acid sequence of SEQ ID NO: 2, HCD2 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 AAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 12; A second antigen-binding domain (D2) comprising three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) contained within HCVR and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained within LCVR, wherein HCDR1 comprises the amino acid sequence of SEQ ID NO: 6, HCD2 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 AAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO:

12. Includes, D1 specifically binds to the first epitope of human MET; and D2 specifically binds to the second epitope of human MET. method.

54. The method according to claim 53, wherein the bispecific antibody is administered in doses of approximately 250 mg, 500 mg, 750 mg, 1000 mg, 1500 mg, or 2000 mg.

55. The method according to claim 54, wherein the bispecific antibody is administered in a dose of approximately 2000 mg.

56. A MET x MET bispecific antibody for use in a method of treating subjects with NSCLC accompanied by MET abnormalities, wherein the efficacy of the MET x MET bispecific antibody is monitored, and the method is (i) Obtain tissue samples and / or liquid samples from the subject, a. On-target MET receptor gene mutations and MET gene silencing (loss of function) that confer resistance to MET tyrosine kinase inhibitors (TKIs) in MET Ex14 Mut patients with preceding TKI Exp; b. TK driver receptor activation selected from TK receptor and ligand gene amplification and TKR activating mutations; and c. Activation gene mutations, TP53 mutations, and cell cycle gene amplifications in pathways selected from the group consisting of the JAK2 / STAT3 pathway, RAS / RAF / MEK / MAPK pathway, and PI3K / AKT / MTOR pathway. Evaluating tissue and / or liquid samples for somatic mutations in one or more genes selected from the group consisting of the following: (ii) Administering MET x MET bispecific antibodies to the target; and (iii) Repeating steps (i) and (ii) throughout the treatment process. Includes, The acquisition of one or more gene mutations is an indication of resistance to therapy and / or a poor prognosis. The MET x MET bispecific antibody is A first antigen-binding domain (D1) comprising three heavy chain complementarity-determining regions (CDRs) (HCDR1, HCDR2, and HCDR3) contained within a heavy chain variable region (HCVR) and three light chain complementarity-determining regions (LCDR1, LCDR2, and LCDR3) contained within a light chain variable region (LCVR), wherein HCDR1 comprises the amino acid sequence of SEQ ID NO: 2, HCD2 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 AAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 12; A second antigen-binding domain (D2) comprising three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) contained within HCVR and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained within LCVR, wherein HCDR1 comprises the amino acid sequence of SEQ ID NO: 6, HCD2 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 AAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO:

12. Includes, D1 specifically binds to the first epitope of human MET; and D2 specifically binds to the second epitope of human MET. Bispecific antibodies.

57. a. On-target MET receptor gene mutations were selected from the group consisting of MET Y1230C, MET D1228H, and MET D1228N; MET gene silence Loss of function was selected from somatic mutations in DNMT3A and TET2; b. TKR activating mutations were selected from the group consisting of EGFR L858R, EGFR G719S, EGFR E709A, EGFR E746_A750del, and EGFR S752_I759del; c. The JAK2 / STAT3 pathway mutation is JAK2 V617F; the RAS / RAF / MEK / MAPK pathway mutations are KRAS G12A / V, GNAS R201H, MKRN-BRAF fusion, BRAF S602Y, RICTOR Amp, and MAP2K1 The group consisting of K57N was selected; PI3K / AKT / MTOR pathway mutations were selected from the group consisting of PIK3CA H1047L, PIK3CA E545K, PIK3CA E542K, PIK3CA N345K, IDH1 R132L, and MTOR E2338Q; PI3K / AKT / MTOR pathway amplification was selected from the group consisting of AKT2 Amp and RICTOR Amp; TP53 mutations were selected from the group consisting of TP53 R280T and TP53 R248Q; cell cycle gene amplification was selected from the group consisting of CDK4 Amp, CDK6 Amp, CCND1 Amp, and CCNE1 Amp. A bispecific antibody for use according to claim 56.

58. A MET x MET bispecific antibody for use in a method of treating NSCLC in a subject, The method is, (i) Obtaining a bodily fluid sample from the subject and determining the amplification of MET in ctDNA from the bodily fluid sample, (ii) Administer the subject with a MET x MET bispecific antibody. Including; Steps (i) and (ii) are repeated once every three weeks. Loss of MET amplification after step (ii) is an indication of sustained response to treatment. The MET x MET bispecific antibody is A first antigen-binding domain (D1) comprising three heavy chain complementarity-determining regions (CDRs) (HCDR1, HCDR2, and HCDR3) contained within a heavy chain variable region (HCVR) and three light chain complementarity-determining regions (LCDR1, LCDR2, and LCDR3) contained within a light chain variable region (LCVR), wherein HCDR1 comprises the amino acid sequence of SEQ ID NO: 2, HCD2 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 AAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 12; A second antigen-binding domain (D2) comprising three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) contained within HCVR and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained within LCVR, wherein HCDR1 comprises the amino acid sequence of SEQ ID NO: 6, HCD2 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 AAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO:

12. Includes, D1 specifically binds to the first epitope of human MET; and D2 specifically binds to the second epitope of human MET. Bispecific antibodies.

59. The bispecific antibody for use according to claim 58, wherein the bispecific antibody is administered intravenously, subcutaneously, or intraperitoneally.

60. The bispecific antibody for use according to claim 58, wherein the bispecific antibody is administered once every three weeks.

61. The bispecific antibody for use according to claim 58, wherein the bispecific antibody is administered three weeks after the immediately preceding dose.

62. A bispecific antibody for use in a method to treat non-small cell lung cancer (NSCLC) in subjects with tumors exhibiting MET abnormalities, to reduce the growth of NSCLC tumors, and / or to induce regression of NSCLC, The method includes administering a 2000 mg intravenous dose of the bispecific antibody to the subject once every three weeks. The bispecific antibody is A first antigen-binding domain (D1) comprising three heavy chain complementarity-determining regions (CDRs) (HCDR1, HCDR2, and HCDR3) contained within a heavy chain variable region (HCVR) and three light chain complementarity-determining regions (LCDR1, LCDR2, and LCDR3) contained within a light chain variable region (LCVR), wherein HCDR1 comprises the amino acid sequence of SEQ ID NO: 2, HCD2 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 AAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 12; A second antigen-binding domain (D2) comprising three heavy chain CDRs (HCDR1, HCDR2, and HCDR3) contained within HCVR and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained within LCVR, wherein HCDR1 comprises the amino acid sequence of SEQ ID NO: 6, HCD2 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 AAS, and LCDR3 comprises the amino acid sequence of SEQ ID NO:

12. Includes, D1 specifically binds to the first epitope of human MET; and D2 specifically binds to the second epitope of human MET. Bispecific antibodies.

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